ovalbumin and Lupus-Erythematosus--Systemic

Topics: Aleutian Mink Disease; Anemia, Hemolytic, Autoimmune; Animals; Antibodies, Antinuclear; Antigen-Antibody Reactions; Collagen Diseases; Disease Models, Animal; Graft vs Host Reaction; Lupus Erythematosus, Systemic; Lymph Nodes; Mice; Ovalbumin; Rabbits; Spleen; Transplantation, Homologous

Caspase-1 is required for nephritis and robust autoantibody development in the pristane model of murine lupus. The objective of this study was to evaluate the immune response and to study the splenic B and T cell populations in wild-type (WT) and caspase-1-/- mice following pristane injection in order to develop an understanding of why absence of caspase-1 is protective in pristane-induced lupus.. Immunization responses to NP-Ficoll and NP-ovalbumin were assessed in WT and caspase-1-/- mice. In vitro IgM and IgG responses to R848 were measured by ELISA. Serum IgM anti-dsDNA and IL-1β were also measured by ELISA. B and T cell populations 2 weeks and 6 months following pristane injection were measured by flow cytometry in WT and caspase-1-/- mice.. Caspase-1-/- mice generate equivalent IgG responses to NP-Ficoll and NP-ova antigens when compared to wild-type mice. Additionally, they secrete IgM and IgG in response to TLR7 activation. Pristane injected WT and caspase-1-/- mice generate robust IgM anti-dsDNA responses. Caspase-1-/- mice have a significant reduction in marginal zone B cell populations compared to WT 6 months after pristane exposure whereas T cell responses are intact in these mice.. Caspase-1-/- mice have intact immune responses but do not develop an expanded marginal zone B cell population in response to pristane-induced lupus. This may be one explanation for reduced IgG autoantibody production in these mice.

Topics: Animals; Antibodies, Antinuclear; B-Lymphocytes; Caspase 1; Cells, Cultured; Disease Models, Animal; Ficoll; Genetic Predisposition to Disease; Imidazoles; Immunization; Immunoglobulin G; Immunoglobulin M; Lupus Erythematosus, Systemic; Mice, Inbred BALB C; Mice, Knockout; Nitrophenols; Ovalbumin; Phenotype; Phenylacetates; Spleen; T-Lymphocytes; Terpenes; Time Factors

Neutrophil extracellular traps (NETs) are web-like structures released by activated neutrophils. Recent studies suggest that NETs play an active role in driving autoimmunity and tissue injury in diseases including rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). The purpose of this study was to investigate if celastrol, a triterpenoid compound, can inhibit NET formation induced by inflammatory stimuli associated with RA and SLE. We found that celastrol can completely inhibit neutrophil oxidative burst and NET formation induced by tumor necrosis factor alpha (TNFα) with an IC50 of 0.34 µM and by ovalbumin:anti-ovalbumin immune complexes (Ova IC) with an IC50 of 1.53 µM. Celastrol also completely inhibited neutrophil oxidative burst and NET formation induced by immunoglobulin G (IgG) purified from RA and SLE patient sera. Further investigating into the mechanisms, we found that celastrol treatment downregulated the activation of spleen tyrosine kinase (SYK) and the concomitant phosphorylation of mitogen-activated protein kinase kinase (MAPKK/MEK), extracellular-signal-regulated kinase (ERK), and NFκB inhibitor alpha (IκBα), as well as citrullination of histones. Our data reveals that celastrol potently inhibits neutrophil oxidative burst and NET formation induced by different inflammatory stimuli, possibly through downregulating the SYK-MEK-ERK-NFκB signaling cascade. These results suggest that celastrol may have therapeutic potentials for the treatment of inflammatory and autoimmune diseases involving neutrophils and NETs.

Topics: Arthritis, Rheumatoid; Cells, Cultured; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Extracellular Traps; Humans; I-kappa B Proteins; Immunoglobulin G; Inflammation; Intracellular Signaling Peptides and Proteins; Lupus Erythematosus, Systemic; MAP Kinase Kinase Kinases; Neutrophils; NF-KappaB Inhibitor alpha; Ovalbumin; Pentacyclic Triterpenes; Phosphorylation; Protein-Tyrosine Kinases; Respiratory Burst; Syk Kinase; Tripterygium; Triterpenes; Tumor Necrosis Factor-alpha

