Inhibit T-cell Function Research Articles

Cytotoxic and Immunologic Effects of Methotrexate in Psoriasis

Based on recent experience that Cyclosporin A, an immunosuppressive drug, produces marked improvement in psoriasis, possible immunomodulatory activities of methotrexate (MTX) have been reviewed to look for alternate mechanisms of MTX action in psoriasis. It is generally considered that the therapeutic results of MTX in psoriasis are related to a direct effect on epidermal cell hyperplasia through inhibition of DNA synthesis. Several studies in the literature now suggest possible effects of MTX on the immune system of psoriatics as well as in animal models that may have some pathogenic similarities to psoriasis. In psoriatics receiving MTX, neutrophil chemotaxis is suppressed, resulting in a possible alteration in the potential pathologic activity of neutrophils commonly found in lesional skin. MTX does improve both psoriatic and rheumatoid arthritis. Animal studies of the latter using adjuvant arthritis and graft vs host disease (GVHD) have indicated several possible mechanisms for MTX that affect these processes. In GVHD, MTX selectively destroys cycling CD8+ cells, and in adjuvant arthritis the activation of macrophages is prevented by inhibition of T-cell function. While MTX generally has not been clinically utilized as an immunomodulatory drug for immunologically related diseases, it may, nonetheless, have selective actions that could be specific for some diseases. MTX and Cyclosporin A could work mechanistically in similar ways but at different steps in the activation of T cells and macrophages. It may be that the major direct effect of MTX on epidermal cell proliferation is complemented or even mediated by subtle immunoregulatory effects on the melange of cells in the affected skin and the systemic immune response.

Inhibition of lymphocyte IL2‐receptor expression by factor VIII concentrate: a possible cause of immunosuppression in haemophiliacs

Factor VIII concentrate inhibits T-cell function in vitro and in vivo. The mechanisms underlying the phenomenon were investigated. Factor VIII concentrate has a direct effect on lymphocytes, uninfluenced by haemophilic monocyte dysfunction, since it inhibited lymphocyte transformation with phorbol myristate acetate, a reaction unaffected by monocyte depletion. Inhibition of lymphocyte transformation by factor VIII concentrate is not corrected by the addition of exogenous IL2, suggesting that it does not inhibit lymphocyte function by suppression of IL2 secretion alone. Factor VIII concentrate causes profound inhibition of IL2-receptor expression (CD25); with an 89% reduction in CD25-positive CD4 cells and a 50% reduction in CD25-antigen molecules per cell. CD8 lymphocytes are similarly affected. Smaller reductions in CD71 and HLA-DR expression are also observed. Down modulation of CD25-antigen may explain the reduced IL2 secretion observed by others, and may be an important cause of immunodeficiency in HIV-seronegative haemophiliacs.

Human immunodeficiency virus type 1 envelope glycoprotein gp120 produces immune defects in CD4+ T lymphocytes by inhibiting interleukin 2 mRNA.

Envelope glycoprotein gp120 of human immunodeficiency virus type 1 (HIV-1) is known to inhibit T-cell function, but little is known about the mechanisms of this immunosuppression. Pretreatment of a CD4+ tetanus toxoid-specific T-cell clone with soluble gp120 was found to exert a dose-dependent inhibition of soluble antigen-driven or anti-CD3 monoclonal antibody-driven proliferative response, interleukin 2 (IL-2) production, and surface IL-2 receptor (IL-2R) alpha-chain expression, all of which were reversed by the addition of exogenous IL-2. mRNA for the gene encoding IL-2 was suppressed by treatment with gp120, but IL-2R gene transcription was not inhibited. Bypass activation of the T-cell clone with phorbol 12-myristate 13-acetate plus ionomycin was unaffected by gp120 pretreatment. Thus, gp120-CD4 interaction interferes with an essential role of the CD4 molecule in signal transduction through the CD3-antigen receptor (Ti) complex. Such a mechanism of gp120-induced immunosuppression, if operative in vivo, could contribute to the depressed specific immune responses associated with HIV infection.

