Boesman-Finkelstein M, Walton N E, Finkelstein R A
Department of Microbiology, School of Medicine, University of Missouri-Columbia 65212.
Infect Immun. 1989 Apr;57(4):1227-34. doi: 10.1128/iai.57.4.1227-1234.1989.
The newly parturient cow secretes large quantities of immunoglobulin G1, a relatively protease- and heat-resistant immunoglobulin, in its colostrum and milk. This study establishes the feasibility of producing protective colostral immunoglobulins by immunizing pregnant cows with cholera toxin (CT), a CT-related enterotoxin from Escherichia coli, and Vibrio cholerae outer membranes (OMs). The OMs were prepared from bacteria grown under iron-replete or iron-deficient (to simulate the in vivo environment) conditions. Immunoglobulins were purified from the colostrum of newly parturient control and immunized cows. The bovine anti-CT and anti-H-LT (CT-related heat-labile enterotoxin produced by diarrheogenic E. coli strains of human origin) antibodies were quantitated by enzyme-linked immunosorbent assays and by neutralization of toxin activity in both Y-1 adrenal cell and infant rabbit assays. The bovine anti-OM antibodies from both high-iron-grown and low-iron-grown vibrios were assessed by bacterial agglutination and by Western blot (immunoblot) analysis of polyacrylamide gel electrophoresis of high-iron-grown and low-iron-grown OMs. To test their protective effect, immunoglobulin preparations were administered orally in infant feeding formula to 6-day-old rabbits. Anti-CT and anti-OM immunoglobulins elicited statistically significant protection against diarrhea in infant rabbits challenged intraintestinally with virulent cholera vibrios.
刚分娩的母牛会在其初乳和乳汁中分泌大量免疫球蛋白G1，这是一种相对耐蛋白酶和耐热的免疫球蛋白。本研究确定了通过用霍乱毒素（CT）、一种来自大肠杆菌的与CT相关的肠毒素以及霍乱弧菌外膜（OMs）对怀孕母牛进行免疫来生产具有保护作用的初乳免疫球蛋白的可行性。OMs是从在铁充足或铁缺乏（模拟体内环境）条件下生长的细菌中制备的。从刚分娩的对照母牛和免疫母牛的初乳中纯化免疫球蛋白。通过酶联免疫吸附测定以及在Y-1肾上腺细胞和幼兔试验中对毒素活性的中和作用，对牛抗CT和抗H-LT（由人类源腹泻性大肠杆菌菌株产生的与CT相关的不耐热肠毒素）抗体进行定量。通过细菌凝集以及对高铁生长和低铁生长的OMs进行聚丙烯酰胺凝胶电泳的蛋白质印迹（免疫印迹）分析，评估来自高铁生长和低铁生长弧菌的牛抗OM抗体。为了测试它们的保护作用，将免疫球蛋白制剂添加到婴儿喂养配方中口服给6日龄的兔子。抗CT和抗OM免疫球蛋白对经肠道用强毒霍乱弧菌攻击的幼兔腹泻产生了具有统计学意义的保护作用。
