Favali Cecilia, Costa Dirceu, Afonso Lilian, Conceição Viviane, Rosato Andréa, Oliveira Fabiano, Costa Jackson, Barral Aldina, Barral-Netto Manoel, Brodskyn Claudia Ida
Laboratório de Imunoparasitologia, Centro de Pesquisas Gonçalo Moniz, Fundação Oswaldo Cruz, Rua Waldemar Falcão, 121, Salvador, Bahia 40295-001, Brazil.
Microbes Infect. 2005 Jan;7(1):86-92. doi: 10.1016/j.micinf.2004.09.015. Epub 2004 Dec 13.
T cell-mediated immunity is critical in resistance against Leishmania parasites, and T cell activation requires signals provided by costimulatory molecules. Herein we evaluated the role of costimulatory molecules on cytokine production and T cell surface molecule expression by peripheral blood mononuclear cells (PBMC) from cutaneous leishmaniasis (CL) patients. PBMC from CL patients were stimulated with soluble Leishmania antigen (SLA, 10 microg/ml), in the presence or absence of soluble CTLA4-Ig to block CD28-B7 interaction or in the presence or absence of anti-human CD40L to block CD40-CD40L interaction. Supernatants were harvested to evaluate tumor necrosis factor alpha (TNF-alpha), interleukin 10 (IL-10), transforming growth factor beta (TGF-beta) and interferon gamma (IFN-gamma) production by ELISA. Cells were harvested after 48 h of culture, stained for specific activation markers and analyzed by flow cytometry. Results show that the blockade of CD28-B7 interaction by CTLA4-Ig downmodulated IFN-gamma, IL-10, and TNF-alpha secretion by PBMC from CL patients. No alteration was detected on either TGF-beta production or the expression of CTLA44 or CD25 on CD4+ and CD8+ T cells. When the CD40-CD40L interaction was blockade using anti-CD40L, we did not observe changes in cytokine production or in surface molecule expression. The blockade of the CD28-B7 interactions by CTLA4-Ig also did not alter cytokine production in volunteers immunized against tetanus toxoid (TT). Taken together, these data suggest that the interaction of CTLA4 and CD28-B7 is a TGF-beta-independent mechanism that specifically downmodulates the immune response in cutaneous leishmaniasis patients.
T细胞介导的免疫在抵抗利什曼原虫寄生虫方面至关重要,而T细胞激活需要共刺激分子提供的信号。在此,我们评估了共刺激分子对皮肤利什曼病(CL)患者外周血单个核细胞(PBMC)细胞因子产生和T细胞表面分子表达的作用。用可溶性利什曼原虫抗原(SLA,10微克/毫升)刺激CL患者的PBMC,存在或不存在可溶性CTLA4-Ig以阻断CD28-B7相互作用,或存在或不存在抗人CD40L以阻断CD40-CD40L相互作用。收集上清液,通过酶联免疫吸附测定法评估肿瘤坏死因子α(TNF-α)、白细胞介素10(IL-10)、转化生长因子β(TGF-β)和干扰素γ(IFN-γ)的产生。培养48小时后收集细胞,用特异性激活标记物染色,并通过流式细胞术进行分析。结果表明,CTLA4-Ig阻断CD28-B7相互作用可下调CL患者PBMC分泌的IFN-γ、IL-10和TNF-α。在TGF-β产生或CD4+和CD8+T细胞上CTLA44或CD25的表达方面未检测到改变。当使用抗CD40L阻断CD40-CD40L相互作用时,我们未观察到细胞因子产生或表面分子表达的变化。CTLA4-Ig阻断CD28-B7相互作用也未改变接种破伤风类毒素(TT)的志愿者的细胞因子产生。综上所述,这些数据表明CTLA4与CD28-B7的相互作用是一种不依赖TGF-β的机制,可特异性下调皮肤利什曼病患者的免疫反应。