Belloc Carlos G, Longhi Silvia A, Peña Clara, Blank Viviana A, Miranda Marta E, Retegui Lilia A
Instituto de Química y Fisicoquímica Biológicas, Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina.
Life Sci. 2007 Jul 26;81(7):553-9. doi: 10.1016/j.lfs.2007.06.013. Epub 2007 Jun 28.
Previous works demonstrated that the monoclonal antibody (MAb) called R7B4 is directed to an epitope shared by receptors for lactogenic and somatogenic hormones as well as interleukins 2 and 6 (IL-2 and IL-6). The MAb inhibited the biological effects of those hormones and cytokines by impairing their binding to receptors. It is known that the receptors for growth hormones (GH), prolactins (PRL), IL-2, and IL-6 pertain to the type I cytokine receptor family, sharing the common motif WSXWS or the homologous F(Y)GEFS. Thus, a set of 34 decapeptides corresponding to diverse receptors containing those sequences were synthesized by the PEPSCAN method and their reactions with MAb R7B4 were measured by ELISA. The MAb significantly recognized 21 peptides, allowing us to establish the consensus sequence HGYWSEWSPE as a portion of the R7B4 epitope. The consensus peptide was synthesized and purified by conventional methods, and its capacity to bind to MAb R7B4 paratope confirmed. Moreover, polyclonal Ab to the peptide elicited in mice were able to inhibit the hGH binding to lactogenic, somatogenic and human specific liver receptors. This fact suggests that the consensus peptide could be used as an immunogen to produce anti-hGH receptor Ab behaving as hormone or cytokine antagonists in certain pathological conditions.
先前的研究表明,名为R7B4的单克隆抗体(MAb)针对的是催乳激素和生长激素以及白细胞介素2和6(IL-2和IL-6)受体共有的一个表位。该单克隆抗体通过损害这些激素和细胞因子与受体的结合来抑制它们的生物学效应。已知生长激素(GH)、催乳素(PRL)、IL-2和IL-6的受体属于I型细胞因子受体家族,共享共同基序WSXWS或同源的F(Y)GEFS。因此,通过PEPSCAN方法合成了一组34个十肽,它们对应于含有这些序列的不同受体,并通过ELISA测量它们与单克隆抗体R7B4的反应。该单克隆抗体能显著识别21种肽,使我们能够确定共有序列HGYWSEWSPE作为R7B4表位的一部分。通过常规方法合成并纯化了共有肽,并证实了其与单克隆抗体R7B4互补位结合的能力。此外,在小鼠体内引发的针对该肽的多克隆抗体能够抑制hGH与催乳激素、生长激素和人类特异性肝脏受体的结合。这一事实表明,共有肽可作为免疫原,用于产生在某些病理条件下表现为激素或细胞因子拮抗剂的抗hGH受体抗体。