Dendritic cells play a crucial role in inducing antigen-specific immunity to pathogens. During host-parasite interaction, host immune response to the parasite molecules is considered essential for recognizing novel antigens for control strategies. Therefore, in the present study, chicken dendritic cells (DCs) (ChDCs), derived from spleens were used to evaluate their capacity to proliferate and differentiate autologous T lymphocytes in response to actin-depolymerizing factor from Eimeria tenella (EtADF). Immunoblot analysis showed that recombinant EtADF protein (rEtADF) was able to interact with rat anti-rEtADF antibodies. The immunofluorescence test confirmed rEtADF binding on ChDCs surface. Flow cytometric analysis revealed that phenotypes for MHCII, CD1.1, CD11c, CD80, and CD86 were increased in ChDCs after rEtADF treatment. qRT-PCR results indicated that ChDCs triggered TLR signaling in response to rEtADF, and suppressed Wnt signaling. Transcript levels of CD83, CCL5, and CCR7 in ChDCs were improved following rEtADF treatment. In addition, rEtADF promoted DC-directed T cell proliferation and differentiation of naïve T cells into CD3⁺/CD4⁺ T cells in DC/T cell co-incubation system. Cytokine analysis of rEtADF-pulsed ChDCs showed increased levels of IL-12 and IFN-γ, while IL-10 and TGF-β remained unchanged. Moreover, rEtADF-treated ChDCs enhanced production of IFN-γ when incubated with T cells, and IL-4 secretion remained unchanged. Our findings indicted that rEtADF could facilitate the polarization of Th1 immune cells by triggering both host DCs and T cells. Our findings provide useful insights into future work aimed at anticoccidial vaccine strategies.

树突状细胞在诱导针对病原体的抗原特异性免疫中起关键作用。在宿主-寄生虫相互作用期间，宿主对寄生虫分子的免疫应答被认为是识别新抗原用于控制策略的必要条件。因此，在本研究中，使用源自脾脏的鸡树突状细胞（DC）（ChDC）来评估其响应艾美球虫中肌动蛋白解聚因子而增殖和分化自体T淋巴细胞的能力（EtADF）。免疫印迹分析表明重组EtADF蛋白（rEtADF）能够与大鼠抗rEtADF抗体相互作用。免疫荧光测试证实了rEtADF在ChDCs表面的结合。流式细胞仪分析显示，rEtADF处理后ChDC中MHCII，CD1.1，CD11c，CD80和CD86的表型增加。qRT-PCR结果表明，ChDCs响应rEtADF触发了TLR信号传导，并抑制了Wnt信号传导。经过rEtADF处理后，ChDC中CD83，CCL5和CCR7的转录水平得到了改善。此外，rEtADF促进DC定向T细胞增殖，并促进幼稚T细胞分化为CD3 ⁺ / CD4 ⁺DC / T细胞共孵育系统中的T细胞。对经rEtADF刺激的ChDC的细胞因子分析显示，IL-12和IFN-γ的水平升高，而IL-10和TGF-β保持不变。此外，rEtADF处理的ChDC在与T细胞孵育时可增强IFN-γ的产生，并且IL-4分泌保持不变。我们的发现表明，rEtADF可以通过触发宿主DC和T细胞来促进Th1免疫细胞的极化。我们的发现为今后针对抗球虫疫苗策略的工作提供了有用的见识。