Potentiation of immunosuppressive effects of cyclosporin A by 1α,25-dihydroxyvitamin D 3

Both cyclosporin A (CsA) and 1α,25-dihydroxyvitamin D 3 (vitamin D 3) inhibit T-cell functions. Because CsA has a dose-related systemic toxicity, there is a need for approaches by which the dosage of CsA could be reduced while maintaining the required immunosuppression. Therefore, we examined for any potentiating effect of vitamin D 3 on CsA-induced suppression of T cell functions in vitro. Peripheral blood mononuclear cells were stimulated with phytohemagglutinin in the presence or absence of various concentrations of CsA (1.6–13.2 μg/ml) or vitamin D 3 (10 −10–10 −7 M) or both together and [ 3H]thymidine incorporation (TdR), interleukin 2 (IL-2) production, IL-2 receptor (IL-2R) expression, and the response to exogenous recombinant IL-2 (rIL-2) on TdR were measured. IL-2 production was measured on CTLL cell line and IL-2R expression with anti-Tac monoclonal antibody using FACScan. There was a dose-dependent inhibition of TdR, IL-2 production, and IL-2R expression by CsA. Vitamin D 3 inhibited TdR and IL-2 production in a dose-dependent manner but had no significant ( P > 0.1) effect on IL-2R expression. Recombinant IL-2 had no effect on CsA-induced inhibition of TdR, whereas rIL-2 completely reversed the vitamin D 3-induced inhibition of TdR. A significantly ( P < 0.05) increased inhibition of TdR and IL-2 production was observed when two agents were used together as compared to expected inhibition by the addition of each agent separately. A consistent synergism was observed between all concentrations of CsA used and vitamin D 3 (10 −8 and 10 −7 M). This study shows that the potentiation of immunosuppressive effects of CsA by vitamin D 3 could be used as an approach to reduce the dosage of CsA in clinical use without compromising immunosuppression, thus preventing or minimizing the dose-related toxic effects of CsA.

Helper and Suppressor T-Cell Function in HIV-Infected Hemophilia Patients

AbstractT-lymphocyte helper and suppressor functions were assessed in 61 hemophilia patients. Twenty one patients were HIV-negative (Group 1), 27 were HIV-positive without having AIDS-related complex (ARC)/AIDS (Group 2), and 13 had ARC/AIDS (Group 3). T, CD4-positive, or CD8-positive T lymphocytes were cocultured with B lymphocytes and pokeweed mitogen for 6 days and immunoglobulin producing cells were assessed in a reverse hemolytic plaque assay. In HIV-infected patients, T cells as well as the CD4-positive T cell subset exhibited reduced helper (P <.01, Group 2; P <.0005, Group 3) and elevated suppressor activity (P < .02, Group 2; P <.005, Group 3), whereas no significant difference was found between HIV-negative patients and controls. The number of CD4-positive cells was not correlated with CD4 cell function. CD4-positive cells showed no helper activity (<10% of control T cells) in 8/11 (73%), but an excessive suppressor activity (>80% suppression of plaque formation) in 6/11 (55%) Group 3 patients. Our results show that defective helper and elevated suppressor functions of T cells in HIV-infected patients are caused not only by a change in the CD4/CD8 cell counts but also by functional abnormalities of the CD4-positive T-cell subset. These abnormal helper and suppressor functions may play a role in the development of the immunodeficiency state of AIDS patients.

Functional immunoregulatory T-cell abnormalities in cystic fibrosis patients

The role of B cells and regulatory T cells in the reduced in vitro IgG synthesis of cystic fibrosis (CF) patients was studied. Intact proportion, proliferation, and differentiation of B cells and reduced suppressor and helper T-cell function were found. To explore the T-cell defects further, CF sera or supernatant derived from Pseudomonas aeruginosa cultures (PA supernatant) was added to the relevant T helper- and suppressor-cell assays. Both CF sera derived from PA-positive patients and PA supernatant interfered with the appearance of interleukin 2 (IL-2) receptors and with the functional enhancement caused by exogenously added IL-2. PA-negative CF patients, however, also had functional T-cell defects and inhibitory sera, but these sera did not affect IL-2 pathways. Thus different serum factors and intrinsic T-cell defects in CF patients are suggested.

