Establishment of HTLV-I-infected cell lines from peripheral blood mononuclear cells of Brazilian patients.
To investigate epidemiological and pathogenetic features of HTLV-I infection, a cohort of carriers has been followed at the USP Teaching Hospital since 1991. This study describes the establishment of cell lines from peripheral blood mononuclear cells (PBMC) of infected subjects. Ex vivo PBMC were cultured with those from a seronegative donor and morphologic evidence of cell transformation was obtained after 90 days with detection of multinucleated cells exhibiting cerebriform nuclei. Integration of HTLV-I proviral DNA and expression of viral antigens was demonstrated in culture by PCR and immunofluorescence. Cell lines were maintained for 240 days, gradually weaned from exogenous IL-2. Immunophenotyping of cell lines on flow cytometry yielded evidence of cell activation. Establishment of HTLV-I-infected cell lines from ex vivo PBMC is feasible and may be useful for studies on lymphocyte phenotypic changes and on mechanisms of HTLV-induced cell proliferation. Moreover they may be used with diagnostic purposes in immunofluorescence tests.
Pannuti
Carolina V
CV
Virology Laboratory (LIM-52), Department of Infectious and Parasitic Diseases, School of Medicine, University of São Paulo, Brazil. segurado@usp.br
Jorge
Maria Lúcia S G
ML
Biasutti
Cláudia
C
Kallás
Esper G
EG
Segurado
Aluisio A C
AA
D000328:Adult; D000956:Antigens, Viral / Q000032:analysis; D001938:Brazil; D002461:Cell Line, Transformed / Q000276:immunology / Q000821:virology*; D002472:Cell Transformation, Viral / Q000276:immunology; D015331:Cohort Studies; D005240:Feasibility Studies; D005260:Female; D019085:Fluorescent Antibody Technique, Direct; D015490:HTLV-I Infections / Q000276:immunology; D015368:Human T-lymphotropic virus 1 / Q000276:immunology / Q000502:physiology; D006801:Humans; D016130:Immunophenotyping; D007963:Leukocytes, Mononuclear / Q000821:virology*; D008875:Middle Aged; D016133:Polymerase Chain Reaction
D000956:Antigens, Viral