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巨型艾美耳球虫Th1类细胞因子刺激性分子EmARM-β对鸡PBMC和T细胞亚群免疫功能的影响

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南京农业大学学报 2024年第1期47卷 9-17页

作者：蒲响林潘仰栋相权珈孙晓婷陆明敏严若峰徐立新李祥瑞宋小凯南京农业大学动物医学院江苏南京210095

[目的]本文旨在探讨巨型艾美耳球虫Th1类细胞因子刺激性分子Armadillo/β-catenin样重复序列蛋白(EmARM-β)对鸡免疫细胞功能的影响。[方法]利用密度梯度离心法和免疫磁珠分选法分别分离出鸡外周血单个核细胞(peripheral blood mononucl... 详细信息

[目的]本文旨在探讨巨型艾美耳球虫Th1类细胞因子刺激性分子Armadillo/β-catenin样重复序列蛋白(EmARM-β)对鸡免疫细胞功能的影响。[方法]利用密度梯度离心法和免疫磁珠分选法分别分离出鸡外周血单个核细胞(peripheral blood mononuclear cell,PBMC)、CD8^(+)T细胞和CD4^(+)CD25^(-)T细胞,之后将EmARM-β重组蛋白(rEmARM-β)分别与上述细胞进行体外共孵育6 h,利用CCK-8试剂和qPCR法分别检测并分析其对上述细胞增殖能力及相关细胞因子分泌水平的影响。[结果]经免疫磁珠分选后,流式细胞术检测鸡CD8^(+)T细胞和CD4^(+)CD25^(-)T细胞的纯度分别为93.01%和88.88%。与对照组相比,rEmARM-β显著促进鸡PBMC的增殖能力,显著上调Th1类细胞因子(ifn-γ和il-2)和促炎性细胞因子(il-1β、il-6和il-17a)mRNA水平;显著促进CD8^(+)T细胞的增殖能力,显著上调其ifn-γ、il-2和tnf-α mRNA水平,显著上调穿孔素、fasl(Fas配体)和fas mRNA水平;能显著促进CD4^(+)CD25^(-)T细胞的增殖能力,显著上调其ifn-γ和il-2 mRNA水平,下调il-4和il-10 mRNA水平。[结论]EmARM-β可以有效促进鸡PBMC和T细胞亚群(CD8^(+)T细胞和CD4^(+)CD25^(-)T细胞)的增殖能力,提升其Th1类细胞因子和促炎性细胞因子的分泌水平,提示EmARM-β具有研发鸡球虫病新型疫苗候选抗原的潜力。

关键词：巨型艾美耳球虫 Th1类细胞因子刺激性分子 EmARM-β T细胞亚群免疫功能

巨型艾美耳球虫感染对鸡Toll样受体的影响

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畜牧与兽医 2024年第2期56卷 76-81页

作者：潘仰栋刁胤轩蒲响林陆明敏徐立新严若峰李祥瑞宋小凯南京农业大学动物医学院江苏南京210095

为了探究巨型艾美耳球虫(Eimeria maxima)感染对宿主局部免疫组织盲肠扁桃体和系统免疫组织(脾脏和外周血)中Toll样受体(Toll like receptor,TLR)的影响,用巨型艾美耳球虫感染雏鸡,感染后第0、3、7和11天,采集鸡盲肠扁桃体、外周血和脾... 详细信息

