concanavalin-a and Lupus-Erythematosus--Systemic

Topics: Animals; Autoantibodies; B-Lymphocytes; Concanavalin A; Disease Models, Animal; Dogs; Feedback; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; T-Lymphocytes, Regulatory

Topics: Animals; Ascorbic Acid; Chediak-Higashi Syndrome; Concanavalin A; Cyclic GMP; Disease Models, Animal; Humans; Immunity, Cellular; Leukocytes; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Prostaglandins

Topics: Animals; Antibodies, Antinuclear; Antigen-Antibody Complex; Autoantibodies; Autoimmune Diseases; B-Lymphocytes; Concanavalin A; Cytotoxicity, Immunologic; HLA Antigens; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; T-Lymphocytes

To observe the effects of Xuebijing injection on dendritic cells (DCs) and T lymphocytes, and the potential mechanisms of its therapeutic effect on systemic lupus erythematosus (SLE).. A widely used mouse model, SLE-prone BLLF1 mice aged 8-10 weeks, was employed. Mice were randomly divided into 4 groups: a normal group, a model group and two treatment groups treated with Xuebijing Injection with a dose of 6.4 mL/kg via intraperitoneal administration for SLE-prone BLLF1 mice aged 8 weeks (treatment A group) and 10 weeks (treatment B group). Renal tissue sections were stained with Masson's trichrome and periodic acid-silver methenamine. Histopathological changes in the kidney were evaluated by a light microscopy. The capacity of the DCs isolated from the spleen to stimulate the T cell proliferation in response to concanavalin A (Con A) was determined.. Compared with the model group, levels of anti-dsDNA antibodies in the two treatment groups decreased remarkablly (P<0.01, P<0.05), and levels of serum creatinine and blood urea nitrogen increased (P<0.01, P<0.05). Pathological changes were found in the kidney in the model group. Histopathological abnormalities were alleviated in the two treatment groups. Treatment with Xuebijing injection also significantly upregulated the expression of CD80, CD86 and major histocompatibility class II by DCs compared with the model group (P<0.05). When splenic T lymphocytes from BLLF1 mice were co-cultured with DCs at ratios of 1:100, 1:150 and 1:200 for 3 and 5 days, the proliferation of T lymphocytes was suppressed compared with the normal group (P<0.05), but this was restored by Xuebijing Injection under the same conditions. In the model group, levels of tumor necrosis factor (TNF)-α in supernatants were significantly elevated compared with the normal group (P<0.01), interleukin-2 levels decreased (P<0.05), while these changes were significantly alleviated in the Xuebijing treatment groups.. Xuebijing Injection alleviated renal injury in SLE-prone BLLF-1 mice. The mechanism might be through influencing T cell polarization mediated by DCs, and Xuebijing Injection might be a potential drug that suppresses immune dysfunction in patients with SLE.

Topics: Animals; Antibodies, Antinuclear; Cell Differentiation; Cell Proliferation; Concanavalin A; Dendritic Cells; Drugs, Chinese Herbal; Injections; Interleukin-2; Kidney; Kidney Function Tests; Lupus Erythematosus, Systemic; Mice; Phenotype; T-Lymphocytes; Tumor Necrosis Factor-alpha

To investigate the expression and effect of farnesoid X receptor (FXR) on systemic lupus erythematosus (SLE) liver dysfunction and indicate its hepatoprotective role and the immunomodulatory property. mRNA and protein levels of FXR were determined on the liver specimens of SLE patients with liver injury as well as MRL/lpr rodent models. The FXR agonist chenodeoxycholic acid (CDCA) was administrated to MRL/lpr mice and the control BALB/C with concanavalin A (ConA)-induced liver injury. Blood samples were taken 0, 4, 8, 12, 16, and 24 h after ConA injection for the detection of serum ALT, AST, IFN-γ, TNF-α, and IL-6. FXR was down-regulated at both mRNA and protein levels in the liver specimens of SLE patients with liver injury as well as MRL/lpr mice. MRL/lpr was more susceptible to ConA than BALB/C indicated by significantly higher levels of aminotransferase and inflammatory cytokines. Activation of FXR by CDCA significantly reduced aminotransferase and inflammatory cytokines IFN-γ, TNF-α, and IL-6 caused by ConA injection in MRL/lpr mice. FXR was down-regulated in SLE patients as well as MRL/lpr lupus models with liver dysfunction. FXR activation ameliorated liver injury and suppressed inflammatory cytokines, thereby showing its protective function in SLE. Our findings raised the promising potential target for the treatment of SLE liver injury.

Topics: Alanine Transaminase; Animals; Anti-Inflammatory Agents; Aspartate Aminotransferases; Case-Control Studies; Chemical and Drug Induced Liver Injury; Chenodeoxycholic Acid; Concanavalin A; Cytoprotection; Disease Models, Animal; Female; Humans; Inflammation Mediators; Interferon-gamma; Interleukin-6; Liver; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; Mice, Inbred MRL lpr; Receptors, Cytoplasmic and Nuclear; RNA, Messenger; Time Factors; Tumor Necrosis Factor-alpha

We sought to evaluate the effects of combined downregulation of CD134 and cytotoxic T lymphocyte-associated antigen 4 (CTLA4) on the autoimmune process of lupus. Concanavalin A (ConA)-induced proliferation, T helper cell cytokine secretion, and anti-double stranded DNA (dsDNA) antibody production were measured in cultures of splenic lymphocytes derived from lupus-prone BXSB mice. Splenocytes from six prednisone-treated and six untreated male lupus-prone BXSB mice, as well as from six syngeneically normal C57BL/6 male mice, were stimulated with ConA. BXSB splenocytes from untreated mice were exposed to anti-CD134L mAb, CTLA4 linked to the Fc portion of IgG1 (CTLA4Ig), or both. The magnitude of splenocyte proliferation and the levels of IFN-gamma, IL-6, and anti-dsDNA antibody were: (1) significantly higher in cultures of ConA-stimulated control and other cells than in unstimulated cells, (2) similar in cultures of normal and BXSB cells treated with anti-CD134 and CTLA4Ig or prednisone and (3) significantly reduced in cultures of ConA-stimulated and unstimulated cells treated with anti-CD134L and CTLA4Ig or prednisone compared with cells treated with CD134L or CTLA4Ig alone. Like corticosteroids, anti-CD134L mAb or CTLA4Ig can inhibit T- and B-cell activation by blocking the CD134-CD134L or CD28/CTLA4-B7 co-stimulatory pathway. The combined immune intervention described herein may prove useful for the treatment of autoimmune diseases such as systemic lupus erythematosus.

Topics: Abatacept; Animals; Antibodies, Antinuclear; Antibodies, Monoclonal; B-Lymphocytes; Cell Proliferation; Cells, Cultured; Concanavalin A; Drug Therapy, Combination; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Gene Expression; Immunoconjugates; Immunoglobulin G; Immunosuppressive Agents; Interferon-gamma; Interleukin-6; Lupus Erythematosus, Systemic; Male; Mice; OX40 Ligand; Prednisone; Receptors, OX40; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Spleen; Th1 Cells; Th2 Cells

Systemic lupus erythematosus (SLE) is characterized by inflammatory and dysregulatory immune responses including overactive B cells, overproduction of proinflammatory cytokines, and T cell hyperactivity. PGE(2) modulates a variety of immune processes at sites of inflammation, including production of inflammatory cytokines. However, the role of PGE(2) in dysregulatory inflammatory and immune responses in lupus remains unclear. We investigated whether PGE(2) mediates production of inflammatory cytokines in pristane-induced lupus BALB/c mice. Our results showed that levels of serum and BAL PGE(2) and LPS-stimulated production of PGE(2) by peritoneal macrophages were remarkably increased in pristane-induced lupus mice compared to healthy controls. Exogenous PGE(2) enhanced production of IL-6, IL-10, and NO but decreased TNF-alpha by macrophages and augmented IFN-gamma, IL-6, and IL-10 by splenocytes from pristane-induced lupus mice compared to healthy controls. Exogenous PGE(2) also enhanced production of IFN-gamma, IL-6, and IL-10 by thymocytes from pristane-induced lupus mice. Indomethacin (Indo), a PGE(2) synthesis inhibitor, greatly inhibited LPS-induced production of IL-6 and IL-10 by macrophages from pristane-induced lupus mice, while enhanced TNF-alpha. Indo remarkably inhibited Con A-increased production of IFN-gamma, IL-6, and IL-10 by splenocytes and thymocytes from pristane-induced lupus mice. Therefore, our findings suggest that endogenous PGE(2) may mediate dysregulation of production of proinflammatory cytokines, such as IL-6, IL-10, and IFN-gamma, and NO in pristane-induced lupus mice.

Topics: Animals; Bronchoalveolar Lavage Fluid; Cells, Cultured; Concanavalin A; Cyclooxygenase Inhibitors; Cytokines; Dinoprostone; Disease Models, Animal; Female; Indomethacin; Inflammation Mediators; Interferon-gamma; Interleukin-10; Interleukin-6; Lipopolysaccharides; Lupus Erythematosus, Systemic; Lymphocytes; Macrophages, Peritoneal; Mice; Mice, Inbred BALB C; Nitric Oxide; Spleen; Terpenes; Thymus Gland; Time Factors; Tumor Necrosis Factor-alpha; Up-Regulation

Bone marrow-derived mesenchymal stem cells (bMSCs) can differentiate into a number of different cell/tissue types, and also possess immunoregulatory functions. The present study was undertaken to elucidate the exact immunoregulatory effects of allogeneic bMSCs on T- and B-lymphocyte proliferation, activation, and function maturation of BXSB mice, which has been considered as a experimental model for human systemic lupus erythematosus (SLE). We determined that bMSCs from BALB/c mice had inhibitory effects on BXSB mice T-lymphocyte proliferation, but no inhibitory effect on their activation. In addition, they had a significant inhibitory and stimulatory effect on IL-4- and IFN-gamma-producing T cells, respectively. Also, bMSCs had inhibitory effects on the proliferation, activation, and IgG secretion of B lymphocytes. In addition, BALB/c bMSCs had an enhancing effect on CD40 expression and inhibitory effects on CD40 ligand (CD40L) ectopic hyperexpression on B cells from BXSB mice.

Topics: Animals; B-Lymphocytes; Bone Marrow Cells; CD40 Antigens; CD40 Ligand; Cell Proliferation; Cells, Cultured; Coculture Techniques; Concanavalin A; Disease Models, Animal; Dose-Response Relationship, Drug; Flow Cytometry; Genetic Markers; Immunoglobulin G; Interferon-gamma; Interleukin-4; Lipopolysaccharides; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Mitogens; Spleen; T-Lymphocytes; Transplantation, Homologous

Neopterin (Neo) and 7,8-dihydroneopterin (H(2)Neo) are produced by human monocyte-derived macrophages upon stimulation with IFN-gamma. Increased amounts of Neo and H(2)Neo in human body fluids are found in many disorders, including viral infections and autoimmune diseases. Recent data suggest that neopterin derivatives may exhibit distinct biochemical functions activating redox-sensitive transcription factors and inducing apoptosis in various cell lines. In this study we investigated the effect of H(2)Neo on human peripheral blood T cells (PBT) from healthy blood donors in comparison with PBT isolated from patients with systemic lupus erythematosus (SLE). H(2)Neo induced apoptosis in healthy PBT in a concentration-dependent manner. In short time culture, a significantly lower ability of PBT isolated from patients with SLE to undergo apoptosis in response to H(2)Neo compared to healthy controls was detected. Our results suggest a possible role of the neopterin derivative H(2)Neo in T cell apoptosis mediated by stimulated monocytes/macrophages.

Topics: Adult; Annexin A5; Antioxidants; Apoptosis; Cells, Cultured; Concanavalin A; Cytokines; Dose-Response Relationship, Drug; Female; Humans; Lupus Erythematosus, Systemic; Male; Middle Aged; Neopterin; Phytohemagglutinins; Pteridines; T-Lymphocytes; Time Factors

We have previously observed that aged lupus-prone (NZB/NZW)Fl (BWF1) mice when infected with Plasmodium chabaudi show an improvement in their clinical lupus-like symptoms. In order to study the mechanisms involved in the long-lasting protective effect of the P. chabaudi infection in lupus-prone mice we analysed specific aspects of the cellular response, namely the profiles of cytokine mRNA expression and cytokine secretion levels in old BWF1 mice, in comparison with uninfected age-matched BWF1 mice and infected or uninfected BALB/c mice. Two months after infection, cells from BWF1 mice were stimulated with concanavalin A (Con A) and demonstrated a recovery of T cell responsiveness that reached the levels obtained with BALB/c cells. Old BWF1 mice showed high levels of interferon-gamma (IFN-gamma) and IL-5 production and correspondingly low levels of IL-2 and IL-4 secretion before infection with P. chabaudi. Infection did not modify the IFN-gamma levels of BWF1 T cells, whereas it considerably increased the secretion of the Th2-related cytokines IL-4, IL-5 and IL-10. In addition, only BWF1 T cells showed increased mRNA expression of tumour necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta). This counter-regulatory cytokine network of infected BWF1 mice may be involved in the improvement of their lupus symptoms. The results of our investigations using the complex model of P. chabaudi infection can be extended and, by using more restricted approaches, it may be possible to explain the multiple regulatory defects of lupus-prone mice.

Topics: Animals; Concanavalin A; Crosses, Genetic; Cytokines; Female; Genetic Predisposition to Disease; Immunoglobulins; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphokines; Malaria; Mice; Mice, Inbred BALB C; Mice, Inbred NZB; Plasmodium chabaudi; RNA, Messenger; T-Lymphocytes

To explore the role of the NF-kappaB/Rel transcription factor family in autoimmunity, we investigated whether peripheral blood mononuclear cells (PBMC) and T cells from the blood of patients with systemic lupus erythematosus (SLE) exhibit abnormal expression of c-rel, both when recently isolated and/or during in vitro activation.. Total RNA and protein extracts were prepared from PBMC and T cells isolated by immunoadsorption with magnetic beads. The relative concentrations of c-rel mRNA and of c-Rel protein were determined by semiquantitative assays of competitive reverse transcriptase-polymerase chain reaction and chemiluminescent immunoblots, respectively. Activity of NF-kappaB/Rel was studied by electrophoretic mobility shift assay of nuclear extracts.. Significantly increased levels of c-rel mRNA were found (1) in PBMC from SLE patients (n = 48; p<0.0000001), even during inactive disease (n = 11; p<0.001), compared to controls (n = 54), and (2) in T cells isolated from a subgroup of these patients (n = 11; p<0.00002) and controls (n = 12). c-Rel protein was found increased in the cytosol but not in the nucleus of PBMC of patients with SLE (n = 12; p<0.02) compared to controls (n = 12). No evidence of NF-kappaB/Rel nuclear activity was detected. In vitro stimulation of T cells by incubating PBMC with concanavalin A showed that less c-Rel entered the nucleus in lupus cells than healthy cells, correlating with lower interleukin 2 production. However, the same stimulating conditions provoked an increase in c-rel mRNA to higher levels in lupus cells from 2 patients compared with 2 controls. Increased levels of both IkappaB alpha and IkappaB beta could account for c-Rel cytosolic retention.. Our data suggest that T cells from patients with SLE possess altered regulatory mechanisms of c-rel expression and nuclear import that might potentially determine conditions for developing autoimmunity. Other cells present in the PBMC could also be affected.

Topics: Adolescent; Adult; Aged; Child; Concanavalin A; Female; Gene Expression Regulation; Humans; Interleukin-2; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Male; Middle Aged; NF-kappa B; Proto-Oncogene Proteins c-rel; RNA, Messenger; T-Lymphocytes

Effects on T lymphocyte mediated pathology, phenotypes, and functions in MRLlpr/lpr mice given mycophenolate mofetil (MMF) (100 mg/kg/day) via drinking water or controls given ip cyclophosphamide (CYC) injections (1.8 mg/mouse/week) or water were described. Both MMF and CYC treatment diminished kidney and large salivary gland perivascular cell infiltrates, reduced profoundly double-negative (DN) T cell frequencies, decreased total lymphocyte number in spleen, and increased in vitro proliferative response to Con A. IFN-gamma and IL-10 in supernatants from Con A stimulated spleen cells were augmented after MMF but not CYC treatment. MMF treatment increased whereas CYC reduced IL-12 in serum. Kidney expressions of IFN-gamma, IL-10, and IL-12 mRNA were unaffected by MMF but decreased by CYC. Our results demonstrate that MMF and CYC suppress perivascular T lymphocyte inflammation by reducing the DN T cell population and by amelioration of T cell function. The varying cytokine patterns suggest different mechanisms of the two drugs.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; B-Lymphocytes; Cell Count; Cell Division; Concanavalin A; Cyclophosphamide; Cytokines; Disease Models, Animal; Female; Hypersensitivity, Delayed; Immunophenotyping; Immunosuppressive Agents; Kidney; Lupus Erythematosus, Systemic; Male; Mice; Mice, Inbred MRL lpr; Mycophenolic Acid; RNA, Messenger; Salivary Glands; Spleen; T-Lymphocytes; Vasculitis

Production of cytokines in unstimulated and mitogen-stimulated cultures were evaluated by ELISPOT in 34 SLE patients with low to moderate disease activity and 23 healthy controls. Significantly reduced production of IFN gamma, IL4 and IL12 and significantly increased production of IL6, IL10 and TNF alpha were found in patients with SLE. Regression analysis revealed that production of all six cytokines tended to decrease with increasing disease activity, but negative correlation with SLEDAI was significant (p < 0.05) only for PHA-stimulated IL4, unstimulated and PHA-stimulated IL10 and SAC-stimulated IL6. Negative correlation of stimulated and unstimulated IL6 and TNF alpha production with anti-DNA antibody levels were also significant.

