Biological activities of binding site specific monoclonal antibodies to prolactin receptors of rabbit mammary gland.
The biological activity of three monoclonal antibodies (mAbs) against the rabbit mammary prolactin (PRL) receptor (M110, A82, and A917) were investigated using explants of rabbit mammary gland. The three mAbs which were all able to inhibit the binding of 125I-ovine prolactin to its receptor had different biological activities. Two mAbs (M110 and A82) were able to prevent the stimulating effect of PRL on casein synthesis when the molar ratio between the mAb and PRL was 100. At a lower concentration, M110 moved the PRL dose-response curve to the right by a factor of 2.4. This mAb was also effective in vivo, reducing milk production in a lactating rabbit, in a similar fashion to the prolactin lowering drug, CB-154. One mAb (A917) was able to mimic the action of PRL on both casein and DNA ([3H]thymidine incorporation) synthesis, whereas the other two mAbs were without any stimulatory effect. For this stimulatory effect to be observed, bivalency of the antibody was essential, since monovalent fragments, which were able to inhibit PRL binding, had no agonistic activity. The ability of the mAbs to induce a down-regulation of receptors was also studied. M110, which was equipotent to PRL in occupation of receptors, induced no down-regulation, while A917, which had full biological activity, induced only a small degree of down-regulation. These studies suggest that the binding domain of the receptor might be relatively complex, since only a part of this domain recognized by the antibody with PRL-like activity was able to induce hormonal action. Alternatively, only those antibodies able to microaggregate the receptors may possess PRL-like activity.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology*; D002364:Caseins / Q000096:biosynthesis; D004247:DNA / Q000096:biosynthesis; D005260:Female; D007457:Iodine Radioisotopes; D008321:Mammary Glands, Animal / Q000032:analysis*; D011388:Prolactin / Q000494:pharmacology; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000032:analysis / Q000276:immunology*; D011981:Receptors, Prolactin
D000911:Antibodies, Monoclonal; D002364:Caseins; D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D011981:Receptors, Prolactin; D011388:Prolactin; D004247:DNA