重组捻转血矛线虫半乳糖结合凝集素免疫山羊皱胃细胞因子表达定位

Localization of Cytokines in Abomasum of Goats Immunized with Recombinant Galectins of Haemonchus Contortus

【目的】检测重组雌、雄捻转血矛线虫半乳糖结合凝集素(rHco-GAL-f/m)免疫山羊皱胃转录细胞因子。【方法】用rHco-GAL-f/m免疫山羊,通过原位杂交法对阴性对照组(A)、攻虫对照组(B)、免疫攻虫组(C)和免疫不攻虫组(D)山羊皱胃中白介素-1β(IL-1β)、白介素-2(IL-2)、白介素-4(IL-4)、白介素-6(IL-6)、干扰素-γ(IFN-γ)和肿瘤坏死因子-α(TNF-α)mRNA转录进行定位,并对上述细胞因子阳性反应细胞数量进行分析。【结果】A组山羊皱胃组织中没有检测到细胞因子,其它3组仅在皱胃黏膜下层中检测到了上述细胞因子。B、C和D组间IL-4和IL-6阳性反应细胞数差异显著(P<0.05),其中C组明显高于其它两组;B、C和D组间IL-1?、IL-2和TNF-α阳性反应细胞数差异显著(P<0.05),其中D组明显高于其它两组;IFN-γ阳性反应细胞数在B组和C组之间差异不显著(P>0.05),D组与B组和C组之间差异显著(P<0.05),且明显高于其它两组。【结论】重组捻转血矛线虫半乳糖结合凝集素能够诱导山羊皱胃局部免疫应答。

[Objective] Cytokines in abomasum of goats immunized with rHco-GAL-flm were detected in this study. [Method] In situ hybridization was used to localize IL-4, IL-6, IL-1B, IL-2,IFN-γ and TNF-α mRNAs in abomasum of the negative control group （A）, challenged control group （B）, vaccinated-challenged group （C） and vaccinated-unchallenged group （D） goats immunized with rHco-GAL-f/m, and the numbers of cytokine mRNA＋ cells in each group were also analyzed. [Result] No cytokines were found in the abomasum of group A goats and IL-4, IL-6, IL-1β, IL-2, IFN-7 and TNF-α were detected in the submucosa layer of abomasum of groups B, C and D goats. The difference of IL-4 and IL-6 mRNA＋ cell numbers in groups B, C and D was significant （P〈0.05） and group C was higher than that of groups B and D. The difference of IL-1B, IL-2 and, TNF-α mRNA＋ cell numbers was also significant （P 〈0.05） among groups B, C and D, and group D was the highest. IFN-γ mRNA＋ cells were not significantly different between groups B and C （P〉0.05）, but IFN-γ mRNA＋cells in group D was significantly different and was higher compared to groups B and C （P 〈0.05）. [ Conclusion ] The findings indicated that the recombinant galectins of H. contortus are capable of inducing local immune responses in abomasum of goats