Asunto(s)
Productos del Gen nef/metabolismo , Infecciones por VIH/metabolismo , VIH-1/metabolismo , Interferón gamma/biosíntesis , Interleucina-2/biosíntesis , Péptidos/metabolismo , Secuencia de Aminoácidos , Células Cultivadas , Productos del Gen nef/síntesis química , Productos del Gen nef/farmacología , Humanos , Péptidos y Proteínas de Señalización Intercelular , Interferón gamma/antagonistas & inhibidores , Interleucina-2/antagonistas & inhibidores , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Datos de Secuencia Molecular , Péptidos/síntesis química , Péptidos/farmacología , ARN Mensajero , Proteínas Oncogénicas de Retroviridae/síntesis química , Proteínas Oncogénicas de Retroviridae/farmacología , Transducción de Señal , Proteínas del Envoltorio Viral/síntesis química , Proteínas del Envoltorio Viral/farmacología , Productos del Gen nef del Virus de la Inmunodeficiencia HumanaRESUMEN
Monoclonal antibodies against a synthetic peptide (aa 138-152) from HIV-1 Nef protein were produced and characterized. Three hybridoma lines producing monoclonal antibodies (MAbs) against the synthetic peptide were generated by fusion between P3-X63 Ag8.653 myeloma cells and BALB/c splenocytes from mice immunized with the synthetic peptide coupled to keyhole limpet hemocyanin (KLH). The hybridomas were screened and selected by ELISA with the peptide coupled to bovine serum albumin (BSA) immobilized to the polystyrene surface and specificity for the peptide was confirmed by competitive ELISA with the peptide free in solution. The reactions of the MAbs with a 5-aa motif (WCYKL) included in the sequence were examined with synthetic peptides and two of the MAbs reacted with the motif. The recognitions of recombinant full-length Nef protein were also tested. One MAb reacted with the protein in both ELISA and dot blot, and one only in dot blot, whereas the last MAb did not recognize the recombinant full-length Nef protein.