We investigated the role of effector CD8 T cells in the pathogenesis of immune glomerular injury. BALB/c mice are not prone to autoimmune disease, but after 12 immunizations with OVA they developed a variety of autoantibodies and glomerulonephritis accompanied by immune complex (IC) deposition. In these mice, IFN-γ-producing effector CD8 T cells were significantly increased concomitantly with glomerulonephritis. In contrast, after 12 immunizations with keyhole limpet hemocyanin, although autoantibodies appeared, IFN-γ-producing effector CD8 T cells did not develop, and glomerular injury was not induced. In β2-microglobulin-deficient mice lacking CD8 T cells, glomerular injury was not induced after 12 immunizations with OVA, despite massive deposition of IC in the glomeruli. In mice containing a targeted disruption of the exon encoding the membrane-spanning region of the Ig μ-chain (μMT mice), 12 immunizations with OVA induced IFN-γ-producing effector CD8 T cells but not IC deposition or glomerular injury. When CD8 T cells from mice immunized 12 times with OVA were transferred into naive recipients, glomerular injury could be induced, but only when a single injection of OVA was also given simultaneously. Importantly, injection of OVA could be replaced by one injection of the sera from mice that had been fully immunized with OVA. This indicates that deposition of IC is required for effector CD8 T cells to cause immune tissue injury. Thus, in a mouse model of systemic lupus erythematosus, glomerular injury is caused by effector CD8 T cells that recognize Ag presented as IC on the target renal tissue.

Topics: Animals; Antigen Presentation; Antigen-Antibody Complex; beta 2-Microglobulin; CD8-Positive T-Lymphocytes; Female; Glomerulonephritis; Immunoglobulin mu-Chains; Interferon-gamma; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; T-Lymphocytes, Cytotoxic

Phosphoinositide-3 kinase (PI3K)-δ and PI3K-γ are preferentially expressed in immune cells, and inhibitors targeting these isoforms are hypothesized to have anti-inflammatory activity by affecting the adaptive and innate immune response. We report on a potent oral PI3K-δ and PI3K-γ inhibitor (IPI-145) and characterize this compound in biochemical, cellular, and in vivo assays. These studies demonstrate that IPI-145 exerts profound effects on adaptive and innate immunity by inhibiting B and T cell proliferation, blocking neutrophil migration, and inhibiting basophil activation. We explored the therapeutic value of combined PI3K-δ and PI3K-γ blockade, and IPI-145 showed potent activity in collagen-induced arthritis, ovalbumin-induced asthma, and systemic lupus erythematosus rodent models. These findings support the hypothesis that inhibition of immune function can be achieved through PI3K-δ and PI3K-γ blockade, potentially leading to significant therapeutic effects in multiple inflammatory, autoimmune, and hematologic diseases.

Topics: Animals; Arthritis; Asthma; Collagen Type II; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Humans; Isoquinolines; Lupus Erythematosus, Systemic; Molecular Structure; Ovalbumin; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Purines; Rats; Rats, Inbred Lew; Rats, Wistar; Structure-Activity Relationship

The association of allergic diseases and disease activity in systemic lupus erythematosus (SLE, lupus) is controversial. The study investigates lupus activity-related differences in the induction of late allergic rhinitis (LAR) in the female NZB×NZW(B/W)F(1) mouse model for lupus. The LAR, which is induced by ovalbumin, was examined during the preactive (before clinical onset) and active (after clinical onset) phases in mice. Induction of LAR was less severe in mice with active lupus in contrast to clinically healthy lupus mice that developed a more severe allergic rhinitis. Inhibition of the development of LAR may be due to reduced eosinophilia and local interleukin-4 secretion during active autoimmune disease. In addition, systemic interferon-γ, but not IL-4, production increased during the active phase, but not the preactive phase. This suggests that the predominating Th1 lineage commitment in mice with active lupus may be responsible for the inhibition of the allergic Th2 response. The present study may shed some light on the controversy of the prevalence of allergic diseases in SLE patients.

Topics: Animals; Antibodies, Antinuclear; Blood Urea Nitrogen; Creatinine; Eosinophilia; Eosinophils; Female; Inflammation; Interferon-gamma; Interleukin-4; Leukocytes; Lupus Erythematosus, Systemic; Lymphocyte Count; Mice; Mice, Inbred BALB C; Mice, Inbred NZB; Nasal Lavage Fluid; Neutrophils; Ovalbumin; Proteinuria; Rhinitis, Allergic; Rhinitis, Allergic, Perennial; Spleen; Th1 Cells; Th2 Cells

