Kooyman F N, Van Kooten P J, Huntley J F, MacKellar A, Cornelissen A W, Schallig H D
Department of Parasitology and Tropical Veterinary Medicine, Utrecht University, The Netherlands.
Parasitology. 1997 Apr;114 ( Pt 4):395-406. doi: 10.1017/s0031182096008633.
Part of the C epsilon 3-C epsilon 4 region of the ovine immunoglobulin E (IgE) gene (nucleotides 1111-1575) was amplified by PCR. The recombinant protein (recIgE1-2) was expressed in E. coli and both monoclonal and polyclonal antibodies were produced. These antibodies recognized recIgE1-2 and native IgE on Western blots and in ELISA. The polyclonal serum showed cross-reactivity with other sheep immunoglobulin classes. The monoclonal antibody was specific for ovine IgE and goat IgE. Infection of sheep with the abomasal nematode Haemonchus contortus resulted in elevated IgE levels in serum 2-4 weeks after infection, as measured by sandwich ELISA using the rabbit polyclonal as capture antibody and the monoclonal antibody against ovine IgE as second antibody. A negative correlation between worm counts and total serum IgE levels at the end of the experiment was found in repeatedly infected sheep. Significant increased levels of excretory-secretory antigens specific IgE levels were found after H. contortus infection. In contrast, no significant changes in 3rd-stage larvae (L3) antigen-specific IgE titre in sera could be detected after infection.
通过聚合酶链反应（PCR）扩增了绵羊免疫球蛋白E（IgE）基因的Cε3 - Cε4区域的一部分（核苷酸1111 - 1575）。重组蛋白（recIgE1 - 2）在大肠杆菌中表达，并产生了单克隆抗体和多克隆抗体。这些抗体在蛋白质免疫印迹法（Western blots）和酶联免疫吸附测定（ELISA）中能识别recIgE1 - 2和天然IgE。多克隆血清与其他绵羊免疫球蛋白类别显示出交叉反应性。单克隆抗体对绵羊IgE和山羊IgE具有特异性。用皱胃线虫捻转血矛线虫感染绵羊后，通过夹心ELISA检测发现，感染后2 - 4周血清中IgE水平升高，该夹心ELISA使用兔多克隆抗体作为捕获抗体，抗绵羊IgE的单克隆抗体作为第二抗体。在反复感染的绵羊中，实验结束时发现虫体计数与血清总IgE水平呈负相关。捻转血矛线虫感染后，发现排泄分泌抗原特异性IgE水平显著升高。相比之下，感染后血清中第三期幼虫（L3）抗原特异性IgE滴度未检测到显著变化。
