Neutralization of primary human immunodeficiency virus type 1 isolates: a study of parameters implicated in neutralization in vitro.
Various studies have reported that primary human immunodeficiency viruses seem to be more refractory to neutralization by HIV-positive sera than T cell line-adapted strains. In this study we also show that adaptation of the HIV-1SF-2 strain, produced in PBMCs, to the cell line CEM-SS renders this isolate sensitive to neutralization by almost all the sera tested. Further neutralization studies should thus focus on the development of an assay involving primary isolates in order to detect antibodies having a neutralizing activity in vivo. Neutralization protocols currently use either an antibody end-point dilution assay, which combines a fixed inoculum of virus with serial dilutions of antibody, or an infectivity reduction assay, which uses serial dilutions of virus with a single dilution of antibody. We have developed an assay designed for studying the neutralization of primary isolates that combines these two approaches. Performing the assay on PBMCs allows all primary isolates to be analyzed, not just those multiplying in T cell lines. The neutralizing titer measured on PBMCs for human HIV-positive sera is low, but reproducible and independent of the virus titer in a given experiment. It can be increased about five-fold by changing the temperature and duration of virus-serum interaction (overnight at 4 degrees C instead of 1 hr at 37 degrees C). These results emphasize the need for a relevant neutralization assay involving primary isolates and primary cells for a better understanding of the role of humoral response in HIV infection.
Moog
C
C
INSERUM U74, Institut de Virologie, Strasbourg, France.
Spenlehauer
C
C
Fleury
H
H
Heshmati
F
F
Saragosti
S
S
Letourneur
F
F
Kirn
A
A
Aubertin
A M
AM
D000595:Amino Acid Sequence; D000818:Animals; D015496:CD4-Positive T-Lymphocytes; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015483:HIV Antibodies / Q000097:blood*; D015699:HIV Envelope Protein gp120 / Q000235:genetics; D006679:HIV Seropositivity / Q000276:immunology; D015497:HIV-1; D006801:Humans; D007106:Immune Sera; D007963:Leukocytes, Mononuclear; D008969:Molecular Sequence Data; D009500:Neutralization Tests / Q000379:methods*; D002679:Pan troglodytes; D010446:Peptide Fragments / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014779:Virus Replication
D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; C059762:HIV envelope protein gp120 (305-321); D007106:Immune Sera; D010446:Peptide Fragments; D012367:RNA, Viral