The relationship between HLA-DR3 and T-cell regulation of immunoglobulin production in primary sclerosing cholangitis

Immunoglobulin production in vitro, its control by concanavalin A-activated suppressor T-cells, and the relationship between abnormalities in nonantigen-specific suppression and histocompatibility antigens have been studied in 20 patients with primary sclerosing cholangitis (PSC). Suppression of IgG, IgM, and IgA synthesis was impaired in 12 patients with PSC alone, but was normal in 8 with PSC and associated ulcerative colitis (UC). The HLA antigens B8 and DR3 were increased in frequency in both groups of patients, but an association between DR3, and to a lesser extent B8, and defective suppressor T-cell function was only observed in the patients with PSC alone. These results not only provide further evidence of an association between HLA DR3 and impaired nonantigen-specific suppression but also indicate the genetic complexity of this association and its specificity, being found in this study in only one subgroup of patients with PSC.

Suppressor T‐cell activity in chronic hepatitis B‐virus infection: Relationship with the presence of HBV‐DNA in serum

Suppressor T-cell activity and allogeneic T-cell response to concanavalin A (ConA) were investigated in 46 patients chronically infected with hepatitis B virus (HBV). Thirty-eight patients had chronic active hepatitis, seven of whom were superinfected with Delta virus, and eight were healthy chronic HBV carriers. T-cell suppressor activity was in the normal range in healthy carriers and in patients negative for serum HBV-DNA, independent of the e antigen status. In contrast, the group of patients positive for HBV-DNA exhibited a significant reduction in suppressor activity. Longitudinal studies in patients who cleared serum HBV-DNA demonstrated that suppressor T-cell activity became normal thereafter. These results suggest a relationship between suppressor T-cell function and the stage of viral replication in individuals with chronic HBV infection.

Cell-cell adhesion mediated by CD8 and MHC class I molecules.

CD4 and CD8 are cell-surface glycoproteins expressed on mutually exclusive subsets of peripheral T cells. T cells that express CD4 have T-cell antigen receptors that are specific for antigens presented by major histocompatibility complex class II molecules, whereas T cells that express CD8 have receptors specific for antigens presented by MHC class I molecules (reviewed in ref. 1). Based on this correlation and on the observation that anti-CD4 and anti-CD8 antibodies inhibit T-cell function, it has been suggested that CD4 and CD8 increase the avidity of T cells for their targets by binding to MHC class II or MHC class I molecules respectively. Also, CD4 and CD8 may become physically associated with the T-cell antigen receptor, forming a higher-affinity complex for antigen and MHC molecules, and could be involved in signal transduction. Cell-cell adhesion dependent CD4 and MHC II molecules has recently been demonstrated. To determine whether CD8 can interact with MHC class I molecules in the absence of the T-cell antigen receptor, we have developed a cell-cell binding assay that measures adhesion of human B-cell lines expressing MHC class I molecules to transfected cells expressing high levels of human CD8. In this system, CD8 and class I molecules mediate cell-cell adhesion, showing that CD8 directly binds to MHC class I molecules.

CD3-associated heterodimeric polypeptides on suppressor hybridomas define biologically active inhibitory cells.

We have investigated the relationship between CD3 expression and the suppressor T-cell function. We have isolated stable clonal cell lines of the F12.23 suppressor T-cell hybridoma that are either CD3+ or CD3-. These lines were subjected to functional assays including inhibition of in vivo hapten-specific delayed-type hypersensitivity responses, in vitro hapten-specific interleukin 2 responses, as well as hapten-specific cytotoxic T-lymphocyte assays. In all assays, the functional suppressor phenotype absolutely correlated with CD3 surface expression. Furthermore, we have immunoprecipitated heterodimeric proteins that share molecular features with some receptor polypeptides previously described. CD3 polypeptides found on the surface of suppressor T cells are phosphorylated after phorbol ester stimulation. Collectively these studies unambiguously define the suppressive supernatant function as a product of CD3+ receptor-bearing T cells.

