Inmunización con hormona liberadora de gonodatropinas / Immunization with gonodatropins-releasing hormone

El uso de la inmunización contra la hormana liberadora de gonadotropinas (GnRH), es un método empleado con fines contraceptivos. Se estudió el efecto de una vacuna de GnHR unida al toxoide diftérico sobre el aparato reproductor de la rata blanca adulta y los niveles plasmáticos de testosterona (T). Se encontró que al tercer mes de haber sido inmunizados los animales con GnHR se produjo una significativa pérdida de peso de los testículos, vesículas seminales y próstata. El peso corporal no se afectó. Al estudio histológico se encontró que en los testículos existía una marcada afectación en la espermatogénesis, llegando en algunos animales a haber solamente células de Sertoli. Se concluye que la combinación empleada de GnHR con el toxoide diftérico, así como el esquema de vacunación empleado afectan profundamente al aparato reproductor de la rata. Desconocemos si el proceso deletéreo inducido por la vacunación es reversible o no

Monoclonal anti-gonadotropin releasing hormone (GnRH) antibodies reacting to sequence of GnRH preferentially recognize to native hormone.

Monoclonal anti-GnRH antibodies (MoAbs) P862, P778, P813 and P764 reacted optimally to native GnRH and poorly to GnRH(OH) of sequence 4-6, 7-10, 4-10 and 1-10. The heptapeptide 4-10 showed maximum reactivity amongst the four peptides tested for immunoreactivity. Sepharose 4B-GnRH(OH)4-10 (H-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-OH) column was therefore used to purify the fraction of MoAb reacting to this sequence. The affinity purified MoAbs (A-MoAbs) were further characterized for their binding to the different sequences and affinity with native GnRH. The binding, cross-reactivity and affinity characteristics of A-MoAbs were comparable with those of MoAbs. Immunoreactivity of A-MoAbs was also observed to be partly regained when GnRH(OH)1-10 was coupled to Lys, Lys-MDP or H-Ala-Ala-Thr-Lys-Pro-Arg-OH. These observations clearly demonstrate that MoAbs were neither contaminated nor were sequence specific but were directed against the conformation of the molecule.

Use of blocking ELISA additivity test and antibody-antibody competition technique for analysis of recognition ability of monoclonal anti-gonadotropin releasing hormone antibodies.

Monoclonal anti-GnRH antibodies reacting to the heptapeptide 4-10 (H-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-OH) were isolated by affinity chromatography using Sepharose 4B-heptapeptide (4-10) column. The ELISA additivity test and antibody-antibody competition techniques were used to study whether the affinity purified MoAb (A-MoAb) fraction recognize the sequence or the conformation of the native hormone. All four A-MoAbs, P862, P778, P764 and P813, were able to recognize the common epitope and did not allow to bind the conventional anti-GnRH antibodies (CoAbs) indicating that the CoAbs were conformation specific. Similarly in antibody-antibody competition technique, all A-MoAbs were able to compete with CoAbs, indicating that MoAbs were generated against the conformation of GnRH involving the entire molecule.