TY - JOUR
T1 - A transcription inhibitor, Actinomycin D, enhances HIV-1 replication through an interleukin-6-dependent pathway
AU - Imamichi, Tomozumi
AU - Conrads, Thomas P.
AU - Zhou, Ming
AU - Liu, Yuxin
AU - Adelsberger, Joseph W.
AU - Veenstra, Timothy D.
AU - Lane, H. Clifford
PY - 2005/12
Y1 - 2005/12
N2 - 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)-β. 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 TNFβ in Sup200 by 85%. Northern blot analysis showed that ActD treatment increased IL-6 transcripts; however, no change was seen in TNFβ transcripts. These results suggest that ActD induces replication of HIV-1 through modulation of cytokine production.
AB - 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)-β. 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 TNFβ in Sup200 by 85%. Northern blot analysis showed that ActD treatment increased IL-6 transcripts; however, no change was seen in TNFβ transcripts. These results suggest that ActD induces replication of HIV-1 through modulation of cytokine production.
KW - Conditioned medium
KW - Cytokines
KW - HIV-1 activation
KW - Transcription inhibitor
UR - http://www.scopus.com/inward/record.url?scp=27944505920&partnerID=8YFLogxK
U2 - 10.1097/01.qai.0000179466.25700.2f
DO - 10.1097/01.qai.0000179466.25700.2f
M3 - Article
C2 - 16280692
AN - SCOPUS:27944505920
SN - 1525-4135
VL - 40
SP - 388
EP - 397
JO - Journal of Acquired Immune Deficiency Syndromes
JF - Journal of Acquired Immune Deficiency Syndromes
IS - 4
ER -