Serial cell-mediated immunological changes in terminal uremic patients on continuous ambulatory peritoneal dialysis therapy.

To investigate the mechanism of immunologic defects in uremia, we examined the cell-mediated immunity in 20 uremic patients with well balanced dietary assessments before continuous ambulatory peritoneal dialysis (CAPD) therapy and the serial changes in cell-mediated immunity 3, 6, 9, and 12 months after therapy. Absolute lymphocyte count, active T, total T, OKT4, OKT8, and B-cell levels were significantly lower in uremic patients than in controls, but progressively increased after CAPD treatment. The lymphoproliferative responses to mitogens decreased and were more evidently suppressed in the presence of autologous plasma. The suppressive effect of autologous plasma could be abolished by indomethacin treatment. The plasma prostaglandin E2 level, which was increased in uremic patients, decreased after CAPD treatment. These results suggest that prostaglandin E2 may play an important role in the suppression of T-cell function. The T-cell response to the stimulation of autologous non-T cells (autologous mixed-lymphocyte reaction; AMLR) in uremic patients was also low in patients before CAPD treatment. The patients' autoreactive T cells could suppress AMLR, and so did the supernatant from AMLR and isolated T-cell culture of uremic patients. These results suggest the existence of suppressor autoreactive T cells which can release suppressor factor(s). However, to our surprise, the autoreactive T-cell proliferation increased, and the suppressive effect of the supernatant was abolished after CAPD therapy. The mechanism for this peculiar phenomenon is not known and deserves further investigation.(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibition of antisuppression by the acute murine graft-versus-host reaction

Profound suppression of both humoral and cell-mediated immunity is a significant systemic effect of graft-versus-host reactions. Although no complete explanation has been advanced for this immunosuppression suppressor cells have been implicated. The data presented in this paper indicate that acute GVH reactions in (C57BL/6J X A/J) F1-hybrid mice induced by the injection of A/J cells severely disrupts the function of the antisuppressor T-cell pathway at both its induction and effector stages. Results show that within 3 weeks of induction of the reaction, Ly1+-T antisuppressor inducer cells lose their ability to generate the serum factor that mediates antisuppression. This factor is normally taken up by and activates Ly2+ T cells which then inhibit suppressor T-cell function. The data also reveal that Ly2+ T cells collected 2 weeks after induction lose their ability to be activated by the antisuppressor factor produced in normal mice. These cells are thus unable to function as antisuppressor effector cells. The uptake of the antisuppressor factor by Ly2+ T cells depends on the expression of Ia antigens on the surface of these cells. Experiments have shown that these antigens are absent from the surface of T cells derived from mice with GVH reactions. This finding may provide an explanation for the inability of these cells to function as antisuppressor effectors. Antisuppression is an important T-cell pathway that is intimately associated with the regulation of immune function. It is possible that the immunosuppression arising in mice with GVH reactions may stem, in part, from unopposed suppressor T-cell activity that results from widespread interference by the reaction with a pathway that normally inhibits suppressor cell activity.

In vitro effect of TP-1 (a calf thymic extract) on suppressor T-cell function of patients with autoimmune chronic active hepatitis

Concanavalin A-activated T-lymphocyte suppression of IgG production was found to be significantly impaired in patients with untreated active autoimmune chronic hepatitis when compared to normals or patients with inactive disease. When the dose-response effect of TP-1, a calf thymic extract, on in vitro suppressor cell activity was assessed, lymphocytes from six out of eight patients with previously reduced suppressor cell function showed a significant improvement, while over a similar range the suppressor cell activity of most normal controls declined. These results support the possibility that defective immunoregulation in patients with autoimmune chronic active hepatitis may be related to a deficiency in thymic hormone levels.