Topics: Adolescent; Adult; Aged; Antibodies, Antinuclear; Biomarkers; Cells, Cultured; China; Concanavalin A; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunosuppressive Agents; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-4; Interleukin-6; Lupus Erythematosus, Systemic; Male; Middle Aged; Muromonab-CD3; Phytohemagglutinins; Staphylococcus aureus; Tumor Necrosis Factor-alpha

The cell surface protein tyrosine phosphatase CD45 is a major target of IgM anti-T cell autoantibodies in systemic lupus erythematosus (SLE). The autoreactive determinants on CD45 are O-linked glycans expressed on activated T cells and certain T cell lines, rather than linear or conformational polypeptide epitopes or N-linked glycans. To identify oligosaccharide structures that may play a role in the functional interactions of CD45 or are candidate target epitopes of SLE anti-CD45 autoantibodies, autoreactive CD45 purified from Jurkat T cells and non-autoreactive CD45 purified from CLL B cells were tested by ELISA for expression of mucin-type O-glycan structures. Monoclonal antibodies (mAbs) directed against blood group A, type 1 H chains, type 2 H chains, T, Le(a), sialylated-Le(a), Le(b), sialylated-Le(c), Le(x), sialylated-Le(x), multi-fucosylated Le(x), Le(y), and sialylated-extended Le(v) failed to react with CD45 from either B cells or T cells. However, mAbs directed against Tn (galNAcalpha1-->O-ser/thr) or sialosyl-Tn (neuNAcalpha2-6gaINAcalpha1-->O-ser/thr) structures reacted with CD45 derived from Jurkat T cells, but not from CLL B cells. Anti-Tn mAbs also reacted in western blotting procedures with CD45 isolated from Jurkat T cells, but did not react with CD45 isolated from CEM, MOLT-3, or PEER T cells; Daudi, Raji, or CLL B cells; or resting or Con A-activated PBL. However, anti-sialosyl-Tn mAbs stained blots of CD45 isolated from Jurkat and CEM T cells and Con A-activated PBL, a pattern of reactivity similar to that of the anti-CD45 autoantibodies. Flow cytometric analyses demonstrated that the sialosyl-Tn epitopes are expressed on a subpopulation of CD4 +/CD8- T cells.

Topics: Antibodies, Monoclonal; Autoantibodies; B-Lymphocytes; Carbohydrates; Cell Line; Concanavalin A; Epitopes; Humans; Immunoglobulin M; In Vitro Techniques; Jurkat Cells; Leukocyte Common Antigens; Lupus Erythematosus, Systemic; Lymphocyte Activation; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Sialic Acids; T-Lymphocyte Subsets; T-Lymphocytes

To investigate the primary autoantigens which contribute to the production of anti-DNA antibodies. These antibodies are serological hallmark and pathogenic factor of systemic lupus erythematosus (SLE).. Nonautoimmune predisposed BALB/c mice were immunized with concanavalin A (Con A) activated, lipopolysaccharide (LPS) activated and nonactivated syngeneic spleen cells. Nuclei and chromatin from activated/nonactivated lymphocytes were isolated and syngeneic mice were immunized. Sera were taken after the third immunization. IgG anti-dsDNA antibody was determined by ELISA (calf thymus DNA treated with S1 nuclease was used as the coated antigen). The glomerular IgG deposition was observed by immunofluorescence one month after the third immunization.. Con A activated T cells and LPS activated B cells induced anti-double stranded (ds) DNA antibody in syngeneic nonautoimmune BALB/c mice and formed the glomerular IgG deposition. Further studies showed that active chromatin isolated from activated lymphocytes induced anti-ds DNA antibody, but not resting chromatin isolated from nonactivated lymphocytes.. Activated lymphocytes and their active chromatin could be the autoimmunogen(s) driving the anti-dsDNA antibodies. The change of chromatin's antigenicity by environmental factors and genetic background may be the common pathway to SLE pathogenesis.

Topics: Animals; Antibodies, Antinuclear; Chromatin; Concanavalin A; Female; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Random Allocation

We have evaluated the production of PRL by human peripheral mononuclear cells (PBMNC) from normal subjects and patients with systemic lupus erythematosus (SLE). Conditioned medium prepared from basal and Con-A-stimulated PBMNC was assessed for the presence of PRL-like by its ability to stimulate growth of PRL-responsive Nb2 rat lymphoma cells. In the presence or absence of Con-A, SLE PBMNC secrete significantly higher (P < 0.001) amounts of bioactive PRL-like species than normal cells. Growth of Nb2 cells by conditioned medium was inhibited with specific antiserum to human PRL. Western blotting using a polyclonal antibody to human PRL revealed a single 60-kDa PRL-like species in both normal and SLE PBMNC extracts, the immunoreactivity of which was preferentially found in SLE subjects. With the use of reverse transcription-PCR an expected 633-bp band was observed, and its similarity to pituitary PRL was further confirmed by Southern blot analysis with human PRL complementary DNA as a probe. We conclude that a high molecular mass PRL-like species is synthesized and secreted by PBMNC, and patients with SLE have an increased secretion of lymphocyte-derived PRL-like material.

Topics: Adolescent; Adult; Animals; Biological Assay; Blotting, Southern; Blotting, Western; Cells, Cultured; Concanavalin A; Culture Media, Conditioned; Female; Gene Expression; Humans; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Lymphoma; Molecular Weight; Polymerase Chain Reaction; Prolactin; Rats; RNA, Messenger

We previously reported that difluoromethylornithine (DFMO), an inhibitor of polyamine biosynthesis, exerted significant beneficial effects on the lifespan and disease expression of MRL-lpr/lpr mice, which spontaneously develop a lupus-like syndrome. Polyamine levels in splenic T-cells of MRL-lpr/lpr mice were significantly higher than those of Balb/c mice. In the present investigation, we examined the role of endogenous polyamines in transmembrane Ca2+ influx, generation of InsP3 and tyrosine phosphorylation of the p56lck protein in concanavalin A-stimulated splenic T-cells. Cytosolic free calcium concentrations ([Ca2+]i) in concanavalin A-stimulated T-cells of MRL-lpr/lpr and Balb/c mice were 250 +/- 25 and 450 +/- 42 nM respectively. Treatment of MRL-lpr/lpr mice with DFMO increased [Ca2+]i to 360 +/- 30 nM (P < 0.05). InsP3 levels of concanavalin A-stimulated MRL-lpr/lpr splenic T-cells were only 20% higher than those of unstimulated controls, whereas those of Balb/c T-cells were 90% higher. DFMO treatment increased InsP3 levels in concanavalin A-treated MRL-lpr/lpr T-cells to 67%. Western-blot analysis showed a 7-fold higher level of p56lck phosphorylation of MRL-lpr/lpr splenic T-cells than that of Balb/c mice. DFMO treatment reduced tyrosine phosphorylation of p56lck of MRL-lpr/lpr mice significantly (P < 0.001). Two-colour flow-cytometric analysis revealed no significant difference in the CD4+/CD8+ ratio in splenic T-cells of MRL-lpr/lpr mice after DFMO treatment. Polyamine levels in splenocytes were significantly reduced by DFMO treatment. These data show that DFMO treatment could alter signal-transduction pathways of splenic T-cells of MRL-lpr/lpr mice. Increased levels of polyamines in T-cells of untreated lpr mice contribute to defective signal-transduction pathways and the pathogenesis of lupus-like symptoms.

Topics: Animals; Autoimmune Diseases; Blotting, Western; Calcium; CD4-CD8 Ratio; Concanavalin A; Eflornithine; Female; Inositol Phosphates; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; Ornithine Decarboxylase; Phosphorylation; Phosphotyrosine; Polyamines; Signal Transduction; Spleen; T-Lymphocytes

(NZB x NZW)F1 (B/W) mice spontaneously develop a lupus-like syndrome characterized by an increased level of autoantibodies in old mice. We analysed the role of T cells in the regulation of anti-DNA antibody production by B cells in vitro as a function of age. In cultures of old mouse T and B cells, IgG and IgM anti-DNA antibodies were synthesized at high levels, in contrast to consistently lower amounts, particularly of IgG, measured in cultures of young mouse cells. Addition of young mouse T cells to old B cells inhibited IgG, but not IgM, anti-DNA production, whereas T cells from old mice stimulated IgG synthesis by young mouse B cells. Addition of supernatants harvested from concanavalin A (Con A)-stimulated T cells to B-cell cultures induced similar effects. Therefore, we evaluated possible modifications of lymphokine synthesis compared to that of the healthy NZW parent. T cells from old mice were able to secrete normal levels of interferon-gamma (IFN-gamma) and interleukin (IL)-10; however, secretion of IL-2 and IL-4 was dramatically decreased. Semi-quantitative polymerase chain reaction analysis of constitutive RNA messengers showed increased IFN-gamma levels in young and old B/W mice, and normal IL-10 mRNA levels in young and higher levels in old mice. Constitutive IL-2 and IL-4 mRNA were detected only after Con A stimulation and their levels decreased in old compared to young B/W mice; in particular IL-2 mRNA was considerably lower in old B/W than in control NZW mice. Taken together, these results suggest that, despite constitutive T-cell abnormalities, young B/W mice are able partially to control their lymphokine production, whereas aged mice exhibit a deficient synthesis, associated with an increased capacity to produce IFN-gamma.

Topics: Aging; Animals; Antibodies, Antinuclear; Autoimmune Diseases; Cells, Cultured; Concanavalin A; Culture Media, Conditioned; Female; Gene Expression; Immunoglobulin G; Immunoglobulin M; Lupus Erythematosus, Systemic; Lymphokines; Male; Mice; Mice, Inbred NZB; RNA, Messenger; T-Lymphocytes

Topics: Blood Cells; Case-Control Studies; Concanavalin A; Dendritic Cells; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; In Vitro Techniques; Interleukin-1; Lupus Erythematosus, Systemic; Lymphocyte Activation; T-Lymphocytes

Cytokine specific ELIspot assays were used to monitor the number of cells secreting IL-4 and IFN gamma in MRL-lpr/lpr mice of different ages. The cytokine repertoire expressed by MRL-lpr/lpr mice was skewed towards the over-production of IFN gamma (and away from IL-4) when compared to that of MRL-+/-/+/- and BALB/c mice. With increasing age and disease severity, the ratio of IFN gamma: IL-4 secreting cells rose in MRL-lpr/lpr mice but decreased in normal animals. This changing ratio of IFN gamma: IL-4 secreting cells correlated with changes in the ratio of IgG2a: IgG1 secreting cells in these mice. Our findings suggest that cytokine secreting CD4+ and CD8+ cells are hyperactivated in young MRL-lpr/lpr mice, and that disease progression is associated with a disruption in the normal ratio of Th1:Th2 production in vivo.

Topics: Age Factors; Aging; Animals; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Concanavalin A; Immunoglobulin G; Immunoglobulin Isotypes; Interferon-gamma; Interleukin-4; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; Mice, Mutant Strains

Splenic T-cells from lupus strain (NZB/W F1, Mrl/lpr) mice lack the ability to respond to concanavalin A (Con A) by secretion of IL-2 and hence expression of IL-2 receptor and proliferation. These defects were found not only in an aged group (> 5 months) of mice in which obvious clinical 'SLE like' symptoms and elevated levels of serum autoantibodies were observed, but also in mice as young as 4-wk. We demonstrate here that the defective mitogenic activation of T-cells from lupus mice is due to the inability of T-helper cells to produce IL-2 and this defect can be restored by exogenous IL-2 in vitro. Con A-induced cell proliferation and IL-2 receptor expression on CD3+ cells from lupus mice occur only in the presence of exogenous IL-2, whereas normal T-cells from BALB/c and CBA control mice are activated by the mitogen and undergo complete cell cycling in the absence of exogenous IL-2, as they are able to secrete sufficient endogenous IL-2. The detection of impaired T-helper function in young lupus mice, before development of overt disease, and the reversible nature of the defect indicate that defective IL-2 activity may be fundamental to the mechanism of development of pathology in SLE.

Topics: Age Factors; Animals; Autoimmune Diseases; Cells, Cultured; Concanavalin A; Female; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred CBA; Mice, Inbred NZB; Receptors, Interleukin-2; T-Lymphocyte Subsets; T-Lymphocytes, Helper-Inducer

We investigated serum levels of interleukin-6 (IL-6), interferon-gamma (IFN-gamma), and tumor necrosis factor alpha (TNF alpha) from patients with systemic lupus erythematosus (SLE) and its various clinical manifestations of disease and from patients with rheumatoid arthritis (RA) and other rheumatic diseases. The serum levels of IL-6 and IFN-gamma were highly elevated from patients with SLE associated with lymphadenopathy (LN) or nephrotic syndrome (NS). On the contrary, the serum levels of TNF alpha were elevated from most patients with SLE associated with thrombocytopenia (TP). However, serum levels of TNF alpha were in the normal range from patients with SLE associated with NS, LN, or central nervous system disease. Of interest, patients with SLE associated with humoral immunodeficiency disorder, hypogammaglobulinemia, had highly elevated levels of serum IL-6. The concanavalin A-stimulated mononuclear cells (MNC) of patients with SLE associated with TP secreted highly elevated levels of TNF alpha compared to other patient groups. We suggest that abnormal production of various cytokines in SLE is an intrinsic defect of MNC and the immune system that may be the key element for a variety of clinical manifestations of this disease.

Topics: Adolescent; Adult; Arthritis, Rheumatoid; Behcet Syndrome; Concanavalin A; Cytokines; Female; Humans; Interferon-gamma; Interleukin-6; Lupus Erythematosus, Systemic; Lymphadenitis; Male; Middle Aged; Nephrotic Syndrome; Polymyalgia Rheumatica; Rheumatic Diseases; Sjogren's Syndrome; Thrombocytopenia; Tumor Necrosis Factor-alpha

The accessory cell (AC) functions of peripheral blood dendritic cells (PDC) in concanavalin A (Con A)-induced T cell proliferation were investigated in patients with systemic lupus erythematosus (SLE). In Con A-induced T cell proliferation using added autologous PDC, AC activity was present in SLE patients (p less than 0.01). The Con A-induced T cell proliferation, however, was significantly lower in SLE patients than in normal subjects (p less than 0.01), and the level of decrease became greater with increased activity of the disease (p less than 0.01). In investigating the Con A-induced T cell proliferation using added allogeneic PDC, we compared T cells from SLE patients + PDC from normal subjects or T cells from normal subjects + PDC from SLE patients with T cells + allogeneic PDC both from normal subjects. The former two combinations showed significantly reduced T cell proliferation in active SLE (p less than 0.01 for each), while no significant differences were found for inactive SLE. The above results suggest that the reduced function of PDC as AC may be involved in reduced Con A-induced T cell proliferation in patients with active SLE.

Topics: Adult; Concanavalin A; Dendritic Cells; Female; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; T-Lymphocytes

Oestrogen is known to accelerate glomerulonephritis and autoantibody production in human and murine systemic lupus erythematosus (SLE). In this study we demonstrate that treatment of castrated autoimmune MRL +/+ mice with physiological doses of oestrogen results in enhanced immunoglobulin and autoantibody production as well as increased deposition of IgG in renal glomeruli. Accelerated development of glomerulonephritis was also evident from the increase of albuminuria. Interestingly, in contrast to these deteriorative effects of oestrogen on immune complex-mediated disease we now show that the lymphocytic infiltrations in the submandibular glands and perivascular lesions in the kidneys were significantly diminished after exposure to oestrogen. This remarkable impact of physiological oestrogen levels on the outcome of SLE in MRL +/+ mice is postulated to be the result of a differential effect on T and B cell-mediated immune responses.

Topics: Animals; Antibodies, Antinuclear; Autoimmunity; Castration; Concanavalin A; Estradiol; Glomerulonephritis; Hemoglobinuria; Immunoglobulins; Kidney; Lupus Erythematosus, Systemic; Male; Mice; Oxazolone; Rheumatoid Factor; Salivary Glands; Sialadenitis; Spleen; Vasculitis

Inbred mouse genomes contain two subclasses of proviruses related to mink cell focus-forming (MCF) retroviruses: polytropic (Pmv), and modified polytropic (Mpmv). To determine whether one of these subclasses is associated with murine lupus, oligonucleotide probes specific for Pmv or Mpmv sequences were used in Northern analyses. Thymus 8.4 kb Mpmv RNA was expressed in five of five lupus-prone strains and crosses and this expression was not affected by genes that retard or accelerate development of lupus. Two of four leukemia-prone strains expressed low levels of such thymic transcripts, but none of 11 control strains did. 8.4 kb Mpmv RNA expression was not induced in thymuses of control mice by the lpr/lpr or gld/gld genotypes (which cause polyclonal immune activation) nor by treatment with mitogens. In contrast to Mpmv, thymic 8.4 kb Pmv expression was poorly associated with autoimmunity: it was easily detected in nearly all strains, and was increased by polyclonal activation in control mice. These studies indicate that the organ-specific thymic 8.4 kb Mpmv expression (a) is characteristic of several genetic backgrounds which predispose to murine lupus, (b) precedes and does not correlate with disease development, (c) is not due to polyclonal activation, and (d) is regulated independently of 8.4 kb Pmv expression.

