Identification and characterization of differentially expressed mRNAs in HIV type 1-infected human T cells.
We used a novel differential display (DD) technique to identify host factors involved in virus replication, pathogenesis, and host response in HIV-1-infected T cells. Thirteen cDNA fragments differentially expressed in HIV-1NL4-3-infected MT-4 cells prior to the occurrence of specific apoptotic cell death were sequenced and identified. Two of seven elevated genes were identical to HIV-1 sequences and the other five were MIP-1alpha, ACTE-III, CD11c, arginase I, and CCR5. The six downregulated genes included prothymosin-a, Jaw-1, proteasome subunit XAPC7, splicing factor 9G8, GA17 protein, and an unknown mRNA. Northern blot and RT-PCR analyses confirmed the altered gene expressions in MT-4 cells as well as in another T cell line, MOLT-4. We also revealed that the amount of MIP-1alpha in culture supernatant of HIV-1-infected cells was increased by more than 15-fold relative to control cells, and the expression of its receptor CCR5 was cooperatively upregulated on the surface of these cells. Furthermore, the upregulation of CD11c after HIV-1 infection was slightly inhibited by blocking the MIP-1alpha-mediated signal transduction. These results indicate that genes altered on HIV-1 infection may be mutually organized and play an important role in HIV-1-induced pathogenesis.
Ryo
A
A
Department of Microbiology, School of Medicine, Tokyo Medical and Dental University, Japan.
Suzuki
Y
Y
Arai
M
M
Kondoh
N
N
Wakatsuki
T
T
Hada
A
A
Shuda
M
M
Tanaka
K
K
Sato
C
C
Yamamoto
M
M
Yamamoto
N
N
D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism / Q000821:virology; D002460:Cell Line; D002478:Cells, Cultured; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D016167:Integrin alphaXbeta2 / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D019402:Macrophage Inflammatory Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D019713:Receptors, CCR5 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000378:metabolism* / Q000821:virology*
D054405:Chemokine CCL3; D054407:Chemokine CCL4; D016167:Integrin alphaXbeta2; D019402:Macrophage Inflammatory Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D019713:Receptors, CCR5