为了探究巨型艾美耳球虫(Eimeria maxima)感染对宿主局部免疫组织盲肠扁桃体和系统免疫组织(脾脏和外周血)中Toll样受体(Toll like receptor,TLR)的影响,用巨型艾美耳球虫感染雏鸡,感染后第0、3、7和11天,采集鸡盲肠扁桃体、外周血和脾脏淋巴细胞,用实时荧光定量PCR检测上述组织中8种鸡TLR的mRNA水平变化。结果显示:TLR4和TLR21 mRNA水平在3种组织中无显著变化;TLR2 mRNA水平在感染后第3天开始在外周血和脾脏中显著下降(P<0.05),在盲肠扁桃体中第3和7天无显著变化,在第11天显著上升(P<0.01)。TLR1、TLR3、TLR5、TLR7和TLR15 mRNA水平在3种组织中显著上升(P<0.05),但应答时间有所差异,具体为TLR1和TLR3 mRNA水平在盲肠扁桃体中在第3天开始上升,而在外周血和脾脏中第7天开始上升;TLR5和TLR15 mRNA水平在外周血和脾脏中第3天开始上升,而在盲肠扁桃体中第7天开始上升。结果表明:巨型艾美耳球虫感染上调宿主3种组织中的TLR1、TLR3、TLR5、TLR7和TLR15 mRNA水平,下调外周血和脾脏中的TLR2 mRNA水平;盲肠扁桃体TLR1和TLR3 mRNA水平上升早于外周血和脾脏,而其TLR5和TLR15 mRNA水平上升晚于外周血和脾脏。本研究为揭示鸡TLR在抗球虫感染中的作用提供依据,也为揭示鸡球虫先天性免疫机制提供参考。

关键词：巨型艾美耳球虫 Toll样受体先天性免疫鸡球虫病

长角血蜱Ykt6蛋白抗东方泰勒虫感染的生物学效应

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中国兽医科学 2024年第3期54卷 386-392页

作者：罗金赵帅阳车丽妍严若锋宋小凯任巧云关贵全殷宏刘光远李祥瑞南京农业大学动物医学院、教育部动物健康与食品安全国际合作联合实验室江苏南京210095 中国农业科学院、兰州兽医研究所、动物疫病防控全国重点实验室、甘肃省动物寄生虫病重点实验室、农业部草食动物疫病重点开放实验室甘肃兰州730046

为了解Ykt6基因在蜱体中影响东方泰勒虫感染的生物学效果,通过长角血蜱实验室培养株和野外株,评估Ykt6蛋白对东方泰勒虫侵染长角血蜱过程中蜱的存活率、繁殖率以及对东方泰勒虫感染率等指标的生物学效果。结果显示,Ykt6基因参与了东方... 详细信息

为了解Ykt6基因在蜱体中影响东方泰勒虫感染的生物学效果,通过长角血蜱实验室培养株和野外株,评估Ykt6蛋白对东方泰勒虫侵染长角血蜱过程中蜱的存活率、繁殖率以及对东方泰勒虫感染率等指标的生物学效果。结果显示,Ykt6基因参与了东方泰勒虫与长角血蜱之间的宿主-寄生虫相互作用。Ykt6基因的表达水平与东方泰勒虫在长角血蜱体内的存活和繁殖能力密切相关,当该基因表达水平降低时,东方泰勒虫的存活率和繁殖能力显著下降。进一步的研究发现,Ykt6基因的变异可能影响东方泰勒虫与长角血蜱之间的相互作用。一些野外长角血蜱个体具有Ykt6基因的突变,导致东方泰勒虫在长角血蜱体内的存活和繁殖能力显著下降。这一发现提示,Ykt6基因的变异可能是东方泰勒虫入侵长角血蜱的一个关键因素。总之,Ykt6基因对东方泰勒虫入侵长角血蜱具有重要影响。通过研究Ykt6基因的功能和变异,可以深入了解东方泰勒虫与长角血蜱之间的相互作用,为控制东方泰勒虫入侵提供新的思路和方法。这将有助于保护人畜健康和生态环境的平衡。

关键词：长角血蜱东方泰勒虫 Ykt6蛋白感染生物学效果

Chimeric DNA vaccines encoding Eimeria acervulina macrophage migration inhibitory factor(***)induce partial protection against experimental Eimeria infection

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Chimeric DNA vaccines encoding Eimeria acervulina macrophage...

中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者：宋小凯 Ruirui Zhang Lixin Xu ruofeng yan 李祥瑞南京农业大学动物医学院

Chimeric DNA vaccines co-expressing Eimeria acervulina macrophage migration inhibitory factor(***) and chicken IL-2(IL-2) or interferon-γ(IFN-γ) were constructed and their efficacies against *** were *** open reading frame(ORF) of *** was cloned from *** merozoites and subcloned into the eukaryotic expression vector pVAX1 with chicken cytokine gene IFN-γ or IL-2 to construct the DNA vaccines ***-IFN-γ,***-IL-2 and *** in vivo transfection of the target genes was detected by use of reverse transcription polymerase chain reaction(RT-PCR) and Western *** were carried out by vaccinating chickens twice with a dose rate of 100 μg *** days post second immunization,all chickens except the unchallenged control group were challenged orally with l×10

关键词： E.acervulina orthologue of macrophage migration inhibitory factor DNA vaccine genetic adjuvants recombinant protein

Transmembrane protein 147(TMEM147):another partner protein of Haemonchus contortus galectin on the goat PBMC

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Transmembrane protein 147(TMEM147):another partner protein o...

