Ko C, Smith C K, McDonell M
Synergen, Inc., Boulder, CO 80301.
Mol Biochem Parasitol. 1990 Jun;41(1):53-63. doi: 10.1016/0166-6851(90)90096-5.
Monoclonal antibodies (Mab) were produced against Eimeria tenella merozoites. A single Mab, LPMC-61, was selected because of its ability to bind to merozoites by indirect immunofluorescence assay (IFA) and to inhibit in vitro sporozoite development. Mab LPMC-61 reacts with an approx. 10-12-kDa merozoite polypeptide in reduced SDS-PAGE, but with an approx. 80-kDa protein in non-reduced SDS-PAGE. The monoclonal recognizes similarly sized polypeptides in E. tenella sporozoites, oocysts and schizonts. A partial cDNA (LPMC-61f) encoding the LPMC-61 antigen was identified from an E. tenella sporozoite cDNA library in bacteriophage lambda gt11. In addition to Mab LPMC-61, the recombinant beta-galactosidase/LPMC-61f fusion protein is recognized by hyperimmune rabbit anti-E. tenella sporozoite serum, rabbit anti-E. tenella merozoite serum, and E. tenella-infected and immune chicken sera. DNA sequencing of LPMC-61f cDNA showed that the putative protein has an unusual tandem, non-perfect repeated sequence, with glutamine comprising about 48% of the predicted amino acids. A hydropathicity plot of the predicted amino acid sequence shows a central hydrophilic region, consisting of the repeated sequences, surrounded by hydrophobic regions on both sides. Since the merozoite stage of avian Eimeria has been implicated in the induction of a protective immune response in chickens, LPMC-61 may be an important immunogen for use as a vaccine against E. tenella.
制备了针对柔嫩艾美耳球虫裂殖子的单克隆抗体（Mab）。选择了一种单克隆抗体LPMC - 61，因为它能够通过间接免疫荧光试验（IFA）与裂殖子结合，并抑制体外子孢子发育。单克隆抗体LPMC - 61在还原型SDS - PAGE中与一种约10 - 12 kDa的裂殖子多肽反应，但在非还原型SDS - PAGE中与一种约80 kDa的蛋白质反应。该单克隆抗体在柔嫩艾美耳球虫子孢子、卵囊和裂殖体中识别大小相似的多肽。从噬菌体λgt11中的柔嫩艾美耳球虫子孢子cDNA文库中鉴定出编码LPMC - 61抗原的部分cDNA（LPMC - 61f）。除了单克隆抗体LPMC - 61外，重组β - 半乳糖苷酶/LPMC - 61f融合蛋白还能被超免疫兔抗柔嫩艾美耳球虫子孢子血清、兔抗柔嫩艾美耳球虫裂殖子血清以及柔嫩艾美耳球虫感染和免疫鸡血清识别。LPMC - 61f cDNA的DNA测序表明，推定的蛋白质具有不寻常的串联、非完美重复序列，谷氨酰胺约占预测氨基酸的48%。预测氨基酸序列的亲水性图谱显示，由重复序列组成的中央亲水区域两侧被疏水区域包围。由于禽艾美耳球虫的裂殖子阶段与鸡体内保护性免疫反应的诱导有关，LPMC - 61可能是一种重要的免疫原，可用于制备抗柔嫩艾美耳球虫疫苗。
