Isolation of human immunodeficiency virus type 2 biological clones from peripheral blood lymphocytes.
Infectious variants of human immunodeficiency virus (HIV) can be isolated from peripheral blood mononuclear cells (PBMC) of infected individuals by propagating the virus in co-cultures with healthy donor PBMC. Standardized culture protocols have been designed specifically for the isolation of HIV type 1 (HIV-1) and proven effective for the isolation of virus from virtually all HIV-1-infected individuals. For the isolation of HIV-2, however, standard HIV-1 culture protocols have been only partially effective. While suitable for the isolation of HIV-2 from PBMC of individuals in advanced stages of disease, they appeared largely inadequate for the isolation of HIV-2 from asymptomatically HIV-2-infected individuals. This chapter describes a variant HIV isolation procedure with culture conditions adapted to the isolation of HIV-2 from PBMC of "healthy" HIV-2-infected individuals, i.e., individuals with high CD4+ T-cell numbers and no detectable viral RNA in plasma. By using a limiting dilution format, several biological clones representing individual HIV variants can be obtained from the PBMC of an infected individual. In addition, the frequency of PBMC infected with HIV-2 variants capable of propagating in vitro can be estimated.
Blaak
Hetty
H
Department of Virology, Erasmus MC, Rotterdam, The Netherlands.
D018414:CD8-Positive T-Lymphocytes / Q000821:virology; D002999:Clone Cells; D018920:Coculture Techniques / Q000379:methods; D015925:Cryopreservation / Q000379:methods; D015498:HIV-2 / Q000302:isolation & purification*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D008214:Lymphocytes / Q000821:virology*; D012367:RNA, Viral / Q000097:blood; D016176:T-Lymphocyte Subsets / Q000821:virology; D019562:Viral Load; D014776:Virus Cultivation
D012367:RNA, Viral