concanavalin-a and Kidney-Failure--Chronic

Sirtuin 1 (SIRT1) depletion in vascular endothelial cells mediates endothelial dysfunction and premature senescence in diverse cardiovascular and renal diseases. However, the molecular mechanisms underlying these pathologic effects remain unclear. Here, we examined the phenotype of a mouse model of vascular senescence created by genetically ablating exon 4 of Sirt1 in endothelial cells (Sirt1(endo-/-)). Under basal conditions, Sirt1(endo-/-) mice showed impaired endothelium-dependent vasorelaxation and angiogenesis, and fibrosis occurred spontaneously at low levels at an early age. In contrast, induction of nephrotoxic stress (acute and chronic folic acid-induced nephropathy) in Sirt1(endo-/-) mice resulted in robust acute renal functional deterioration followed by an exaggerated fibrotic response compared with control animals. Additional studies identified matrix metalloproteinase-14 (MMP-14) as a target of SIRT1. In the kidneys of Sirt1(endo-/-) mice, impaired angiogenesis, reduced matrilytic activity, and retention of the profibrotic cleavage substrates tissue transglutaminase and endoglin accompanied MMP-14 suppression. Furthermore, restoration of MMP-14 expression in SIRT1-depeleted mice improved angiogenic and matrilytic functions of the endothelium, prevented renal dysfunction, and attenuated nephrosclerosis. Our findings establish a novel mechanistic molecular link between endothelial SIRT1 depletion, downregulation of MMP-14, and the development of nephrosclerosis.

Topics: Acute Kidney Injury; Animals; Cellular Senescence; Concanavalin A; Down-Regulation; Endothelium, Vascular; Exons; Extracellular Matrix; Fibrosis; Folic Acid; Gene Expression Regulation, Enzymologic; Human Umbilical Vein Endothelial Cells; Kidney; Kidney Failure, Chronic; Male; Matrix Metalloproteinase 14; Mice; Mice, Mutant Strains; Mice, Transgenic; Neovascularization, Physiologic; Nephrosclerosis; Regeneration; Sirtuin 1; Vasodilation

The in vitro response of peripheral blood mononuclear cells (PBMCs) to the suppressive effects of calcineurin inhibitors is known to correlate with the clinical efficacy of drugs used in renal transplantations. The present study was conducted to examine the differences of PBMC responses to calcineurin inhibitors between chronic renal failure (CRF) patients awaiting renal transplantation and cirrhosis patients awaiting liver transplantation. The study included 99 CRF patients awaiting renal transplantation and 27 cirrhosis patients awaiting liver transplantation. Twenty milliliters of venous blood was taken 1-7 days before transplantation. The in vitro drug concentrations giving 50% inhibition of PBMC blastogenesis stimulated with concanavalin A (IC(50)s) were calculated. The suppressive effects of tacrolimus against PBMC blastogenesis were more than 10-100 times stronger than those of cyclosporine. The median IC(50) value for cyclosporine against the CRF PBMCs was not significantly different from the median IC(50) value against the cirrhosis PBMCs. In contrast, tacrolimus sensitivity in cirrhosis PBMCs is approximately seven times higher than that in CRF PBMCs. The median IC(50) value for tacrolimus against cirrhosis PBMCs was significantly lower and therefore the effect was stronger in comparison to the CRF PBMCs (p < 0.001). These data suggest that the PBMCs of cirrhosis patients, in comparison to those of CRF patients, are highly sensitive to the suppressive effect of tacrolimus. However, PBMC sensitivity to cyclosporine was not significantly different between the CRF and cirrhosis patients. These observations raise the possibility that treatment with tacrolimus, rather than cyclosporine, may therefore be a better choice to reduce the risks of allograft rejection in liver transplantation.

Topics: Aged; Calcineurin; Calcineurin Inhibitors; Concanavalin A; Cyclosporine; Enzyme Inhibitors; Female; Humans; Immunosuppressive Agents; Kidney Failure, Chronic; Kidney Transplantation; Liver Cirrhosis; Liver Transplantation; Lymphocyte Activation; Male; Middle Aged; Tacrolimus

