Wunderlin E, Palmer D G
Department of Agriculture, Western Australia, South Perth, Australia.
Vet Immunol Immunopathol. 1995 Nov;49(1-2):169-75. doi: 10.1016/0165-2427(95)05448-f.
A bioassay for the measurement of an ovine eosinophil differentiation factor (EDF) was developed. Recombinant murine interleukin-5 (rmIL-5) and supernatant from concanavalin A stimulated lymphocytes induced eosinophil differentiation and proliferation in ovine bone marrow cell culture, which was determined by an eosinophil peroxidase assay (EPO). The assay was used to measure the production of an ovine eosinophil differentiation factor by lymphocytes following experimental infection of sheep with Trichostrongylus colubriformis. Sixteen sheep were immunised and challenged with T. colubriformis and grouped into responders (n = 10) and non-responders (n = 6) based on the lower 99% confidence limit of the worm burden of the unimmunised control group (n = 10). Total worm burdens of all animals were established 10 days after initial challenge. Peripheral blood lymphocytes at Day 0 (pre-challenge values) and Day 9 (post-challenge values) from all sheep and lymphocytes of mesenteric lymph nodes from eight sheep were stimulated with third stage larval antigen of T. colubriformis. The resulting lymphocyte conditioned medium (LyCM) was tested for EDF by its ability to induce growth and differentiation of eosinophils in liquid bone marrow cell cultures. No significant difference was found between the animal groups. No correlation was found between EDF activity and total worm numbers.
开发了一种用于测量绵羊嗜酸性粒细胞分化因子（EDF）的生物测定法。重组鼠白细胞介素-5（rmIL-5）和伴刀豆球蛋白A刺激的淋巴细胞的上清液可诱导绵羊骨髓细胞培养中的嗜酸性粒细胞分化和增殖，这通过嗜酸性粒细胞过氧化物酶测定法（EPO）来确定。该测定法用于测量绵羊经蛇形毛圆线虫实验感染后淋巴细胞产生的绵羊嗜酸性粒细胞分化因子。16只绵羊用蛇形毛圆线虫进行免疫和攻击，并根据未免疫对照组（n = 10）虫负荷的99%置信下限分组为反应者（n = 10）和无反应者（n = 6）。在初次攻击后10天确定所有动物的总虫负荷。用蛇形毛圆线虫的第三期幼虫抗原刺激所有绵羊在第0天（攻击前值）和第9天（攻击后值）的外周血淋巴细胞以及8只绵羊肠系膜淋巴结的淋巴细胞。通过其在液体骨髓细胞培养物中诱导嗜酸性粒细胞生长和分化的能力，对所得淋巴细胞条件培养基（LyCM）进行EDF检测。动物组之间未发现显著差异。未发现EDF活性与总虫数之间存在相关性。
