Serologic analysis of a murine chemokine, TCA3.
The TCA3 gene encodes a proinflammatory cytokine related to IL-8 and MCP-1 (JE). TCA3 mRNA is produced by T cells in response to stimulation with Ag or Con A. Glycosylated and nonglycosylated forms of rTCA3 produced by transfected Chinese hamster ovary cells were purified. rTCA3 was used to immunize hamsters and produce mAb. Four mAb were characterized; each was specific for TCA3, failing to react with the closely related molecule MCP-1 (JE). Western blotting also demonstrated that all four mAb specifically recognize rTCA3, native TCA3 produced by activated mouse T cells, and the nonglycosylated TCA3 core protein. A sensitive and specific capture ELISA for TCA3 quantitation was developed, allowing measurement of TCA3 levels in the supernatants from activated murine splenocytes and T cell clones. In addition, two mAb demonstrated dose-dependent neutralization of TCA3 inflammatory activity. Thus, the results directly demonstrate that TCA3 is expressed in an activation-specific manner by mouse T cells. The data also provide characterization of mAb reagents specific for this new cytokine.
Luo
Y
Y
Department of Pathology, Harvard Medical School, Boston, MA 02115.
Laning
J
J
Dorf
M E
ME
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D054402:Chemokine CCL1; D019742:Chemokines, CC; D006224:Cricetinae; D003412:Cricetulus; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D000939:Epitopes; D005260:Female; D006031:Glycosylation; D051379:Mice; D008815:Mice, Inbred Strains; D054401:Receptors, CCR8; D013601:T-Lymphocytes / Q000378:metabolism
D000911:Antibodies, Monoclonal; C065971:CCL1 protein, human; C489319:Ccr8 protein, mouse; D054402:Chemokine CCL1; D019742:Chemokines, CC; D016207:Cytokines; D000939:Epitopes; D054401:Receptors, CCR8