Recognition by anti-idiotypic anti-thyrotropin-releasing hormone (TRH) antibody of rat TRH receptors.
We asked whether anti-idiotypic antibodies raised against anti-TRH antibody could bind to TRH receptors in the rat anterior pituitary and brain. Six rabbits were immunized with IgG from a rabbit anti-TRH antiserum. One anti-idiotypic antibody caused strong, dose-dependent inhibition in anti-TRH antibody-binding to [125]I-TRH. This inhibition was not observed after treatment with goat anti-rabbit IgG antibody. The anti-idiotypic anti-TRH antibody significantly immunoprecipitated digitonin-solubilized pituitary TRH receptors. When eluates of digitonin-solubilized membranes which were adsorbed by either an anti-idiotypic anti-TRH IgG-, normal rabbit IgG-linked affinity column or control column were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and visualized by silver stain, only the former eluate showed two bands under nonreducing conditions; one corresponded to a molecular weight marker of 200K, the other to 100K. Western blotting analysis with an anti-idiotypic anti-TRH antiserum showed a single band of molecular weight 56K under reducing conditions. The present study indicates that one can make anti-idiotypic antibodies that specifically recognize TRH receptors.
Satoh
T
T
First Department of Internal Medicine, Gunma University, School of Medicine, Maebashi, Japan.
Mori
M
M
Murakami
M
M
Iriuchijima
T
T
Yamada
M
M
Kobayashi
I
I
Kobayashi
S
S
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology* / Q000378:metabolism; D000918:Antibody Specificity; D001667:Binding, Competitive; D015153:Blotting, Western; D001923:Brain Chemistry; D007114:Immunization; D008297:Male; D010903:Pituitary Gland, Anterior / Q000737:chemistry; D011233:Precipitin Tests; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains; D017981:Receptors, Neurotransmitter / Q000276:immunology*; D018025:Receptors, Thyrotropin-Releasing Hormone; D012995:Solubility; D013973:Thyrotropin-Releasing Hormone / Q000276:immunology*
D000888:Antibodies, Anti-Idiotypic; D017981:Receptors, Neurotransmitter; D018025:Receptors, Thyrotropin-Releasing Hormone; D013973:Thyrotropin-Releasing Hormone