中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会

作者： yan Li Chen Yuan ruofeng yan Xiaokai Song Lixin Xu Xiangrui Li College of Veterinary Medicine Nanjing Agricultural University

Recombinant galectins of male and female Haemonchus contortus(r Hco-gal-m/f) have been recognized as significant regulators of the functions of goat peripheral blood mononuclear cells(PBMC). In previous research, transmembrane protein 63A(TMEM63A) was identified as a partner protein in the regulation associated with H. contortus infection. However, in the identification of binding partners for galectins of male and female H. contortus(Hco-gal-m/f) by yeast two-hybrid(YTH) screening, it was found that the transmembrane protein 147(TMEM147) could also bind to Hco-gal-m/f. In this study, the functions of TMEM147 in the regulations of H. contortus galectin on the goat PBMC were investigated. To identify Hco-gal-m/f-interacting proteins, a yeast two-hybrid system to detect interactions was used. Co-immunoprecipitation and immunoblotting were used to validate the interaction between recombinant galectins of male H. contortus(r Hco-gal-m) and candidate binding protein. The localization of TMEM147 in PBMC was explored by immunofluorescence in confocal imaging studies. Flow cytometry was used to determine the distribution of TMEM147 in T cells, B cells and monocytes in PBMC. The modulatory effects of rH co-gal-m and TMEM147 on cell proliferation, phagocytosis, nitric oxide production, migration, apoptosis and cytokine mRNA transcription were observed by co-incubation of r Hco-gal-m and knockdown of the tmem147 gene. In this research, it was demonstrated that TMEM147 could bind to r Hco-gal-m/f. Immunofluorescence assays showed that TMEM147 was localized to the cell membrane and within the cell membrane in goat PBMC. Flow cytometric analysis revealed that TMEM147 was expressed in all B cells and monocytes in goat PBMC. However, 3.8 % of T cells did not express this protein. Knockdown of the tmem147 gene using RNA interference(RNAi) showed that the interaction of galectin with TMEM147 mainly mediated cell proliferation, cell apoptosis, transcription of interleukin-10(IL-10) an

关键词： Galectin Goat PBMC Haemonchus contortus Member receptor TMEM147

Nanoparticles Entrapped ADP-Ribosylation Factor 1 of Haemonchus contortus Enhances the Immune Responses in ICR Mice

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Nanoparticles Entrapped ADP-Ribosylation Factor 1 of Haemonc...

2021中国畜牧兽医学会兽医寄生虫学分会第16次学术研讨会

作者： Muhammad Waqqas Xu Lixin Song Xiaokai yan ruofeng Li Xiangrui College of Veterinary Medicine Nanjing Agricultural University

ADP-ribosylation factor 1(HcARF1) is one of the Haemonchus contortus(***)excretory/secretory proteins involved in modulating the immune response of goat peripheral blood mononuclear cells(PBMC).Here,we evaluated the immunogenic potential of recombinant HcARF1(rHcARF1) in Institute of Cancer Research(ICR) ***,rHcARF1 was entrapped in poly(D,L-lactide-co-glycolide)(PLGA) and chitosan(CS) nanoparticles(NP) and injected into mice as a ***-six ICR mice were assigned randomly into seven groups,with eight animals in each group,and they were vaccinated *** the end of the experiment(14 th day),the blood and the spleen were collected from euthanized mice to detect lymphocyte proliferation,cytokine analysis,and the production of antigen-specific *** electron microscope was used to determine the size,morphology,and zeta potential of *** cytometry was performed,which presented the increase percentages of CD4+T cells(CD3 e+CD4+),CD8+T cells(CD3 e+CD8+) and dendritic cells(CD1 1 c+CD83+,CD1 1 c+CD86+) in mice vaccinated with rHcARF1+PLGA *** analysis show raised humoral(Immunoglobulin(Ig)G1,IgG2 a,IgM) and cell-mediated immune response(Interleukin(IL)-4,IL-12,and IL-17,and Interferon(IFN)-gamma) induced by rHcARF1+PLGA *** groups that were treated with the antigen-loaded NP yield higher lymphocyte proliferation than the control *** on these results,we could propose that the rHcARF1 encapsulated in NP could stimulate a strong immune response in mice rather than administering alone against the infection of ***.