Topics: Animals; Base Sequence; Blotting, Northern; Concanavalin A; Genes, env; Lupus Erythematosus, Systemic; Mice; Mice, Inbred Strains; Molecular Sequence Data; Oligonucleotide Probes; Retroviridae; RNA, Viral; Species Specificity; Spleen; Thymus Gland

We have previously demonstrated that Captopril, an inhibitor of angiotensin converting enzyme (ACE), ameliorates experimental systemic lupus erythematosus in inbred MRL lpr/lpr (MRL/l) mice. In contrast, Enalapril, another ACE inhibitor with antihypertensive properties but lacking a thiol group, did not show similar beneficial effects. To better understand the mode of action of captopril in the autoimmune disease we have evaluated its immunomodulatory properties with special emphasis on antigen-specific and polyclonal B- and T-cell activation. The results obtained strongly suggest that Captopril exerts its immunomodulatory effects through stimulation of T-lymphocytes.

Topics: Angiotensin-Converting Enzyme Inhibitors; Animals; Antibody Formation; B-Lymphocytes; Captopril; Concanavalin A; Enalapril; Interleukin-2; Lupus Erythematosus, Systemic; Mice; Mice, Inbred Strains; T-Lymphocytes

MRL/l mice develop progressive, virulent autoimmune disease that has many of the features of systemic lupus erythematosus. Prophylactic treatment of MRL/l mice with syngeneic photoinactivated autoimmune splenocytes improves survival and inhibits the fulminant hyperproliferation of abnormal T cells and the production of high titer anti-DNA antibody invariably found in untreated mice. The proliferation of Thy 1+ splenic T cells was significantly decreased, and prolonged retention of the response to T-cell mitogen was found in treated mice. Treatment with unmodified cells induced a partial inhibition of disease features which did not prolong survival rates. These results suggest that phototherapy potentiates a normal immunoregulatory process which enables suppression of the development of abnormal cell populations in young MRL/l mice with relatively intact immune systems.

Topics: Animals; Autoantibodies; Autoimmune Diseases; Concanavalin A; DNA; Female; Light; Lipopolysaccharides; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocyte Transfusion; Lymphocytes; Lymphoid Tissue; Mice; Mice, Inbred Strains; Phenotype; Spleen; Survival Analysis; Transplantation, Isogeneic

Quantitative changes of concanavalin A (Con A)-reactive proteins in serum samples obtained from rats with induced inflammation and from patients with inflammatory and autoimmune diseases were examined by use of lectin blots. Treatment of rats with a single dose of fermented yeast to induce inflammation caused an extensive increase in Con A-reactivity. These changes were time dependent and were similar in both sexes of the animals. When we examined serum samples obtained from patients with various inflammatory disorders for their Con A-reactive proteins as compared with normal donors, we noted that the Con A-reactivity increased in patients with rheumatoid arthritis and systemic lupus erythematosus. Among all the glycoproteins examined by lectin blots with use of Con A, a set of five proteins was selected for detailed analysis by densitometric scanning. These included alpha 2-macroglobulin, P-150, P-95, P-40, and P-35, of Mr 180,000, 150,000, 95,000, 40,000, and 35,000, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Densitometric scanning analysis of the lectin blots revealed that the Con A-reactivity of these proteins increased during inflammation. Because alpha 2-macroglobulin is not an acute-phase protein in humans, an increase in Con A staining of this protein suggested that altered glycation is associated with autoimmune diseases. Thus, study of changes in Con A-reactive proteins in human sera may facilitate our understanding of the etiology and pathophysiology of autoimmune diseases.

Topics: alpha-Macroglobulins; Animals; Arthritis, Rheumatoid; Autoimmune Diseases; Blood Proteins; Collodion; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Female; Glycoproteins; Inflammation; Kinetics; Lupus Erythematosus, Systemic; Male; Molecular Weight; Rats; Rats, Inbred Strains

Lymphocytes of autoimmune mice have been reported to have defective IL-2 production and proliferation in response to the mitogen concanavalin A. We have examined transcription of lymphokine genes in Con A stimulated spleen cells from both autoimmune and normal mice and found that IL-2, IL-4 and gamma-interferon (IFN gamma) were induced in all mice tested. Spleen cells were taken from young (pre-disease) or old (clinically active) MRL/lpr (lpr) and male BXSB autoimmune mice and from their normal counterparts (MRL/n, BXSB females, BALB/c and DBA/2) and stimulated with Con A. Con A induced production of IL-2, IL-4 and IFN gamma message and protein, and kinetics of induction did not vary significantly among the strains. However, in old lpr mice, levels of IL-2 protein and mRNA were about 10-fold lower than in other strains; IL-4 protein and mRNA were decreased approximately three-fold; and IFN gamma mRNA was readily detected in unstimulated cells and low but detectable levels of protein were produced constitutively. In contrast, little or no defect in IL-2 or IL-4 transcription or secretion were seen in male BXSB mice and no constitutive IFN gamma transcription was seen in this strain. These data indicate that all three lymphokine genes are activated by Con A in autoimmune mice, even though Con A-induced proliferation is defective in these mice. Furthermore, autoimmune mouse strains vary in terms of lymphokine expression: male BXSB mice display a normal lymphokine profile, whereas lpr mice show a marked imbalance of lymphokines compared to normal controls.

Topics: Animals; Autoimmune Diseases; Cell Division; Concanavalin A; Genes; Interferon-gamma; Interleukin-2; Interleukin-4; Interleukins; Lupus Erythematosus, Systemic; Lymphocytes; Lymphokines; Mice; Mice, Inbred Strains; Spleen; Transcription, Genetic

Using a modified model, 16 patients with systemic lupus erythematosus were examined for the activity of nonspecific Con A-induced suppressors with the aid of different test systems. The suppressors of both patients and donors were employed. The proliferation of the test culture lymphocytes appeared to be activated. The immunogenetic abnormalities indicated seem likely to be related to extreme activity of the countersuppressor cells blocking the function of the suppressors, correlating with the clinical picture of the disease.

Topics: Adolescent; Adult; Concanavalin A; Female; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mitomycins; T-Lymphocytes, Regulatory

Mice homozygous for the lpr gene develop a lymphoproliferative disorder due to expansion of a subset of CD4-CD8- T cells. Triggering of the T-cell receptor in these lpr T cells does not lead to translocation of protein kinase C or phosphorylation of CD3, interleukin-2 production, or proliferation, whereas a combination of phorbol ester and calcium ionophore does. Stimulation with concanavalin A or anti-CD3 induces phosphoinositide hydrolysis. The rise in inositol bisphosphate, inositol triphosphate, and inositol tetrakisphosphate, identified by HPLC, is similar in +/+ and lpr T cells. The concentration of cytoplasmic free calcium ([Ca2+]i), however, under basal and stimulated conditions is significantly lower in lpr T cells. The lower basal [Ca2+]i may explain why induction of proliferation with phorbol ester and calcium ionophore requires a higher concentration of ionophore in these cells than in normal T cells. The lower [Ca2+]i obtained on stimulation may contribute to the activation defect of CD4-CD8- lpr T cells.

Topics: Animals; Antigens, Differentiation; Calcium; Concanavalin A; Dose-Response Relationship, Drug; Inositol Phosphates; Lupus Erythematosus, Systemic; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Mutant Strains; Phosphatidylinositols; T-Lymphocytes

In 24 patients with systemic lupus erythematosus the proliferating response of lymphocytes of the peripheral blood stream to mitogens PHA, ConA and PWM was examined. The spontaneous incorporation of 3H-thymidine was increased in two patients. The mean response to PHA and PWM was reduced, while the response to ConA was within the normal range. In the active stage of the disease the lymphocyte response was in general lower in the stage of low activity. There was no correlation between the number of CD4 and CD8 lymphocytes nor the CD4/CD8 index and the lymphocyte response to mitogens. In systemic lupus erythematosus the response to PWM was lower than in rheumatoid arthritis, while in rheumatoid arthritis the response to PHA and ConA was lower than in systemic lupus erythematosus.

Topics: Concanavalin A; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Phytohemagglutinins; Pokeweed Mitogens

Topics: Arthritis, Rheumatoid; C-Reactive Protein; Carbohydrates; Concanavalin A; Genetic Variation; Humans; Immunoelectrophoresis; Lupus Erythematosus, Systemic; Orosomucoid; Rheumatic Diseases; Spondylitis, Ankylosing

Levels of tumor necrosis factor/cachectin (TNF alpha) assessed by ELISA were similar in the supernatant of cultures from peripheral blood mononuclear cells (PBMC) from either active or inactive systemic lupus erythematosus (SLE) patients and controls stimulated with mitogen alone. When PBMC were stimulated with mitogen plus phorbol 12-myristate-13-acetate (PMA), the amount of TNF alpha was significantly decreased in culture supernatants from active patients or from the entire group of SLE patients studied. However, spontaneous synthesis of TNF alpha in nonstimulated cultures was increased in the SLE patients. Interferon-gamma (IFN-gamma) enhanced TNF alpha synthesis in cultures from either SLE patients or controls stimulated by concanavalin A (Con A) alone, but depressed the high production of TNF alpha by normal PBMC stimulated with Con A plus PMA. IFN-gamma enhanced TNF alpha production in response to Con A plus PMA in 2 of 3 SLE patients.

Topics: Concanavalin A; Enzyme-Linked Immunosorbent Assay; Humans; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Lymphocyte Activation; Phytohemagglutinins; Pokeweed Mitogens; Tumor Necrosis Factor-alpha

The properties of two types of K+ channels in murine T lymphocytes are described on the basis of whole-cell and isolated-patch recordings using the gigohm-seal technique. Type l (standing for "lpr gene locus" or "large") channels were characterized mainly in T cells from mutant MRL/MpJ-lpr/lpr mice, in which they are present in large numbers. Type n ("normal") K+ channels are abundant and therefore most readily studied in concanavalin A-activated T cells from four strains of mice, MRL-+/+, CBA/J, C57BL/6J, and BALB/c. Type l channels, compared with type n, are activated at potentials approximately 30 mV more positive, and close much more rapidly upon repolarization. Type l channels inactivate more slowly and less completely than type n during maintained depolarization, but recover from inactivation more rapidly, so that little inactivation accumulates during repetitive pulses. Type l channels have a higher unitary conductance (21 pS) than type n (12 pS) and are less sensitive to block by external Co++, but are 100-fold more sensitive to block by external tetraethylammonium (TEA), with half-block of type l channels at 50-100 microM TEA compared with 8-16 mM for type n. TEA blocks both types of channels by reducing the apparent single channel current amplitude, with a dose-response relation similar to that for blocking macroscopic currents. Murine type n K+ channels resemble K+ channels in human T cells.

Topics: Animals; Cations; Concanavalin A; Electric Conductivity; Humans; Ion Channels; Kinetics; Lupus Erythematosus, Systemic; Membrane Potentials; Mice; Mice, Inbred Strains; Mice, Mutant Strains; T-Lymphocytes; Tetraethylammonium Compounds

Concanavalin A (Con A)-activated T lymphocytes from patients with active, but not inactive, systemic lupus erythematosus (SLE) failed to express normal suppressor activity, regardless of the phenotype of CD4+ or CD8+. Con A-activated CD4+ or CD8+ T lymphocytes from the SLE patients and from normal controls were further separated into two populations, using the autologous erythrocyte rosette technique. One population very rich in cells capable of forming rosettes with autologous erythrocytes from the active patients showed the same degree of suppressor activity, as did that from normal controls; the CD4+ or CD8+ population poor in autorosetting cells derived from Con A-activated T lymphocytes from both the controls and patients did not express suppressor activity. Moreover, when autorosetting T cells from the active patients and nonrosetting cells from the same patients were mixed at a normal ratio (4:6), normal suppressor activity could be restored. It was notable that the frequency of autorosette-forming cells was markedly reduced in the Con A-activated T lymphocytes from the active, but not inactive, SLE patients, regardless of the phenotype of CD4+ or CD8+. These findings indicate the presence of a normally functioning suppressor T cell population in patients with active SLE. It seems that the lack of suppressor T cell function in patients with active SLE is due to the dilution of a few normal suppressor T cells by large numbers of nonsuppressor T lymphocytes.

Topics: Adult; Concanavalin A; Female; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Rosette Formation; T-Lymphocytes; T-Lymphocytes, Regulatory

We studied the suppressor cell activity induced by concanavalin A (Con A) in 9 patients with acute febrile juvenile rheumatoid arthritis (JRA). The suppressor activity of JRA patients was higher than that of normal controls. However, the activity was significantly reduced by treating Con A-activated cells with mitomycin C (MMC) (P less than 0.05). On the other hand, the suppressor activity of normal controls and systemic lupus erythematosus (SLE) patients was not affected by MMC treatment. Two of 5 SLE patients showed low activity even before MMC treatment. The addition of the culture supernatant of Con A-stimulated peripheral blood mononuclear cells from a normal donor restored the induction of suppressor activity of JRA which was decreased by MMC treatment. The results indicated that patients with acute febrile type of JRA had reduced MMC resistant suppressor cell activity and that this was due to a defect in the ability of the cells to produce soluble factors needed to induce MMC resistant suppressor cells.

Topics: Arthritis, Juvenile; Autoantibodies; Concanavalin A; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Reference Values; T-Lymphocytes, Regulatory

We have previously characterized the human B cell response to trinitrophenol (TNP)-Brucella abortus (Ba) response as being T cell independent. In this report we examine the role of monocytes in the TNP-Ba antibody response of human peripheral blood mononuclear cells (PBMC). Depletion of monocytes by sequential adherence to plastic and Sephadex G-10 passage did not result in decreased plaque-forming cell responses to TNP-Ba, suggesting that monocytes were not required. On the contrary monocytes were probably inhibitory because their removal resulted in enhanced responses. This was confirmed by showing that adding monocytes back reconstituted the inhibition. When interferon-gamma (IFN-gamma), a potent activator of monocytes, was added to TNP-Ba-driven PBMC cultures, marked inhibition (greater than 90%) of the responses ensued. This IFN-gamma-mediated suppression was monocyte dependent because it was completely abrogated by monocyte, but not T cell depletion. Previously, we described a concanavalin A (Con A), T cell inhibition pathway of the TNP-Ba response. Both the Con A and IFN-gamma pathways were tested for their ability to inhibit systemic lupus erythematosus (SLE) patient responses to TNP-Ba. The B cell response of SLE patients was inhibitable by both pathways. In all of the patients, the inhibition was complete (greater than 95%) when IFN-gamma was added to the cultures. In the presence of Con A, greater than 95% inhibition was observed in six of 10 patients, the remainder being inhibited to a lesser extent. Thus the hyperactive B cells from SLE patients can be down-regulated, particularly in the presence of IFN-gamma.

Topics: B-Lymphocytes; Binding, Competitive; Brucella abortus; Concanavalin A; Hemolytic Plaque Technique; Humans; Interferon-gamma; Kinetics; Lupus Erythematosus, Systemic; Lymphocyte Activation; Macrophage Activation; Monocytes; Nitrobenzenes; Prostaglandins; Trinitrobenzenes

An increased proportion of Ia positive circulating T cells was observed in patients with systemic lupus erythematosus (SLE) (17.2 +/- 5.8%, n = 20, p less than 0.005), compared with normal subjects (3.2 +/- 0.9%, n = 10) by the analysis with flow cytometry using OKIa1. There was a significant difference (p less than 0.05) in the proportion of Ia positive T cells (Ia +T cells) between inactive patients (19.6 +/- 5.8%, n = 12) and active patients (13.6 +/- 3.7%, n = 8). The kinetic analysis in normal subjects revealed that Ia expression on T cells stimulated with Concanavalin A (ConA) was significantly increased on day 4 (10.2 +/- 6.2%, n = 8). However, when stimulated with native DNA from calf thymus (nDNA), Ia expression was not increased (1.4 +/- 1.6%, n = 8). In SLE patients, initial Ia expression faded out within 4 days without stimulus (3.9 +/- 4.0%, n = 10). In spite of the decline of Ia expression with stimulation of both Con A and nDNA, the initial proportion of Ia positive T cells was relatively well maintained with the stimulation of nDNA (16.2 +/- 4.7%). Further analysis of T-cell subpopulations in SLE patients revealed that both OKT8- (T4) and OKT4- (T8) cells expressed Ia antigens. Interestingly, in 4 out of 5 patients with active disease, the incidence of Ia expression in OKT4- cells was increased with nDNA stimulation. Thus, in patients with SLE, circulating Ia positive T cells might be activated in vivo, and be implicated in the development of autoimmunity to DNA.

Topics: Antibodies, Monoclonal; Cells, Cultured; Concanavalin A; DNA; Female; Histocompatibility Antigens Class II; Humans; Kinetics; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; T-Lymphocytes

To investigate the role of interleukin 2 (IL-2) in systemic lupus erythematosus (SLE) mononuclear cells (MNC) of 68 SLE patients were tested for their ability to produce and also to respond to IL-2. Cells were collected monthly over an one year period. IL-2 production by MNC was measured under various conditions after optimal and suboptimal stimulation. Although we found a large variation in IL-2 production by individual MNC preparations no statistical significant differences were found between normal and SLE cells. To study IL-2 responsiveness, proliferation of MNC was studied under conditions where endogenous IL-2 production is limiting. Addition of IL-2 resulted in a four- to eight-fold enhancement of proliferative responses. However also in this respect no differences were found between SLE patients and healthy controls. Thus, in this group of SLE patients no abnormalities in IL-2 production or response could be demonstrated.

Topics: Adult; Aged; Concanavalin A; Female; Humans; Interleukin-2; Lupus Erythematosus, Systemic; Male; Middle Aged; T-Lymphocytes; Time Factors

Elevated production of anti-DNA antibody in patients with systemic lupus erythematosus (SLE) is a central problem in the pathogenesis of tissue injury. In the present study, we attempted to manipulate anti-DNA antibody production through the antigen-cytotoxic drug conjugates, DNA-daunorubicin complexes. The effect of DNA-daunorubicin complexes was determined by examining SLE lymphocytes for spontaneous in vitro production of anti-DNA antibody. These complexes, at 2 micrograms/ml, suppressed anti-DNA antibody production, but not total IgG production, which suggests that specific suppression of anti-DNA antibody production was achieved at this concentration. We believe that the DNA-daunorubicin complexes affected mainly B cells, since such suppression was obtained by treating B cells, as well as B plus T cells. Furthermore, the complexes had no effect on the proliferative responses of SLE T cells to DNA, phytohemagglutinin, or concanavalin A. These results indicate that DNA-daunorubicin complexes may have the potential for selectively suppressing anti-DNA antibody production in patients with SLE.

