Down-modulation of TCR/CD3 surface complexes after HIV-1 infection is associated with differential expression of the viral regulatory genes.
We have investigated the mechanism(s) involved in progressive abrogation of CD3-gamma gene expression after HIV-1 or HIV-2 infection. A comparison of intracellular virus expression with T cell receptor surface density, revealed both high and low levels of viral p24 antigen in the TCR/CD3(hi), TCR/CD3(lo), and TCR/CD3(-) cells. Furthermore, in non-productively infected cells expressing the multiply spliced, virally encoded tat, rev, and nef regulatory gene transcripts, the same progressive loss of surface TCR/CD3 complexes was observed. We treated HIV-1-infected cells with antisense (AS) phosphorothioate oligodeoxynucleotides (P-OdN) targeted to the viral regulatory genes. All of the HIV-1 sequence-specific AS-P-OdN's inhibited intracellular p24 antigen expression in a time- and dose-dependent manner; although, blocking p24 expression alone was not sufficient to modulate TCR/CD3 surface density. Only Tat-AS and Nef-AS were able to delay TCR/CD3 down-modulation on receptor-positive cells or drive receptor up-regulation on receptor-negative cells. In contrast, Rev-AS accelerated TCR/CD3 loss on receptor-positive cells. RT-PCR revealed that Tat-AS and Nef-AS reduce the level of tat, nef, and rev transcripts, while Rev-AS increases the level of tat and nef transcripts in infected cells. Thus, when intracellular conditions favor expression of tat and/or nef in the absence of rev, CD3-gamma gene transcripts and TCR/CD3 surface density are down-modulated.
Willard-Gallo
K E
KE
Laboratory of Experimental Hematology, University of Brussels (ULB), Brussels, Belgium. kwillard@ulb.ac.be
Furtado
M
M
Burny
A
A
Wolinsky
S M
SM
D017252:CD3 Complex / Q000378:metabolism*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D005434:Flow Cytometry; D015967:Gene Expression Regulation, Viral; D015693:Gene Products, nef / Q000235:genetics / Q000378:metabolism*; D015694:Gene Products, rev / Q000235:genetics / Q000378:metabolism*; D015696:Gene Products, tat / Q000235:genetics / Q000378:metabolism*; D016655:HIV Core Protein p24 / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000378:metabolism / Q000821:virology; D013997:Time Factors; D054311:nef Gene Products, Human Immunodeficiency Virus; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus
D017252:CD3 Complex; D015693:Gene Products, nef; D015694:Gene Products, rev; D015696:Gene Products, tat; D016655:HIV Core Protein p24; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D012367:RNA, Viral; D011948:Receptors, Antigen, T-Cell; D054311:nef Gene Products, Human Immunodeficiency Virus; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus