Hendrikx W M
Vakgroep Infectieziekten en Immunologie, afdeling Helminthologie en Entomologie, Faculteit der Diergeneeskunde, Rijks Universiteit Utrecht.
Tijdschr Diergeneeskd. 1990 Dec 1;115(23):1092-101.
In the first part of the study, various tissues of L3 larvae and adult Haemonchus contortus were examined by indirect fluorescence to investigate their antigenicity. L3 larvae were studied in immunoblots as well. The following tissues contained antigens: the sheath and cuticle of L3 larvae; and the cuticle, muscles, epithelium and glandular tissue of the oesophagus and sheath of the spicules of adult nematodes. On the cuticle of exsheathed L3 larvae, we detected by light-microscope slightly raised patches: places where the cuticle differs from the surrounding cuticular morphology. We suggest that the patches are comparable to those found on exsheathed L3 larvae of the horse. Whether the patches produce excretory or secretory antigens or both is unknown, however. Somatic soluble extracts of L3 and adult worms were found to be complex mixtures of proteins by polyacrylamide gel electrophoresis. Results in immunoblots of L3 larvae suggested that the region between the 42 kDa protein and the 18 kDa protein may contain antigens specific for H. contortus. We injected rabbits with the 42-25 kDa fraction of the L3 larvae. The rabbit sera were probed in immunoblots with the total somatic soluble L3 extract for the presence of antibodies. Antibodies reacted with antigens in the 42-25 kDa fraction, however, cross-reactions occurred with antigens in the L3 fractions that were not injected into the rabbits. We found that antibodies against phosphorylcholine were induced and suggest that these cause some of the cross-reactions. In the second part of the study sheep were infected orally with H. contortus and their lymphocyte responses towards L3 antigen and mitogen were measured in a lymphocyte stimulation test. Lymphocyte responses to L3 antigen are expressed in a stimulation index. Two groups of worm-free raised sheep were inoculated (sheep older than 1 year and lambs that were 4 to 6 months old) as well as a group of hyperimmunized sheep. Some sheep were inoculated only once; others were inoculated and then challenged. Lymphocytes of sheep that were hyperimmunized replicated rapidly in the test resulting in significantly higher stimulation indices than those recorded for the lymphocytes of all other sheep. After inoculation the enhanced response further increased. We explain these lymphocyte responses as a result from the immunological memory induced during the hyperimmunization. Lymphocytes of sheep over 1 year old only respond in the test when sheep are infected.(ABSTRACT TRUNCATED AT 400 WORDS)
在研究的第一部分,通过间接荧光法检测了捻转血矛线虫L3幼虫和成虫的各种组织,以研究其抗原性。还对L3幼虫进行了免疫印迹研究。以下组织含有抗原:L3幼虫的鞘和角质层;成虫线虫的角质层、肌肉、上皮以及食管和交合刺鞘的腺组织。在脱鞘L3幼虫的角质层上,我们通过光学显微镜检测到略微隆起的斑块:即角质层与周围角质层形态不同的部位。我们认为这些斑块与在马的脱鞘L3幼虫上发现的斑块类似。然而,这些斑块是否产生排泄性或分泌性抗原或两者都产生尚不清楚。通过聚丙烯酰胺凝胶电泳发现,L3幼虫和成虫的虫体可溶性提取物是蛋白质的复杂混合物。L3幼虫免疫印迹的结果表明,42 kDa蛋白和18 kDa蛋白之间的区域可能含有捻转血矛线虫特异性抗原。我们用L3幼虫的42 - 25 kDa组分注射兔子。用L3幼虫的总虫体可溶性提取物在免疫印迹中检测兔血清中抗体的存在。抗体与42 - 25 kDa组分中的抗原发生反应,然而,也与未注射到兔子体内的L3组分中的抗原发生交叉反应。我们发现诱导了针对磷酸胆碱的抗体,并认为这些抗体导致了一些交叉反应。在研究的第二部分,绵羊经口感染捻转血矛线虫,并在淋巴细胞刺激试验中测量它们对L3抗原和有丝分裂原的淋巴细胞反应。淋巴细胞对L3抗原的反应以刺激指数表示。两组无蠕虫饲养的绵羊(1岁以上的绵羊和4至6个月大的羔羊)以及一组超免疫绵羊进行了接种。一些绵羊只接种一次;其他绵羊接种后再进行攻击。超免疫绵羊的淋巴细胞在试验中迅速增殖,导致刺激指数显著高于所有其他绵羊淋巴细胞的记录值。接种后增强的反应进一步增加。我们将这些淋巴细胞反应解释为超免疫过程中诱导的免疫记忆的结果。只有当1岁以上的绵羊被感染时,其淋巴细胞才会在试验中产生反应。(摘要截断于400字)