Dumont F J, Coker L Z, Habbersett R C, Treffinger J A
J Immunol. 1985 Apr;134(4):2357-65.
The patterns of cellular and strain reactivity of a monoclonal antibody (6C3 MAb) derived from the fusion of SP2/0 cells with splenocytes from rats immunized against MRL/MpJ-lpr/lpr T cells were characterized by using flow cytofluorometry (FCF) analysis. This MAb was found to stain 70 to 90% of T cells of mice with the lpr/lpr genotype and 20 to 60% of T cells of congenic +/+ strains. Dual-parameter FCF analysis of Lyt-2 vs 6C3 expression revealed the existence of several Lyt-2- and Lyt-2+ T cell subsets, one of which (Lyt-2- bright 6C3+) was expanded in lpr/lpr-bearing mice. The 6C3 MAb stained only 2 to 5% normal thymocytes but reacted with 40 to 50% bone marrow (BM) cells. A strain survey demonstrated the expression of the 6C3 antigen on peripheral T cells (and BM cells) of all strains examined, with the exception of NOD, NZB/B1NJ, and ST/bJ. Interestingly, in the positive strains, two types of 6C3 staining patterns of T cells were observed: bimodal or trimodal. Study of BXH and CXB recombinant inbred (RI) strains demonstrated that the bimodal and trimodal 6C3 patterns are associated with the Ly-6.1 and Ly-6.2 phenotypes, respectively. Linkage of 6C3 expression with the Ly-6 locus was confirmed by using the congenic C3H.B6-Ly-6b strain. Moreover, the 6C3 staining of T cells in Ly-6.2 strains was reduced by preincubation with the H9/25 and SK-142-446 MAb, which are known to recognize Ly-6.2-associated antigens. Therefore, the 6C3 MAb appears to detect a frame-work determinant on an Ly-6-linked antigen that is absent from T cells of NOD, NZB, and ST/bJ mice. Analysis of (NZB x C58) NX8 RI strains demonstrated a correlation between the lack of 6C3 expression on T cells and unresponsiveness in autologous mixed lymphocyte reaction (a property of NZB/B1NJ mice). The 6C3 MAb should prove useful for further genetic and biochemical analysis of the Ly-6 locus and its product(s), and for the delineation of functional subsets of T cells and BM cells in normal and lpr/lpr-bearing mice.
利用流式细胞荧光测定法（FCF）分析，对源自SP2/0细胞与免疫抗MRL/MpJ-lpr/lpr T细胞的大鼠脾细胞融合产生的单克隆抗体（6C3单克隆抗体）的细胞和菌株反应模式进行了表征。发现该单克隆抗体可对70%至90%的lpr/lpr基因型小鼠的T细胞以及20%至60%的同基因+/+菌株的T细胞进行染色。对Lyt-2与6C3表达进行双参数FCF分析，揭示了几个Lyt-2和Lyt-2+ T细胞亚群的存在，其中一个亚群（Lyt-2-亮6C3+）在携带lpr/lpr的小鼠中有所扩增。6C3单克隆抗体仅对2%至5%的正常胸腺细胞进行染色，但与40%至50%的骨髓（BM）细胞发生反应。一项菌株调查表明，除了NOD、NZB/B1NJ和ST/bJ之外，在所有检测的菌株的外周T细胞（和BM细胞）上均有6C3抗原的表达。有趣的是，在阳性菌株中，观察到T细胞的两种6C3染色模式：双峰或三峰。对BXH和CXB重组近交（RI）菌株的研究表明，双峰和三峰6C3模式分别与Ly-6.1和Ly-6.2表型相关。通过使用同基因C3H.B6-Ly-6b菌株，证实了6C3表达与Ly-6基因座的连锁关系。此外，用已知可识别Ly-6.2相关抗原的H9/25和SK-142-446单克隆抗体进行预孵育后，Ly-6.2菌株中T细胞的6C3染色减少。因此，6C3单克隆抗体似乎检测到了NOD、NZB和ST/bJ小鼠的T细胞中不存在的Ly-6连锁抗原上的一个框架决定簇。对（NZB×C58）NX8 RI菌株的分析表明，T细胞上缺乏6C3表达与自身混合淋巴细胞反应中的无反应性之间存在相关性（这是NZB/B1NJ小鼠的一个特性）。6C3单克隆抗体应有助于对Ly-6基因座及其产物进行进一步的遗传和生化分析，以及描绘正常和携带lpr/lpr的小鼠中T细胞和BM细胞的功能亚群。
