Leelayuwat C, Hollingsworth P, Pummer S, Lertmemongkolchai G, Thom G, Mullberg J, Witt C, Kaufman J, Degli-Esposti M A, Cosman D, Dawkins R
Department of Clinical Immunology, Faculty of Associated Medical Sciences, Khon Kaen University, Thailand.
Clin Exp Immunol. 1996 Dec;106(3):568-76. doi: 10.1046/j.1365-2249.1996.d01-862.x.
PERB11 (MIC) is a gene family possessing multiple copies located within the MHC. Structurally, PERB11 is related to the MHC class I, neonatal IgG Fc receptor (FcRn) and Zn-alpha 2-glycoprotein molecules. The MHC class I family is complex in terms of its genomic arrangement, expression and function, and available evidence suggests that the PERB11 family may be similarly complex. We have adopted an approach to study the expression of such complex gene families by immunizing with multiple peptides and by screening the resulting antibodies against a large range of tissues. The amino acid sequences of PERB11.1 and PERB11.2 as well as those of other related molecules were analysed and compared. Peptides were chosen for immunization based upon (i) loop formation within the equivalent known structure of the MHC class I molecules; (ii) immunogenicity by computer analysis; and (iii) evolutionary relationships. Antibodies in serum from immunized rabbits bound to three out of six peptides used for immunization. ELISA and immunoprecipitation demonstrated binding both to the peptides and to the PERB11.2 recombinant protein. By immunofluorescent staining of various tissues of several species, the three antisera generated overlapping profiles of activity. These included reactions with kidney, small and large intestine, oesophagus, testis, ovary and human neutrophils. This is the first description of antibodies induced by the PERB11 peptides. The extreme complexity of these profiles requires further investigation, but may be explained in terms of antibodies against diverse products of the PERB11 gene family and/or related molecules.
PERB11(MIC)是一个位于主要组织相容性复合体(MHC)内具有多个拷贝的基因家族。从结构上看,PERB11与MHC I类、新生儿IgG Fc受体(FcRn)和锌-α2-糖蛋白分子相关。MHC I类家族在基因组排列、表达和功能方面都很复杂,现有证据表明PERB11家族可能同样复杂。我们采用了一种方法来研究此类复杂基因家族的表达,即通过用多种肽进行免疫,并针对大量组织筛选产生的抗体。对PERB11.1和PERB11.2以及其他相关分子的氨基酸序列进行了分析和比较。基于以下几点选择用于免疫的肽:(i)MHC I类分子等效已知结构内的环形成;(ii)通过计算机分析的免疫原性;(iii)进化关系。免疫兔血清中的抗体与用于免疫的六种肽中的三种结合。酶联免疫吸附测定(ELISA)和免疫沉淀显示既与肽结合,也与PERB11.2重组蛋白结合。通过对几种物种的各种组织进行免疫荧光染色,产生的三种抗血清显示出重叠的活性谱。这些包括与肾脏、小肠和大肠、食管、睾丸、卵巢以及人类中性粒细胞的反应。这是首次描述由PERB11肽诱导产生的抗体。这些谱的极端复杂性需要进一步研究,但可能可以用针对PERB11基因家族和/或相关分子的不同产物的抗体来解释。