Hiv-1 Latent Human Jurkat T Cell Model And The Chinese Epidemic Strains Of Hiv-1 B / C Recombinant Subtype Infection Of Human Umbilical Cord Blood Hematopoietic Progenitor Cells

High activity anti-retroviral therapy (HAART) effectively suppresses the replication of human immunodeficiency virus (HIV) and slows down the progress of AIDS. However, it can't eradicate HIV because of the presence of latent reservoirs. Therefore, establishment of HIV latent infection models is urgently needed to explore the mechanisms of postintegration latency and the model can be used to screen the drugs to eradicate HIV latency. On the other hand, the issues that whether or not hematopoietic progenitor cells (HPCs) can be infected by HIV-1 (especially by Chinese HIV-1 B/C subtype) are waiting for elucidated. Exploration about such problems will contribute to establish a HIV-1 latent infection model on HPCs.In the first part of present study, HIV-1 pseudoviruses which express green fluorescent protein (GFP) were used to infect Jurkat cells. Potential latent clones were screened by the detections of reactivation of GFP as well as the expression of p24 antigen before and after stimuli.106 TCID50 pseudovirus were used to infect Jurkat cells followed by sorting out GFP- cell population. Under the stimulation of 200 nM TSA or 10 ng/ml PMA, GFP were expressed in some original GFP" cells and these GFP reactivated cell population was isolated by Flow Cytometry. Several clones were acquired by limiting dilution. Further test was performed by HIV p24 Elisa assay and 5 HIV-1 latent clone lines were identified ultimately. Alu-LTR nest-PCR assay also demonstrated that HIV-1 genome had been integrated in the genome of these clone lines. It suggested that we had successfully established a HIV-1 pseudovirus latent infection in vitro model.The second part of present study focused on the possibility of HIV-1 B/C subtype to infect Hematopoietic Progenitor Cells (HPCs) and the potential mechanisms. CD34+ HPCs were isolated from Umbilical Cord Blood which came from healthy pregnant women, the purity ranged in 95～99%, and isolated HPCs had directional differentiation ability. And then different dosages (50 TCID50,200 TCID50,1000 TCID50) of HIV-1 B/C subtype pseudovirus were used to infect. In different time-point, Elisa assay was used to detect p24 antigen in supernatant of cell culture. Cell genome DNA was also extracted to detect integration of HIV-1 genome by Alu-LTR nest PCR. The data suggested HIV-1 subtype B/C may infect HPCs. Furthermore, our study demonstrated that HIV-1 related receptors including CD4, CCR5 and CXCR4 which expressed on CD34+ HPCs may provide a clue for potential mechanisms of the infection.