title: Isolation of human immunodeficiency virus type 2 biological clones from peripheral blood lymphocytes.
abstract: 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.
authors.affiliation: Department of Virology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Hetty
authors.initials: H
authors.lastname: Blaak
mesh_terms: 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
chemical_list: D012367:RNA, Viral