Plasma cell differentiation antigen 1 (PC-1) is an inhibitor of insulinreceptor tyrosine-kinase. PC-1 content is elevated in muscle and adipose tissue from insulin-resistant subjects and its elevation correlates with in vivo insulin resistance. It is known that insulin resistance in uraemia may be improved with erythropoietin (EPO) and vitamin D therapy. Therefore, in this study the effects of human recombinant EPO and 1-alpha-D3 treatments on lymphocyte PC-1 expression in patients with end-stage renal failure on haemodialysis (HD) were investigated.. Lymphocyte basal, concanavalin A (Con A), and phorbol-12-myristate13-acetate (PMA)-stimulated PC-1 activity were investigated in HD patients before and after a two-month treatment with subcutaneous EPO (15 patients, 2000-3000 U thrice weekly) or oral 1-alpha-D3 (14 patients, 2 mug thrice weekly). Twenty-nine patients (16 men and 13 women), aged 22-68 years (49+/-7 years), on HD from 13 to 112 months, and 30 healthy controls participated in the study. None was obese and all had normal fasting plasma glucose.. A two-month EPO treatment produced a 41% haematocrit increase, with a rise in haemoglobin from 6.51+/-0.18 g/dL to 9.69+/-0.14 g/dL. Basal lymphocyte PC-1 activity in HD patients was found to be significantly increased (P<0.005) over the level in healthy controls. Treatment of patients with EPO decreased unstimulated lymphocyte PC-1 activity to values significantly lower than before the treatment (P<0.001). Lymphocyte Con A and PMA-stimulated PC-1 activity in patients on HD was found to be slightly increased over the level in healthy controls, but significantly reduced (P<0.005 and 0.05, respectively) after the EPO treatment. A two-month pulse oral 1-alpha-D3 treatment increased haematocrit by 21% and raised haemoglobin from 7.11+/-0.32 g/dL to 8.80+/-0.39 g/dL. This treatment normalized serum alkaline phosphatase activity and slightly reduced serum parathyroid hormone concentration. PC-1 in unstimulated and PMA-stimulated lymphocytes was unchanged, but significantly decreased (P<0.05) in Con A-stimulated lymphocytes after 1-alpha-D3 treatment. Fasting plasma glucose was not changed by the treatment.. An increased lymphocyte PC-1 activity over control was found in HD patients. A two-month EPO therapy significantly decreased PC-1 activity to the control values, suggesting that an effect on PC-1 expression could be implicated in the amelioration of insulin resistance in uraemic patients treated with EPO. Treatment with pulse oral 1-alpha-D3 had an effect only on PC-1 of Con A-transformed lymphocytes of haemodialysed patients and requires further investigation.

Topics: Adjuvants, Immunologic; Administration, Oral; Adult; Aged; Blood Glucose; Carcinogens; Concanavalin A; Diabetes Complications; Diabetes Mellitus, Type 2; Erythropoietin; Female; Humans; Hydroxycholecalciferols; Insulin Resistance; Kidney Failure, Chronic; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Phosphoric Diester Hydrolases; Pyrophosphatases; Recombinant Proteins; Renal Dialysis; Tetradecanoylphorbol Acetate

Alkaline phosphatases (ALPs) originating from different organs are frequently detected in the serum and urine of patients with renal failure.. We investigated the characteristics of ALPs in the serum and urine of 108 patients with chronic renal failure (CRF) and of 106 healthy control subjects.. In polyacrylamide gel electrophoresis, three atypical ALP bands in serum of patients were designated as atypical-s1, -s2 and -s3, respectively. In contrast, five atypical bands (u1, u2, u3, u4 and u5) were detected in the urine of patients. The atypical ALPs were electrophoretically isolated and assayed to determine their biochemical properties, i.e., neuraminidase sensitivity, heat stability, reactivity to anti-intestinal or anti-tissue nonspecific ALP antibodies, molecular sizes and sugar chain heterogeneities. From these results, we found that atypical-s1 and -s2 were the intestinal-type ALP, while s3 was the tissue-unspecific type ALP. Atypical-u1, -u2 and -u3 were high-molecular type ALPs, which we suggested as the ones that originated from the intestine. Atypical-u4, a tissue-unspecific type ALP, was detected with considerable frequency in the urine of patients. In patients with CRF, the appearance of these atypical ALPs was accompanied by a deterioration of the creatinine clearance.. The appearance of atypical ALPs in the serum and urine of patients with CRF may be a useful marker for renal disease.

Topics: Adult; Aged; Alkaline Phosphatase; Chromatography, Affinity; Concanavalin A; Electrophoresis; Female; Humans; Isoenzymes; Kidney Failure, Chronic; Male; Molecular Weight; Reference Values; Renal Insufficiency

Induction of apoptosis by immunosuppressive agents such as glucocorticoids (GCs) and cyclosporine (CsA) in cultured lymphoid cells has been suggested. However, there are few studies which demonstrate the induction of apoptosis by these agents in the activation process of human peripheral blood mononuclear cells (PBMCs). Here we show that potent immunosuppressive GCs and CsA induce apoptosis in concanavalin A (con A)-activated human PBMCs. In this study, GCs and CsA suppressed human PBMC-blastogenesis when activated by con A in a dose-dependent manner, where healthy PBMCs treated with > 100 ng/ml of each immunosuppressive agent exhibited a DNA-ladder structure in electrophoretic analysis. In three chronic renal failure (CRF) patients, dose-dependency of the PBMC-apoptosis induction was confirmed by our quantification of fragmented DNA using ELISA. Furthermore, the enrichment of DNA fragmentation was significantly associated with the rate of PBMC-blastogenesis when treated with GCs or CsA (r = -0.466, P < 0.01). These results suggested that suppression of the mitogen-induced PBMC-blastogenesis by the immunosuppressive agents should be correlated with the induction of apoptosis.

