Isolation, characterization, and transmission of human T-lymphotropic virus types I and II in culture.
Highly sensitive coculture methods were developed both for isolation of human T-lymphotropic virus types I and II (HTLV-1 and HTLV-II) from infected individuals and for productive infection of lymphoid cells. Mitogen-activated peripheral blood mononuclear cells (PBMC) from 13 HTLV-I- and 20 HTLV-II-positive specimens were cocultured with an equal number of mitogen-activated PBMC from HTLV-seronegative individuals, and culture supernatants were tested for the presence of soluble p24gag antigens at weekly intervals for 4 weeks. Eleven of 13 (85%) HTLV-I and 14 of 20 (70%) HTLV-II cultures were positive for p24 antigens. None of the 17 HTLV-seroindeterminate or six HTLV-seronegative specimens were positive for the presence of p24 antigen. The isolation rates for HTLV-I and HTLV-II by an alternative whole-blood lysis procedure were comparable to those obtained by standard PBMC cultures. Furthermore, cocultivation of PHA-stimulated PBMC from healthy donors with lethally irradiated HTLV-I- and HTLV-II-infected cell lines (SP and Mo-T, respectively) resulted in productive viral infection, as reflected by the appearance of p24gag antigens concomitant with specific genomic amplification of HTLV proviral DNA after 3 weeks of cocultivation. Thus, the cocultivation technique provides a highly sensitive and specific procedure both for HTLV isolation and for infection of target cells.
Kitamura
K
K
Retrovirus Diseases Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333.
Rudolph
D L
DL
Goldsmith
C
C
Folks
T M
TM
Lal
R B
RB
D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers; D015683:Gene Products, gag / Q000032:analysis; D015490:HTLV-I Infections / Q000382:microbiology*; D015491:HTLV-II Infections / Q000382:microbiology*; D015368:Human T-lymphotropic virus 1 / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification*; D015367:Human T-lymphotropic virus 2 / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007962:Leukocytes / Q000382:microbiology; D008214:Lymphocytes / Q000382:microbiology; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000382:microbiology / Q000648:ultrastructure; D015682:Retroviridae Proteins, Oncogenic / Q000032:analysis; D014776:Virus Cultivation / Q000379:methods*
D017931:DNA Primers; D015683:Gene Products, gag; D015682:Retroviridae Proteins, Oncogenic; C047061:internal protein p24, Human T-lymphotropic virus 2; C060954:p24 protein, Human T-lymphotropic virus 1