A macaque adherent cell line that expresses human CD4 is susceptible to SIV: utility for assessing neutralizing antibody.
A macaque CD4 + adherent cell line was generated by stable expression of the human CD4 gene in a rhesus macaque mammary tumor cell line, CMMT. The resulting cell line CMMT/CD4 expressed surface CD4 and was sensitive to infection by a wide range of isolates of simian immunodeficiency virus (SIV) of different subgroups, but was not susceptible to infection with HIV-1. The CMMT/CD4 cell line was used to develop a microassay for measurement of neutralizing antibody in plasma of SIV-infected or immunized animals. Single infected cells could be detected in a monolayer of CMMT/CD4 by immunoperoxidase and a 90% reduction in the number of positive cells was used as a measure of neutralizing activity of two-fold plasma dilutions. This assay had comparable sensitivity to methods based upon detecting a reduction in reverse transcriptase activity of SIV, reduction of viral antigen, or inhibition of cytopathic effect.
Goldstein
S
S
Immunodeficiency Viruses Section, LID, NIAID, NIH Twinbrook Facility, Rockville, MD 20852, USA.
Hague
B
B
Montefiori
D
D
Hirsch
V M
VM
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D015704:CD4 Antigens / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D002448:Cell Adhesion; D003588:Cytopathogenic Effect, Viral; D015497:HIV-1 / Q000472:pathogenicity; D006801:Humans; D008253:Macaca mulatta; D009500:Neutralization Tests; D015302:Simian Immunodeficiency Virus / Q000276:immunology / Q000472:pathogenicity*; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000276:immunology / Q000821:virology*
D000914:Antibodies, Viral; D015704:CD4 Antigens