Bendixsen T, Emery D L, Rothwell T L
CSIRO Division of Animal Health McMaster Laboratory, Glebe, New South Wales, Australia.
Int J Parasitol. 1991 Dec;21(8):883-9. doi: 10.1016/0020-7519(91)90162-z.
The cultivation of bone marrow was used to quantitate the levels of eosinophil differentiation factors (EDF) produced in conditioned medium (CM) by incubation of mesenteric lymph node cells (MLNC) with mitogens or specific antigens from the intestinal nematode parasite, Trichostrongylus colubriformis. In liquid cultures with 20 units ml-1 recombinant murine interleukin-5 (IL-5), bone marrow cells (BMC) from either normal or infected donors contained less than 5% eosinophils and differentiated to greater than 50% eosinophils over 2-3 weeks. Conditioned medium from 3-4 week infected donors produced between 20 and 50% eosinophils when donor MLNC were stimulated with the specific antigen preparation SP3, but macrophages predominated when using CM from MLNC incubated with Concanavalin A (ConA). CM from MLNC of challenged donors incubated with SP3 produced 30-70% eosinophils in BMC assays, with highest levels induced by CM from high responder (HR) donors. Marrow from parasitized or normal donors gave rise to comparable proportions of eosinophils. CM was also produced from LNC of donors given protein or parasite antigens in adjuvant where between 28 and 35% eosinophils were produced in culture. There were no differences between activities attributable to the antigen, but Freund's complete adjuvant induced earlier differentiation of BMC than alum-induced CM. The results confirm that high levels of EDF activity are specifically induced by parasitic infection, and can also be produced by intraperitoneal and subcutaneous inoculation of adjuvanted antigens. Consistent with the greater eosinophilia exhibited by HR guinea pigs to infection with T.colubriformis L3, their MLNC also produced the highest levels of EDF activity.
通过将肠系膜淋巴结细胞（MLNC）与丝裂原或来自肠道线虫寄生虫——蛇形毛圆线虫的特异性抗原一起孵育，利用骨髓培养来定量条件培养基（CM）中产生的嗜酸性粒细胞分化因子（EDF）水平。在含有20单位/毫升重组鼠白细胞介素-5（IL-5）的液体培养物中，来自正常或感染供体的骨髓细胞（BMC）含有不到5%的嗜酸性粒细胞，并在2至3周内分化为超过50%的嗜酸性粒细胞。当用特异性抗原制剂SP3刺激供体MLNC时，来自感染3至4周供体的条件培养基在BMC检测中产生20%至50%的嗜酸性粒细胞，但当使用与伴刀豆球蛋白A（ConA）孵育的MLNC的CM时，巨噬细胞占主导。在BMC检测中，用SP3孵育的受攻击供体的MLNC的CM产生30%至70%的嗜酸性粒细胞，其中高反应者（HR）供体的CM诱导的水平最高。来自寄生或正常供体的骨髓产生相当比例的嗜酸性粒细胞。还从在佐剂中给予蛋白质或寄生虫抗原的供体的LNC产生CM，在培养中产生28%至35%的嗜酸性粒细胞。抗原所致的活性之间没有差异，但弗氏完全佐剂诱导BMC的分化比明矾诱导的CM更早。结果证实，高水平的EDF活性是由寄生虫感染特异性诱导的，也可通过腹腔内和皮下接种佐剂化抗原来产生。与HR豚鼠对蛇形毛圆线虫L3感染表现出的更强嗜酸性粒细胞增多一致，它们的MLNC也产生最高水平的EDF活性。