T cell hyperactivation and complement consumption are prominent features of the immunopathology of systemic lupus erythematosus. Although complement activation is secondary to autoantibodies that form immune complexes (ICs), the trigger for alterations in human peripheral blood T cells is poorly understood. To study the impact (on T cells) of several types of preformed ICs and terminal complement complex, also referred to as C5b-9, we incubated these immune reactants with peripheral blood naive CD4(+) T cells as well as Jurkat cells and analyzed their effects on cellular behavior. We first assembled the C5b-9 in situ on the membrane and observed its assembly primarily on a single site where it promoted aggregation of membrane rafts and recruitment of the CD3 signaling complex. However, C5b-9 alone did not initiate proliferation or commencement of downstream signaling events associated with T cell activation. When T cells were treated with ICs together with nonlytic C5b-9, changes associated with T cell activation by possible antigen engagement then occurred. T cell antigen receptor signaling proteins, including ζ-chain, ZAP-70, Syk, Src, and Lck, were phosphorylated and organized in a synapse-like structure. The cytoskeleton formed F-actin spindles and a distal pole complex, resulting in a bipolar distribution of phosphorylated ezrin-radixin-moesin and F-actin. Furthermore, ICs and nonlytic C5b-9 induced T cell proliferation and IFN-γ production. These results raise the possibility that ICs and the nonlytic C5b-9 modulate T cell-mediated responses in systemic lupus erythematosus and other related chronic inflammatory disorders.

Topics: Actins; CD3 Complex; CD4-Positive T-Lymphocytes; Cell Proliferation; Complement Membrane Attack Complex; Complement System Proteins; Cytoskeleton; Humans; Immune System; Interferon-gamma; Jurkat Cells; Lupus Erythematosus, Systemic; Microscopy, Confocal; Neurofibromin 2; Ovalbumin; Phosphorylation; Signal Transduction; T-Lymphocytes

Autoantibody-mediated diseases like myasthenia gravis, autoimmune hemolytic anemia and systemic lupus erythematosus represent a therapeutic challenge. In particular, long-lived plasma cells producing autoantibodies resist current therapeutic and experimental approaches. Recently, we showed that the sensitivity of myeloma cells toward proteasome inhibitors directly correlates with their immunoglobulin synthesis rates. Therefore, we hypothesized that normal plasma cells are also hypersensitive to proteasome inhibition owing to their extremely high amount of protein biosynthesis. Here we show that the proteasome inhibitor bortezomib, which is approved for the treatment of multiple myeloma, eliminates both short- and long-lived plasma cells by activation of the terminal unfolded protein response. Treatment with bortezomib depleted plasma cells producing antibodies to double-stranded DNA, eliminated autoantibody production, ameliorated glomerulonephritis and prolonged survival of two mouse strains with lupus-like disease, NZB/W F1 and MRL/lpr mice. Hence, the elimination of autoreactive plasma cells by proteasome inhibitors might represent a new treatment strategy for antibody-mediated diseases.

Topics: Animals; Boronic Acids; Bortezomib; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; Mice, Inbred MRL lpr; Mice, Inbred NZB; Models, Immunological; Nephritis; Ovalbumin; Plasma Cells; Protease Inhibitors; Proteasome Inhibitors; Pyrazines; Time Factors

Previous studies have shown the importance of somatic mutations and arginine residues in the complementarity-determining regions (CDRs) of pathogenic anti-double-stranded DNA (anti-dsDNA) antibodies in human and murine lupus, and in studies of murine antibodies, a role of mutations at position 53 in V(H) CDR2 has been demonstrated. We previously demonstrated in vitro expression and mutagenesis of the human IgG1 monoclonal antibody B3. The present study was undertaken to investigate, using this expression system, the importance of the arginine residue at position 53 (R53) in B3 V(H).. R53 was altered, by site-directed mutagenesis, to serine, asparagine, or lysine, to create 3 expressed variants of V(H). In addition, the germline sequence of the V(H)3-23 gene (from which B3 V(H) is derived) was expressed either with or without arginine at position 53. These 5 new heavy chains, as well as wild-type B3 V(H), were expressed with 4 different light chains, and the resulting antibodies were assessed for their ability to bind to nucleosomes, alpha-actinin, cardiolipin, ovalbumin, beta(2)-glycoprotein I (beta(2)GPI), and the N-terminal domain of beta(2)GPI (domain I), using direct binding assays.. The presence of R53 was essential but not sufficient for binding to dsDNA and nucleosomes. Conversely, the presence of R53 reduced binding to alpha-actinin, ovalbumin, beta(2)GPI, and domain I of beta(2)GPI. The combination B3 (R53S) V(H)/B3 V(L) bound human, but not bovine, beta(2)GPI.. The fact that the R53S substitution significantly alters binding of B3 to different clinically relevant antigens, but that the alteration is in opposite directions depending on the antigen, implies that this arginine residue plays a critical role in the affinity maturation of antibody B3.