Topics: Apoptosis; Concanavalin A; Cyclosporine; DNA; Dose-Response Relationship, Drug; Electrophoresis; Glucocorticoids; Humans; Immunosuppressive Agents; Kidney Failure, Chronic; Leukocytes, Mononuclear; Lymphocyte Activation; Mitogens; Stimulation, Chemical

A positive correlation between successful kidney transplantation, few rejection episodes, greater susceptibility to infection and morbidity in patients with high tissue levels of aluminium (Al) indicate that the metal may play a role in the immune response. The aim of this study was to determine if experimental aluminium intoxication could result in significant changes in lymphocyte activity in uraemic and nonuraemic rats.. Lewis rats were divided into four groups: normals (N), nephrectomized control (U), and Al-treated (N + Al) and nephrectomized Al-treated (U + Al), which received a cumulative dose of 30 mg Al over a 4-week period. Al quantification, histology, histochemical analysis and immunological assays were performed after Al intoxication.. High tissue levels of Al and positive histochemical staining in bones were seen in Al-treated rats. Bone histology revealed osteomalacia in U + Al rats. No statistical differences were observed in mixed lymphocyte cultures from controls and Al-treated rats, whereas U and Al-treated rats showed a decrease in lymphoproliferative response to mitogen and natural killer cell cytotoxic activity. A decreased helper T lymphocyte: cytotoxic T lymphocyte cell ratio and a reduction in interleukin-2 production were observed only in the U + Al group. A reduced number of total T lymphocytes was detected in the spleens of all Al-treated rats.. These findings suggest that aluminium toxicity may contribute to immunological impairment in chronic renal failure.

Topics: Aluminum; Animals; Bone and Bones; Cell Division; Cells, Cultured; Concanavalin A; Cytotoxicity, Immunologic; Immunity, Cellular; Interleukin-2; Kidney Failure, Chronic; Lymphocyte Activation; Lymphocyte Count; Male; Nephrectomy; Osteomalacia; Rats; Rats, Inbred Lew; T-Lymphocytes; T-Lymphocytes, Cytotoxic; T-Lymphocytes, Helper-Inducer; Uremia

The changes in mitogen-induced lymphocyte response occurring in nine patients undergoing chronic hemodialysis and treated with 1,25 dihydroxyvitamin D3 (0.5 micrograms/day) were investigated. Prior to treatment the stimulation indices (SI) in patients with 15.4 +/- 4.2 for phytohemagglutinin (PHA) and 7.2 +/- 0.7 for conconavalin A (Con A). In the controls, stimulation indices were 44.4 +/- 13.5 and 20.2 +/- 5.3 for PHA and Con A, respectively. Following treatment, the stimulation indices increased to 36.9 +/- 6.2 for PHA (p < 0.05) and 18.6 +/- 3.9 for Con A (p > 0.05) indicating the beneficial effect of oral 1,25 dihydroxyvitamin D3 treatment on lymphocyte function in patients with chronic renal failure.

Topics: Administration, Oral; Adolescent; Adult; Calcitriol; Calcium; Case-Control Studies; Concanavalin A; Drug Therapy, Combination; Female; Humans; Kidney Failure, Chronic; Lymphocyte Activation; Male; Middle Aged; Phosphates; Phytohemagglutinins; Renal Dialysis