Topics: Antibodies, Antinuclear; Concanavalin A; Daunorubicin; DNA; DNA Adducts; Humans; In Vitro Techniques; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocytes; Phytohemagglutinins

The relationship of lectin-dependent cell-mediated cytotoxicity (LDCC) to interleukin-2 (IL-2) production was studied in healthy subjects and in patients with systemic lupus erythematosus (SLE). Profoundly depressed levels of LDCC were elicited by peripheral blood mononuclear cells (PBMC) from nine patients with active SLE in comparison to LDCC from seven controls, and eleven inactive SLE donors, using 3H-TdR-prelabelled adherent HEP-2 cells as targets in a 24 h assay with 25 micrograms/ml Con A. In parallel experiments, no individual correlation was found between LDCC activity and IL-2 production for healthy or SLE subjects. Further, no major differences were detected in IL-2 release when the three groups of donors were compared, a tendency observed at the Con A doses (5 and 25 micrograms/ml) and incubation times (24, 48, and 72 h) used to induce IL-2 production. In additional studies, impaired Con A-induced blastogenesis was noted for PBMC from active SLE patients in comparison to the PBMC from the controls or patients with inactive SLE. While strong individual correlation was obtained between blastogenesis and IL-2 secretion in controls and patients with inactive SLE, no such relationship was found in patients with active SLE. While addition of exogenous IL-2 to the cytotoxicity assay considerably enhanced LDCC by healthy donors it failed to improve LDCC by patients with active SLE. These data suggest that depressed LDCC and Con A-induced blastogenesis of patients with active SLE may not be related to impaired IL-2 production but rather to an inherent dysfunction of the effector lymphocytes, including their unresponsiveness to IL-2.

Topics: Concanavalin A; Cytotoxicity, Immunologic; Dose-Response Relationship, Immunologic; Female; Humans; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocyte Activation; Time Factors

Isoniazid (INH) and hydralazine (HYD) are transglutaminase (TGase, E.C.2.3.2.13.) substrates containing catalytically recruitable hydrazyl groups. Since they can be expected to inhibit TGase-mediated cell functions by competing with physiological substrates, their effect upon allogeneically and lectin-induced proliferation of mononucleocytes and upon zymosan-induced chemiluminescence of phagocytes was studied. Both compounds inhibited chemiluminescence in a dose-dependent manner. ID50 of HYD was consistently below 20 microM, while that of INH was above 120 microM. Proliferation of immunocompetent cells was suppressed by HYD with an ID50 of 60 microM, INH was inhibitory only above 5000 microM. Analogs of both compounds not containing hydrazyl groups proved to be inactive. Control experiments indicated that inhibition is not due to toxicity or lipophilicity of the compounds, structural analogs lacking a hydrazyl moiety were inactive. It is suggested that, in vivo, HYD interferes with signal-induced TGase-dependent leucocyte functions essential for immunologic stability, and that the resultant dysregulation with disruption of self tolerance contributes to the HYD promoted lupus-like syndrome.

Topics: Concanavalin A; Humans; Hydralazine; In Vitro Techniques; Isoniazid; Leukocytes; Luminescent Measurements; Lupus Erythematosus, Systemic; Lymphocyte Activation; Models, Biological; Syndrome; Transglutaminases

To better define the relationship between suppressor cell function and number and disease expression, the immunoregulatory profiles of 12 probands with systemic lupus erythematosus (SLE) and 34 of their asymptomatic family members were studied, using concanavalin A induced suppressor cells for functional analysis. SLE family members as a whole showed no impairment of mean suppressor levels, although 7 of 34 had altered suppression of DNA synthesis and 5 of 34 had altered suppression of IgG synthesis. Ratios of OKT4/T8 T cell subsets showed no difference between the study population, although 3 SLE family members had an increased ratio (greater than 2 SD) relative to controls. The 12 family members with either altered suppressor cell number or function had higher antibody levels to dRNA (Poly A . U) than did those with normal suppressor function and number. The results demonstrate that altered suppressor cell number and function occur in certain asymptomatic family members of SLE patients and may be weakly associated with markers of a preceding RNA viral infection.

Topics: Complement System Proteins; Concanavalin A; DNA; Female; Humans; Immunoglobulin G; Immunoglobulins; Lupus Erythematosus, Systemic; Male; Polynucleotides; T-Lymphocytes, Regulatory; Virus Diseases

Accessory function of monocytes for T-cell activation was studied in patients with systemic lupus erythematosus (SLE). Nylon column-purified T cells alone were not activated to proliferate by stimulation with concanavalin A (Con A), but the addition of dish-adherent monocytes restored the T-cell response in a dose-dependent manner (accessory function). This accessory function is mediated by HLA-DR-positive monocytes. This accessory function of monocytes was markedly impaired in SLE patients. The dysfunction of monocytes was marked in an active stage of SLE but not in an inactive stage of SLE. Furthermore, SLE T cells were not fully activated with Con A in the presence of normal monocytes, suggesting that both monocyte and T-cell functions were impaired in SLE patients. The dysfunction of SLE monocytes was due to neither the development of suppressor monocytes nor the overproduction of prostaglandins, because SLE monocytes did not suppress the accessory function of normal monocytes and indomethacin did not restore the dysfunction of SLE monocytes. The percentage of HLA-DR-positive cells in a monocyte population was markedly decreased in active SLE patients and moderately decreased in inactive SLE patients. Thus, the impairment of accessory function of monocytes in SLE patients seems to be derived from a decrease in HLA-DR-positive monocytes. These results suggest that the dysfunction of HLA-DR-positive monocytes plays an important role in the pathogenesis of SLE.

Topics: Autoantibodies; Concanavalin A; Histocompatibility Antigens Class II; HLA-DR Antigens; Humans; Leukocyte Count; Lupus Erythematosus, Systemic; Lymphocyte Activation; Monocytes; T-Lymphocytes, Regulatory

Alfalfa sprouts can induce systemic lupus erythematosus (SLE) in monkeys. This property of alfalfa sprouts has been attributed to their non-protein amino acid constituent, L-canavanine. Occurrence of autoimmune hemolytic anemia and exacerbation of SLE have been linked to ingestion of alfalfa tablets containing L-canavanine. In this report we show that L-canavanine has dose-related effects in vitro on human immunoregulatory cells, which could explain its lupus-inducing potential. These effects include: 1) diminution of the mitogenic response to both phytohemagglutinin and concanavalin A but not to pokeweed mitogen, as determined in both thymidine incorporation and cell cycle studies, and 2) abrogation of concanavalin A-induced suppressor cell function, which results in increased release of both IgG and DNA binding activity into supernatants by cells from normal subjects and SLE patients. These immunoregulatory effects of L-canavanine may explain the induction or exacerbation of SLE by alfalfa.

Topics: Adult; Autoantibodies; B-Lymphocytes; Canavanine; Cell Cycle; Cell Survival; Concanavalin A; DNA; Humans; Immunoglobulin G; Lupus Erythematosus, Systemic; Lymphocyte Activation; Medicago sativa; Plant Lectins; T-Lymphocytes; T-Lymphocytes, Regulatory

The proliferation of peripheral blood lymphocytes, cultured with Con A, can be inhibited by ionizing radiation. Lymphocytes from patients with conditions associated with autoimmunity, such as rheumatoid arthritis, systemic lupus erythematosus and polymyositis, are more radiosensitive than those from healthy volunteers or patients with conditions not associated with autoimmunity. The nuclear material isolated from the lymphocytes of patients with autoimmune diseases is, on average, lighter in density than the nuclear material from most healthy controls. This difference in density is not related to increased sensitivity to ionizing radiation but the degree of post-irradiation change in density (lightening) is proportional to the initial density, i.e. more dense nuclear material always shows a greater upward shift after radiation. The recovery of preirradiation density of nuclear material, 1 h after radiation exposure, taken as an indication of DNA repair, correlates with the radiosensitivity of lymphocyte proliferation (Con A response); failure to return to pre-irradiation density being associated with increased sensitivity of proliferative response. These results require extension but, taken with previously reported studies of the effects of DNA methylating agents, support the idea that DNA damage and its defective repair could be important in the aetio-pathogenesis of autoimmune disease.

Topics: Adult; Aged; Arthritis, Rheumatoid; Autoimmune Diseases; Cell Division; Cell Nucleus; Concanavalin A; DNA; DNA Repair; Humans; Lupus Erythematosus, Systemic; Lymphocytes; Middle Aged; Myositis; Radiation Tolerance

The effect of thymopoietin penta- (TP-5), tetra-(TP-4), and tripeptides (TP-3) was studied on the depressed lectin-dependent cell-mediated cytotoxicity (LDCC) against adherent HEp-2 target cells by peripheral blood mononuclear cells from patients with active systemic lupus erythematosus (SLE). LDCC activity was evaluated by detachment from the monolayer of 3H-thymidine-prelabelled HEp-2 cells in the presence of concanavalin A (Con A). While 10(-5)M TP-3 moderated the depression, 10(-5) M TP-5 strongly enhanced LDCC activity in SLE patients up to the normal level. On the other hand, LDCC activity by normal donors was not influenced by TP-3 and TP-5. TP-4 had no major effect either in control or in SLE patients. In parallel experiments none of the thymopoietin peptides affected the Con A-induced suppressor activity on the blastogenesis of lymphocytes. A selective immunostimulatory effect of TP-3 and TP-5 on the generation of LDCC effector cells in patients with SLE is suggested.

Topics: Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; In Vitro Techniques; Lupus Erythematosus, Systemic; Lymphocyte Activation; Peptide Fragments; Thymopentin; Thymopoietins; Thymus Hormones

Zymosan-stimulated neutrophils from 6 patients with untreated, active systemic lupus erythematosus (SLE), from patients with bacterial infections, and from healthy controls, were studied for production of oxygen intermediates (O-2, H2O2, OH . and chemiluminescence) and lysosomal enzymes. Oxygen intermediate production was highest in neutrophils from active SLE patients, while lysosomal enzyme release was highest in neutrophils from patients with bacterial infections. SLE neutrophils, upon culture with autologous or normal lymphocytes, markedly reduced the number of surviving OKT4+ cells and the proliferative response of the surviving cells to mitogens; a reduction was also observed amongst the surviving lymphocytes in the proportion of total T cells and OKT8+ cells, and in the generation of Con A induced suppressor activity. When superoxide dismutase and catalase were included in the neutrophil-lymphocyte co-cultures, the number of T- and OKT8+ cells, and the suppressor activity were restored but not completely (60-75%), the lymphocyte mitogenic response and number of OKT4+ cells were less well restored (40-50%). When lymphocytes were co-cultured with neutrophils from healthy or infected subjects, there was a mild decrease in mitogenic responses and OKT4+ cells, while the suppressor T-cell activity was markedly enhanced. These results were not affected by scavengers. These results suggest that in SLE, reduced T-lymphocyte subpopulations and altered immunoreactivity may be partially due to excessive production of oxygen intermediates and probably other factors by stimulated neutrophils; these results further suggest that in all the subjects, diseased or healthy, neutrophils generate unidentified factors other than oxygen intermediates that reduce the generation of OKT4+ cells and lymphocyte mitogenic responses, and that potentiate suppressor T-cell activity.

Topics: Adolescent; Adult; Concanavalin A; Female; Humans; In Vitro Techniques; Lupus Erythematosus, Systemic; Lymphocyte Activation; Neutrophils; Oxygen; Superoxides; T-Lymphocytes, Regulatory

Interleukin-1 (IL-1) is a monocyte product with diverse amplifying effects on immune cell reactions. We have studied 16 untreated SLE patients to determine the production of IL-1 by their monocytes under the stimulus of E. Coli lipopolysaccharide (LPS) or phorbol myristate acetate (PMA) and measured by the capacity of their supernatants to augment normal autologous mixed lymphocyte cultures (AMLR) or to replace accessory cells in Con A-induced proliferation of T lymphocytes. Concurrently, we studied the response of T lymphocytes from these same patients to IL-1 by its capacity to increase the percentage of stable E rosette forming cells and by the enhancement of T cell proliferation in AMLR. Monocytes from SLE patients produced significantly less IL-1 activity than those of age matched controls, regardless of the stimulus (LPS or PMA), as well as of the indicator system. All patients with active disease and seven of the 10 patients with inactive disease had decreased production of IL-1 activity as determined by at least one method. Response of T lymphocytes from SLE patients to IL-1 produced by normal monocytes was also found decreased as compared to normals. This defect was more marked in the T cells from patients with active than in those of patients with inactive disease. These findings indicate that the immunoregulatory disturbance that SLE patients have encompasses monocytes as well as T and B lymphocytes and suggest that the defect is either multicentric or originates in the stem cell.

Topics: Adult; Cell Division; Concanavalin A; Female; Humans; Interleukin-1; Lipopolysaccharides; Lupus Erythematosus, Systemic; Lymphocyte Culture Test, Mixed; Monocytes; Rosette Formation; T-Lymphocytes; Tetradecanoylphorbol Acetate

To determine whether patients with systemic lupus erythematosus (SLE) and active nephritis have more profound defects in suppressor cell activity, we studied concanavalin A (Con A)-induced suppressor cell activity (SCA) in 12 patients with lupus nephritis (LN) and 11 patients with chronic mesangial proliferative glomerulonephritis (CGN) without renal insufficiency. The levels of Con A-induced SCA were decreased in patients with LN compared with those in normal controls and those in CGN patients and lower in LN patients with the nephrotic syndrome (NS) than in those without NS. In contrast, the mean responses of Con-A-induced SCA in CGN patients with or without NS did not differ from normal subjects. These findings may lend further insight into the understanding of the immunoregulatory defect in LN.

Topics: Adolescent; Adult; Cell Communication; Concanavalin A; Female; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Nephritis; T-Lymphocytes, Regulatory

The role of OKT4+ and OKT8+ T cell subsets was studied in depressed lectin-dependent cell-mediated cytotoxicity (LDCC) against adherent HEp-2 target cells by peripheral blood mononuclear cells (PBMC) from patients with active systemic lupus erythematosus (SLE). LDCC activity was evaluated by detachment from the monolayer of 3H-TdR-prelabelled HEp-2 cells in a 24 hr assay at 50:1 effector-target cell ratio in the presence of 25 micrograms/ml concanavalin A (Con A). Decreased levels of LDCC were performed by all studied effector cell populations of SLE patients, including both OKT4+ and OKT8+ T cell fractions. LDCC by isolated OKT8+ T cells was superior to that by OKT4+ and unfractionated T lymphocytes from all healthy and SLE subjects. This suggests that the defect of LDCC activity in SLE did not affect the inherently higher LDCC effector activity of OKT8+ to OKT4+ cells. In parallel studies a reduced proliferation of PBMC in response to Con A and failure of OKT8+ T cells to suppress Con A-induced blastogenesis was observed in patients with SLE.

Topics: Adult; Antibodies, Monoclonal; Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; In Vitro Techniques; Lupus Erythematosus, Systemic; Lymphocyte Activation; Middle Aged; T-Lymphocytes; T-Lymphocytes, Cytotoxic; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

Plasma levels of C3 and C3d in 30 healthy donors and 58 patients with systemic lupus erythematosus were determined by rocket immunoelectrophoresis and Con-A affinoimmunoelectrophoresis. Significantly decreased levels of C3 were found in approximately 50% of the SLE sera. Slight increases of free C3d were found in 55%, and marked increases in 9% of the SLE sera; the remaining SLE sera did not show increased C3d levels. When results were expressed as quotients of plasma C3d/C3, a significant pattern emerged and 70% of the SLE sera clearly fell outside the normal range. 10% of the SLE sera had normal C3d/C3 indices but had markedly depressed C3 levels. We conclude that in cases where total serum C3 is not markedly reduced (less than 60% of normal), the C3d/C3 quotient may be a more sensitive parameter for assessing in vivo complement activation than C3 or C3d determinations alone.

Topics: Complement C3; Complement C3d; Concanavalin A; Enzyme Activation; Humans; Immunoelectrophoresis, Two-Dimensional; Lupus Erythematosus, Systemic; Reference Values

The ability of peripheral blood lymphocytes from patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and Sjogren's syndrome (SS) to produce interleukin 2 (IL-2) and respond to it in-vitro was examined. Phytohemagglutinin-stimulated lymphocytes from over half of the SLE patients exhibited a decreased ability to produce IL-2 while their concanavalin A-generated blast cells responded normally to exogenous IL-2. Lymphocytes from RA patients not only produced less IL-2 than normals (P less than 0.001), but also responded poorly to exogenous IL-2 (P = 0.011). These abnormalities did not correlate with the patient's age, sex, duration of disease, or disease activity. Production of and response to IL-2 was widely varied among patients with SS and not different from controls. The decreased response of RA lymphocytes to IL-2 may result from a smaller number of cell surface IL-2 receptors since IL-2 adsorption to RA cells was lower than to either SLE or normal cells. These data suggest that IL-2-related abnormalities may play a role in the disordered immunoregulation characteristic of RA and perhaps of SLE.