Topics: Amino Acid Sequence; Antiphospholipid Syndrome; Arginine; Binding Sites; DNA Primers; Germ-Line Mutation; Humans; Immunoglobulin G; Immunoglobulin Heavy Chains; Immunoglobulin Variable Region; Lupus Erythematosus, Systemic; Molecular Sequence Data; Mutagenesis, Site-Directed; Mutation; Ovalbumin; Reproducibility of Results

Secreted IgA plays a pivotal role in the mucosal immunity to maintain the front line of body defense. We found that the level of fecal IgA was dramatically decreased in aged (NZB x NZW)F(1) (BWF(1)) mice developing lupus nephritis, whereas levels in similarly aged New Zealand Black (NZB) and New Zealand White (NZW) mice remained unchanged compared with young mice. The number of cells obtained from Peyer's patches was markedly decreased in aged BWF(1) mice. Aged BWF(1) mice showed increased susceptibility to pathogenic bacterial infection. Furthermore, oral administration of OVA failed to inhibit secondary IgG response induced by systemic immunization, suggesting defective oral tolerance in aged BWF(1) mice. A significant amount of orally administered OVA was incorporated directly into the intestinal lamina propria in aged BWF(1) mice whereas it was mainly localized in subepithelial domes and interfollicular region in Peyer's patches in young mice. T cells obtained from renal and pulmonary lymph nodes of aged BWF(1) mice that had been orally administered with OVA showed an Ag-specific T cell proliferation, whereas those from young BWF(1), aged NZB, and aged NZW mice did not. Interestingly, aerosol exposure to OVA of aged BWF(1) mice, which had been orally administered with the same Ag, provoked an eosinophil infiltration in the lung. These results demonstrate that mucosal immunity in the gut is impaired and oral Ags induce systemic sensitization instead of oral tolerance in the development of murine lupus.

Topics: Administration, Inhalation; Administration, Oral; Aging; Animals; Antigens; Cell Movement; Crosses, Genetic; Eosinophilia; Escherichia coli Infections; Female; Genetic Predisposition to Disease; Hydrazines; Immune Tolerance; Immunity, Mucosal; Immunoglobulin A; Intestinal Mucosa; Lupus Erythematosus, Systemic; Lupus Nephritis; Lymphocyte Activation; Mice; Mice, Inbred NZB; Ovalbumin

Sle3 is an NZM2410-derived lupus susceptibility locus on murine chromosome 7. Congenic recombination has resulted in a novel mouse strain, B6.Sle3, associated with serum antinuclear autoantibodies (ANAs), T cell hyperactivity, and elevated CD4/CD8 ratios. An OVA-specific TCR transgene was used as a tool to demonstrate that Sle3 facilitated heightened T cell expansion in vitro, and in vivo, following antigen challenge. Indeed, continued T cell expansion was noted even in response to a tolerogenic signal. However, these phenotypes did not appear to be T cell intrinsic but were dictated by hyperstimulatory B6.Sle3 APCs. Importantly, B6.Sle3-derived DCs and macrophages appeared to be significantly more mature/activated, less apoptotic, and more proinflammatory and were better at costimulating T cells in vitro, compared with the B6 counterparts. Finally, the adoptive transfer of B6.Sle3-derived DCs into healthy B6 recipients elicited increased CD4/CD8 ratios and serum ANAs, 2 cardinal Sle3-associated phenotypes. We posit that their heightened expression of various costimulatory molecules, including CD80, CD106, I-A, and CD40, and their elevated production of various cytokines, including IL-12 and IL-1beta, may explain why Sle3-bearing DCs may be superior at breaching self tolerance. These studies provide mechanistic evidence indicating that intrinsic abnormalities in DCs and possibly other myeloid cells may dictate several of the phenotypes associated with systemic lupus, including ANA formation and T cell hyperactivity.

Topics: Adoptive Transfer; Amino Acid Sequence; Animals; Antibodies, Antineutrophil Cytoplasmic; Antigen-Presenting Cells; CD4-CD8 Ratio; Cytokines; Dendritic Cells; In Vitro Techniques; Lupus Erythematosus, Systemic; Lymphocyte Activation; Macrophages; Mice; Mice, Mutant Strains; Mice, Transgenic; Molecular Sequence Data; Neutrophils; Ovalbumin; Peptides; Receptors, Antigen, T-Cell; Self Tolerance; T-Lymphocytes

Ly-6 proteins appear to serve cell adhesion and cell signaling function, but the precise role of Ly-6A.2 in CD4+ T lymphocytes is still unclear. Overexpression of Ly-6A.2 in T lymphocytes has allowed us to analyze the influence of elevated Ly-6A.2 expression on T cell function. In this study we report reduced proliferation of CD4+ T cells overexpressing Ly-6A.2 in response to a peptide Ag. Moreover, the Ly-6A.2-overexpressing CD4+ cells generated elevated levels of IL-4, a key factor that propels the differentiation of naive CD4+ T cells into Th2 subset. The hyporesponsiveness of Ly-6A.2 transgenic CD4+ T cells is dependent on the interaction of Ly-6A.2 T cells with the APCs and can be reversed by blocking the interaction between Ly-6A.2 and a recently reported candidate ligand. Overexpression of Ly-6A.2 in CD4+ T cells reduced their Ca(2+) responses to TCR stimulation, therefore suggesting effects of Ly-6A.2 signaling on membrane proximal activation events. In contrast to the observed Ag-specific hyporesponsiveness, the Ly-6A.2 transgenic CD4+ T cells produced IL-4 independent of the interactions between Ly-6A.2 and the candidate Ly-6A.2 ligand. Our results suggest that 1) interaction of Ly-6A.2 with a candidate ligand regulates clonal expansion of CD4+ Th cells in response to an Ag (these results also provide further functional evidence for presence of Ly-6A.2 ligand on APC); and 2) Ly-6A.2 expression on CD4+ T cells promotes production of IL-4, a Th2 differentiation factor.

