Monoclonal antibodies and chemiluminescence immunoassay for detection of the surface protein of human T-cell lymphotropic virus.
Monoclonal antibodies (MAbs) raised against human T-cell lymphotropic virus type I (HTLV-I) recognized five distinct antigenic domains of viral env gene-encoded proteins. By using recombinant env proteins and synthetic peptides as mapping antigens, it was determined that the most immunogenic region represented a central portion of the retroviral surface protein (domain 2; amino acids 165 to 191). However, only a single MAb was able to react strongly with native viral proteins. This antibody (clone 6C2) was directed to an epitope within domain 4 (amino acids 210 to 306) of the retroviral env gene and reacted with envelope proteins in both HTLV-I and HTLV-II, as determined by immunoprecipitation, solid-phase binding, and immunoblotting. No reactivity against envelope components of other human retroviruses, including human immunodeficiency virus types 1 and 2, was present. Flow cytometry data demonstrated that MAb 6C2 reacted with cell lines chronically infected with HTLV-I or HTLV-II and also with surface antigens expressed on fresh adult T-cell leukemia cells, following up-regulation with interleukin-2. By a chemiluminescence immunoassay procedure, picogram amounts of viral surface protein could be detected in the unconcentrated supernatants of HTLV-infected cell lines and in diagnostic cultures. Levels of env and gag proteins released by cells into culture supernatants were not directly related to percent expression of cell surface viral-coat proteins. Further, the molar ratio of p19 to gp46 in conditioned media varied from strain to strain, possibly reflecting differences in viral assembly or packaging mechanisms. MAb 6C2 will be of value in characterizing the biochemical and immunological behavior of retroviral env gene proteins and in studying the interaction of HTLV-I and HTLV-II with their receptors.
Papsidero
L D
LD
Cellular Products, Inc., Buffalo, New York.
Dittmer
R P
RP
Vaickus
L
L
Poiesz
B J
BJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D015485:Deltaretrovirus Antigens / Q000032:analysis* / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D015686:Gene Products, env / Q000032:analysis* / Q000235:genetics / Q000276:immunology*; D015752:Genes, env; D015486:HTLV-I Antigens / Q000032:analysis / Q000235:genetics; D015487:HTLV-II Antigens / Q000032:analysis / Q000235:genetics; D015368:Human T-lymphotropic virus 1 / Q000235:genetics; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008163:Luminescent Measurements; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D015485:Deltaretrovirus Antigens; D000939:Epitopes; D015686:Gene Products, env; D015486:HTLV-I Antigens; D015487:HTLV-II Antigens; D010455:Peptides