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猫和羊免疫球蛋白ε重链cDNA的部分序列以及重组IgE片段特异性抗体在不同物种间的比较结合研究

Partial sequences of feline and caprine immunoglobulin epsilon heavy chain cDNA and comparative binding studies of recombinant IgE fragment-specific antibodies across different species.

作者信息

Griot-Wenk M E, Obexer-Ruff G, Fluri A, Marti E

机构信息

Institute of Animal Breeding, Bremgartenstrasse 109a, University of Bern, 3012, Bern, Switzerland.

出版信息

Vet Immunol Immunopathol. 2000 Jun 30;75(1-2):59-69. doi: 10.1016/s0165-2427(00)00183-5.

Abstract

Parts of the feline and caprine IgE epsilon heavy chain cDNA (third and fourth constant domains, IgEf3/4) were cloned, sequenced, and expressed to raise antibodies (Abs). The DNA and derived protein sequences of the feline recombinant IgEf (rIgEf) shared high homology with the analogous canine parts (81% at the nucleotide and 71% at the protein levels) and the caprine with the ovine ones (95%/84%), respectively. The polyclonal Abs raised in chickens against the feline and caprine rIgEf3/4 were subjected to a comparative binding study utilizing an ELISA including rIgEf and specific Abs to these rIgEf from dog and horse (rIgEf2 and rIgEf3/4) and sheep (rIgEf3/4). All but the ovine-specific rIgEf3/4 Ab were polyclonal, which had been raised in chickens, and bound to most applied rIgEf; the ovine-specific monoclonal mouse Ab recognized only in addition to ovine rIgEf3/4 the closely related caprine rIgEf3/4. Significant, positive correlations were detected between binding reactions of the polyclonal Abs in ELISA and percentage protein sequence homology (p<0.01). Thus, the newly described feline and caprine IgE nucleotide sequences and corresponding Abs represent useful tools for further species-specific and comparative allergy and disease-associated research.

摘要

克隆、测序并表达了猫和羊的IgE ε重链cDNA的部分序列(第三和第四恒定结构域,IgEf3/4),以制备抗体(Abs)。猫重组IgEf(rIgEf)的DNA和推导的蛋白质序列分别与犬的类似部分(核苷酸水平81%,蛋白质水平71%)以及羊的山羊对应部分(95%/84%)具有高度同源性。利用酶联免疫吸附测定(ELISA)对鸡体内产生的针对猫和羊rIgEf3/4的多克隆抗体进行了比较结合研究,该ELISA包括rIgEf以及针对犬和马(rIgEf2和rIgEf3/4)和羊(rIgEf3/4)的这些rIgEf的特异性抗体。除羊特异性rIgEf3/4抗体外,所有抗体均为在鸡体内产生的多克隆抗体,且能与大多数应用的rIgEf结合;羊特异性单克隆小鼠抗体除了能识别羊rIgEf3/4外,还能识别密切相关的山羊rIgEf3/4。在ELISA中多克隆抗体的结合反应与蛋白质序列同源百分比之间检测到显著的正相关(p<0.01)。因此,新描述的猫和羊IgE核苷酸序列以及相应抗体是进一步进行物种特异性和比较性过敏及疾病相关研究的有用工具。

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