HIV-1感染恒河猴外周血单个核细胞的生物学特性和C2-V3区基因序列变化

目的探讨艾滋病病毒1型(HIV-1)感染恒河猴外周血单个核细胞(PMBCs)，并在其中传代的可能性。方法用HIV-1感染原代猴PBMCs，采用酶联免疫吸附试验(ELISA)方法测定培养0、2、3、7天时上清液中的HIV-1P24抗原，以观察HIV-1在猴细胞中的复制情况，比较HIV-1分别在人和猴细胞感染的复制动力学。同时采用聚合酶链反应(PCR)方法扩增HIV-1的enV基因，并对其C2-V3区进行序列分析。结果HIV-1毒株SF33、89．6、ⅢB感染猴PBMCs后的第3天，P24抗原达到最高，随着培养时间的延长，P24抗原逐渐降低。将SF33、ⅢB、89．6、YNl9四个毒株分别感染猴PBMCs并盲传3代后，P24抗原测定结果均为阴性。提取盲传各代细胞脱氧核糖核酸(DNA)进行PCR扩增，结果显示，SF33、ⅢB、YN19、89．6感染猴细胞的第一代和89．6感染猴细胞的第二代，均可以检测到HIV-1前病毒核酸。比较原毒株enV基因C2-V3区的序列发现，传代后的基因序列SF33有5个碱基改变外，其它HIV-1只有0～2个核苷酸改变。与SF33感染猴PBMCs的表现相反，其感染人PBMCs时。HIV-1的复制一直维持在较高的水平。结论应用3个实验株、1个临床分离株HIV-1，虽都能够感染恒河猴外周淋巴细胞，但均难以继代，提示能否用恒河猴作为HIV-1研究的动物模型，尚待进一步的体内外研究。

Infection of rhesus macaque''''s PBMCs with HIV-1 in vitro-related biological features and C2-V3 sequence

Objective HIV-1 strains were tested for their ability to infect,replicate and passage in rhesus macaque's peripheral blood mononuclear cells(PBMCs).Methods The infection and replication of HIV-1 strains in rhesus macaque PBMCs in vitro were observed by detecting HIV-1 P24 in the culture supernatant on the day 0,2,3,7 of post-infection, using enzyme-linked immunosorbent assay(ELISA);and the viral replication in macaque's PBMCs was compared with human PBMCs.HIV-1 C2-V3 region of infected PBMs was amplified by polymerase chain reaction(PCR) and followed with gene sequence.Results SF33,89.6 and IIIB could be replicated efficiently in rhesus macaque's PBMCs.The HIV-1 P24 appeared the highest in 3 days after infection and decreased gradually afterwards.HIV-1 P24 could not be detected in the culture supernatant when SF33,IIIB,89.6 and YN19 were blindly passaged in Macaque's PBMs for 3 passages.However,HIV-1 proviral DNA could be amplified in passage 1 of SF33,IIIB,89.6 and YN19 and passage 2 of 89.6 in Macaque's PBMs.Comparing with C2-V3 region of original HIV-1 strain,there was less changes(0-2 base)in C2-V3 region after HIV-1 was passaged in macaque's PBMCs,except for SF33(5 bases change).Contrary to SF33 infection in macaque's PBMCs,the viral replication of SF33 infection in human PBMCs maintained high level after HIV-1 replication.Conclusion Although three laboratory HIV-1 strains and one HIV-1 isolates can infect rhesus macaque's PBMs ,passage of HIV-1 strains can be hardly achieved in rhesus macaque's PBMC,suggesting that further in vivo/in vitro studies are needed to clarify whether rhesus macaques may act as a relevant animal model for HIV-1.