Topics: Absorption; Adult; Arthritis, Rheumatoid; Concanavalin A; Female; Humans; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Receptors, Immunologic; Receptors, Interleukin-2

Palmerston North (PN) mice, a newly recognized model of systemic lupus erythematosus, were compared with autoimmune hybrid NZB/NZW mice in a study designed to examine spleen cell responsiveness to T-cell and B-cell mitogens. Modest reductions of responses to phytohemagglutinin (PHA) and concanavalin A (Con A) were noted in PN females after 24 weeks of age; these responses were reduced significantly in NZB/NZW females. In contrast, male PN and NZB/NZW mice responded actively to PHA and Con A throughout the first year of life. Responses to lipopolysaccharide were not affected by age or sex. Anti-DNA antibody levels, blood urea nitrogen, and glomerular histology were analyzed to determine if autoantibody production or renal failure correlated with suppressed mitogenic responsiveness. These factors, examined singly and together, were not as important as age. In this system, age and sex did not influence spleen cell responses to mitogens in normal CD-1 mice. Age and sex were of minimal importance in determining responses to T-cell mitogens in the recently defined PN model of autoimmunity. In contrast, age and sex exerted strong influences upon responses to PHA and Con A in the NZB/NZW model of lupus.

Topics: Age Factors; Animals; Autoantibodies; B-Lymphocytes; Concanavalin A; DNA; Female; Glomerulonephritis; Lipopolysaccharides; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Mice; Phytohemagglutinins; Sex Factors; Spleen; T-Lymphocytes

In patients with rheumatoid arthritis (RA) a significantly decreased autologous mixed lymphocyte reaction (AMLR) was observed which varied widely and did not correlate with disease activity, clinical course or treatment schedules. When supernatants of AMLR cultures were tested for the presence of soluble factors no differences were found in regard to the suppression of allogeneic and mitogen induced lymphocyte proliferation. Furthermore both test groups failed to produce detectable amounts of interferon during the course of the AMLR, in contrast to the allogeneic situation were both RA patients and normal controls exhibited a similar interferon activity in the culture supernatants.

Topics: Adult; Aged; Antigens; Arthritis, Rheumatoid; Autoantigens; Concanavalin A; Humans; In Vitro Techniques; Interferons; Isoantigens; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Middle Aged; T-Lymphocytes

Isoprinosine (IPS) is a new anti-viral agent which appears to have immunomodulatory activities which include its ability to enhance the in vitro blastogenic responses of normal lymphocytes to mitogens. The present study compares the effects of IPS on the in vitro immune functions of peripheral blood mononuclear cells (PBMC) from systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) patients with its effects on PBMC from normal controls. Each mitogen (Con A, PHA or PWM) was used at its optimal concentration with a range of IPS concentrations (0-25 micrograms/ml). PHA-induced blastogenesis by PBMC from all three groups was enhanced by IPS at or above 5 micrograms/ml. The Con A-induced responses of SLE lymphocytes were significantly enhanced over controls by IPS (P less than 0.02 at 5 micrograms/ml) while those of RA lymphocytes were not. IPS had little effect on PWM-induced blastogenesis by RA lymphocytes but did enhance the blastogenic responses of SLE lymphocytes (P less than 0.01 at 5 micrograms/ml). In contrast, the characteristically high immunoglobulin synthesis by SLE lymphocytes was decreased by IPS. The mechanism responsible for these effects is not known but IL-2 production by patient lymphocytes in vitro which was low for both RA (P less than 0.01) and SLE (P less than 0.02) increased significantly (P less than 0.05) when SLE lymphocytes were cultured with IPS. These data identify IPS as an agent for the study of aberrant immune regulation in autoimmune diseases and suggest that it may have potential therapeutic value in SLE.

Topics: Adult; Arthritis, Rheumatoid; Autoimmune Diseases; Cells, Cultured; Concanavalin A; Humans; Immunoglobulins; Inosine; Inosine Pranobex; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocytes; Middle Aged; Mitosis; Phytohemagglutinins

Con A induced suppressor cell assay was further characterized in this study. Significant lower mitogenic responses of autologous fresh cells cocultured with Con A-activated cells were found when compared with the response of cocultures of responders plus control cells preincubated in medium alone. However, Con A-activated cells did not express a real suppression, since no difference was found between responses of fresh responders alone and responses of cocultures with Con A-activated cells. So, Con A activation seemed to block the expression of the enhancement provided by control cells to autologous responders, rather than to induce a real suppressor activity. The induction of the Con A suppressor cell activity required cell proliferation but it was not proportional to the degree of DNA synthesis. Monocyte depleted cell populations exhibited lower Con A suppressor cell activity compared to unfractionated cells, suggesting a cooperative role of monocytes in the Con A induction step. The expression of this activity was not due to a cytotoxicity against the responders and was dependent on the number of activated cells added. Mononuclear cells from active lupus erythematosus (SLE) patients, but not from inactive SLE patients differed from normal controls in showing a significant loss of suppression index.

Topics: Adult; Concanavalin A; Dose-Response Relationship, Drug; Humans; Lupus Erythematosus, Systemic; Middle Aged; Mitomycin; Mitomycins; Monocytes; T-Lymphocytes, Regulatory

Patients with systemic lupus erythematosus (SLE) have decreased precursors of cytotoxic/suppressor T lymphocytes in their peripheral blood, as determined by monoclonal antibodies. To determine whether decrease of the cytotoxic or the suppressor parts (or both) of this subpopulation of T lymphocytes is being reflected by this peripheral mononuclear cell (MNC) abnormality, a series of experiments was conducted in which both the suppressive function (concanavalin A induced and spontaneous) and the generation of cytotoxic responses against alloantigens were tested. Cytotoxic responses were consistently diminished while suppressor capacity of MNC from patients with SLE (measured on several assays of normal T- and B-lymphocyte functions) was comparable to that of MNC from normal individuals. The defect in cytotoxic responses to alloantigens by MNC from SLE patients persisted following secondary stimulation in mixed-leukocyte cultures; the cytotoxic responses were not amplified and remained well below the responses of normal MNC. These experiments indicate that the decreased peripheral population of cytotoxic/suppressor lymphocytes in SLE patients represents low or absent precursors of cytotoxic cells rather than of precursors of suppressor cells.

Topics: Antibodies, Monoclonal; Complement C3; Concanavalin A; Humans; Isoantigens; Lupus Erythematosus, Systemic; Lymphocyte Culture Test, Mixed; Phenotype; Rosette Formation; T-Lymphocytes; T-Lymphocytes, Cytotoxic; T-Lymphocytes, Regulatory

Pregnant systemic lupus erythematosus (SLE) patients with inactive disease were found to have normal spontaneously generated suppressor-cell function and slightly higher concanavalin A-induced suppressor function as compared to matched normal pregnant and nonpregnant females. In six SLE patients studied sequentially throughout pregnancy and postpartum, suppressor functions were found to fall sharply within the first week after delivery. One of these patients had been studied before she became pregnant and found to have a decreased suppressor function. Nonpregnant SLE patients had both suppressor functions diminished despite their disease being similarly inactive. This group was also the only one to have decreased responses in autologous mixed-lymphocyte cultures. Both pregnant and nonpregnant SLE patients had decreased absolute numbers of total lymphocytes, T cells, and their subpopulations, but the proportions of these cells were similar in all four groups. Despite this apparent normalcy of immune regulation, pregnant SLE patients had higher levels of Clq-binding immune complexes than did nonpregnant ones. Functional T-cell abnormalities found in SLE patients tend to be corrected by pregnancy. This may explain in part the disease remissions that occur in them during the second half of pregnancy.

Topics: Concanavalin A; Female; Humans; Immunoglobulin G; Immunosuppression Therapy; Lupus Erythematosus, Systemic; Lymphocyte Culture Test, Mixed; Postpartum Period; Pregnancy; Pregnancy Complications; Receptors, Fc; Rosette Formation; T-Lymphocytes; T-Lymphocytes, Regulatory

The loss of suppressor T-cell function results in an abundant production of autoantibodies in systemic lupus erythematosus (SLE). As a cause of this suppressor T-cell defect, anti-T-cell antibody seems to be of prime importance. On the other hand, anti-T-cell antibodies can be detected in various other autoimmune diseases, but their functional characteristics have not been determined. In the present study, the functional characteristics of anti-T-cell antibody from a selected subgroup of patients with ulcerative colitis (UC) were compared with those from patients with SLE. Anti-T-cell antibody from the patients with SLE reacted with a T8 subset, resulting in a suppressor defect, whereas anti-T-cell antibody from the UC patients reacted primarily with a T4 subset. Functionally, SLE- T cells failed to proliferate in response to concanavalin A, whereas UC- T cells from UC patients failed to proliferate in response to phytohaemagglutinin. In the Ig synthesis system, both SLE- and UC- T cells increased Ig production of B cells. Since UC+ T cells did not contribute to the generation of Con-A-inducible suppressor activity, we believe that serum from the selected subgroup of patients with UC reacted with the inducer T-cell subset.

Topics: Adult; Antibodies, Monoclonal; Antibody-Dependent Cell Cytotoxicity; Autoantibodies; Colitis, Ulcerative; Complement System Proteins; Concanavalin A; Female; Humans; Lupus Erythematosus, Systemic; Lymphocyte Culture Test, Mixed; Male; Middle Aged; Phytohemagglutinins; Pokeweed Mitogens; T-Lymphocytes

B cell hyperactivity, a feature common to all lupus-prone murine strains, may be caused by hyperresponsiveness to, overproduction of, or bypassing of certain signals required for B cell activation, proliferation, and differentiation. In this study, we have compared the responses of B cells from three lupus-prone strains of mice (BXSB males, MRL and NZB/W females) and normal strains in a number of assays for which two or more signals are required to obtain a response. In medium to low density cultures of B cells from BXSB and NZB/W but not MRL/l lupus mice, the cells' proliferation induced by bacterial lipopolysaccharide (LPS) or anti-mu antibody was much higher than that of B cells from normal controls. At low B cell density, polyclonal activation by these substances and subsequent Ig secretion were dependent on accessory signals present in supernatants of concanavalin A-treated normal lymphocytes (CAS) or on the MRL/l proliferating T cell-derived B cell differentiation factor (L-BCDF) in both lupus-prone and immunologically normal mice. However, the responses of B cells from BXSB and NZB/W, but not MRL/l, mice to these accessory signals were higher than those of normal mice. Ig synthesis by fresh B cells of BXSB and NZB/W mice cultured in the absence of mitogens but in the presence of CAS or L-BCDF was higher than by similar cells from other strains, suggesting an increased frequency of B cells activated in vivo in these two autoimmune strains of mice. The patterns of IgG subclass secretion in response to LPS (without added CAS or L-BCDF) were abnormal in all lupus strains, with a predominance of IgG2b and/or IgG2a and low levels of IgG3, contrary to normal B cells for which IgG3 synthesis predominated. However, IgG1 synthesis in vitro by autoimmune and normal B cells alike was highly dependent on T cell-derived soluble mediators. Antigen-specific responses to SRBC in vitro of B cells from all lupus strains, like those of B cells from normal strains, required a minimum of three signals (antigen, LPS, T cell-derived antigen nonspecific helper factors). Yet, once triggered, B cells of BXSB and NZB/W mice gave higher responses than those of the other strains. We conclude that B cells of lupus mice have signal requirements similar to those of normal mice. Nevertheless, B cells of BXSB and NZB/W, but not MRL/l, lupus mice hyperrespond or process some accessory signals abnormally.

Topics: Animals; Antibodies; B-Lymphocytes; Cell Differentiation; Cell Division; Cells, Cultured; Concanavalin A; Female; Immunoglobulin G; Immunoglobulin mu-Chains; Immunoglobulins; Lipopolysaccharides; Lupus Erythematosus, Systemic; Male; Mice; Mice, Inbred NZB; Mice, Inbred Strains

Lymph node and spleen cells of the autoimmune MRL/Mp-lpr/lpr mouse strain spontaneously produce (in the absence of mitogenic stimulation) a factor(s) that induces B cell differentiation. This factor is not produced by the congenic MRL/n mouse strain that lacks the lpr gene or by normal mouse strains. However, lymphoid cells of the B6-lpr/lpr (B6/1) strain also produce a B cell differentiation factor. Although the factor acts on resting B cells, its effect is greatly magnified by activating the B cells with anti-mu or lipopolysaccharide. MRL/l mice begin producing the factor as early as 1 mo of age but levels increase with age and appearance of lymphoproliferation. Cell depletion studies reveal that this factor is produced by T cells of the Lyt-1+2-phenotype. Because of its association with the lpr/lpr genotype, we term this B cell differentiation factor L-BCDF. Functional analysis of L-BCDF reveals that it acts regardless of cell density in culture and in the absence of interleukin 2 (IL-2). In fact, the increase in the production of L-BCDF by MRL/1 T cells with aging occurs concomitantly with a marked decrease in their ability to produce IL-2. No T cell replacing factor activity or B cell growth factor-like activity can be detected in MRL/l-derived supernatants. L-BCDF induces both IgM and IgG synthesis in lipopolysaccharide-activated B cells; however, it has a greater effect on IgG secretion. In particular, the production of IgG1, IgG2a, and IgG2b are markedly enhanced in the presence of L-BCDF. The spontaneous production of L-BCDF by T cells of SLE mice of lpr/lpr genotype suggests an association of this factor with autoimmunity.

Topics: Aging; Animals; Autoimmune Diseases; B-Lymphocytes; Concanavalin A; Female; Genotype; Growth Substances; Immunoglobulin G; Immunoglobulin M; Interleukin-2; Interleukin-4; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphokines; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Mutant Strains; T-Lymphocytes

The cytotoxic activities of individual lupus sera were demonstrated against T gamma cells, T gamma (-) cells and B cells. Immunoglobulin classes were also determined in these antilymphocyte antibodies (ALA) in SLE. T gamma-specific ALA and B-cell-specific ALA were almost equally distributed as regards IgG-dominant type, mixed IgG and IgM type and IgM dominant type, while IgM dominant type was predominant in T gamma (-)-specific ALA. Only the IgG type of T gamma-specific ALA among these ALA was significantly associated with clinical parameters, including hypocomplementemia, elevated immune complex levels and high anti-double-stranded (ds) DNA titres. Serial studies on ALA in a typical case of SLE were performed. Active clinical signs were associated with elevated cytotoxicities of IgG type of T gamma-specific ALA and with relatively low cytotoxicities of the IgM type, whereas after massive corticosteroid therapy, these signs disappeared in tact with the reduction in cytotoxic activities of IgG type of the ALA and the relative rise in cytotoxicities of IgM type of the ALA. These results suggested that IgG class of T gamma-specific ALA seemed to play a major role among several types of ALA in SLE.

Topics: Antibodies, Antinuclear; Antigen-Antibody Complex; Antilymphocyte Serum; B-Lymphocytes; Complement System Proteins; Concanavalin A; Cytotoxicity, Immunologic; DNA; Humans; Immunoglobulin G; Immunoglobulin M; Lupus Erythematosus, Systemic; Staphylococcal Protein A; T-Lymphocytes

We studied the distribution of autologous rosette forming cells (ARFC) in the peripheral blood of 30 healthy adult donors, 30 patients with IgA nephropathy, 20 patients with primary glomerular diseases, eight patients with systemic diseases and 25 patients with other renal diseases. The mean percentages of ARFC were markedly reduced in the IgA nephropathy patients compared with the healthy adult donors. The values for ARFC were even more significantly reduced in IgA nephropathy patients compared with patients with primary glomerular diseases, systemic diseases and other renal diseases. This means that the immunoregulatory abberation in IgA nephropathy primarily involves T cells.

Topics: Adult; Concanavalin A; Erythrocytes; Humans; IgA Vasculitis; Immunoglobulin A; Kidney Diseases; Kidney Glomerulus; Lupus Erythematosus, Systemic; Lymphocyte Activation; Rosette Formation; T-Lymphocytes

Spontaneous cytotoxicity mediated by natural killer (NK) cells is impaired in several human diseases including systemic lupus erythematosus (SLE). The precise mechanism(s) by which NK activity is suppressed in patients with SLE is generally unknown. The present study was designed to focus on cellular defects per se in NK cells from patients with SLE. It was observed that the usual enhancing effect of interferon (IF) and IF inducers was markedly impaired in SLE patients. Of 24 SLE patients studied, 17 had significantly decreased NK activity relative to controls. NK activity had a significant negative correlation with clinical activity score (r = -0.56, P less than 0.005) but was not correlated with corticosteroid dose, antinuclear antibody titers, total hemolytic complement (CH50), or sedimentation rate. Furthermore, significant depressions in NK activity correlated with variations in disease activity in six patients followed serially. Depressed NK function could not be reversed by prolonged in vitro incubation at 37 degrees C or with protease treatment. Furthermore, depressed NK activity was not altered by removal of glass adherent cells nor was a suppression of NK activity in normal controls seen by the addition of SLE peripheral mononuclear cells. No reversal of depressed activity to normal levels was seen by the addition of indomethacin nor did the supernatants from SLE cell cultures cause a suppression of normal NK function. NK activity in SLE patients did not respond normally to IF inducers (poly-I:C and concanavalin A) even if the SLE patients had normal NK function. The response of SLE cells to exogenous IF was also impaired. The number of effector-target conjugates was quantitated with several target cells (K562, Yac-1, Fravel) in SLE patients and controls. A significant correlation between the proportion of glass nonadherent mononuclear cells that formed effector-target conjugates with these various targets and the magnitude of NK lysis was observed. However, SLE and normal subjects had equal numbers of effector-target conjugates independent of NK function. Release of a soluble cytotoxic factor was induced with concanavalin A, and was markedly impaired in SLE patients relative to normal controls. Thus, impaired NK cell function in SLE does not appear to be related to cell-mediated suppressive mechanisms or to the deletion of effector cells; rather, the decreased NK activity may be related to an impaired release of a soluble cytotoxic factor.

