Employment of antipeptide antisera to distinguish two closely related cytokines induced in human T cells.
AT464 and AT744 are two cytokines encoded by mitogen-induced genes from human T lymphocytes. These proteins belong to a family of structurally related chemotactic proteins associated with inflammation. By expression of their full length cDNAs in COS cells and in baculovirus-infected Sf9 cells, we found that pAT464 and pAT744 cDNAs represent secreted polypeptides of a molecular mass of about 8,000 and 11,000 Da, respectively. Biochemical characterization of these proteins has been persued through polyclonal antisera, which were derived to synthetic peptides. Using these sera for Western blotting analyses the recombinant AT464 and AT744 proteins could be detected as secreted products from transfected COS cells and from baculovirus-infected Sf9 cells. Similarly the native proteins could be detected in the supernatants of activated human peripheral blood T cells. The recombinant and the T cell-secreted AT464 and AT744 proteins appear to be identical as judged by their mobility by SDS-PAGE and by their reactivity with the rabbit polyclonal antisera.
Zipfel
P F
PF
Bernhard Nocht Institute for Tropical Medicine, Hamburg, FRG.
Wharton
K A
KA
Villalobos
J
J
Irving
S G
SG
Kelly
K
K
Siebenlist
U
U
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D016207:Cytokines / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D006801:Humans; D007106:Immune Sera; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection
C076908:AT 464; C076909:AT 744; D016207:Cytokines; D007106:Immune Sera; D010455:Peptides; D011994:Recombinant Proteins; D004247:DNA