The understanding of human immunodeficiency virus type 1 (HIV-1) pathogenesis, in vivo, requires the ability to precisely quantitate HIV-1 proviral DNA and RNA in a variety of tissues and secretions. Although HIV- 1 has long been known to infect semen and cervical/vaginal secretions of HIV-1-seropositive individuals, the molecular mechanisms affecting the sexual transmission of HIV-1, in vivo, are poorly defined. Understanding the factors which affect the infectivity of HIV- 1-seropositive individuals is of critical importance. Utilizing two novel molecular techniques, developed in our laboratory, the precise HIV-1-proviral load in the cellular components of semen and cervical/vaginal secretions, and the HIV- 1 RNA levels in the genital secretions shall be determined. An in situ polymerase chain reaction (PCR) for HIV-1 DNA, which allows amplification of a segment of the HIV-1 genome in intact cells, and a quantitative HIV-1 reverse transcriptase PCR technique will be utilized. Total HIV-1 RNA, in both cell-associated and cell-free fractions, of these genital secretions shall be evaluated. As well, an aberrant pattern of HIV- l RNA, previously correlated with restricted viral replication, will be investigated in the cellular components of these genital secretions. HIV-1 proviral and HIV-1 RNA total levels and patterns, in genital secretions, will be correlated with other clinical markers of HIV- 1 infection and initial mode of HIV-1 infection. As well, the levels of HIV- l proviral-harboring cells and HIV-l RNA in genital secretions shall be followed, longitudinally, through disease progression and correlated with clinical parameters of genital inflammation and various treatment modalities using anti-retroviral agents. In addition, levels of HIV-1 proviral-containing cells in the peripheral blood will be assessed, utilizing in situ PCR, and correlated with HIV-l RNA and DNA levels in genital secretions. The cellular tropism of HIV- l strains and the presence of defective viral genomes, in genital secretions of select cohorts of patients, will be assessed via cloning and sequencing techniques. The hypotheses, that macrophage-tropic strains correlate with increased sexual transmission while partially defective viruses lead to aberrant viral RNA patterns and possibly decreased transmissibility, will be tested. To further assess correlation of molecular parameters, described above, with clinical infectivity of HIV-l-seropositive individuals, couples both discordant and concordant for HIV-1 infection will be evaluated. In these ways, it is suggested that a more precise understanding of the mechanisms involved in controlling the sexual transmissibility of HIV-1 will be gained.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI033810-01A2
Application #
2068861
Study Section
AIDS and Related Research Study Section 1 (ARRA)
Project Start
1994-02-01
Project End
1998-01-31
Budget Start
1994-02-01
Budget End
1995-01-31
Support Year
1
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Thomas Jefferson University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
061197161
City
Philadelphia
State
PA
Country
United States
Zip Code
19107
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Shaheen, F; Sison, A V; McIntosh, L et al. (1999) Analysis of HIV-1 in the cervicovaginal secretions and blood of pregnant and nonpregnant women. J Hum Virol 2:154-66
Zhang, H; Dornadula, G; Beumont, M et al. (1998) Human immunodeficiency virus type 1 in the semen of men receiving highly active antiretroviral therapy. N Engl J Med 339:1803-9
Dornadula, G; Zhang, H; Bagasra, O et al. (1997) Natural endogenous reverse transcription of simian immunodeficiency virus. Virology 227:260-7
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Zhang, H; Dornadula, G; Alur, P et al. (1996) Amphipathic domains in the C terminus of the transmembrane protein (gp41) permeabilize HIV-1 virions: a molecular mechanism underlying natural endogenous reverse transcription. Proc Natl Acad Sci U S A 93:12519-24
Zhang, H; Dornadula, G; Wu, Y et al. (1996) Kinetic analysis of intravirion reverse transcription in the blood plasma of human immunodeficiency virus type 1-infected individuals: direct assessment of resistance to reverse transcriptase inhibitors in vivo. J Virol 70:628-34
Zhang, H; Duan, L X; Dornadula, G et al. (1995) Increasing transduction efficiency of recombinant murine retrovirus vectors by initiation of endogenous reverse transcription: potential utility for genetic therapies. J Virol 69:3929-32
Pomerantz, R J; Trono, D (1995) Genetic therapies for HIV infections: promise for the future. AIDS 9:985-93

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