Recombinant human erythropoietin (rHuEpo) is now widely employed in correction of the anemia of end stage renal disease (ESRD). Recent reports suggest that rHUEpo, in addition to its effect on CFU-E and burst-forming-unit-erythroid (BFU-E), may stimulate granulocyte/macro-phage production and pluripotential progenitors of the myeloid and monocyte lineage. Furthermore, there is now data which demonstrate that ESRD patients who received rHuEpo have enhanced cytokine production. Taken together, these observations suggest that the administration of rHuEpo may augment the diminished immune response of renal failure patients. To evaluate the effects of rHuEpo therapy on cell-mediated immunity in hemodialysis patients, a prospective controlled study was conducted. Two parameters of immune function were tested. One was natural killer cell (NK) activity, and the other proliferation in response to the T cell mitogen concanavalin A (Con-A). NK activity of the ESRD patients was comparable with that of normal controls at the start of the study and was unaffected by rHuEpo therapy. In contrast to this, anemic ESRD patients initially demonstrated impaired mitogen driven proliferation (initial stimulation index (S.I.) = 42.5 +/- 11.9) which significantly improved following rHuEpo therapy (final S.I. = 73.3 +/- 14.7, p < 0.05). The later value exceeded the mitogen response in less anemic ESRD patients who did not receive rHuEpo (initial S.I. = 60.7 +/- 16.5, final S.I. = 61.0 +/- 16.7), but did not reach values seen in normal controls. The data suggest that rHuEpo therapy may be associated with enhanced immune responses in patients with ESRD.

Topics: Aged; Concanavalin A; Erythropoietin; Female; Humans; Immunity, Cellular; Kidney Failure, Chronic; Killer Cells, Natural; Male; Middle Aged; Prospective Studies; Renal Dialysis; T-Lymphocytes

Effects of hemodialysate of patients with chronic renal failure (CRF) on blastogenesis of human peripheral blood lymphocytes in the presence of concanavalin A or phytohemagglutinin was investigated. Hemodialysates from 11 CRF patients significantly promoted lymphocyte blastogenesis when compared with control dialysis fluids (p less than 0.01). The strongest activity (39% promotion of the lymphocyte blastogenesis) was observed with a hemodialysate obtained at 0.5 h after beginning dialysis. The activity decreased thereafter. On the contrary, the blastogenesis-promoting activity in plasma decreased significantly after hemodialysis (p less than 0.01, n = 11). To further confirm the presence of low-molecular-weight factor(s) in CRF, ultrafiltration of plasma obtained from CRF and healthy subjects was conducted using a membrane filter with a molecular cutoff of 3,000 D. The filtrate of CRF plasma significantly promoted lymphocyte blastogenesis when compared to that of healthy subjects (p less than 0.01). Heat treatment (100 degrees C, 40 min) did not abolish the activity of the hemodialysate. None of the drugs taken by the patients nor creatinine accumulated in CRF promoted the lymphocyte blastogenesis. Chromatographic analysis of a hemodialysate demonstrated several peaks which were absent in the control dialysis fluid. These results showed the presence of a novel lymphocyte-stimulating factor(s) in CRF, which is heat stable and has a low molecular weight (less than 3,000 D).

Topics: Allopurinol; Anabolic Agents; Androstanols; Calcitriol; Cells, Cultured; Concanavalin A; Creatinine; DNA Replication; Female; Furosemide; Humans; Kidney Failure, Chronic; Lectins; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Nifedipine; Reference Values; Renal Dialysis; Thymidine; Time Factors

Blood transfusion induces immunosuppression by a mechanism which is probably multifactorial and, in large part, obscure. In this study cellular immune suppression was examined in patients with end-stage renal disease (ESRD) following a single blood transfusion (BT). OKT8+ cells proliferated within four hours, their count increasing significantly, and reached their peak after four days. Since OKT4+ cells tended to decrease, the OKT4/OKT8 ratio declined significantly from the fourth day. Evidence in support of the above results is offered in studies on ability of concanavalin A reagent (ConA)-activated T cells to form autorosettes with human red blood cells (RBC); these T cells have been shown to function as suppressor effector cells. Autorosetting cells (RFC) proliferated in a similar pattern to OKT8+ cells. Moreover, the percentage of rise at one and two weeks of OKT8+ and RFC correlated directly, suggesting that the proliferating OKT8+ cells are in large part a subset of suppressor cells. Functional studies were performed to establish the suppressor activity of T cells before and at various times after blood transfusion. A significant increase in T cell ConA-induced suppressor activity was found one week after BT, peaking at two weeks following BT. Monocytes increased significantly from day 4 following BT, but prostaglandin E2 (PGE2) started to rise immediately, peaking at day 4. PGE2 might induce suppression by induction of suppressor cells or by direct suppression of interleukin-2. Both monocytes and suppressor cells respond immediately, but they probably induce suppression most effectively four days or more after BT.

Topics: Adult; Aged; Blood Transfusion; Concanavalin A; Female; Graft Survival; Humans; Kidney Failure, Chronic; Kidney Transplantation; Male; Middle Aged; Rosette Formation; T-Lymphocytes, Regulatory

The ability of cultured peripheral-blood mononuclear cells (PBMC) to release interleukin-1 alpha and beta (IL-1 alpha, IL-1 beta) in response to concanavalin A (con A) was investigated in patients with chronic renal failure (CRF) and in renal transplant recipients. Mean IL-1 alpha level released by PBMC of healthy subjects (n = 42), CRF patients (n = 42), or transplants 2 months after operation (n = 69) was 152 +/- 103, 110 +/- 80, or 154 +/- 87 pg/5 x 10(5) cells/ml culture, respectively. IL-1 alpha release from PBMC of recipients 2 months after renal transplantation was significantly higher than that of CRF patients (p less than 0.05). Mean IL-1 beta level released by PBMC of healthy subjects (n = 34), CRF (n = 30), or transplants (n = 55) was 223 +/- 159, 135 +/- 129, or 276 +/- 155 pg/5 x 10(5) cells, respectively. Similar to IL-1 alpha, the level in CRF was significantly lower than that in healthy subjects (p less than 0.05). A time course study indicated that the ability of PBMC from transplants to release IL-1 alpha and beta promptly decreased following the operation, possibly owing to prednisolone and ciclosporin immunosuppressive therapy. However, after maintaining a low level for 2-3 weeks, IL-1 release from PBMC gradually increased thereafter. The results were consistent with known characteristics of decreased immunity in CRF states, and further suggested that the decreased ability of PBMC to release IL-1 alpha and beta in response to con A in CRF patients is recovered 2 months after renal transplantation.

Topics: Adult; Aged; Concanavalin A; Humans; In Vitro Techniques; Interleukin-1; Kidney Failure, Chronic; Kidney Transplantation; Leukocytes, Mononuclear; Middle Aged; Time Factors

The total concentration and concanavalin A (ConA)-dependent microheterogeneity of alpha-1 acid glycoprotein (AAG) were studied in thirty hemodialyzed uremic patients and eighteen non-dialyzed uremic patients, by comparison with healthy volunteers. Serum concentrations of AAG were significantly higher in the non-dialyzed uremic (1.27 +/- 0.47 g/l) and hemodialyzed patients (1.29 +/- 0.33 g/l) than in the volunteers (0.79 +/- 0.09 g/l). The proportions of strongly ConA-reactive AAG fractions were also higher in non-dialyzed uremic (16.7%) and hemodialyzed patients (18.5%) than in volunteers (14.1%). These data may be related to an increase in bi-antennary glycans, as observed in patients on peritoneal dialysis, together with a probable change in sialylation. AAG serum levels and microheterogeneity were similar in non-dialyzed and hemodialyzed patients and did not appear in the dialyzed patients to depend on the type of dialysis membrane used, i.e. cuprophan (CU), cellulose acetate (CA), hemophan (HE), polyacrylonitrile (PAN), and polysulfon (PS), in spite of differences in biocompatibility. In patients dialyzed with CA membranes, there was a distinct decrease in the ConA non-reactive fraction (38.0%) and an increase in ConA slightly-reactive (42.2%) and strongly-reactive (19.7%) fractions. Differences in AAG serum levels and ConA reactivity between patients dialyzed with CA and PAN membranes seem to justify further investigations of other acute-phase reactants and immunological parameters.

Topics: Concanavalin A; Humans; Immunoelectrophoresis, Two-Dimensional; Kidney Failure, Chronic; Male; Membranes, Artificial; Middle Aged; Orosomucoid; Renal Dialysis

Normal peripheral blood mononuclear cells (PBMC) were primed with sera from 22 predialytic uremic patients, normal sera, concanavalin A, or both. Their suppressive activity was subsequently tested on fresh phytohemagglutinin-stimulated allogeneic responders (i.e., genetically unrelated to either primed cell or sera donors). Uremic serum induced suppressor cell activity in the normal PBMC. No correlation was found between serum urea/creatinine levels and their effects on suppressor cell activity. The induced activity, expressed as percent suppression, was similar to that induced by concanavalin A. In PBMC primed with both concanavalin A and uremic serum, an additive suppressive effect was evident. The suppressor subset(s) induced by uremic serum proved to belong to adherent cell population. Addition of indomethacin or catalase to responder systems did not abolish the suppressive effects, thus suggesting a mechanism of action other than prostaglandin or hydrogen peroxide release.

Topics: Adult; Aged; Concanavalin A; Female; Humans; Kidney Failure, Chronic; Male; Middle Aged; T-Lymphocytes, Regulatory; Uremia

Topics: Adult; Concanavalin A; Female; Humans; Kidney Failure, Chronic; Lymphocyte Activation; Male; Renal Dialysis; T-Lymphocytes; T-Lymphocytes, Regulatory

The effect of blood transfusion on humoral immunity in chronic renal failure was studied by examining immunoglobulin production in vitro, in patients awaiting renal transplantation. Pokeweed mitogen (PWM) induced IgG plaque formation was normal in non-transfused uraemic patients while both spontaneous and Staphylococcus aureus Cowan I (SAC) induced immunoglobulin production were reduced. Five to ten units of third party blood transfusion reduced PWM-driven B cell differentiation, but had no effect on SAC-induced plaque formation, while spontaneous production of immunoglobulin was either enhanced or unaffected. As it is known that the response to SAC is less affected by suppressor T cell activity than that to PWM, these differences in the inhibitory effects of blood transfusion on B cell differentiation are further evidence that transfusion may act by increasing suppressor T cell activity.

