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Updated: Jul 14, 2025
Nucleofection and In Vivo Propagation of Chicken Eimeria Parasites
Published on: February 14, 2020
Inhibitory effect of Eimeria maxima IFN-γ inhibitory molecules on the immune function of T cell subsets in chickens
Xianglin Pu1, Yangdong Pan1, Quanjia Xiang1
1Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China.
Abstract:
It has been reported that infection of chicken coccidian could inhibit the production of Th1 cytokine IFN-γ, thereby evading clearance by the host immune system. The present study aimed to have a further investigation into the effects of Eimeria maxima IFN-γ inhibitory molecules (EmHPSP-2 and EmHPSP-3) on the immune function of chicken peripheral blood mononuclear cells (PBMC) and various T cell subsets. First, separated PBMC or sorted T cell subsets were used for incubation with recombinant proteins of EmHPSP-2 (rEmHPSP-2) and EmHPSP-3 (rEmHPSP-3). Subsequently, the effects of rEmHPSP-2 and rEmHPSP-3 on proliferative capacity, nitric oxide (NO) release and mRNA levels of cytokines of the above cells were detected. The sorting purity of CD8+, CD4+ CD25-, CD4+, and CD4+ CD25+ T cells was 93.01, 88.88, 87.04, and 81.26%, respectively. The NO release of PBMC was significantly inhibited by rEmHPSP-2 and rEmHPSP-3. The proliferation of PBMC and CD4+ T cells was significantly inhibited by rEmHPSP-2 and rEmHPSP-3, whereas CD8+, CD4+ CD25-, and CD4+ CD25+ T cells was significantly promoted by the 2 proteins. The 2 proteins significantly downregulated interferon-gamma (IFN-γ) mRNA level, upregulated the transcriptional levels of interleukin-10 (IL-10) and transforming growth factor-beta1 (TGF-β1) in PBMC. IFN-γ and IL-2 transcriptional levels were markedly inhibited in CD8+ T cells. IFN-γ transcriptional level was significantly inhibited, but IL-4 was promoted by rEmHPSP-2 and rEmHPSP-3 in CD4+ CD25- T cells. Meanwhile, the inhibitory effects of rEmHPSP-2 and rEmHPSP-3 on the transcriptional levels of IFN-γ and IL-2 were more obvious in CD4+ T cells containing CD25+ cells compared with the CD25+ cells depletion group. It was found that IL-10, TGF-β1, and cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) mRNA levels were significantly upregulated upon stimulation of chicken CD4+ CD25+ T cells by proteins. This study is not only of great significance to clarify the immune evasion mechanism of chicken coccidia, but also provides candidate antigen molecules for development of a novel vaccine against chicken coccidiosis.
Insights
Eimeria maxima proteins EmHPSP-2 and EmHPSP-3 inhibit chicken immune responses, downregulating IFN-γ and upregulating IL-10/TGF-β1. These molecules offer potential for a novel coccidiosis vaccine.
Area of Science:
- Immunology
- Parasitology
- Veterinary Medicine
Background:
- Chicken coccidiosis, caused by Eimeria species, can evade host immunity by inhibiting Th1 cytokine production, such as interferon-gamma (IFN-γ).
- Eimeria maxima secretes IFN-γ inhibitory molecules, including EmHPSP-2 and EmHPSP-3, which are implicated in parasite immune evasion.
Purpose of the Study:
- To investigate the effects of Eimeria maxima IFN-γ inhibitory molecules (EmHPSP-2 and EmHPSP-3) on chicken peripheral blood mononuclear cells (PBMC) and T cell subsets.
- To elucidate the immunomodulatory mechanisms employed by these molecules during Eimeria infection.
Main Methods:
- Peripheral blood mononuclear cells (PBMC) and sorted T cell subsets (CD8+, CD4+ CD25-, CD4+, CD4+ CD25+) were incubated with recombinant EmHPSP-2 (rEmHPSP-2) and rEmHPSP-3.
- Assessed the impact of these proteins on cell proliferation, nitric oxide (NO) release, and cytokine mRNA expression (IFN-γ, IL-2, IL-4, IL-10, TGF-β1, CTLA-4).
Main Results:
- rEmHPSP-2 and rEmHPSP-3 significantly inhibited NO release and proliferation in PBMC and CD4+ T cells, while promoting proliferation in CD8+, CD4+ CD25-, and CD4+ CD25+ T cells.
- These proteins downregulated IFN-γ mRNA levels and upregulated IL-10 and TGF-β1 mRNA in PBMC. In CD8+ T cells, IFN-γ and IL-2 transcription were inhibited. In CD4+ CD25- T cells, IFN-γ was inhibited while IL-4 was promoted.
- Upregulation of IL-10, TGF-β1, and CTLA-4 mRNA was observed in chicken CD4+ CD25+ T cells upon stimulation with rEmHPSP-2 and rEmHPSP-3.
Conclusions:
- EmHPSP-2 and EmHPSP-3 play a significant role in the immune evasion mechanisms of chicken coccidia.
- These molecules modulate T cell responses and cytokine production, offering potential as candidate antigens for developing a novel vaccine against chicken coccidiosis.
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