The major population of PHA-stimulated PBMC infected by R5 or X4 HIV variants after a single cycle of infection is predominantly composed of CD45RO+CD4+ T lymphocytes.
PHA-stimulated peripheral blood mononuclear cells (PBMCs) are widely used for investigating replication and neutralization of HIV primary isolates in vitro. The objective of this study was to identify the T lymphocyte subset(s) that are found infected after one replication cycle by either R5- or X4-HIV-1 variants in PHA-stimulated PBMCs from healthy donors. Infected T lymphocytes were detected by intracellular p24 staining and characterized by cell surface immunophenotyping using flow cytometry. The predominant lymphocyte subset expressing p24 after 24 h of infection with either R5 or X4 HIV-1 strains was found to exhibit mainly the memory CD45RO phenotype, a greater percentage of CD62L(+)CD45RO(+) central memory T lymphocytes was infected with X4 HIV strains. Although some CD45RA(+) lymphocytes were also infected, these cells co-expressed CD45RO(+). The proportion of lymphocytes expressing CD4 and CD4/CD45RO decreased by 20% after 24 h of infection. A 2-fold decrease of CD4(+)CD8(+) T lymphocytes could also be recorded, even though this subset accounted for less than 5% of total lymphocytes in control cultures. Moreover, CD4(+)CD8(+) T cells further decreased by 90% after 4 days of infection, a time at which they scored p24(+). Therefore, our results indicate that the in vitro infection system of PHA-stimulated PBMC utilized in neutralization assays provides an appropriate model for the study of infected CD45RO(+) lymphocytes but not CD45RA(+) lymphocytes.
Holl
V
V
Institut de Virologie, EA 3770, Université Louis Pasteur, Strasbourg, France.
Schmidt
S
S
Aubertin
A-M
AM
Moog
C
C
D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology; D014644:Genetic Variation; D015658:HIV Infections / Q000276:immunology*; D015497:HIV-1 / Q000145:classification / Q000235:genetics* / Q000472:pathogenicity*; D015234:HLA-A Antigens / Q000097:blood; D015235:HLA-B Antigens / Q000097:blood; D015236:HLA-C Antigens / Q000097:blood; D006801:Humans; D017493:Leukocyte Common Antigens / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000821:virology*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology; D012016:Reference Values
D015234:HLA-A Antigens; D015235:HLA-B Antigens; D015236:HLA-C Antigens; D016693:Receptors, Antigen, T-Cell, alpha-beta; D017493:Leukocyte Common Antigens