Topics: Adult; Antibody-Producing Cells; Antigens, Bacterial; Blood Transfusion; Concanavalin A; Female; Hemolytic Plaque Technique; Humans; Immunoglobulin G; Kidney Failure, Chronic; Male; Middle Aged; Pokeweed Mitogens; Staphylococcus aureus; Uremia

Suppressor cell function was assessed in 13 patients with chronic renal failure not on dialysis, 10 patients on regular hemodialysis and 11 renal transplant patients by means of the Concanavalin A inducible Suppressor Cell Assay. The mean Suppression Index (S.I.) in the undialysed patients (0.76 +/- 0.39) and those of the dialysed patients (0.72 +/- 0.18) were not significantly different from the 25 healthy controls (0.68 +/- 0.22). However the S.I. in the renal transplant patients (1.11 +/- 0.41) was significantly different from the normal controls as well as the undialysed and dialysed patients. Although there was no correlation between S.I. and age, post transplant follow up period, serum creatinine concentration, prednisolone or azathioprine dosage it is likely that the cause of the impaired suppressor cell function was due to the immunosuppressive agents used.

Topics: Adult; Concanavalin A; Female; Humans; Kidney Failure, Chronic; Kidney Transplantation; Male; Renal Dialysis; T-Lymphocytes, Regulatory

Concanavalin A (Con A)-induced proliferative responses by mononuclear leukocytes (MNCs) from patients with chronic renal failure (CRF) who were undergoing maintenance hemodialysis were studied with serum-free media to analyze leukocyte function independent of either uremic or normal serum factors. When we increased the number of cultured MNCs by 2.5- to fivefold over that normally used in microcultures and reduced the mitogen concentration, Con A induced proliferative responses 10- to 1000-fold higher than those in unstimulated cultures. In 12 of 15 patients with CRF observed, Con A-induced MNC responses were significantly depressed as compared with those in age- and sex-matched controls. Responses in 10 of these 12 patients with CRF improved significantly immediately after dialysis, but the improvement was only temporary. With MNCs from patients with CRF before dialysis, removal of adherent cells significantly improved their responses to Con A. Similar increases with adherent cell depletion were not found either in cultures of control MNCs or in patient MNCs after dialysis. Indomethacin, a cyclooxygenase inhibitor, added to unseparated MNCs from patients before dialysis, significantly increased responses in 13 of 15 patients. This effect of indomethacin was found less frequently in MNC cultures from normal persons or from patients with CRF after dialysis. Nonadherent lymphocytes from patients with CRF were not abnormally sensitive to inhibition by exogenously added prostaglandin E2. We conclude that MNCs from most patients with CRF have depressed reactivity when cultured without serum and that responses improve temporarily after dialysis. Adherent cells are largely responsible for inhibiting lymphocyte responses, and monocyte-released cyclooxygenase products appear to mediate much of this suppression.

Topics: Cell Adhesion; Cell Division; Concanavalin A; Culture Media; Dinoprostone; Humans; Indomethacin; Kidney Failure, Chronic; Lymphocyte Activation; Mitogens; Monocytes; Prostaglandin-Endoperoxide Synthases; Prostaglandins E; Renal Dialysis

Suppressor cell activity after concanavalin A induction was studied in peripheral blood mononuclear cells of patients undergoing long-term hemodialysis. Suppression both of the mixed lymphocyte reaction and of allogeneic cells stimulated with phytohemagglutinin was significantly higher with peripheral blood mononuclear cells from patients undergoing hemodialysis than with cells from control subjects. Expression of the Ia antigen on T lymphocytes (associated with immunologic activation) was studied by staining with monoclonal antibodies and two-color fluorescence analysis in a computer-linked cytofluorograph. In unstimulated cells, there was no significant difference between the patients and control subjects. After concanavalin A induction, the percentage of T4, and particularly of T8, cells expressing the Ia antigen was significantly higher in the group undergoing hemodialysis. The functional suppression seen after concanavalin A induction in the mixed lymphocyte reaction was significantly reduced by treatment with OKT8 monoclonal antibody and complement; in phytohemagglutinin cultures, both OKT8 and OKIa*1 antibodies were effective. The reduced in vitro response of uremic lymphocytes may thus be a consequence of increased suppressor activity associated with the T8-positive, Ia-positive subset of T cells.