Topics: Animals; Antigen Presentation; Antigen-Presenting Cells; Antigens; Antigens, Ly; Calcium; CD4-Positive T-Lymphocytes; Cells, Cultured; Cytokines; Interleukin-4; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Mice, Inbred MRL lpr; Mice, Transgenic; Ovalbumin; Peptide Fragments; Receptors, Antigen, T-Cell; Spleen

The expression of a transgene encoding the I-E alpha chain prevents a lupus-like autoimmune syndrome in BXSB mice. However, it had not been elucidated whether the E alpha d transgene-mediated protective effect results from I-E expression or from the generation of I-E alpha chain-derived peptides (E alpha peptide) displaying high affinity for the I-Ab molecule. To address this question, two different BXSB lines expressing the transgene at low or high levels were crossed with lupus-prone MRL mice; this resulted in three types of (MRL x BXSB)F1 mice, differing in the expression levels of I-E molecules and of E alpha peptides presented by I-Ab molecules. Comparative analysis of these three (MRL x BXSB)F1 mice as well as several BXSB transgenic lines showed that the E alpha d transgene-mediated protection paralleled the expression levels of E alpha peptide presented by I-Ab molecules, but not of I-E molecules on B cells. In addition, use of transgenic and nontransgenic double bone marrow chimeras showed a selective activation of nontransgenic B cells during I-Ab-restricted T cell-dependent immune responses, while both transgenic and nontransgenic B cells were comparably activated during T cell-independent responses. These results favor a model of autoimmunity prevention based on competition for antigen presentation, in which excessive generation of E alpha peptides prevents, because of their high affinity to the I-A molecules, activation of potential autoreactive T and B cells.

Topics: Amino Acid Sequence; Animals; Crosses, Genetic; Female; Gene Dosage; Histocompatibility Antigens Class II; Lupus Erythematosus, Systemic; Male; Mice; Mice, Transgenic; Molecular Sequence Data; Ovalbumin; Peptides; Protein Binding; Radiation Chimera; Transgenes

Procainamide (PA) is the drug most commonly associated with the induction of autoantibodies and drug-related lupus (DRL). While the majority of these patients express autoantibodies, antibodies to the parent drug and metabolites, PA-hydroxylamine (PAHA) or nitroso-PA (NOPA), have not been reported in humans. Hapten-carrier conjugates were prepared using human hemoglobin (HgB) or autologous rabbit erythrocytes with PAHA or NOPA. PA was conjugated to rabbit serum albumin (RSA) or egg albumin (OVA) via diazotization and condensation methods. Rabbits were immunized with hapten conjugates in Freund's adjuvant. These hapten-carrier compounds (5-10 micrograms/ml) were used as test antigens for antibodies in sera from the rabbits and 40 patients on chronic PA treatment. 10 SLE patients, 33 elderly and 20 young normal controls by ELISA. Type I and II collagens were also used as test antigens for human sera. Sera from rabbits immunized with the PA compounds had elevated IgG antibody values to PA, PAHA and NOPA, but no autoantibodies. Absorption of the rabbit sera with the PA compounds reduced the antibody levels; ssDNA and histones failed to inhibit the total binding values. Mean binding to PA-OVA was 0.95 +/- 0.41 for PA patients and 1.37 +/- 0.26 standard error of means (S.E.M.) in the SLE patients compared to 0.37 +/- 0.14 S.E.M. in the normal sera (P < or = 0.05); similar binding values to PAHA-HgB and NOPA-HgB were also observed. Sixty-eight percent of the PA patients had antibodies to type II collagen. Elevated binding values to PA compounds were inhibited by absorption of human sera with ssDNA or total histones; absorption with PA or PAHA had no significant effect. These findings suggest that sera from PA patients containing high titers of autoantibodies cross-react in vitro with unrelated antigens.

