Regulation of T cell response to leishmania antigens by determinants of histocompatibility leukocyte class I and II molecules

It has been shown that HLA class I molecules play a significant role in the regulation of the proliferation of T cells activated by mitogens and antigens. We evaluated the ability of mAb to a framework determinant of HLA class I molecules to regulate T cell proliferation and interferon gamma (IFN-g) production against leishmania, PPD, C. albicans and tetanus toxoid antigens in patients with tegumentary leishmaniasis and healthy subjects. The anti-major histocompatibility complex (MHC) mAb (W6/32) suppressed lymphocyte proliferation by 90 percent in cultures stimulated with aCD3, but the suppression was variable in cultures stimulated with leishmania antigen. This suppression ranged from 30-67 percent and was observed only in 5 of 11 patients. IFN-g production against leishmania antigen was also suppressed by anti-HLA class I mAb. In 3 patients IFN-g levels were suppressed by more than 60 percent, while in the other 2 cultures IFN-g levels were 36 and 10 percent lower than controls. The suppression by HLA class I mAb to the proliferative response in leishmaniasis patients and in healthy controls varied with the antigens and the patients or donors tested. To determine whether the suppression is directed at antigen presenting cells (APCs) or at the responding T cells, experiments with antigen-primed non-adherent cells, separately incubated with W6/32, were performed. Suppression of proliferation was only observed when the W6/32 mAb was added in the presence of T cells. These data provide evidence that a mAb directed at HLA class I framework determinants can suppress proliferation and cytokine secretion in response to several antigens

Cytokine profile and pathology in human leishmaniasis

The clinical spectrum of leishmaniasis and control of the infection are influenced by the parasite-host relationship. The role of cellular immune responses of the Th1 type in the protection against disease in experimental and human leishmaniasis is well established. In humans, production of IFN-gamma is associated with the control of infection in children infected by Leishmania chagasi. In visceral leishmaniasis, an impairment in IFN-gamma production and high IL-4 and IL-10 levels (Th2 cytokines) are observed in antigen-stimulated peripheral blood mononuclear cells (PBMC). Moreover, IL-12 restores IFN-gamma production and enhances the cytotoxic response. IL-10 is the cytokine involved in down-regulation of IFN-gamma production, since anti-IL-10 monoclonal antibody (mAb) restores in vitro IFN-gamma production and lymphoproliferative responses, and IL-10 abrogates the effect of IL-12. In cutaneous and mucosal leishmaniasis, high levels of IFN-gamma are found in L. amazonensis-stimulated PBMC. However, low or absent IFN-gamma levels were observed in antigen-stimulated PBMC from 50 percent of subjects with less than 60 days of disease (24 + 26 pg/ml). This response was restored by IL-12 (308 + 342 pg/ml) and anti-IL-10 mAb (380 + 245 pg/ml) (P<0.05). Later during the disease, high levels of IFN-gamma and TNF-alpha are produced both in cutaneous and mucosal leishmaniasis. After treatment there is a decrease in TNF-alpha levels (366 + 224 pg/ml before treatment vs 142 + 107 pg/ml after treatment, P = 0.02). Although production of IFN-gamma and TNF-alpha might be involved in the control of parasite multiplication in the early phases of Leishmania infection, these cytokines might also be involved in the tissue damage seen in tegumentary leishmaniasis.

The absence of gamma-interferon production of S. mansoni antigens in patients with schistosomiasis

No gamma-interferon production was observed in peripheral blood mononuclear cells (PBMC) cultures from 45 patients living in an endemic area of schistosomiasis in Brazil following in vitro stimulation with schistosomula or adult worm antigens from Schistosoma mansoni (4.9 +/- 24 and 1.0 +/- 3.4 pg/ml, respectively). This immunological abnormality was observed in patients both with a high degree of infection (> or = 400 eggs/g feces) and with a low degree of infection (< 400 eggs/g feces), and was independent of the degree of natural exposure to infection. This absence of gamma-interferon production was antigen specific since high levels of this cytokine were detected in the same patients when their cells were stimulated with PPD (247 +/- 179 pg/ml) or PHA (408 +/- 328 pg/ml). In two of four subjects cured of a previous S. mansoni infection and currently living outside the endemic area, gamma-IFN was produced when their PBMC were stimulated with adult worm antigen (75 +/- 2.5 pg/ml).

Characterization of T cell responses to purified leishmania antigens in subjects infected with Leishmania chagasi

T cell responses to lipophosphoglycan-associated protein (LPG-AP) and the rgp63 antigens were studied in subjects with either asymptomatic L. chagasi infection or cured visceral leishmaniasis. The [3H]-thymidine uptake of lymphocytes stimulated with LPG-AP and rgp63 (mean +/- SD) was 14275 +/- 5048 and 3523 +/- 1678 cpm, respectively, for subjects with asymptomatic L. chagasi infection and 20046 +/- 5102 and 5086 +/- 3500 cpm, respectively, for subjects cured of visceral leishmaniasis. The responses to LPG-AP in both asymptomatic and cured visceral leishmaniasis were higher (P < 0.01) than those observed with rpg63. LPG-AP induced IFN-gamma production in all subjects studied, while rgp63 did not induce lymphocyte proliferation or IFN-gamma production in the majority of the subjects tested. IFN-gamma levels in cultures stimulated with LPG-AP were 103 +/- 81 pg/ml in individuals with asymptomatic L. chagasi infection and 127 +/- 123 pg/ml in subjects cured of visceral leishmaniasis. IFN-gamma levels in cultures stimulated with LPG-AP from subjects with asymptomatic L. chagasi infection were comparable to those observed in subjects cured of visceral leishmaniasis (P > 0.05). These data indicate that LPG-AP is recognized and induces T cell proliferation and IFN-gamma production in subjects with protective immune response against Leishmania chagasi.

Gamma interferon production by lymphocytes from children infected with L. chagasi

The present study was performed to evaluate the ability of lymphocytes from 18 children living in an endemic area of visceral leishmaniasis (VL) to produce gamma-interferon. These children had no previous history of VL and were considered to be infected with Leishmania chagasi based on leishamnial seroconversion. The gamma IFN levels were determined by radioimmunoassay on supernatants of lymphocyte cultures (3 x 10**6/ml). stimulate with PHA (final dilution 1:10) and Leishamnia chagasi antigen (10µg/ml). The gamma-IFN production by lymphocytes from seroconverting children stimulated with PHA (178 ñ 151 U/ml) and Leishmania chagasi (47 ñ 77 U/ml) was significantly higher than that observed in visceral leishmaniasis. For clinical follow-up, these 18 seroconverting children were divided into three groups: asymptomatic infection (N=4); self-healing subclinical illness (N=9), and sublinical infection progressing to VL (N=5). Gamma IFN levels inchildren with either asymptomatic or subclinical infection (65 ñ 85 U/ml were significantly higher (P < 0.003) than those observed in children progressing to VL (9 ñ 6 U/ml). The data demonstrate that there is an association between gamma IFN levels and the clinical course of Leishmania infection