Topics: Adult; Aged; Antibodies, Monoclonal; Concanavalin A; Humans; Kidney Failure, Chronic; Lymphocyte Activation; Middle Aged; Renal Dialysis; T-Lymphocytes; T-Lymphocytes, Regulatory

Stimulation of human lymphocytes with concanavalin A (Con A) resulted in variable lymphokine responses as indicated by factors inhibiting macrophage migration (MIF) or stimulating macrophage migration (MStF), or resulted in negligible responses. These responses were consistent for a given individual when repeated after several months. MIF responses were observed more frequently than MStF responses in patients with renal failure who had demonstrable alloantibodies. MStF responses were statistically associated with the presence of HLA DR1 antigens in patients with renal failure and two separate groups of healthy individuals, while MIF responses were associated with DR7 in the three groups studied. There was no correlation between immunoglobulin allotypes and lymphokine responses. These results suggest that lymphokine responses to Con A are indicators of nonspecific immunological responsiveness and are influenced by genes associated with the major histocompatibility complex.

Topics: Adolescent; Adult; Cell Migration Inhibition; Concanavalin A; Female; Histocompatibility Antigens Class II; HLA-DR Antigens; HLA-DR1 Antigen; HLA-DR7 Antigen; Humans; Immunoglobulin Allotypes; Isoantibodies; Kidney Failure, Chronic; Lymphokines; Macrophage Migration-Inhibitory Factors; Macrophage-Activating Factors; Male; Middle Aged; T-Lymphocytes

An experimental model in rats was developed to define the nature of humoral and cellular factors that contribute to impaired immune responsiveness in chronic renal failure. Addition of uremic rat serum to both normal and uremic lymphocytes significantly suppressed cellular responses, the suppression being more pronounced with uremic lymphocytes. Lymphocytes from uremic rats were only marginally less responsive than normal lymphocytes to concanavalin A stimulation when normal rat serum was added to the cultures, indicating that the cellular factors in impairment were less important than humoral ones. Antibody formation in rat splenocyte cultures to bovine serum albumin was suppressed by addition of uremic serum, but the response to sheep erythrocytes was unaffected. Thus the effect on antibody response in uremic animals is dependent upon the antigen tested.

Topics: Animals; Antibody Formation; Cells, Cultured; Concanavalin A; Enzyme-Linked Immunosorbent Assay; Erythrocytes; Kidney Failure, Chronic; Lymphocyte Activation; Male; Rats; Rats, Inbred Lew; Serum Albumin, Bovine; Sheep; Spleen; Uremia

Suppressor cell activity was studied in 30-non-transfused renal dialysis patients using the Concanavalin A enhancement method. Suppressor cell numbers were estimated in 19 patients by the T suppressor (T gamma) rosette technique and in 15 patients using a monoclonal antibody (OKT8) specific for T suppressor cells. The role of prostaglandin secreting monocytes in the observed suppression was investigated in 6 patients using the prostaglandin secreting inhibitor flurbiprofen. Both suppressor cell numbers and activity were either normal or low compared with normal healthy controls. Flurbiprofen at concentrations of 10(-4) and 10(-6) M caused a decrease in activity in both patients and normal controls. The results indicate that the increased suppressor cell activity and number reported in dialysis patients may be due to the effect of blood transfusion previously administered to the patients.

Topics: Adult; Concanavalin A; Female; Flurbiprofen; Humans; Immunity, Cellular; Kidney Failure, Chronic; Leukocyte Count; Lymphocyte Activation; Male; Middle Aged; Monocytes; Prostaglandins; Renal Dialysis; T-Lymphocytes, Regulatory

Suppressor cells were assayed by numerical and functional tests in adults on chronic hemodialysis. Peripheral blood mononuclears (PBM) were classified as total T-cells by E-rosettes and by the monoclonal antibody OKT3, as T-cell subsets by OKT4 (inducer/helper T-cells) and OKT8 (cytotoxic/suppressor T-cells) and as B-cells by the presence of surface immunoglobulin. The suppressive effect of PBM pretreated with either Concanavalin A (Con A), sodium periodate, or serum rich in immune complexes, on normal homologous phytohemagglutinin (PHA) lymphocyte transformation, was determined. Usual tests of T-cell function were not done. T lymphopenia was due to significant diminution (P less than 0.002) in numbers of OKT4+ cells in patients (516 +/- 44 cells/mm3, mean +/- sem) as compared to controls (906 +/- 96 cells/mm3). The number of OKT8+ cells in patients was not different from normal although their percentage (45 +/- 4%) was slightly higher than controls (36 +/- 5%) (P less than 0.10). Suppressor activity using only a suboptimal dose of Con A (5 micrograms/ml), was significantly lower (P less than 0.002) in uremic patients (36 +/- 12%) than in controls (67 +/- 7%). An important finding was that no significant correlations were detected between the numerical and functional assays of suppression used or between any of these immunological tests and biochemical parameters studied. The implications of these results for immunoparesis in uremia are discussed with particular reference to the discordance between marker and functional assays of suppressor cells.