Topics: Adult; Aged; Cells, Cultured; Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; Indomethacin; Interferon Inducers; Interferon Type I; Killer Cells, Natural; Lupus Erythematosus, Systemic; Middle Aged; Poly I-C

Serum obtained from patients with spontaneous systemic lupus erythematosus or from patients treated at least 3 months with procainamide could specifically inhibit Con A mitogenesis of cultured peripheral blood mononuclear cells obtained from procainamide-treated patients or from normal donors. Transfer of procainamide treated patients to N-acetylprocainamide eliminated the blocking factor from their serum. The blocking factor is not procainamide itself since adding the drug to normal serum and only slight effects on mitogenesis of normal peripheral blood mononuclear cells. These data suggest that systemic lupus erythematosus and procainamide-induced lupus may differ in the relative reversibility of a regulatory defect associated with a Con A responsive population of peripheral blood mononuclear (PBM) cells.

Topics: Acecainide; Antibodies; Binding, Competitive; Concanavalin A; Humans; Immunoglobulin G; In Vitro Techniques; Lupus Erythematosus, Systemic; Lymphocyte Activation; Procainamide

Addition of prostaglandin E2 (PGE2) to blood mononuclear cell cultures containing pokeweed mitogen (PWM) enhances plasma cell (PC) differentiation measured by intracytoplasmic immunoglobulin 7 days later. T-cell mitogenesis to concanavalin A is inhibited using the same concentrations of PGE2. PGE2 failed to enhance the PC differentiation of lymphocytes from patients with systemic lupus erythematosus (SLE). Indomethacin, on the other hand, either had no effect or suppressed PC differentiation. The data is discussed in terms of the effect of PGE2 on human suppressor T-cell function.

Topics: Adult; Cell Differentiation; Concanavalin A; Dinoprostone; Humans; Indomethacin; Lupus Erythematosus, Systemic; Middle Aged; Plasma Cells; Pokeweed Mitogens; Prostaglandins E; T-Lymphocytes, Regulatory

Topics: Antibody-Producing Cells; Cells, Cultured; Concanavalin A; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Pokeweed Mitogens; T-Lymphocytes, Regulatory

Suppressor cell activity (SCA) was analyzed in 8 patients with idiopathic membranous nephropathy (MN) and in 11 patients with chronic proliferative glomerulonephritis (CGN). We have assessed the ability of peripheral blood lymphocytes (PBL) stimulated by concanavalin A (Con A) to inhibit the proliferative response on normal allogenic lymphocytes by both Con A and phytohemagglutinin (PHA). It was found that the MN patients with nephrotic syndrome (NS) had significantly increased levels of suppression index (SI) when compared to the values obtained with normal controls. In contrast, the mean suppression values in the PBL from MN patients in remission and CGN patients with or without NS, whether the mitogen used was Con A or PHA, were similar to those of the control subjects. Thus, the majority of MN patients wih NS demonstrated an alteration in Con-A-induced SCA. The possible significance of these phenomena in the pathophysiology of MN is discussed.

Topics: Adult; Chronic Disease; Concanavalin A; Dose-Response Relationship, Immunologic; Female; Glomerulonephritis; Humans; Immunoglobulin A; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Middle Aged; T-Lymphocytes, Regulatory

Human peripheral blood lymphocytes (PBL) are activated to become suppressor T cells (S-T-C) by incubation with Concanavalin-A (Con-A). This has become the standard method for evaluation of suppressor function in patients. S-T-C function has been found to be impaired in several autoimmune diseases, including systemic lupus erythematosus (SLE). Using this assay, we have investigated suppressor-cell function in 2 autoimmune disorders, bullous pemphigoid (BP) and pemphigus vulgaris (PV), studying 6 patients from each group. Three patients with active SLE (positive controls), and 11 normal donors (negative controls) were also included. None of these patients had received systemic therapy with the exception of 2 patients with PV who were treated with gold in the past. PBL from these patients were incubated with and without 40 micrograms/ml Con-A for 72 hr to generate suppressor cells. Both groups of PBL were then irradiated wih 1500 r cobalt. Co-cultures were set up in sextuplicate using normal PBL as responders. Responder PBL were stimulated with 0.5, 1.0, and 2.0 micrograms/ml of phytohemagglutin (PHA) and 5.0, 10.0, and 20.0 micrograms/ml of Con-A. Cultures were pulsed on day 3 with 3H-thymidine and harvested on day 4. Data were analyzed using Student's t-test. S-T-C function was found to be significantly impaired in SLE vs normal (p = 0.0316). No statistically significant difference was seen in BP (p = 0.5883) and PV (p = 0.0921) as compared with normals. A defect in suppressor cell function may still be present in patients with PV and BP for the defect may be antigen-specific and therefore remain undetected by the Con-A suppressor assay.

Topics: Cobalt; Concanavalin A; Culture Techniques; Fluorescent Antibody Technique; Humans; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocytes; Pemphigoid, Bullous; Pemphigus; Phytohemagglutinins; Skin Diseases, Vesiculobullous; T-Lymphocytes, Regulatory

Topics: Adult; Antibody-Producing Cells; B-Lymphocytes; Concanavalin A; Cytotoxicity Tests, Immunologic; Female; Hemolytic Plaque Technique; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Male; Middle Aged; Pokeweed Mitogens; T-Lymphocytes, Regulatory

The pathogenesis of procainamide-induced autoantibody production is unknown. To test the effect of procainamide on the immune system, we studied in vitro suppressor cell function and immunoglobulin G (IgG) secretion in 11 patients who developed autoantibodies while taking procainamide. The procainamide group was compared with patients with spontaneous systemic lupus erythematosus (n = 15) and a normal control population (n = 40). Impaired in vitro suppressor cell function was found in 11 of 14 patients with spontaneous systemic lupus erythematosus but in none of the patients taking procainamide. However, total in vitro IgG secretion was significantly increased in the procainamide group with regard to the control and systemic lupus erythematosus groups. There was a direct correlation between the circulating anti-SS DNA antibody titer and in vitro IgG secretion. Furthermore, T cells isolated from the procainamide-treated patients stimulated IgG secretion by normal allogeneic peripheral blood lymphocytes. The added T cells did not affect in vitro suppressor cell function. We postulate that autoantibody production in patients taking procainamide is due to enhanced helper T cell function and not to impaired suppression. However, the development of clinical disease requires the participation of additional genetic or immunologic factors.

Topics: Adolescent; Adult; Aged; Antibodies, Antinuclear; Autoantibodies; Concanavalin A; DNA, Single-Stranded; Female; Humans; Immunoglobulin G; In Vitro Techniques; Lupus Erythematosus, Systemic; Male; Middle Aged; Pokeweed Mitogens; Procainamide; Radioimmunoassay; T-Lymphocytes; T-Lymphocytes, Regulatory

Topics: Adolescent; Adult; Chronic Disease; Concanavalin A; Female; Glomerulonephritis; Humans; Lupus Erythematosus, Systemic; Male; Middle Aged; Phytohemagglutinins; T-Lymphocytes, Regulatory

Suppressor-cell activity of 26 SLE patients suffering from active disease was compared to that of 15 healthy controls. ConA-induced and spontaneous suppression was evaluated. The mitogen-driven proliferation of normal allogeneic cells was significantly impaired by ConA-induced as well as spontaneous suppressor cells. However, no difference in suppressor-cell activity could be demonstrated between SLE patients and controls.

Topics: Concanavalin A; Humans; Immune Tolerance; Lupus Erythematosus, Systemic; T-Lymphocytes, Regulatory

Natural killer (NK) cell activity was studied in 23 patients with systemic lupus erythematosus (SLE). The overall NK activity was lower in patients with SLE than in normal female individuals. Patients with clinically active SLE disease had slightly lower NK activity than the patients with inactive disease. Other clinical parameters as well as treatment status did not correlate with NK activity. Interferon (IFN) enhanced the NK activity of normal individuals and of 11 SLE patients, while it did not enhance in the remaining 12 patients. The patients whose NK activity was enhanced by beta-IFN had significantly higher initial activity than those who did not respond to beta-IFN. Furthermore, peripheral mononuclear cells (MNC) from IFN responders produced gamma-IFN after stimulation with concanavalin A (Con A) in titres comparable to those of normals. In contrast, peripheral MNC from beta-IFN non-responders failed to produce significant titres of gamma-IFN after stimulation with Con A. These results indicate that certain patients with SLE have low NK activity, which is generally paralleled by an inability to respond to exogenous beta-IFN and by blunted production of gamma-IFN after stimulation with Con A.

Topics: Cells, Cultured; Concanavalin A; Female; Humans; Interferon Type I; Interferon-gamma; Killer Cells, Natural; Leukemia, Myeloid; Leukocytes; Lupus Erythematosus, Systemic

NZB mice produce a natural thymocytotoxic autoantibody (NTA) capable of specifically injuring thymocytes and T cells. NTA-reactive antigen (NTA-A) shows a different density distribution among T cells, and partial killing with NTA and complement can eliminate T cells bearing NTA-A in high density. Thy-1 antigen is similar to NTA-A in this respect. To determine the effects of NTA and anti-Thy-1 on distinct functional subsets of T cells, Con A-induced suppressor T cell (Con A-Ts) activity against the allogeneic mixed lymphocyte reaction (MLR), responding T cell (TMLR) activity in the allogeneic MLR, and Con A-induced cytotoxic T cell (Con A-Tc) activity were examined simultaneously in BALB/c spleen cells before and after partial elimination of NTA- and anti-Thy-1-sensitive T cells. Treatment with NTA and complement resulted in a marked reduction in Con A-Ts activity, a significant increase in TMLR-activity and a slight and inconstant decrease in Con A-Tc activity. Since Con A-generated T's were much less sensitive to NTA, the NTA-sensitive T cells involved in Con A-Ts activity appear to be precursors or promoters of the Con A-Ts. In contrast, the precursors of Con A-Tc seem to relatively resistant to NTA. The increase in TMLR activity caused by NTA suggests the possibility that NTA is less cytotoxic for TMLR and cytotoxic for some suppressor T cells in allogeneic MLR. The monoclonal anti-Thy-1 antibody showed no such preferential cytotoxic effects on the three T cell functions. The NTA-sensitive T cells, in contrast to anti-Thy-1-sensitive T cells, were reduced gradually during Con A stimulation. All these findings indicate that NTA-A not only differs from Thy-1 antigen but that it appears to be a unique T cell antigen.

Topics: Animals; Antilymphocyte Serum; Autoantibodies; Concanavalin A; Cytotoxicity, Immunologic; Female; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred NZB; T-Lymphocytes

Systemic lupus erythematosus (SLE) is characterized by increased numbers of circulating B cells activated polyclonally to secrete immunoglobulin. Because T cells secrete, or shed, various factors that are functionally important in regulating immunoglobulin production by B cells, a reverse hemolytic plaque assay was developed to quantitate such activated T cells. In this technique, we used a rabbit antiserum raised to supernatants of concanavalin-A--stimulated human lymphocytes. The relevant antigenic specificity of this antiserum is directed toward the shed surface membrane determinant(s) preferentially expressed on activated T cells. Freshly isolated peripheral blood mononuclear cells from 14 SLE patients contained more than 10 times the number of endogenously activated T cells than cells from normal subjects. Within the SLE group, plaque-forming T cells were particularly increased in patients with active disease. By linear regression analysis, a significant positive correlation was revealed between such activated T cells and immunoglobulin-secreting B cells, also measured by a reverse plaque assay (r = 0.83). It appears that both activated B cells and T cells circulate in increased numbers in SLE. Additional investigation will be required to define the molecular nature of the T cell product(s) being measured and to clarify the relationship of these findings to the immunoregulatory abnormalities in this disorder.

Topics: Animals; B-Lymphocytes; Concanavalin A; Hemolytic Plaque Technique; Humans; Immunoglobulin G; Lupus Erythematosus, Systemic; Pokeweed Mitogens; Rabbits; T-Lymphocytes

Topics: Antilymphocyte Serum; Cell Membrane; Concanavalin A; Hemolytic Plaque Technique; Humans; Lupus Erythematosus, Systemic; Phytohemagglutinins; Pokeweed Mitogens; T-Lymphocytes

Topics: Adolescent; Adult; Aged; Child; Concanavalin A; DNA; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Middle Aged; Mitogens; Phytohemagglutinins; Pokeweed Mitogens; Prednisone

Natural cell-mediated cytotoxicity /NCMC/ was evaluated using human adherent 3H-thymidine-prelabelled HEp-2 epipharynx carcinoma cells as targets at 50:1 effector-target cell ratio in a 24 hr assay. For lectin-dependent cell-mediated cytotoxicity /LDCC/ studies cultures contained also 25/micrograms/ml concanavalin A /Con A/. Peripheral blood mononuclear cells /PBMC/ of nine patients with active systemic lupus erythematosus /SLE/ failed to exert NCMC or LDCC against HEp-2 targets. In contrast, an increased adherence /decreased detachment from the monolayer/ of HEp-2 target cells was observed in the presence of PBMC from SLE patients that was further promoted by the addition of Con A during LDCC assay.

Topics: Adolescent; Adult; Cell Adhesion; Cell Line; Cell Survival; Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; Immunity, Cellular; Killer Cells, Natural; Lupus Erythematosus, Systemic; Middle Aged; Pharyngeal Neoplasms

Forty-eight cases of systemic lupus erythematosus (SLE) were studied for lymphocyte subpopulations, and 42 cases were studied for lymphocyte response to mitogen stimulation. A decreased percentage of thymus-derived cells (T cells) and an increased percentage of bursa-equivalent derived cells (B cells), null cells (N cells), double labelled cells (D cells) were found in non-treated cases of SLE. There was no significant difference in these lymphocyte subpopulations in the treated cases in comparison with the normal control. There was an inverted linear relationship between T cells and N cells (p less than 0.001). No such relationship was found between B cells and N cells. The lymphocyte transformation in response to phytohemagglutinin (PHA), pokeweed mitogen (PWM) and concanavalin A (Con A) was expressed in three ways: (1) the net count of transformed data, (2) the difference between the square roots of the isotope incorporation in the stimulated and the non-stimulated cultures (Dsq), and (3) the stimulation index (SI). There were significantly decreased lymphocyte transformation in response to PHA, PWM stimulation in both the non-treated and the treated cases of SLE when results were expressed as net count and Dsq. But decreased counts in response to Con A was only found in non-treated cases. In contrast, no significantly lowered response to all three mitogens was found when data were expressed as stimulation index. Simultaneous study of the percentages of T cells, B cells, N cells, and D cells might be helpful in assessing the clinical activity of SLE and monitoring the effect of therapy. In the study of lymphocyte response to mitogen stimulation, Con A response was a more sensitive indicator of disease activity. The stimulation index was not a good method to demonstrate the result of lymphocyte response to mitogen stimulation in cases of SLE.

Topics: Concanavalin A; Humans; Immunity, Cellular; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mitogens; Mitosis; Phytohemagglutinins; Pokeweed Mitogens

Topics: Adult; Animals; Chronic Disease; Concanavalin A; Female; Hepatitis; Humans; Immunoglobulin Fc Fragments; Lupus Erythematosus, Systemic; Male; Middle Aged; Phytohemagglutinins; T-Lymphocytes; T-Lymphocytes, Regulatory

The T cell mitogen concanavalin A (Con A) was added to lymphocytes pre-cultured for 24 hours in vitro in tissue culture medium. Delayed addition of the mitogen resulted in an enhanced lymphocyte activation measured by incorporation of radioactive precursors of DNA. T cells isolated by rosetting with sheep erythrocytes also exhibited this enhancement. In a limited study, lymphocytes from both atopic patients and those with systemic lupus erythematosus (SLE) showed little or no enhanced responsiveness to Con A. The possibility that some patients with atopy and SLE possess defective suppressor T cells is discussed in the light of these data.

Topics: Adult; Animals; Blood; Cattle; Concanavalin A; Female; Fetus; Humans; Hypersensitivity, Immediate; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Middle Aged; T-Lymphocytes; T-Lymphocytes, Regulatory; Time Factors

Lysosome proteins derived from peripheral blood granulocytes of patients with multiple sclerosis (MS), lupus erythematosus (LE), rheumatoid arthritis (RA) and recurrent uveitis cause the impairment of Con A-induced suppressor cell activity in two-step culture in vitro. This effect is independent of lysosome protease activity. Such a phenomenon, observed in vitro, may cause disturbances in suppressor cell function in the course of the above diseases.

Topics: Adult; Arthritis, Rheumatoid; Autoimmune Diseases; Concanavalin A; Female; Humans; Isoflurophate; Lupus Erythematosus, Systemic; Male; Middle Aged; Multiple Sclerosis; Neutrophils; Proteins; Recurrence; T-Lymphocytes, Regulatory; Uveitis

The lysosomal factors obtained from PMNL of healthy persons and patients with multiple sclerosis, systemic lupus erythematosus and other neurological diseases inhibited in vitro the generation of Con A-induced suppressor cell activity. The results obtained proved that these factors from granulocytes of MS-relapse and active LE have a stronger inhibitory effect on suppressor cell activity than on healthy ones.

