中文摘要：

目的：研究Ag85B蛋白促PPD^＋人外周血单个核细胞的增殖活性。方法：用PCR技术扩增Ag85B基因，构建重组pcDNA3-Ag85B，并将其转染B16细胞株，用RT—PCR鉴定阳性细胞克隆，用Western blotting鉴定其在阳性细胞克隆内的表达，用^3H—TdR掺入法来检测Ag85B的促增殖活性。结果：获得了阳性B16细胞克隆，并鉴定在阳性细胞克隆内有Ag85B成熟蛋白表达；此阳性细胞克隆对PPD^＋人的PBMC具有较明显地促PBMC增殖活性。结论：转染了pcDNA3-Ag85B的B16阳性细胞克隆对PPD^＋人的PBMC具有较明显地促PBMC增殖活性，为Ag85B诱发抗肿瘤免疫应答及其机制的研究奠定了基础。

英文摘要：

Objective： To study the proliferation - activity of Ag85B protein stimulating the PBMCs of PPD^＋ human donors, Methods： The gene encoding Ag85B mature protein was amplified by polymerase chain reaction （PCR）, and recombinant vector （pcDNA3） was reconstructed and transfected into B16 cell lines. The positive cell clones by RT - PCR and its protein expression was measured by Western blotting. The PBMCs＇ proliferation was analyzed by ：^3H - TdR after the stimulation of positive cell clones in the PBMCs of PPD ^＋ and PPD^- human donors. Results： The positive B16 cell clones were obtained and identified by Western blotting the Ag85B mature protein expressed in the clones of positive cells, which could stimulate the PBMCs＇ proliferation - activity in PPD^＋ human donors, Conclusion： These modified B16/pcDNA3 - Ag85B cells significantly enhanced the proliferation of PBMCs of PPD^＋ human donors, which provided foundation to the study of Ag85B antitumor immune activity.