不同嗜性HIV复制与树突状细胞成熟状态关系的研究

Study on the relationship between replication of HIV-1 with different tropism and the developmental state of dendritic cells

目的 探讨不同嗜性HIV-1在人树突状细胞（dendriticcell，DC）内复制与DC成熟状态的关系。方法 采用MACS磁珠分选法纯化出CD14^＋细胞，加入粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素-4体外培养7d，得到未成熟DC；再加入肿瘤坏死因子-α继续培养3d，得到成熟DC。感染不同嗜性HIV-1病毒株后，检测培养上清p24含量，观察HIV-1复制能力。结果 受T嗜性HIV-1攻击后，未成熟DC及成熟DC培养上清p24抗原含量随培养时间延长无明显增加。受M嗜性HIV-1攻击后，未成熟DC培养上清p24抗原含量随培养时间延长明显增加，成熟DC培养上清p24抗原含量不随培养时间延长明显增加。结论 T嗜性HIV-1不能在未成熟DC和成熟DC内复制，M嗜性HIV-1能在未成熟DC内复制但不能在成熟DC内复制，提示HIV-1能否在DC内复制与HIV-1嗜性和DC成熟状态有关。

Objective To detemine the relationship between replication of HIV-1 primary isolates with M-tropic or T-tropic and differentiation state of dendritic cells（DC）. Methods CD14^＋ cells were isolated from peripheral blood mononuclear cells of healthy donors. The purified CD14^＋ cells were cultured in the presence of granulocytemacrophage colony-stimulating factor（GM-CSF） and intedukin-4 （IL-4） for 7 days. Cells were then cultured for fur- ther 3 days in the presence of TNF-α to get mature DC. After infection with different tropic HIV- 1 primary isolates, the level of p24 antigen in the culture supematant was detected to observe the dynamic alteration of HIV- 1 replication. Results The level of p24 antigen in culture supematant of immature DC and mature DC was not significantly increased after exposure to T-tropic HIV-1. The level of p24 antigen in culture supematant of immature DC increased significantly after exposure to M-tropic HIV-1, but rather than mature DC. Conclusion T-tropic HIV-1 do not replicate in immature and mature DC. M-tropic HIV-1 could replicate in immature DC, but not in mature DC. The data indicated that whether HIV-1 replicate in DC depended on the tropism of HIV-1 and the differential stage of DC.