Topics: Adult; Aged; Concanavalin A; Humans; Long-Term Care; Lupus Erythematosus, Systemic; Middle Aged; Multiple Sclerosis; Nervous System Diseases; Neutrophils; Proteins; T-Lymphocytes, Regulatory

We have recently described that human autologous rosette-forming (Tar) cells have the characteristics of postthymic precursor cells. Herein we report that we found circulating Tar cells significantly diminished in 32 patients with untreated systemic lupus erythematosus (SLE) as compared to 32 age/sex matched controls. Pretreatment of peripheral blood mononuclear cells (MNC) from SLE patients with serum from young normal adults or wtih serum thymic factors (FTS) increased their percentages of Tar cells significantly but reached near normal values in only 3 patients with inactive disease. Patients and normal subjects had similar percentages of Tar cells binding peanut-agglutinin. Characteristic functions of postthymic precursor cells are feedback inhibition and generation of suppressor cells which we studied in systems where we depleted or added Tar cells to Tmu and B cells, or MNC, respectively, using as indicators the production of immunoglobulins measured in culture supernatants or 3H-thymidine incorporation. We found both functions diminished in SLE patients despite using the presence of a qualitative as well as quantitative defect. In two SLE patients studied both of these functions corrected partially when their Tar cells were pretreated with FTS. In 20 SLE patients we studied Tgamma and Tmu cells as well as Concanavalin-A-induced, spontaneously-expanded suppression and found Concanavalin-A-induced, spontaneously-expanded suppressor function and Tgamma cells diminished. However only the reduction of Tgamma and of spontaneously-expanded suppressor function were found to relate to disease activity. On the other hand, Tmu cells were found to be similar in numbers in SLE patients and normal controls.

Topics: Adult; Concanavalin A; Feedback; Female; Humans; Lupus Erythematosus, Systemic; Male; Receptors, Fc; Rosette Formation; T-Lymphocytes; Thymic Factor, Circulating; Thymus Hormones

21 patients with Sjögren's syndrome (sicca syndrome) with either glandular or extraglandular involvement, but without other connective tissue diseases, were studied with regard to immunoregulatory T-cell subpopulations, B-cell function, and suppressor cell capabilities. Patients with isolated glandular disease as well as patients with extraglandular disease had normal absolute numbers of total lymphocytes, T cells, and B cells. However, 9 of 11 patients with extraglandular disease and only 3 of 10 patients with glandular disease had decreased relative proportions of T cells bearing receptors for the Fc portion of immunoglobulin (Ig)G (T(G)) which was explained by a factor that blocked the expression of the IgG Fc receptor on T(G) cells. This blockage was reversible since the factor could be removed by trypsinizing the T cells before T(G) determination. Serum from patients with abnormal proportions of T(G) cells, but not serum from patients with normal proportions of T(G) cells, blocked the expression of the IgG Fc receptor on normal T cells. The serum factor upon fractionation over Bio-Gel A 1.5 columns as well as over staphylococcal protein A-Sepharose 4B columns was found diffusely within the IgG fraction, and not in the IgM fraction. Neither patients with glandular nor patients with extraglandular disease manifested increased numbers of in vivo-activated circulating lymphocytes as determined by spontaneous anti-trinitrophenyl (TNP) plaque-forming cells (PFC). However, patients with glandular disease had reduced numbers of pokeweed mitogen-induced anti-sheep erythrocyte PFC (P < 0.01) as compared with normals and patients with glandular disease. Of note was the fact that despite the modulation of T(G) subpopulation by the serum factor in patients with extra-glandular disease, these patients manifested normal concanavalin A-generated suppressor cells of pokeweed mitogen-induced PFC responses in allogeneic co-cultures. This was unlike the suppressor cell defect previously described in this system with systemic lupus erythematosus patients. The discrepancy was attributed both to the fact that the T(G) defect was reversible and to the fact that concanavalin A-generated suppressor cells are not limited to the T(G) subset. Thus, these studies have demonstrated reversible abnormalities in T(G) cells in patients with extraglandular Sjögren's syndrome which are not associated with suppressor cell defects. The discrepancy between these findings and the immuno-reg

Topics: Adult; Aged; B-Lymphocytes; Concanavalin A; Female; Humans; Immunity; Immunoglobulin G; In Vitro Techniques; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Middle Aged; Mitogens; Receptors, Fc; Sjogren's Syndrome; T-Lymphocytes; T-Lymphocytes, Regulatory; Trypsin

We have investigated in vitro the magnitude, nature, and regulation of spontaneous and mitogen-induced Ig secretion by splenic lymphocytes from several autoimmune murine strains (NZB, NZB X W, MRL/l BXSB) and appropriate, normal mice. All autoimmune strains had increased numbers of mature splenic B lymphocytes, which secreted and/or contained Ig, compared to age-matched normal strains. In NZB and NZB X W mice, the high frequency of mature B cells was apparent early in life, whereas in MRL/l and BXSB mice it was first noted shortly before the clinical onset of disease. Spleen cells from young autoimmune mice of all four strains secreted predominantly IgM, but with aging and the appearance of disease, the cells switched to IgG secretion predominantly. In contrast, spleen cells from normal mice were predominantly IgM, but with aging and the appearance of disease, the cells switched to IgG secretion predominantly. In contrast, spleen cells from normal mice were predominantly IgM secretors throughout the animals' lives. Approximately 15% of the total Ig-secreting cells in older NZB, NZB X W, and MRL mice were committed to secretion of anti-ssDNA antibodies. In both autoimmune and normal spleen cells, the B-cell population alone contained fewer secreting cells than the total lymphocyte population, indicating that T cells were required to achieve maximal levels of plaque-forming cells. Spleen cells of NZB and NZB X W mice had a greater response to lipopolysaccharide (LPS) than other autoimmune and normal strains. Responsiveness to LPS, as measured by the frequency of induced Ig-secreting cells, was considerably diminished with age and onset of disease in all autoimmune but not in normal strains. LPS-induced Ig secretion by B cells of autoimmune and normal mice was subject to regulation by splenic T cells. No significant differences were observed between concanavalin-A (Con A) stimulated spleen cells from young and older autoimmune mice and normal control strains in effectively suppressing spontaneous and LPS-induced Ig secretion. Moreover, B cells from autoimmune mice and from normal strains were equally receptive to Con A-induced suppressor signals. T cells from young and older NZB and BXSB mice added to a standard number of B cells from syngeneic young mice provided equal help in enhancing LPS-induced Ig secretion, and this help in turn was equivalent to that provided by T cells from normal mice of the same H-2 haplotype. The exception was the MRL/l strain; T

Topics: Animals; Autoantibodies; Autoimmune Diseases; B-Lymphocytes; Cell Differentiation; Concanavalin A; Female; Immunoglobulins; Lipopolysaccharides; Lupus Erythematosus, Systemic; Male; Mice; Mice, Inbred NZB; Spleen; T-Lymphocytes; T-Lymphocytes, Regulatory

Topics: Cell Division; Cell Separation; Concanavalin A; Hemolytic Plaque Technique; Humans; Immunoglobulin Fc Fragments; Immunoglobulin G; Immunoglobulins; Isoantigens; Lupus Erythematosus, Systemic; Mitogens; Mitomycins; Phagocytes; Pokeweed Mitogens; Rosette Formation; T-Lymphocytes, Regulatory; Time Factors

Topics: Adult; B-Lymphocytes; Concanavalin A; Humans; Lupus Erythematosus, Systemic; Lymphocyte Culture Test, Mixed; Macrophages; T-Lymphocytes

Topics: Adult; Concanavalin A; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; T-Lymphocytes, Regulatory

The in vitro antibody response of peripheral blood lymphocytes (PBL) from 19 patients with untreated systemic lupus erythematosus (SLE) was compared with that of 20 control patients and 44 normal subjects. Trinitrophenyl polyacrylamide beads (TNP-PAA) were used to induce IgM anti-TNP plaque-forming cells. SLE patients displayed a markedly depressed, and in most instances virtually absent, response. This was not due to an unusual kinetics of the response; nor could it be induced by preincubation of SLE patients' PBL. In co-cultures of SLE patients and normal PBL, the former, with few exceptions, did not exert a suppressive effect. In four patients the anti-TNP response of either unfractionated or T-depleted SLE PBL could be restored by T cells from a normal individual. Conversely in three of these patients, SLE T cells could not support the response of normal B cells, suggesting a T helper cell defect in SLE PBL. Concanavalin A (Con A)-induced suppressor cells of the antibody response could be assayed by two approaches: (a) in responder SLE patients, by the direct addition of Con A to TNP-PAA-stimulated cultures; (b) in seven patients by transfer of Con A-activated cells to the responding culture of a normal allogeneic donor. In both cases SLE PBL were able to exert a suppressive effect to the same extent as normal PBL.

Topics: Adult; Antibody-Producing Cells; Concanavalin A; Female; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Middle Aged; T-Lymphocytes; T-Lymphocytes, Regulatory

Topics: Adult; Concanavalin A; Humans; Immunoglobulins; Lupus Erythematosus, Systemic; Lymphokines; Middle Aged; Spleen; Suppressor Factors, Immunologic; T-Lymphocytes; T-Lymphocytes, Regulatory

Levamisole-treated mononuclear cells from normal volunteers or from patients with systemic lupus erythematosus (SLE), when cultured with normal allogeneic lymphocytes, did not significantly influence the ability of the latter cells to proliferate when stimulated with either phytohemagglutinin (PHA) or concanavalin A (Con A). Levamisole, furthermore, did not influence the ability of Con A to induce suppressor cell function from normal mononuclear cells. When mononuclear cells from patients with SLE were treated with therapeutic concentrations of levamisole in the presence of Con A, the drug was unable to restore the ability of the Con A to induce normal suppressor cell function. SLE cells treated with a high concentration of levamisole (60 micrograms/ml) in the presence of Con A did show some suppressor cell activity on normal autologous mononuclear cells activated by Con A, but not by PHA. Although the significance of this latter finding is unclear it appears that levamisole at therapeutic concentrations does not influence in vitro suppressor cell function of normal or SLE mononuclear cells.

Topics: Concanavalin A; Humans; Levamisole; Lupus Erythematosus, Systemic; Lymphocytes; T-Lymphocytes, Regulatory

T lymphocytes suppressing in vitro polyclonal immunoglobulin production can be activated by allogeneic stimuli or concanavalin A (Con A). These cells are deficient in systemic lupus erythematosus (SLE). In order to investigate the cellular requirements for this effect, we studied IgM and IgG biosynthesis by (a) normal and SLE lymphocytes cultured with pokeweed mitogen (PWM) and mitomycin C-blocked allogeneic normal or SLE lymphocytes, and (b) by normal and irradiated lymphocytes cultured with PWM and with Con A-pretreated normal and irradiated allogeneic and autochthonous blocked lymphocytes. Results showed that blocked allogeneic normal cells or blocked Con A-pretreated, normal or irradiated, autochthonous or allogeneic cells served as potent stimuli for suppression of immunoglobulin biosynthesis by normal responder cells, but not by SLE responder cells. This suggests that Con A stimulates a radioresistant suppressor inducer cell which in turn activates a radiosensitive, proliferation-dependent suppressor effector cell; the latter can also be activated by allogeneic stimulation and is deficient in SLE.

Topics: Cells, Cultured; Concanavalin A; Humans; Immunoglobulin G; Immunoglobulin M; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocytes; Mitomycins; Pokeweed Mitogens; T-Lymphocytes, Regulatory

Decreases in numbers and functions of SLE T lymphocytes as a form of lymphoreceptopathy were investigated using T lymphocytes and sera obtained from both SLE patients and healthy subjects with respect to the effects of suppressor T cells and Con A induced suppressor factor against antibody formation, changes in the membrane microviscosity and the ultrastructure of membrane associated particles (MAPs) of T lymphocytes. The lymphoreceptopathy of SLE T lymphocytes was induced by decreased microviscosity and abnormalities of distribution, size and density of MAPs on the T lymphocyte membrane which can be modulated by serum factors consisting mainly of IgG as T cell membrane binding antibody and partially of IgM as cytotoxic antibody.

Topics: Antibody Formation; Concanavalin A; Humans; Immunoglobulin G; Immunosuppression Therapy; Leukocyte Count; Lupus Erythematosus, Systemic; Receptors, Antigen, B-Cell; T-Lymphocytes; Viscosity

Eighty percent of 31 untreated patients with systemic lupus erythematosus (SLE) had abnormalities in their spontaneously expanded and/or Con-A-induced suppressor cell function, but the association of defects detected with both systems was only 68%. Loss of spontaneous suppression related positively to disease activity (r = 0.641) and the number of T gamma cells (r = 0.624) whereas Con-A-induced suppression correlated negatively with disease activity (r = -0.456) and the number of T gamma cells (r = 0.089). Incubation of mononuclear cells from SLE patients in antiribonucleoprotein IgG caused further loss of suppression in some, but not all, instances. The suppressor cell dysfunction found in SLE may result from diverse mechanisms, including a basic defect in the generation of suppressor cells and the abrogation of suppressor function by autoantibodies.

Topics: Adult; Animals; B-Lymphocytes; Chickens; Complement C3; Concanavalin A; Hemolytic Plaque Technique; Humans; Immunoglobulin G; Lupus Erythematosus, Systemic; Mitogens; Phagocytes; Rabbits; Rats; Sheep; T-Lymphocytes; Time Factors

A naturally occurring T-lymphocytotoxic autoantibody (Hu-NTA) in serum from a patient with systemic lupus erythematosus (SLE) showed a differential cytotoxic effect on functionally different T cell subsets as did natural thymocytotoxic autoantibody (NTA) of NZB mice. When the normal peripheral blood lymphocytes were treated, in the presence of complement, with Hu-NTA at a dilution that eliminated 25 to 30% of Hu-NTA-sensitive T cells, there was a marked reduction or a total depletion in the ability of resultant cells to show Con A-activated suppression on the proliferative response of responder cells in mixed lymphocyte reaction. The treatment of PBL in the same manner also resulted in a marked reduction in its responsiveness to Con A and PHA. However, the responder cells to allogeneic stimulator cells in MLR were found to be much more resistant to the cytotoxicity of Hu-NTA than other functional T cell subsets tested. These results suggest that Hu-NTA is responsible for the selective loss of certain functional T cell subsets including suppressor T cells in patients with SLE.

Topics: Antilymphocyte Serum; Autoantibodies; Concanavalin A; Humans; Lupus Erythematosus, Systemic; Lymphocyte Culture Test, Mixed; Phytohemagglutinins; T-Lymphocytes

Topics: Animals; B-Lymphocytes; Concanavalin A; Humans; Immunoglobulin M; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; T-Lymphocytes, Regulatory

Antibodies to T cells present in the plasma of patients with active systemic lupus erythematosus (SLE) plus complement are able to eliminate concanavalin A-induced suppressor function for the proliferative responses of T cells to allogeneic lymphocytes (MLR) and of B cells to pokeweed mitogen (PWM). Such antibodies were found to be effective in eliminating suppressor function only when T cells were treated before activation; there was no effect when treatment was performed after activation. These studies indicate that the antibodies preferentially interact with a T cell necessary for the generation of suppressor cells, rather than with mature, activated suppressor cells. Studies of individual SLE patients indicate that the same defects observed in SLE T cells were induced in normal T cells by plasma from that patient. Such observations suggest that many T-cell defects associated with active SLE may not be intrinsic T-cell abnormalities, but, rather, secondary effects of anti-T-cell antibodies. Studies of the T-cell subpopulations responsible for suppression of the MLR and PWM responses indicate that only T gamma cells (T cells bearing receptors for the Fc portion of immunoglobulin [Ig]G) acted as precursors of suppressor cells for the MLR, whereas both T gamma and T non-gamma cells (T cells not bearing receptors for the Fc portion of IgG) could be activated to suppress the PWM response. Consistent with this observation, SLE anti-T-cell antibodies that preferentially killed T gamma cells preferentially eliminated suppressor cells for the MLR.

Topics: Autoantibodies; Autoimmune Diseases; Cells, Cultured; Concanavalin A; Humans; Immunoglobulin G; Lupus Erythematosus, Systemic; Pokeweed Mitogens; Receptors, Concanavalin A; Receptors, Fc; Receptors, Mitogen; T-Lymphocytes, Regulatory

Effect of anti-lymphocyte antibody of active systemic lupus erythematosus (SLE) on lymphocyte function was examined. Lymphocytes from normal individuals treated with anti-lymphocyte antibody and complement exhibited marked inhibition of response to concanavalin A (Con A), while the response of lymphocytes to phytohaemagglutinin M (PHA-M) and pokeweed mitogen (PWM) was slightly affected. In mixed lymphocyte culture response, both stimulator and responder cells were insensitive to anti-lymphocyte antibody. Treatment of sensitized lymphocytes with anti-lymphocyte antibody and complement caused a dose-dependent suppression of blastogenic response to purified protein derivatives (PPD). No effect, however, was noted on migration-inhibitory factor (MIF)-producing cells. In PWM-driven Ig synthesis, T lymphocytes lacking the anti-lymphocyte antibody-reactive T-cell subset enhanced PWM-driven Ig synthesis of autologous B lymphocytes. Con-A-induced suppressor function of lymphocytes was abolished by the treatment with anti-lymphocyte antibody and complement. The present study demonstrated that lymphocytes from normal individuals after treatment with anti-lymphocyte antibody and complement showed similar immunological reactivities with lymphocytes from active SLE, indicating that those anti-lymphocyte antibodies could play an important role in defective suppressor cell function.