Reversal of antigen-induced resistance to collagen arthritis by cyclophosphamide

Treatment of rats with intravenous injection of 1 mg of soluble native type II collagen induced resistance against the subsequent induction of active arthritis by type II collagen immunization. This resistant state was accompanied by suppressed antibody response and delayed-type hypersensitivity (DTH) skin reaction to type II collagen. However, pretreatment of rats with 20 mg/kg of cyclophosphamide (CY), an agent reputed to damage suppressor T-cell function, 2 days before intravenous injection of soluble type II collagen abrogated the antigen-induced resistance against the subsequent induction of active arthritis. The DTH skin reaction to type II collagen was completely restored and the antibody response to type II collagen was significantly though not completely restored by CY pretreatment. These results provide evidence that antigen-induced resistance to collagen arthritis is mediated, at least in part, under the control of CY-sensitive events.

The protective effect of hepatic dysfunction on vascularized allograft survival

Recent studies have demonstrated that hepatic dysfunction, induced by experimental biliary ligation (EBL), impairs lymphocytic responsiveness to PHA stimulation in vitro and to cellular antigens in vivo. This suppression appears to be selective for T-cell mechanisms while B-cell-mediated functions remain intact. The purpose of this study was to determine whether coexisting hepatic insufficiency could exert a protective effect on vascularized or nonvascularized allograft survival in the transplanted recipient. Female Wistar-Furth ( Rtl w ) 225 g rats were assigned randomly to three groups: EBL, sham operation (Sham) and normal control (NC). Fourteen days following operation animals received heterotopic cardiac or skin allografts from Buffalo ( Rtl b ) donors. Cardiac and skin graft survival was determined daily, rejection was confirmed histologically, and technical failures were omitted from analysis. Allograft survival was expressed as median survival time ± SEM. Serum total bilirubin (mean ± SEM) was significantly elevated at Day 14 in EBL animals compared to Sham and NC groups (15.1 ± 1.0 vs 0.1 ± 0 and 0.2 ± 0.1 mg/dl, respectively, P < 0.01). Median cardiac allograft survival time by Probit was 10.6 ± 2.6 vs 5.6 ± 0.7 and 6.0 ± 0.9 days, respectively ( P < 0.03). Skin graft survival (mean and range) was similar in all groups. These results demonstrate that EBL in the rat suppresses T-cell function and significantly prolongs vascularized allograft survival, but not skin allograft survival across the Rtl histocompatibility barrier. The mechanism whereby coexisting hepatic dysfunction exerts a protective effect on vascularized allograft survival warrants further investigation.

Cytotoxic and immunoregulatory function of intestinal lymphocytes in Chlamydia trachomatis proctitis of nonhuman primates.

To study the role of natural killer cells and immunoregulatory T cells in the pathogenesis of proctitis due to Chlamydia trachomatis (L2 serovar), lymphocytes were obtained from the rectal mucosa and other sites of nonhuman primates and studied by using phenotypic and functional assays. In animals with lymphogranuloma venereum (LGV) proctitis, the percentage of lymphocytes with the natural killer cell phenotype (Leu-11+) was not significantly higher at any site in LGV infection, and natural killer cell function of lymphocytes isolated from the rectum was lower during LGV infection. This was not due to the suppressive effect of factors in serum, rectal lymphocytes, or LGV elementary bodies. In studies of regulatory T cells, the Leu-3+/Leu-2+ ratio was lower in the peripheral blood and the spleen during LGV infection, but the ratio did not decrease in lamina propria T cells. Both peripheral blood and rectal lymphocytes had higher helper T-cell function for polyclonal immunoglobulin G (IgG) synthesis in pokeweed mitogen-stimulated cultures 2 weeks following LGV infection. Increased suppressor T-cell function for pokeweed mitogen-stimulated IgG synthesis was found only in the peripheral blood of animals 2 weeks after infection, but not in isolated rectal lymphocytes. These results indicate that in LGV proctitis natural killer cells are not an important component of the inflammatory infiltrate at the site of infection, and helper T-cell function increases in peripheral blood and rectal lymphocytes.

