FULL-LENGTH SEQUENCE CHARACTERIZATION OF INFECTIOUS HIV-1 LONG-TERM NONPROGRESSOR VIRUS

Tiffany Ethridge¹, M. Cloyd, Ph.D.^1,2, G. Sahu, Ph.D.², S. Sahu², J. Huang, Ph.D.

¹Department of Pathology, ²Department of Microbiology, UTMB

Background: Although most patients with HIV-1 infection develop AIDS within 10 years, approximately five percent of infected individuals remain relatively healthy for 15 years or longer. These long-term nonprogressor (LTNP) patients have been shown to exhibit a low viral load and develop a strong anti-HIV immune response. LTNP viruses have also been shown to replicate more efficiently in macrophages than T-cells. The full-length sequence of LTNP viruses has yet to be characterized in detail. Most LTNP sequences previously analyzed have been derived directly from PBMCs, which means the sequenced virus may or may not be infectious. Objective: The objective is to characterize infectious long-term nonprogressor viruses in detail by analyzing the full-length sequences of six infectious LTNP virus isolates and comparing them with sequences obtained from progressor viruses. Methods: I am growing virus from 6 different LTNP patients in VB cells. I will isolate chromosomal DNA from infected VB cells, amplify the entire viral sequence in two parts, and create recombinant molecular clones of the LTNP virus isolates. Next I will test to ensure the clones are capable of producing infectious virus using transient transfection and infectivity assays and test growth of these cloned viruses in macrophages and T-cells to verify cell tropism. Clones will then be completely sequenced and these sequences compared with those from standard HIV strains isolated from rapid progressors in the HIV database. Supported by NIH grants R01-AI062453 and R21-AI066999, Grad School of Biomedical Sciences.