Pharmaceutical Sciences Faculty Publications
A Transcription Inhibitor, Actinomycin D, Enhances HIV-1 Replication Through an Interleukin-6-Dependent Pathway
Document Type
Article
Publication Date
12-1-2005
Journal Title
Journal of Acquired Immune Deficiency Syndromes
ISSN
1525-4135
Volume
40
Issue
4
First Page
388
Last Page
397
DOI
10.1097/01.qai.0000179466.25700.2f
PubMed ID
16280692
Abstract
We previously demonstrated that Actinomycin D (ActD) enhanced HIV-1 replication in the MT-2 cell, a human T-cell leukemia virus type-1-infected cell line. The MT-2 cell is known to produce multiple cytokines spontaneously. In this study, we investigated the impact of ActD on the cytokine production from MT-2 cells and HIV-1 replication in a latently infected cell line, U1. MT-2 cells were pulse-treated with 0 or 200 nM of ActD, and culture supernatants were collected 3 days after incubation. Supernatants from untreated cells (Sup0) induced HIV-1 replication by 150-fold in U1 cells. Culture supernatants from ActD-treated cells (Sup200) enhanced HIV-1 replication by 1200-fold. A combination of a sequential chromatographic approach and mass spectrometric analysis identified that the HIV-inducing factors in Sup200 were interleukin (IL)-6 and tumor necrosis factor (TNF)-beta. Quantitative analysis revealed that ActD treatment increased the concentration of IL-6 in Sup200 by 600% compared with that in Sup0 but decreased the amount of TNFbeta in Sup200 by 85%. Northern blot analysis showed that ActD treatment increased IL-6 transcripts; however, no change was seen in TNFbeta transcripts. These results suggest that ActD induces replication of HIV-1 through modulation of cytokine production.
Keywords
Blotting, cell line, chromatography, dactinomycin, HIV-1, mass spectrometry, RNA, messenger, transcription, genetic, virus replication
Recommended Citation
Imamichi, Tomozumi; Conrads, Thomas P.; Zhou, Ming; Liu, Yuxin; Adelsberger, Joseph W.; Veenstra, Timothy D.; and Lane, H. Clifford, "A Transcription Inhibitor, Actinomycin D, Enhances HIV-1 Replication Through an Interleukin-6-Dependent Pathway" (2005). Pharmaceutical Sciences Faculty Publications. 414.
https://digitalcommons.cedarville.edu/pharmaceutical_sciences_publications/414