Mamedov I Z, Britanova O V, Chkalina A V, Staroverov D B, Amosova A L, Mishin A S, Kurnikova M A, Zvyagin I V, Mutovina Z Y, Gordeev A V, Khaidukov S V, Sharonov G V, Shagin D A, Chudakov D M, Lebedev Y B
Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.
Autoimmunity. 2009 Sep;42(6):525-36. doi: 10.1080/08916930902960362.
Ankylosing spondylitis (AS) is commonly characterized by clonal expansions of T cells. However, these clonal populations are poorly studied and their role in disease initiation and progression remains unclear. Here, we performed mass sequencing of TCR V beta libraries to search for the expanded T cell clones for two AS patients. A number of clones comprising more than 5% of the corresponding TCR V beta family were identified in both patients. For the first time, expanded clones were shown to be stably abundant in blood samples of AS patients for the prolonged period (1.5 and 2.5 years for two patients, correspondingly). These clones were individually characterized in respect to their differentiation status using fluorescent cell sorting with CD27, CD28, and CD45RA markers followed by quantitative identification of each clone within corresponding fraction using real time PCR analysis. Stable clones differed in phenotype and several were shown to belong to the proinflammatory CD27 - /CD28 - population. Their potentially cytotoxic status was confirmed by staining with perforin-specific antibodies. Search for the TCR V beta CRD3 sequences homologous to the identified clones revealed close matches with the previously reported T cell clones from AS and reactive arthritis patients, thus supporting their role in the disease and proposing consensus TCR V beta CDR3 motifs for AS. Interestingly, these motifs were also found to have homology with earlier reported virus-specific CDR3 variants, indicating that viral infections could play role in development of AS.
强直性脊柱炎（AS）通常以T细胞的克隆性扩增为特征。然而，这些克隆群体的研究较少，它们在疾病发生和发展中的作用仍不清楚。在此，我们对两名AS患者的TCR Vβ文库进行了大规模测序，以寻找扩增的T细胞克隆。在两名患者中均鉴定出了一些占相应TCR Vβ家族5%以上的克隆。首次发现，在AS患者的血液样本中，扩增的克隆在较长时间内（两名患者分别为1.5年和2.5年）稳定存在。使用CD27、CD28和CD45RA标记通过荧光细胞分选对这些克隆的分化状态进行了个体表征，随后通过实时PCR分析对相应组分内的每个克隆进行了定量鉴定。稳定的克隆在表型上有所不同，其中一些被证明属于促炎的CD27 - /CD28 - 群体。用穿孔素特异性抗体染色证实了它们潜在的细胞毒性状态。搜索与已鉴定克隆同源的TCR Vβ CDR3序列，发现与先前报道的AS和反应性关节炎患者的T细胞克隆密切匹配，从而支持了它们在疾病中的作用，并提出了AS的TCR Vβ CDR3共有基序。有趣的是，还发现这些基序与早期报道的病毒特异性CDR3变体具有同源性，表明病毒感染可能在AS的发展中起作用。
