Cukrová V, Neuwirtová R, Bartůnková J, Jonásová A, Cermák J, Homolková H, Malíková I
Ujstav hematologie a krevní transfuze, Praha.
Cas Lek Cesk. 2006;145(8):647-52; discussion 653.
Successful therapy with ATG and cyclosporine A in some myelodysplastic syndrome (MDS) patients led us to study the existence of T cells attacking autologous hemopoietic cells. In our study, we attempted to give the direct prove of autoreactive T cells in MDS (autoreactivity analysis). Simultaneously, we analysed the capacity of MDS patients to respond to allogeneic cells from unrelated individuals (alloreactivity analysis).
Autoreactive lymphocytes directed against own bone marrow mononuclear cells were analysed using the modification of cell mediated cytotoxic reaction. With one exception we did not confirm the presence of autoreactive T cells among 10 patients examined. Analysis of alloreactivity was performed by means of standard cell mediated cytotoxic reaction and mixed lymphocyte reaction. Surprisingly, the cytotoxic response to allogeneic cells was negative in 11 MDS patients from 16 analysed. When comparing refractory anaemia (RA) and refractory anaemia with ring sideroblasts (RARS) patients, the proportion of negative results was higher in RA (78 %) than in RARS (40 %). In mixed lymphocyte reaction, the response of MDS cells to allogeneic cells of unrelated individual was positive in all tested patients. The preliminary testing of TNF and IFNgamma secretion examined in supernatants of effector cells showed impaired levels of both cytokines in RA and normal levels in RARS in accordance with the findings achieved in alloreactivity analysis.
Autoreactive T cells were not found in MDS patients using our experimental arrangement. Analysis of alloreactivity showed the defect in effector--cytotoxic--phase of cell mediated cytotoxic reaction in the majority of MDS patients. The initial phase of this reaction represented in vitro by mixed lymphocyte reaction gave normal results. The possible reasons of disturbed alloreactivity and its relevance to immunity in MDS are commented in discussion.
抗胸腺细胞球蛋白(ATG)和环孢素A在部分骨髓增生异常综合征(MDS)患者中治疗成功,这促使我们研究攻击自体造血细胞的T细胞的存在情况。在我们的研究中,我们试图直接证明MDS中自身反应性T细胞的存在(自身反应性分析)。同时,我们分析了MDS患者对来自无关个体的异基因细胞的反应能力(同种异体反应性分析)。
使用改良的细胞介导细胞毒性反应分析针对自身骨髓单个核细胞的自身反应性淋巴细胞。在所检测的10例患者中,除1例例外,我们未证实存在自身反应性T细胞。通过标准的细胞介导细胞毒性反应和混合淋巴细胞反应进行同种异体反应性分析。令人惊讶的是,在16例接受分析的MDS患者中,有11例对异基因细胞的细胞毒性反应为阴性。比较难治性贫血(RA)和伴有环形铁粒幼细胞的难治性贫血(RARS)患者时,RA患者(78%)的阴性结果比例高于RARS患者(40%)。在混合淋巴细胞反应中,所有受试患者的MDS细胞对无关个体的异基因细胞的反应均为阳性。对效应细胞上清液中肿瘤坏死因子(TNF)和γ干扰素(IFNγ)分泌的初步检测显示,根据同种异体反应性分析的结果,RA患者中这两种细胞因子的水平受损,而RARS患者中这两种细胞因子的水平正常。
使用我们的实验方法未在MDS患者中发现自身反应性T细胞。同种异体反应性分析显示,大多数MDS患者在细胞介导细胞毒性反应的效应 - 细胞毒性阶段存在缺陷。该反应的初始阶段在体外以混合淋巴细胞反应表示,结果正常。讨论中对同种异体反应性受干扰的可能原因及其与MDS免疫的相关性进行了评论。