Chang J C, Smith L R, Froning K J, Kurland H H, Schwabe B J, Blumeyer K K, Karasek M A, Wilkinson D I, Farber E M, Carlo D J, Brostoff S W
Immune Response Corporation, Carlsbad, Calif, USA.
Arch Dermatol. 1997 Jun;133(6):703-8.
We previously demonstrated a clonal dominance in the V beta 13.1 messages isolated from the lesional CD8+ T cells of psoriasis vulgaris, which suggested an interaction of V beta 13.1+ CD8+ T cells with skin antigens.
To determine whether the clonality observed accurately reflected a clonal population of infiltrating T cells or was skewed by an overabundance of messages from a small number of cells, and to extend our study of V beta gene usage by lesional CD8+ T cells to 9 new patients.
Case study.
Patients were enrolled at the Psoriasis Research Institute in Palo Alto, Calif, and samples were analyzed at The Immune Response Corporation in Carlsbad, Calif.
For the 2 previous patients, skin samples were sorted directly for V beta 13.1+ T cells, for which the T-cell receptors were sequenced. For the 9 new patients, CD8+ T cells were sorted and their T-cell receptor V beta gene usage measured using semiquantitative polymerase chain reaction with V beta-specific primers.
The directly sorted V beta 13.1+ T cells exhibited clonal dominance in both patients. The dominant V beta 13.1 clone in each patient was the same as that found in the previous 2 biopsy specimens for which CD8+ T cells were sorted. Additionally, in 8 of the 9 new patients examined, we again found a preferential usage of V beta 3 and/or V beta 13.1 genes by the lesional CD8+ T cells.
The clonality, which was found in the V beta messages of the sorted CD8+ T cells, accurately reflects the dominance of these clones in the infiltrating T cells. Moreover, the persistence in the same patient of the same clone for as long as 15 months and the overrepresentation of V beta 3 and/or V beta 13.1 in lesional CD8+ T cells in the new patients examined support the pathogenic role of T cells bearing these V betas.
我们之前证实,从寻常型银屑病皮损处CD8⁺ T细胞中分离出的Vβ13.1信息存在克隆优势,这提示Vβ13.1⁺ CD8⁺ T细胞与皮肤抗原之间存在相互作用。
确定观察到的克隆性是否准确反映浸润性T细胞的克隆群体,或者是否因少数细胞的信息过多而产生偏差,并将我们对皮损处CD8⁺ T细胞Vβ基因使用情况的研究扩展到9名新患者。
病例研究。
患者在加利福尼亚州帕洛阿尔托的银屑病研究所入组,样本在加利福尼亚州卡尔斯巴德的免疫反应公司进行分析。
对于之前的2名患者,直接对皮肤样本进行Vβ13.1⁺ T细胞分选,并对其T细胞受体进行测序。对于9名新患者,分选CD8⁺ T细胞,并使用Vβ特异性引物通过半定量聚合酶链反应测量其T细胞受体Vβ基因的使用情况。
直接分选的Vβ13.1⁺ T细胞在两名患者中均表现出克隆优势。每名患者中占主导地位的Vβ13.1克隆与之前分选CD8⁺ T细胞的2份活检标本中发现的克隆相同。此外,在检查的9名新患者中的8名中,我们再次发现皮损处CD8⁺ T细胞优先使用Vβ3和/或Vβ13.1基因。
在分选的CD8⁺ T细胞的Vβ信息中发现的克隆性准确反映了这些克隆在浸润性T细胞中的优势。此外,同一克隆在同一患者中持续长达15个月,以及在新检查患者的皮损处CD8⁺ T细胞中Vβ3和/或Vβ13.1的过度表达支持了携带这些Vβ的T细胞的致病作用。