Cellular control of nuclear import of HIV

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Date
2001-08-22T00:00:00Z
Authors
Rastogi, Ruchi
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Abstract
To further define the relationship between T cell activation and the nuclear import of HIV pre-integration complex (PIC), this thesis examines the role of topoisomerase II (topoII), and enzyme induced and involved in a major remodeling of nucleus during T cell activation. The negatively selected highly purified resting CD4+ human T cells, free of monocytes, were infected with HIVIIIB and stimulated with a combination of anti-CD3 and anti-CD28 monoclonal antibodies (mAbs) in the presence or absence of different topoII inhibitors (VM-26, mAMSA, novobiocin, ICRF 187) that act through different mechanisms and block the activities of both topoII_ and topoIIB isozymes. Specific mRNA expression was monitored by RT-PCR, IL-2 production by ELISA and the expression of IL-2R_ by flow cytometry. Fully reverse-transcribed viral genome was monitored by PCR amplification and the nuclear migration of PIC was monitored by semi-quantitative PCR assays of 2-LTR circles. The entry of cells into S-phase was monitored by 3H-thymidine incorporation. (Abstract shortened by UMI.)
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