Immunoregulatory T-lymphocyte functions in patients with small cell lung cancer

The present study was performed to elucidate the differences in immune status between patients with small cell lung cancer (SCLC) and those with non-small cell lung cancer. The study group consisted of 18 patients with SCLC and 15 with non-SCLC. Two healthy volunteers and 13 patients with benign disease were also included in the present study as the non-cancer control. In the non-SCLC group, although not statistically significant, the percentages of both OKT3+ and OKT4+ T-lymphocytes in the peripheral blood lymphocytes (PBL) were slightly decreased, associated with a slight increase in the percentage of OKT8+ T-cells, and a slight decrease in the OKT4+ to OKT8+ T-cell ratio. In contrast, the PBL of the SCLC patients showed significantly lower proliferative responses to phytohemagglutinin and human recombinant interleukin 2 than did the PBL of both the SCLC patients and the noncancer control group. The ability of PBL to produce lymphokines (interleukin 2 and macrophage activating factor) was significantly impaired in the SCLC group but not in the non-SCLC group. These results suggest that suppression of helper T-cell functions and/or potentiation of suppressor T-cell functions should occur in patients with SCLC.

Identification of a monoclonal antibody specific for a murine T3 polypeptide.

A monoclonal antibody (145-2C11) specific for the murine T3 complex was derived by immunizing Armenian hamsters with a murine cytolytic T-cell clone. The antibody is specific for a 25-kDa protein component (T3-epsilon) of the antigen-specific T-cell receptor. It reacts with all mature T cells and can both activate and inhibit T-cell function. These results identify T3-epsilon as a cell surface protein involved in the transduction of activation signals.

Decreased T-cell-mediated suppression of IgM-rheumatoid factor synthesis in rheumatoid arthritis

Circulating B-cell precursors specific for rheumatoid factor (RF) are present in normal subjects but spontaneous in vitro synthesis of RF occurs only in rheumatoid arthritis (RA). The regulatory role of RF-specific suppressor T cells in this process was studied in pokeweed mitogen-stimulated in vitro cultures of peripheral blood mononuclear cells. Addition of graded numbers of suppressor T8(+) cells from RA patients to normal B cells resulted in consistently less suppression of IgM and RF synthesis than that achieved by normal suppressor T cells. A preculture systen was then used to probe for RF-specific suppressor precursor lymphocytes. RA T-cell populations failed to generate normal levels of RF-specific suppression during in vitro culture for 4 days. Incubation with human-aggregated IgG (HaIgG) resulted in an increase in RF-specific suppression to normal levels. The data indicate that induction and full expression of RF-specific suppressor T-cell function is blocked in vivo in RA but can be overcome in vitro by incubation with HaIgG.

Immunological changes after hyposensitization in house-dust-sensitive asthmatic children: a review.

Patients with allergic diseases are characterized by the presence of elevated total serum IgE and specific IgE antibodies against a variety of environmental allergens. To explore the causes for augmented IgE antibody production and the working mechanisms of hyposensitization (HS), a series of studies has been conducted on house-dust-sensitive, newly diagnosed, and hyposensitized asthmatic children and normals. The specific IgE and IgG antibodies were measured by radioallergosorbent test; the lymphoproliferative capability was measured by 3H-thymidine incorporation; the allergen-specific suppressor activity was determined by the extent of house-dust-activated, interleukin-2 (IL-2)-expanded lymphocytes to suppress the allergen-induced proliferation of autologous mononuclear cells (MNC); and IL-2 was produced by stimulating MNC with allergen or phytohemagglutinin (PHA) and quantitated by its capability to support the proliferation of mouse IL-2-dependent cytotoxic T-cell line. The results showed: 1) HS was effective in 90% of patients in terms of decreased attacks and medication taken; 2) the patients were defective in suppressor T-cell function for IgE production; 3) HS was able to restore the regulatory T-cell function and increase the production of IgG-blocking antibody; and 4) IL-2 production may be used as an indicator for initiation and discontinuation of HS. Therefore, hyposensitization is an effective and specific treatment for allergic bronchial asthma and can partially correct an immunoregulatory aberration in atopic individuals.