Topics: Adult; Aged; Aged, 80 and over; Animals; Antibody Specificity; Autoantibodies; Collagen; Female; Haptens; Hemoglobin A; Humans; Lupus Erythematosus, Systemic; Male; Middle Aged; Ovalbumin; Procainamide; Rabbits; Serum Albumin

We used synthetic peptides to analyze the human natural antibody response to V beta determinants. A major determinant recognized by IgM and IgG autoantibodies of clinically healthy individuals as well as those suffering from rheumatoid arthritis (RA) was a peptide corresponding to the first complementarity-determining region (CDR1). The natural IgM response of RA patients to this synthetic autoepitope was significantly elevated relative to that shown by healthy individuals. The levels of IgM reactivity to determinants corresponding to this region decreased with increasing age. By contrast, IgG autoantibodies to certain V beta CDR1 peptides increased markedly with age. In order to determine whether the CDR1 V beta determinant might be involved in immunization, we immunized rabbits with a human peptide that is greater than 80% identical to the homologous sequence derived from a rabbit V beta gene. As a control, the rabbits were immunized with a peptide of equal length derived from the N-terminus of the human V beta chain. Like humans, rabbits tended to have high levels of autoantibodies to the CDR1 peptide but not to the N-terminal segment. Following immunization, the rabbits produced strong IgG responses to the N-terminal peptide. By contrast, immunization with the CDR1 peptide resulted in levels of IgG antibody less than or equal to the natural activity in unimmunized rabbits. These studies indicate that the CDR1 region of Tcr V beta is a widely recognized autoantigenic portion of the Tcr that most probably functions as a regulatory epitope in man and other species.

Topics: Adult; Aged; Aged, 80 and over; Aging; Amino Acid Sequence; Arthritis, Rheumatoid; Autoantibodies; Autoimmune Diseases; Dose-Response Relationship, Immunologic; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoglobulin G; Immunoglobulin M; Immunoglobulin Variable Region; Immunoglobulins, Intravenous; Lupus Erythematosus, Systemic; Male; Middle Aged; Molecular Sequence Data; Ovalbumin; Receptors, Antigen, T-Cell, alpha-beta; Sequence Alignment; Sequence Homology, Amino Acid

B cells from autoimmune NZB mice were transferred into unmanipulated non-autoimmune NZB.xid mice. The number of antibody-producing cells against various antigens in recipient mice was monitored at varying time after cell transfer using ELISPOT assay. NZB B cells producing antibody against all antigens we examined were able to proliferate in NZB.xid mice, which supports the idea of polyclonal B cell activation. However, anti-DNA producing cells proliferated most rapidly, and anti-BrMRBC producing cells proliferated more rapidly than B cells of other antigenic specificities. The percentage of anti-DNA producing cells in total immunoglobulin-producing cells increased over time whereas the percentage of anti-ovalbumin producing cells kept the same level. This indicates directly the preferential proliferation of NZB anti-DNA producing cells in NZB.xid mice. The result shows the responsibility of antigen-specific stimulation or activation on autoimmunity in the context of polyclonal B cell activation.

Topics: Actins; Animals; Antibodies, Antinuclear; Autoimmunity; B-Lymphocytes; Cell Division; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Erythrocytes; Immunization, Passive; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Ovalbumin; Spleen; Thymus Gland

Topics: Administration, Oral; Age Factors; Animals; Caseins; Dietary Proteins; Female; gamma-Globulins; Humans; Immune Tolerance; Immunization; Injections, Intraperitoneal; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Ovalbumin

Primary oral antigen exposure normally induces mucosal immunity and an active suppression of the systemic immune response. Patients with systemic lupus erythematosus (SLE) have increased antibodies to bovine gamma-globulin (BGG), which suggested a possible failure of oral tolerance in SLE. We examined this possibility in murine lupus. NZB/W females were fed BGG or saline and were subsequently immunized ip. Primary and secondary responses were assessed. At 1 month of age the mice tolerized normally in response to feeding with BGG but, at 4 months of age, not only did they not tolerize, the mice fed BGG had a 5- to 7-fold higher response to parenteral immunization than did the saline-fed mice. Control strain mice tolerized normally at both ages (a 5- to 10-fold lower response). Conversely, when fed ovalbumin, NZB/W females tolerized normally at both 1 and 4 months of age, and patients with SLE had normal levels of antibody to this antigen. However, we also found increased levels of antibodies to bovine casein in SLE patients, and found that NZB/W mice failed to orally tolerize with this antigen at either 1 or 4 months of age. Thus, the failure of oral tolerance in the NZB/W mice appears to be antigen specific and age dependent and, at least with respect to these three antigens, appears to parallel the antibody patterns seen in human SLE.

