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The Individual Contribution of Transcription Factors Mobilized Following T-cell Receptor (TCR) or Mitogenic Activation in the Reactivation of HIV from Latency

Hokello, Joseph Francis
http://rave.ohiolink.edu/etdc/view?acc_num=case1267065851

Abstract Details

2010, Doctor of Philosophy, Case Western Reserve University, Molecular Virology.
Human Immunodeficiency Virus latency remains the single greatest obstacle to successful HIV eradication. The precise molecular mechanisms utilized by HIV to emerge from latency are poorly understood although it is believed that NF-kappaB is the major transcription factor utilized. While NF-kappaB may be the major factor involved in HIV transcription, additional transcription factors including NF-AT and AP-1 have also been implicated in various studies to regulate HIV transcription albeit the individual contribution of each of these factors in HIV LTR-mediated transcription are unknown. In this dissertation, we determined the individual contribution NF-kappaB, NF-AT and AP-1 transcription factors in the reactivation of HIV from latency following TCR or mitogenic activation using Jurkat T-cell clones harboring single latent HIV proviruses. In order to tease out the contribution of each factor in HIV transcription, we utilized MAPK inhibitor PD98059 and Cyclosporin A to block AP-1 and NF-AT, respectively. Following TCR co-activation, NF-kappaB, NF-AT and AP-1 exhibit unique nuclear induction levels and kinetics. Each factor becomes available in the nucleus when it is required to modulate transcription. For instance, NF-kappaB exhibits two distinct nuclear entry cycles during which latent HIV proviruses are transcribed while AP-1 becomes available during latter time points where AP-1 c-Fos synergizes with NF-kappaB to modulate HIV transcription elongation which is inhibited by PD98059. Certain control cellular genes are transcribed during the first or second NF-kappaB induction cycles. Following TCR co-activation, NF-AT inhibits LTR activation through competitive binding with NF-kappaB. However, AP-1 c-Fos synergizes with NF-AT to activate HIV transcription in the absence of NF-kappaB following selective TCR activation. Mitogenic PHA activation of Jurkat T-cell clones induces NF-kappaB p65 and AP-1 as the major transcription factors during which AP-1 c-Fos synergizes with p65 homodimers in the absence of p50 to activate HIV transcription. Induction of NF-kappaB, NF-AT and AP-1 through co-stimulation of Jurkat T-cell clones using anti-CD3 antibody and TNF-alpha (replaces anti-CD28 antibody) combination to mimic classic TCR activation using anti-CD3 and anti-CD28 antibodies does not permit AP-1 c-Fos to synergize with NF-kappaB or NF-AT to modulate HIV transcription. We are utilizing these approaches to study HIV latency in primary CD4+ T-lymphocytes.
Jonathan Karn, Ph.D. (Advisor)
David McDonald, Ph.D. (Committee Chair)
Carlos Subauste, M.D. (Committee Member)
Eric Arts, Ph.D. (Committee Member)
Koh Fujinaga, Ph.D. (Committee Member)
270 p.
Biomedical Research; Molecular Biology; Virology
T-cell Receptor Signaling; Reactivation of HIV Latency; HIV transcription regulation

Recommended Citations

Citations

  • Hokello, J. F. (2010). The Individual Contribution of Transcription Factors Mobilized Following T-cell Receptor (TCR) or Mitogenic Activation in the Reactivation of HIV from Latency [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1267065851

    APA Style (7th edition)

  • Hokello, Joseph. The Individual Contribution of Transcription Factors Mobilized Following T-cell Receptor (TCR) or Mitogenic Activation in the Reactivation of HIV from Latency. 2010. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1267065851.

    MLA Style (8th edition)

  • Hokello, Joseph. "The Individual Contribution of Transcription Factors Mobilized Following T-cell Receptor (TCR) or Mitogenic Activation in the Reactivation of HIV from Latency." Doctoral dissertation, Case Western Reserve University, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=case1267065851

    Chicago Manual of Style (17th edition)

case1267065851
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© 2010, all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.