固相PF18-3单抗促进亚适量ConA诱导的胸腺细胞活化后凋亡

Enhancement of sub optimal doses of Con A-mediated activation and apoptosis of thymocytes by immobilized PF18-3 McAb

目的 研究PF18 3单抗识别分子和胸腺细胞活化及凋亡的关系。方法 应用3H TdR掺入实验及流式细胞术 (FACS)技术 ,分析了胸腺细胞与固相单抗PF18 3共育后细胞活化过程中3H TdR掺入、细胞周期及DNA亚二倍体变化、早期活化及凋亡分子的表达。结果 固相单抗PF18 3有弱的促胸腺细胞3H TdR掺入和促进CD2 5的表达 ,当有亚适量ConA存在时作用更明显。与对照组相比 ,PF18 3及亚适量ConA共育后 ,2 4、48h时活细胞数量减少。细胞周期分析 ,G0 /G1期细胞相对减少 ,S期相对增多 ,但G2 /M期无明显变化。凋亡相关分析 ,共育后 2 4、48h时DNA亚二倍体阳性细胞及 12h时AnnexinV结合阳性细胞增多。结论 PF18 3单抗识别分子能协同亚适量ConA对胸腺细胞的活化 。

Objective To investigate the relationship between PF18 3 McAb recognized molecule and thymocyte activation induced apoptosis. Methods Use 3H TdR uptake test and FACS technique to analyze 3H TdR uptake, cell cycle, DNA hypodiploid and expression of early activation or apoptosis molecule. Results Immobilized PF18 3 McAb increased 3H TdR uptake and CD25 expression on thymocytes when costimulated with sub optimal doses of Con A. However, in contrast to control group, the number of viable cells decreased. Cell cycle analysis in thymocyte cultures showed that the proportion of cells at G0/G1 phase decreased, cells at S phase increased relatively, but no obvious change at G2/M phase. Apoptosis connected analysis showed that both DNA hypodiploid cell (at 24-48h) and AnnexinV binding positive cells (at 72h) increased. Conclusion The molecule recognized by PF18 3 McAb enhanced sub optimal doses of Con A to induce activation of thymocytes and early cell death by apoptosis. Subject words Thymocyte; Monoclonal antibody; Apoptosis