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Unveiling the immunomodulatory properties of Haemonchus contortus adhesion regulating molecule 1 interacting with goat T cells

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Unveiling the immunomodulatory properties of Haemonchus cont...

2021中国畜牧兽医学会兽医寄生虫学分会第16次学术研讨会

作者： Lu Mingmin Xu Lixin Song Xiaokai yan ruofeng Li Xiangrui College of Veterinary Medicine Nanjing Agricultural University

Background:Gastrointestinal nematodes could release excretory-secretory(ES)proteins into the host environment to ensure their *** ES proteins act as immunomodulators to suppress or subvert the host immune response via the impairment of immune cell functions,especially in chronic *** our preliminary study,Haemonchus contortus adhesion-regulating molecule 1(HcADRM1) was identified from *** ES proteins(HcESPs) that interacted with host T cells via liquid chromatography-tandem mass spectrometry ***,little is known about HcADRM1 as an ES protein which may play a pivotal role at the parasite-host ***:Based on bioinformatics approaches,multiple amino acid sequence alignment was conducted and the evolutionary relationship of HcADRM1 with ADRM1 orthologues was *** RT-qPCR and immunohistochemistry assays,temporal transcriptional and spatial expression profiles of HcADRM1 were *** immunostaining approaches integrated with immunological bioassays,the immunomodulatory potentials of HcADRM1 on goat T cells were ***:We hereby demonstrated that HcADRM1 with immunodiagnostic utility was a mammalian ADRM1 orthologue abundantly expressed at all developmental stages of *** the implications of ADRM1 proteins in cell growth,survival and development,we further investigated the immunomodulatory property of HcADR1 as an individual ES protein acting at the parasitehost *** rHcADRM1 stimuli notably suppressed T cell viability,promoted intrinsic and extrinsic T cell apoptosis,inhibited T cell proliferation and induced cell cycle arrest at G1 ***,rHcADRM1 stimuli exerted critical controls on T cell cytokine secretion profiles,predominantly by restraining the secretions of interleukin(IL)-4,IL-10 and ***:Importantly,HcADRM1 protein may have prophylactic potential for *** vaccine ***,these findings ma

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Molecular cloning of enolase from Trichinella spiralis and the protective immunity in mice

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Molecular cloning of enolase from Trichinella spiralis and t...

第五届全国人畜共患病学术研讨会

作者： Xuliang Zhang Lixin Xu Xiaokai Song Xiangrui Li ruofeng yan College of Veterinary Medicine Nanjing Agricultural University

Trichinella spiralis,the main pathogen of trichinosis,infects a wide range of mammalian hosts and is one of the most widespread parasites *** parasites,glycolysis is the most important way to generate *** studies showed that some enzymes involved in the glycolytic pathway play roles in regulation the host *** this paper,enolase from T.

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A DNA vaccine co-expressing Trichinella spiralis MIF and MCD-1 with murine ubiquitin induces partial protective immunity in mice

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A DNA vaccine co-expressing Trichinella spiralis MIF and MCD...

第五届全国人畜共患病学术研讨会

作者： Tang Fang Xu Lixin yan ruofeng Song Xiaokai Li Xiangrui College of Veterinary Medicine Nanjing Agricultural University

Co-expression of Trichinella spiralis macrophage migration inhibitory factor(TsMIF)with *** cystatin-like domain protein(TsMCD-1)in a DNA vaccine induces a Thl immune response and partial protection against *** *** present study evaluated whether co-expression of mouse ubiquitin(Ub)with TsMIF and TsMCD-1 might improve the immune response against *** *** of BALB/c mice were immunized twice at 2-week

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