Biological actions of monoclonal antibodies to bovine lutropin receptor.
Monoclonal antibodies (Mabs) were produced against bovine Lutropin receptor (LH-R). Antibodies were detected by an enzyme linked immunosorbant assay (ELISA). Hybridomas were subcloned to achieve monoclonality. Ascites were developed in Balb/c mice. Hybridoma supernatants were purified by ammonium sulfate precipitation and chromatography on hydroxylapatite columns. LH-R antibodies showed upto 50% inhibition of 125I-labeled hCG binding to bovine luteal cell membranes and up to 80% inhibition of testosterone production by hCG stimulated mouse Leydig cells. LH-R antibodies were predominantly IgM isotype. Purified antibodies showed a 78-kDa band, in SDS-PAGE, as the heavy chain of the immunoglobulin. LH-R antibodies were localized specifically in the thecal and luteal cells of the rat ovaries as well as in the Leydig cell of mouse testes. Injections of the LH-R antibody caused a constant estrus in normal rats. One month after the cessation of the injections the animals returned to normal estrus cycle and fertility. Pregnant mice injected with LH-R antibodies produced only 3 viable pregnancies and 10 pups, as compared to 8 pregnancies and 45 pups born to normal controls. LH-R antibodies also caused, approximately, a 50% reduction in testosterone production in normal male rats. These observations indicate a high degree of specificity of the Mab to LH-R and their potential in studies on gonadal function.
Pal
R
R
National Institute of Immunology, New Delhi, India.
Talwar
G P
GP
Saxena
A
A
Sam
M G
MG
Rathnam
P
P
Saxena
B B
BB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D002417:Cattle; D002852:Chromatography, Ion Exchange; D003338:Corpus Luteum / Q000378:metabolism; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006825:Hybridomas / Q000276:immunology; D007132:Immunoglobulin Isotypes / Q000032:analysis; D007295:Inositol Phosphates / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010053:Ovary / Q000378:metabolism; D051381:Rats; D011974:Receptors, LH / Q000096:biosynthesis / Q000276:immunology*; D015398:Signal Transduction; D013737:Testis / Q000276:immunology
D000911:Antibodies, Monoclonal; D007132:Immunoglobulin Isotypes; D007295:Inositol Phosphates; D011974:Receptors, LH