Analysis of 1pr-dnT Cell Function in Sle-Prone Mouse, MRL/1pr.

本研究研究了MRL/1pr小鼠t细胞表面抗原的表达以及干细胞或早期再成熟t细胞特异性的功能特征。如前所述，MRL/1pr淋巴结细胞Lyt-2阴性，L3T4阴性，分别是细胞毒性和辅助t细胞亚群的常规表面抗原。采用IgM型抗LyT-2和L3T4细胞毒抗体与兔补体，纯化LyT-2（-）、L3T4（-）双阴性t细胞，并进行进一步实验。发现1pr双阴性t细胞Pgp-1和B220抗原阳性，这是早期t细胞的标志。然后我们开发了单克隆抗体识别干细胞t细胞或非常不成熟的t细胞。这些单克隆抗体被命名为Sca-1和Sca-2，用于检测1pr双阴性细胞的表面抗原。我们的研究结果表明，1pr-双阴性t细胞表达Sca-1，但不表达Sca-2，这表明1pr-双阴性t细胞属于t细胞谱系，它们确实处于早熟状态。在接下来的实验中，我们研究了可能是1pr双阴性细胞产物的生长因子。纯化后的1pr双阴性t细胞在RPMI 1640培养基中培养于96孔板中。纯化后的上清液采用凝胶过滤技术进行分析。我们发现上清含有类似肿瘤生长因子β的生长因子。需要进一步的研究来阐明1pr双阴性t细胞的功能。

We investigated the expression of T-cell surface antigens and the functional characteristics specific for stem cells or early remature T-cells in MRL/1pr mouse in the present study.As previously reported, MRL/1pr lymph node cells was Lyt-2 negative, L3T4 negative which were conventional surface antigens of cytotoxic and helper T-cell subsets, respectively. Using IgM type cytotoxic antibodies against LyT-2 and L3T4 with rabbit complement, Lyt-2 (-), L3T4 (-) double negative T-cells were purified, and subjected to the further experiments. The 1pr-double negative T-cells were found that they were positive for Pgp-1 and B220 antigens, which were markers of premature T-cells. We then developed monoclonal antibody which recognize stem T-cells or very immature T-cells. These monoclonal antibodies, designated as Sca-1 and Sca-2, were used for the detection of the surface antigens of 1pr-double negative cells. Our findings indicated that 1pr-double negative T-cells expressed Sca-1 but not Sca-2, suggesting that 1pr-double negative T-cells were on the T-cell lineage, and they were really premature state. In the next experiments we investigated the growth factors which might be the products of 1pr-double negative cells. The purified 1pr-double negative T-cells were cultured in 96-well plates in RPMI 1640 media. The supernatants were analyzed after purification by gel-filtration technique. We found that the supernatant contained growth factors which had function like tumor growth factor beta. Further study will be needed to clarify the function of 1pr-double negative T-cells.