Topics: Administration, Oral; Age Factors; Animals; Antibody Formation; Antigens; BCG Vaccine; Caseins; Female; Humans; Immune Tolerance; Lupus Erythematosus, Systemic; Mice; Mice, Inbred Strains; Ovalbumin; T-Lymphocytes, Regulatory

The investigation described here is concerned with the T cell regulation of the antigen-specific antibody response which has been studied in patients suffering from systemic lupus erythematosus (SLE). Apart from the fact that T helper cell activity was found to be less efficient, it appeared that the peripheral blood leucocytes (PBL) of patients in an active stage of the disease did not contain the suppressor precursor cells, which functions as the target cell for the inductive signal of T mu+ suppressor inducer cells. The absence of the suppressor precursor cells in SLE patients coincided with the absence of T gamma+ suppressor effector cells. Characterization of the (post-thymic) precursor cells (derived from normal donors) with the aid of monoclonal antibodies of the OKT series and several other markers pointed out that this population contains OKT4+ as well as OKT8+ cells. Further experiments demonstrated that the cells are capable of rosetting with autologous erythrocytes, and do not bear Fc receptors for IgM or IgG. Considering the various findings as a whole the conclusion is warranted that the post-thymic suppressor precursor T cell can differentiate into a suppressor effector cell only after interaction with T suppressor inducer cells.

Topics: Antibodies, Monoclonal; Antibody Formation; Antibody Specificity; Hemolytic Plaque Technique; Humans; Lupus Erythematosus, Systemic; Ovalbumin; Phenotype; T-Lymphocytes, Regulatory

A technique is described which allows the antibodies of circulating immune complexes to be isolated as their F(ab')2 fragments. The method is based on the precipitation of the complexes by the sequential addition of conglutinin and anti-conglutinin, and the subsequent digestion of these precipitates by pepsin. Using this technique it has been possible to show antibodies to Epstein-Barr (EB) virus antigens in the immune complexes of patients with Burkitt's lymphoma and to microbial antigens in two patients with nephritis. By substituting DNAase for pepsin it has also been possible to show antibodies to DNA-containing nuclear antigens in the serum of patients with systemic lupus erythematosus.

Topics: Antibodies, Viral; Antigen-Antibody Complex; Antigens, Bacterial; Burkitt Lymphoma; Collectins; Fluorescent Antibody Technique; Glomerulonephritis; Herpesvirus 4, Human; Humans; Immunodiffusion; Immunoglobulin Fab Fragments; Lupus Erythematosus, Systemic; Methods; Ovalbumin; Pepsin A; Serum Globulins

Antigen-specific suppressor cell activity of peripheral blood mononuclear cells was investigated in twenty-nine patients with systemic lupus erythematosus (SLE) and sixteen normal, age- and sex-matched healthy controls. Suppressor cell activity was generated by priming peripheral blood mononuclear cells with high dose antigen (ovalbumin) and adding the washed primed or control (unprimed) cells to autologous optimally stimulated target plaque-forming cell (PFC) cultures. The ability of the primed cells to interfere with an optimal ovalbumin-specific PFC response in the target cultures was used as a measure of antigen-specific suppressor cell activity. The results demonstrated reduced suppressor cell activity in the SLE patients relative to controls--46.8 +/- 3.6% vs 63 +/- 2.4% suppression respectively (P less than 0.01). Consistent with reduced suppressor cell activity was an increase in the plaque-forming cell response to ovalbumin in patients relative to controls (880 +/- 73 vs 763 +/- 102 PFC/10(6) cells respectively [P = 0.10]). No correlation was demonstrated between suppressor cell activity in SLE patients and disease activity or therapy.

Topics: Adult; Cells, Cultured; Epitopes; Female; Hemolytic Plaque Technique; Humans; Lupus Erythematosus, Systemic; Male; Middle Aged; Ovalbumin; T-Lymphocytes, Regulatory

A new assay for the detection of circulating immune complexes, the polyethylene glycol precipitation-complement consumption assay (PEG-CC), is described. The test is both simple and sensitive, and exhibits a high degree of specificity. Immune complexes are first isolated from serum by precipitation in 2.5% polyethylene glycol (PEG) and concentrated. They are then assayed functionally by measuring their ability to fix complement using a sensitive kinetic assay for total haemolytic complement. The test can detect aggregated IgG in serum at concentrations around 6.0 micrograms/ml (about 2.0 micrograms absolute. Using DNA-anti-DNA and ovalbumin-anti-ovalbumin immune complexes prepared in vitro, antigen concentrations less than 0.5 micrograms/ml can be detected. Interference by endogenous complement, polyanions and other factors in test sera has been virtually eliminted by the design of the assay. The increased specificity of the PEG-CC test for immune complexes, should prove useful in the diagnosis and monitoring of immune complex-mediated diseases.