Topics: Adolescent; Adult; Antibodies, Monoclonal; Antigen-Antibody Complex; Concanavalin A; Female; Humans; Kidney Failure, Chronic; Lymphocyte Activation; Male; Middle Aged; Periodic Acid; Phytohemagglutinins; Rosette Formation; T-Lymphocytes, Regulatory; Uremia

Topics: Blood Transfusion; Concanavalin A; Humans; Kidney Failure, Chronic; Lymphocyte Activation; Renal Dialysis; Rosette Formation; T-Lymphocytes, Regulatory

In 8 patients with chronic peritoneal dialysis before and after 24-hour peritoneal dialysis examinations on the cellular immunity were performed (lymphocyte blast transformation with PHA and Con A, ADCC). There was no difference to the normal findings in healthy control persons. Blast transformation decreased during peritoneal dialysis, no changes were visible with ADCC. Compared with reports in literature concerning regular hemodialysis therapy the state of cellular immunity in chronic peritoneal dialysis seemed to be superior.

Topics: Antibody-Dependent Cell Cytotoxicity; Concanavalin A; Female; Humans; Immunity, Cellular; Kidney Failure, Chronic; Lymphocyte Activation; Male; Peritoneal Dialysis; Phytohemagglutinins

A protein component present in normal human urine has been found on the surface of epidermal cells and lymphocytes. This protein, called urinary acidic antigen (UA), can not be detected in concentrated fractions of normal human serum by double immunodiffusion, suggesting that it is quickly cleared from the circulation. It is readily detected, however, in sera of patients with renal failure. Although it can be eliminated from the cell surface by repeated washings with PBS, it was shown to cap with anti-UA-specific antiserum. Anti-UA suppresses PWM-induced proliferation, but not the lymphocyte response to PHA, Con A, or allogeneic cells. Thus UA appears to have a specific relationship to the pokeweed response. Whether it is a structural component of the PWM receptor is uncertain.

Topics: Animals; Antigens, Surface; Concanavalin A; Epidermis; Fluorescent Antibody Technique; Glycoproteins; Humans; Immunoglobulin G; Kidney Failure, Chronic; Leukocytes; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphocytes; Phagocytes; Pokeweed Mitogens; Rabbits; Rosette Formation; Spleen

The enhanced in vitro proliferative response of peripheral mononuclear cells (PMNC) when pre-cultured for 24 hours prior to the addition of Concanavalin-A has been used as an indirect parameter of suppressor cell activity in healthy subjects and in patients undergoing chronic haemodialysis. The proliferative response of PMNC from haemodialysis patients is lower than that of control subjects when Con-A is added at the initiation of culture but approximates to that of normal subjects when the addition of Con-A was deferred for 24 hours. The Suppressor Indices of PMNC from haemodialysis patients were at least as great and sometimes greater than those of control subjects but the absolute T-cell counts were lower in haemodialysis patients than in controls. These results suggest that the relative energy of haemodialysis patients is partly attributable to T-cell depletion but this is accompanied by retention, and in some cases augmentation, of suppressor cell activity.

Topics: Adult; Concanavalin A; Female; Humans; Kidney Failure, Chronic; Leukocyte Count; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Renal Dialysis; T-Lymphocytes; T-Lymphocytes, Regulatory

Lymphocyte blastogenic transformation in response to plant lectins and allogenic cells was studied in patients with nonuremic, far-advanced, chronic renal failure and in healthy controls. Cell cultures were studied in the presence of normal sera, patient's sera, and with media of different buffering capacities. Minimal blastogenic depression was observed when patient's lymphocytes were cultured in indifferent plasma with effective bicarbonate buffering compared with the use of pooled patient's plasma or HEPES buffer. Fresh plasma in culture depressed concanavalin A (Con A) blastogenesis. The data suggest that, under optimal conditions, lymphocytes from patients with chronic severe renal insufficiency are more responsive to stimuli than previously reported and as a group are near normal control values. Further, the defect observed may be a result of intracellular acidosis.

Topics: Adult; Concanavalin A; Humans; Kidney Failure, Chronic; Lectins; Lymphocyte Activation; Male; Mitogens; Renal Dialysis