Topics: Antibody-Dependent Cell Cytotoxicity; Antilymphocyte Serum; Complement System Proteins; Concanavalin A; Humans; Immunoglobulins; Leukocyte Migration-Inhibitory Factors; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Phytohemagglutinins; Pokeweed Mitogens; Rosette Formation; T-Lymphocytes; T-Lymphocytes, Regulatory

Circulating antibodies that could be responsible for the suppressor thymus-derived (T)-cell dysfunction in active systemic lupus erythematosus (SLE) were investigated. Sera from 14 active and inactive SLE patients were compared with a pool of 22 normal sera. All sera were adsorbed with a pool of normal platelets to exclude antihistocompatibility leukocyte antigen antibodies; with AB erythrocytes to exclude isohemagglutinins; and with a pool of normal bone marrow-derived (B) lymphocytes, monocytes, and neutrophils to deplete anti-B-cell antibodies, Fc-receptor antibodies, and antibodies directed against neutrophils or monocytes. Sera from active SLE patients were capable of inhibiting the activation of normal, blood lymphocytes by concanavalin A to become suppressor cells. The latter were assayed by coculturing the concanavalin A-activated cells with autologous lymphocytes, which were then activated with either phytohemagglutinin for proliferative response or with pokeweed mitogen for B-cell immunoglobulin (Ig) synthesis and secretion. Specific incorporation of cultures with phytohemagglutinin showed a value of 67+/-13 (mean+/-SD) for suppressor cells treated with adsorbed, active SLE sera. This value was significantly different (P < 0.001) from that of cells treated with the inactive SLE sera or with the pool of normal sera. Similar findings were seen with respect to the B-cell target parameters. Cytoplasmic Ig and IgG in supernates of cultures with pokeweed mitogen showed values of 17+/-5% and 717+/-134 ng/culture, respectively, for suppressor cells treated with the adsorbed, active SLE sera. This was significantly different from those treated with the inactive SLE sera or with the pool of normal sera. The antisuppressor-cell factor was shown to be IgG, complement independent, not cytotoxic, active at 37 degrees C and at room temperature, but not at 4 degrees C, and adsorbable with T cells. Suppressor T-cell antibody in sera of active SLE patients could be responsible for the observed suppressor T-cell dysfunction seen in active SLE. The mechanisms responsible for the induction of the antisuppressor-cell antibody are unknown.

Topics: Antibodies; Concanavalin A; Humans; Immunoglobulins; Lupus Erythematosus, Systemic; T-Lymphocytes

Patients with systemic lupus erythematosus (SLE) produce excessive amounts of autoantibodies. It has also been demonstrated in several systems that such patients have a relative loss of suppressor thymus-derived (T) cells that inhibit the immune response. This loss of suppressor cells has been suggested as one of the causes of the excessive production of antibodies in patients with SLE. In the present report we have tested the hypothesis that anti-T-cell antibodies found in the plasma of some patients with SLE preferentially kill suppressor cells. T cells from normal individuals can be activated by concanavalin A to develop suppressor cell activity. We therefore cultured normal T cells together with concanavalin A in the presence of plasma or plasma fractions from patients with SLE. We found that plasma from patients with active SLE, in which anti-T-cell antibodies were present, inhibited the development of suppressor activity in such cultures. In contrast, plasma from other active patients and patients with inactive SLE, in which no anti-T-cell antibodies could be detected, failed to block the development of such suppressor activity. Absorption of the plasma that contained anti-T-cell antibodies with T cell, but not non-T cells, could eliminate the suppressor-inhibiting activity of the SLE plasma that contained anti-T-cell antibodies. The immunoglobulin (Ig)M, but not the IgG, fraction of the plasma was shown to possess the inhibiting property and complement was found to be necessary for the effect of such anti-T-cell antibodies. We also demonstrated that exposure of normal T cells to such anti-T-cell antibodies and complement did not affect another population of T cells that could proliferate in response to mitogens.Thus, certain patients with SLE have in their plasma an antibody of the IgM class that can selectively eliminate a population of T cells capable of developing suppressor function. The loss of suppressor T cells in patients with SLE may be the result of the effects of such antibody activity in vivo.

Topics: Adult; Autoantibodies; Complement System Proteins; Concanavalin A; Humans; Immunoglobulin M; Lupus Erythematosus, Systemic; Lymphocyte Activation; T-Lymphocytes

Topics: Adolescent; Adult; Antibodies, Antinuclear; Concanavalin A; DNA; Humans; Immunosuppression Therapy; Lupus Erythematosus, Systemic; Middle Aged; T-Lymphocytes

We tested the hypothesis that abnormalities of central immune function are genetically controlled in patients with systemic lupus erythematosus. We used an in vitro suppressor-cell assay to evaluate central immunoregulation in 15 patients, 50 of their clinically healthy family members and 41 normal persons. Impaired suppressor-cell function was found in 11 patients; there was no correlation between disease activity and test results. Abnormal suppressor-cell activity was also found in 13 first-degree relatives, 12 of whom were women. We found no correlation between results of the suppressor-cell assay and the presence or absence of lymphocytotoxic antibodies in the relatives. Impaired suppressor-cell function cannot by itself explain the pathogenesis of systemic lupus erythematosus. Our results support the hypothesis that certain abnormalities of suppressor cells are genetic markers. We propose that the development of systemic lupus erythematosus requires the participation of at least two functionally distinct classes of genes.

Topics: Adolescent; Adult; Cells, Cultured; Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; Immunoglobulin G; Lupus Erythematosus, Systemic; Lymphocytes; Male; Middle Aged; Pedigree; T-Lymphocytes

Normal human T cells proliferate vigorously when stimulated with autologous non-T cells. This autologous mixed lymphocyte reaction (MLR) between T and non-T cells was defective in patients with active systemic lupus erythematosus (SLE). In contrast, T cells and non-T cells from active SLE patients behaved normally as responding and stimulating cells, respectively, in the allogeneic MLR. The etiology of the impaired autologous MLR was further examined by studying the functional capacity of subsets of stimulating or responding cells. B cells, L cells, and monocytes from active SLE patients failed to stimulate autologous T cells but these cells effectively stimulated allogeneic T cells. Fc(IgG)+ T cells from active patients were unable to respond in both the autologous and allogeneic MLR; their Fc(IgG)-T cells responded well in the allogeneic but not in the autologous MLR. The Fc(IgG)+ T cells, but not the Fc(IgG)- T cells, from inactive SLE patients also failed to respond in the both autologous and allogeneic MLR. These studies indicate that patients with SLE have functionally defective Fc(IgG)+ T cells and a defective autologous MLR, both of which may contribute to impaired regulation of immune functions.

Topics: Adult; Autoantigens; Binding Sites; Concanavalin A; Female; Humans; Immunoglobulin Fc Fragments; Immunosuppression Therapy; Isoantigens; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphocytes; Male; Receptors, Antigen, B-Cell; T-Lymphocytes

Normal mononuclear leukocytes were incubated with serum from patients with active systemic lupus erythematosus (SLE) and healthy subjects and then studied on lymphoproliferative tests. Serum from SLE patients that contained an autoantibody to a subpopulation of thymus-derived (T) lymphocytes inhibited suppressor T-cell activity induced with concanavalin A. These sera did not inhibit lymphoproliferative responses or suppression by monocytoid cells. Mitogen-activated suppressor cells were not inhibited with serum from SLE patients or healthy subjects lacking T-cell autoantibody. This abnormality may contribute to the altered immune response that occurs with SLE.

Topics: Adult; Autoantibodies; Binding, Competitive; Concanavalin A; Humans; Immunity; Immunosuppression Therapy; Lupus Erythematosus, Systemic; Lymphocyte Activation; T-Lymphocytes

To characterize the cell(s) responsible for the suppressor-cell dysfunction in active systemic lupus erythematosus (SLE), we fractionated blood mononuclear cells into thymus-derived (T), bone marrow-derived (B), and monocyte-depleted populations. Various cell populations from active SLE, inactive SLE, or normals, were activated with Concanavalin A, washed, and then co-cultured with active SLE cells. Soluble immune response suppressor (SIRS) from culture supernates of the activated cells was also used for the possible correction of the suppressor-cell dysfunction. Suppression was tested by enumerating DNA-binding cells by radioautography and by quantitating anti-DNA antibody in culture supernates by radioimmunoassay; and immunoglobulin was tested in cells and supernates by the immunofluorescence and the immunofluor techniques, respectively. Except for the numbers of DNA-binding cells, which were not suppressed, all the other three parameters in co-cultures with cells from active SLE patients were suppressed by Concanavalin A-activated cells (P < 0.001), or by SIRS (P < 0.05) from normals or inactive SLE patients. Concanavalin A-activated autologous or allogeneic active SLE cells and nonactivated cells from active or inactive SLE failed to suppress the various B-cell functions. Nonactivated normal cells suppressed levels of anti-DNA and immunoglobulin in supernates (P < 0.05). In characterizing the cells responsible for the suppressor dysfunction, it was clear from the results that T cells responsive to Concanavalin A activation are deficient in active SLE and fail to generate SIRS. On the other hand, monocytes from active SLE patients are responsive to signals from the activated T cells of normals or inactive SLE donors. Because SIRS suppresses active SLE cells in vitro, it might be considered therapeutically for the in vivo modulation of SLE.

Topics: Concanavalin A; Humans; Immunity, Cellular; Immunosuppression Therapy; Lupus Erythematosus, Systemic; Lymphocyte Activation; Monocytes; T-Lymphocytes

The mechanism of poor lymphocyte transformation to mitogens was studied in selected patients with rheumatoid arthritis and systemic lupus erythematosus. Low lymphocyte response to PHA and Con-A in media containing autologous and homologous sera was usually associated with poor response to the calcium ionophore A23187, which induces blastogenesis by a different mechanism. The low lymphocyte response to mitogens in patients with rheumatoid arthritis could be restored by in vivo treatment with the anthelmintic drug, levamisole. The present findings suggest that intrinsic defects are responsible for the decreased cellular response in patients with rheumatoid arthritis and systemic lupus erythematosus.

Topics: Adolescent; Adult; Arthritis, Rheumatoid; Calcimycin; Concanavalin A; Humans; Lectins; Levamisole; Lupus Erythematosus, Systemic; Lymphocyte Activation; Middle Aged

Immunological reactivity in patients with SLE was studied in vitro trinitrobenzene sulphonate (TNP) specific antibody formation by peripheral blood lymphocytes. Lymphocytes from patients with SLE could produce an increased number of TNP-specific plaque-forming cells (PFC), while no such response could be seen in normal controls. Co-culture of lymphocytes from active SLE patients and normal controls was performed with TNP-Horse red blood cells (TNP-HRBC). The number of PFC by B lymphocytes from active SLE patients was suppressed by T lymphocytes from normal controls. On the other hand, the number of PFC by B lymphocytes from normal controls was increased by T lymphocytes from active SLE patients. Co-culture of lymphocytes from identical twins discordant for SLE was also performed, and the same results were obtained. We further examined the effects of Con A on antibody formation. Con A-treated T lymphocytes from a normal control markedly suppressed TNP-specific PFC by peripheral lymphocytes from active SLE patients. However, Con A-treated T lymphocytes from an active SLE patient did not suppress TNP-specific PFC by lymphocytes from another active SLE patient. These results suggest that active SLE patients showed a loss of suppressor T-lymphocyte function.

Topics: Adolescent; Adult; Antibody Formation; Concanavalin A; Diseases in Twins; Female; Hemolytic Plaque Technique; Humans; Immunosuppression Therapy; Lupus Erythematosus, Systemic; Male; Middle Aged; Rosette Formation; T-Lymphocytes

Topics: Adolescent; Adult; Cell Separation; Cells, Cultured; Child; Concanavalin A; Female; Hemolytic Plaque Technique; Humans; Lupus Erythematosus, Systemic; Lymphopenia; Male; Middle Aged; Pokeweed Mitogens; T-Lymphocytes

Spleen cells from normal mice were cultured with Concanavalin A to produce an immunosuppressive supernate. This supernate was used to treat the lupus-like autoimmune disease of NZB/NZW mice. Such treated mice lived significantly longer than did controls, but only if treatment was initiated early in the course of the illness.

Topics: Age Factors; Animals; Autoimmune Diseases; Cell Extracts; Cells, Cultured; Concanavalin A; Evaluation Studies as Topic; Female; Immunosuppressive Agents; In Vitro Techniques; Lupus Erythematosus, Systemic; Mice; Mice, Inbred Strains; Molecular Weight; Spleen; Stimulation, Chemical; Time Factors; Tissue Extracts

Patients with systemic lupus erythematosus lacked suppressor T cell function. Suppressor cell activity was induced in cells from many of these patients by incubation with thymosin or cultured thymic epithelium. These results suggest that thymic manipulation may be a useful therapeutic modality in this disease.

Topics: Adolescent; Adult; Concanavalin A; Culture Techniques; Epithelium; Female; Humans; Immunotherapy; Lupus Erythematosus, Systemic; Male; Middle Aged; T-Lymphocytes; Thymosin; Thymus Gland; Thymus Hormones

Normal peripheral blood mononuclear cells demonstrated increased DNA synthesis and secretion of newly synthesized protein when suboptimal concentrations of Concanavalin A (Con A) were added to the cultures after 24-h incubation in vitro. Cells stimulated by Con A, 1 mug/ml, after 24-h incubation demonstrated 3.0 times more tritiated thymidine incorporation, and 4.4 times more 14C-amino acid incorporation into newly synthesized secreted protein, than cells stimulated at 0 h (P less than 0.001). The acquisition of increased responsiveness was not abrogated by washing and resuspending the cells in fresh medium. Since the increased responsiveness could be inhibited by the addition to the cultures of small numbers of cells previously activated by Con A it is suggested that the enhanced reactivity acquired in culture represents the loss of a subpopulation of suppressor cells that modulate the T-lymphocyte response. Cells from nine patients with active, untreated systemic lupus erythematosus demonstrated normal responses to optimal concentrations of Con A added at 0 h, but an impaired response to Con A, 1 mug/ml. When these cells were incubated for 24 h, a significant increased response to Con A was not observed. This observation suggests that patients with active SLE lack circulating suppressor cells. When seven SLE patients were again studied after corticosteroid therapy had led to clinical improvement, the response to Con A, 1 mug/ml, added after 24-h incubation was similar to that observed in normal controls, suggesting that suppressor function in SLE returns as disease activity declines.

Topics: Adolescent; Adult; Concanavalin A; Culture Media; Humans; Immunosuppression Therapy; Lectins; Lupus Erythematosus, Systemic; Lymphocytes; Monocytes; Streptodornase and Streptokinase

Attempts were made to isolate a virus from systemic lupus erythematosus patients, using lymphocyte cultures prepared from peripheral blood. Both cocultivation with VERO cells and fusion experiments in which lysolecithin and Concanavalin A were employed as fusing agents failed to reveal any presence of virus. Con A proved to be useful for heterokaryon formation in primate cells, fusion levels varying from 14-21% as shown by autoradiography.

Topics: Cell Fusion; Cell Line; Concanavalin A; Cytopathogenic Effect, Viral; Lupus Erythematosus, Systemic; Lymphocytes

Topics: Adolescent; Adult; Arthritis, Rheumatoid; Autoimmune Diseases; Concanavalin A; Humans; Lectins; Lupus Erythematosus, Systemic; Lymphocytes; Middle Aged

On the basis of the previous study, on the cell interaction between malignant tumor cells and other cells, especially with lymphocytes, the present study was carried out by investigating cell to cell interaction of human malignant tumor cells and human lymphoblastoid cells such as T-cell (MOLT-4 cell) and B-cell (Burkitt lymphoma cell). As a result it has been revealed that live lymphoblastoid cells were not adhered on the cell surface of the tumor cells, nor is it ingested by tumor cells, but in thepresence of HVJ (Sendai virus: 2,000 H.A. units) it adheres slightly on the cell surface of tumor cell but no cell fusion of tumor cells and lymphoblastoid cells is observable. On the other hand, the tumor cell as well as T-cell and B-cell all have receptors to concanavalin A (Con. A) on their cell surfaces, and they show a marked cell binding such as tumor cell and T-cell, tumor cell and B-cell, and there can be observed a marked phagocytosis of lymphoblastoid cells by tumor cells. Moreover, the tumor cells that have phagocytized lymphoblastoid cells undergo a marked cell destruction within 4 hours of cell-binding and phagocytosis, which is especially prominent in the case of phagocytosis of E.B cell by tumor cell.

Topics: Animals; Antigen-Antibody Reactions; B-Lymphocytes; Cell Line; Cell Transformation, Neoplastic; Chickens; Concanavalin A; Cricetinae; Cytotoxicity Tests, Immunologic; Erythrocytes; Female; Glutaral; Humans; Immune Adherence Reaction; Leukemia, Lymphoid; Lupus Erythematosus, Systemic; Lymphocytes; Neoplasms; Ovarian Neoplasms; Parainfluenza Virus 1, Human; Phagocytosis; Sheep; T-Lymphocytes; Teratoma

The response of peripheral blood lymphocytes to the phytomitogens, PHA, Con A, and PWM, was evaluated in 30 SLE patients and in 30 age, sex, and race-matched controls using dose and time responses. The proliferative response to the three phytomitogens was not depressed in this group of subacute and chronic SLE patients. Active lupus nephritis and a slow acetylator phenotype were associated with a decreased lymphocyte response. The incidence of a slow acetylator phenotype in spontaneous SLE was 68%. In interpreting the lymphocyte response to phytomitogens, the importance of a clear definition of the SLE group under study, the activity of the disease, and treatment status are emphasized.

Topics: Adolescent; Adult; Aged; Concanavalin A; Female; Humans; Immunity, Cellular; Lectins; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Nephritis; Sulfamethazine

Topics: ABO Blood-Group System; Acetamides; Animals; beta-Aminoethyl Isothiourea; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Separation; Complement System Proteins; Concanavalin A; Cysteine; Cytotoxicity Tests, Immunologic; Dithiothreitol; Dose-Response Relationship, Drug; Erythrocytes; Glutathione; Hemagglutination Tests; Hemoglobinuria, Paroxysmal; Humans; Hydrogen-Ion Concentration; Hypersplenism; Immune Adherence Reaction; Isoantibodies; Leukemia, Lymphoid; Lupus Erythematosus, Systemic; Lymphocytes; Mercaptoethanol; Microscopy, Electron, Scanning; Sheep; Sulfhydryl Compounds