Topics: Animals; Antigen-Antibody Complex; Chemical Precipitation; Complement System Proteins; DNA; Edetic Acid; Female; Freezing; Humans; Immunoassay; Immunoglobulin G; Lupus Erythematosus, Systemic; Macromolecular Substances; Male; Ovalbumin; Polyethylene Glycols; Rabbits; Temperature

An assay for circulating immune complexes is described which uses radiolabelled bovine conglutinin as ligand and polyethylene glycol precipitation as the technique for separating bound ligands. The technique is simple to perform and gives good sensitivity detecting artificial immune complexes. Its use in detecting complexes in systemic lupus erythematosus and Burkitt's lymphoma is described and it is compared with the Clq binding assay also performed with polyethylene glycol. It is suggested that the simultaneous performance of polyethylene glycol assays using radiolabelled Clq and radiolabelled conglutinin may be an advantageous method for screening sera for the presence of immune complexes.

Topics: Animals; Antigen-Antibody Complex; Binding Sites; Burkitt Lymphoma; Cattle; Chemical Precipitation; Complement Fixation Tests; DNA; Edetic Acid; Humans; Iodine Radioisotopes; Lipopolysaccharides; Lupus Erythematosus, Systemic; Ovalbumin; Polyethylene Glycols; Rabbits; Solubility

The corneo-scleral changes described are highly characteristic and may be the first signs of an underlying systemic disorder so that otherwise healthy patients who present with limbal guttering and scleral disease must be continuously monitored with this association in mind. The clinical and histological features of limbal guttering in connective tissue disorders strongly suggest that a local antigen-antibody reaction triggers off a number of biochemical and cellular responses which combine to produce lysis of scleral and corneal collagen, although immune complexes have not so far been demonstrated in these eyes. Modes of therapy aimed at one particular chain of events have varying degrees of success, as indeed does more blunderbuss treatment with steroids, anti-inflammatory drugs, or cytotoxic agents. The early stages of the ocular lesion in the rabbit are now being studied, as is the immunological basis for its production. It is hoped that further work with the animal model will lead to a deeper understanding of the pathogenesis of these conditions, which will turn provide both ophthalmologists and rheumatologists with more scientific guidelines for treatment.

Topics: Animals; Arthritis, Rheumatoid; Collagen Diseases; Cornea; Corneal Diseases; Disease Models, Animal; Granulomatosis with Polyangiitis; Humans; Lupus Erythematosus, Systemic; Ovalbumin; Polyarteritis Nodosa; Rabbits

Topics: Animals; Antigens; Antimetabolites; Cell Migration Inhibition; Cells, Cultured; DNA; Erythrocytes; Guinea Pigs; Humans; Hypersensitivity, Delayed; Immune Sera; Immunity, Cellular; Immunization, Passive; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocytes; Macrophage Migration-Inhibitory Factors; Macrophages; Myelin Basic Protein; Ovalbumin; Peptides; Phagocytosis; Rabbits; Sheep; Tuberculin

Topics: Animals; Antibodies, Antinuclear; Antigen-Antibody Complex; Electrophoresis; Female; Fluorescent Antibody Technique; Glomerulonephritis; Graft vs Host Disease; Immune Sera; Immunization; Immunoglobulin G; Kidney Glomerulus; Lupus Erythematosus, Systemic; Male; Mice; Microscopy, Electron; Nephritis; Ovalbumin; Rabbits

Topics: Animals; Antibodies, Anti-Idiotypic; Antigen-Antibody Reactions; Autoimmune Diseases; Complement Fixation Tests; Disease Models, Animal; Female; Hemagglutination Tests; Hypersensitivity, Immediate; Lupus Erythematosus, Systemic; Male; Mice; New Zealand; Ovalbumin; Polysaccharides; Proteins; Species Specificity

Topics: Antigen-Antibody Complex; Chromatography; Chromatography, Gel; Complement System Proteins; Edetic Acid; Glomerulonephritis; Humans; Immune Sera; Immunodiffusion; Lupus Erythematosus, Systemic; Molecular Weight; Nephritis; Ovalbumin; Radioimmunoassay

Young adult mice (30 to 43 days old) of autoimmune NZB and BIW F(1) strains failed to develop immunologic tolerance when treated with ultracentrifuged bovine gamma globulin and then challenged 12 days later. By contrast, weanling NZB and B/ W F(1) mice (18 to 25 days) as well as weanling C3H mice (16 to 19 days) became tolerant and had no serum antibody 12 days after challenge. The C3H mice remained unresponsive, but NZB and B! W F(1) mice produced antibody between days 27 and 41. The rapid loss of previously established tolerance to foreign antigens could have its parallel in the loss of tolerance to autoantigens with subsequent development of lupus nephritis and Coombs' positive hemolytic anemia in these animals.

Topics: Anemia, Hemolytic, Autoimmune; Animals; Animals, Newborn; Antibody Formation; Autoantibodies; Cattle; gamma-Globulins; Immune Tolerance; Lupus Erythematosus, Systemic; Mice; Nephritis; Ovalbumin