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Protective immunity against acute toxoplasmosis in BALB/c mice induced by a DNA vaccine encoding Toxoplasma gondii 10kDa excretory-secretory antigen(TgESA10)

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Protective immunity against acute toxoplasmosis in BALB/c mi...

第五届全国人畜共患病学术研讨会

作者： Shuai Wang YuJian Wang XinChao Liu ZhengChao Zhang XiaoNi Sun Javaid Ali Gadahi TingQi Liu Ibrahim A.Hassan LiXin Xu ruofeng yan XiaoKai Song Xiangrui Li 南京农业大学动物医学院

Toxoplasma gondii 10kDa excretory-secretory antigen(TgESAlO)is involved in the early stages of host *** aim of this study was to evaluate the immune protective efficacy of a DNA vaccine encoding TgESAlO gene against acute *** infection in *** gene sequence encoding TgESAlO was inserted into the eukaryotic expression vector pVAX1,and the efficacy of

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Actin-depolymerizing factor from Eimeria tenella promotes immunogenic function of chicken dendritic cells

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Actin-depolymerizing factor from Eimeria tenella promotes im...

2021中国畜牧兽医学会兽医寄生虫学分会第二届青年科学家学术论坛

作者： Shakeel Ahmed Lakho Muhammad Haseeb Jianmei Huang Zhang yang Muhammad Waqqas Hasan Muhammad Tahir Aleem Muhammad Ali Memon XiaoKai Song ruofeng yan Lixin Xu 李祥瑞 College of Veterinary Medicine Nanjing Agricultural University

Dendritic cells play a crucial role in inducing antigen-specific immunity to *** host-parasite interaction,host immune response to the parasite molecules is considered essential for recognizing novel antigens for control ***,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 *** immunofluorescence test confirmed rEtADF binding on ChDCs *** *** revealed that phenotypes for MHCII,CD1.1,CD11 c,CD80,and CD86 were increased in ChDCs after rEtADF ***-PCR results indicated that ChDCs triggered TLR signaling in response to rEtADF,and suppressed Wnt *** levels of CD83,CCL5,and CCR7 in ChDCs were improved following rEtADF *** addition,rEtADF promoted DC-directed T cell proliferation and differentiation of naive T cells into CD3/CD4T cells in DC/T cell co-incubation *** analysis of rEtADF-pulsed ChDCs showed increased levels of IL-12 and IFN-γ,while IL-10 and TGF-β remained ***,rEtADF-treated ChDCs enhanced production of IFN-γ when incubated with T cells,and IL-4 secretion remained *** findings indicted that rEtADF could facilitate the polarization of Th1 immune cells by triggering both host DCs and T *** findings provide useful insights into future work aimed at anticoccidial vaccine strategies.

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Poly(D,L-lactide-co-glycolide) delivery system improve the protective efficacy of recombinant antigen TA4 against Eimeria tenella infection

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Poly(D,L-lactide-co-glycolide) delivery system improve the p...

2021中国畜牧兽医学会兽医寄生虫学分会第二届青年科学家学术论坛

作者： Jianmei Huang Siying Chen Zhouyang Zhou Xiaoting Sun Muhammad Haseeb Shakeel Ahmed Lakho yang Zhang Jiabin Liu Muhammad Ali A Shah Xiaokai Song Lixin Xu ruofeng yan 李祥瑞 College of Veterinary Medicine Nanjing Agricultural University Departure of Parasitology and Microbiology PMAS Arid Agriculture University

Eimeria tenella is a protozoan parasite endemic in chickens and is one of the causative agents of avian *** aim of this research was to determine if poly(D,L-lactide-co-glycolide)(PLGA) nanoparticles carrying recombinant TA4 protein of ***(rEtTA4) could improve the level of protective immunity against *** *** TA4 protein was expressed and ***(D,L-lactide-co-glycolide) loaded with rEtTA4(PLGA-rEtTA4) nanoparticles was prepared and was delivered to 2-week-old layer chickens via intramuscular *** injected with PBS and PLGA nanoparticles were served as control *** rEtTA4 and PLGA-rEtTA4 nanoparticles induced changes of serum cytokines,IgY levels,and T lymphocytes subpopulation,and the protective efficacy against *** challenge was *** showed that both rEtTA4 and PLGA-rEtTA4 vaccination groups induced significantly higher levels of specific EtTA4 IgY antibody and IL-17 and higher proportion of CD8 T ***,no significant differences were observed in the proportion of CD4 T lymphocytes compared with the PBS *** immunized with rEtTA4 and PLGA-rEtTA4 prominently increased the BW gains and decreased oocyst output compared with chickens immunized with PBS and PLGA after oral challenge with ***(D,L-lactide-co-glycolide) encapsulated rEtTA4 nanoparticles-immunized chickens significantly induced higher levels of interferon gamma,IL-6,and IL-17 and a little bit higher proportion of CD8+T lymphocytes compared with rEtTA4 subunit vaccine-immunized ***,PLGA encapsulated rEtTA4 nanoparticles appeared to have great potential to enhance the immune response and improved the protective efficacy against *** *** results provided available protective subunit vaccine rEtTA4 and PLGA loaded with rEtTA4 nanoparticles against coccidiosis and suggested that PLGA nanoparticles could be an effective adjuvant to enhance the protective efficacy

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Nanoparticles(PLGA and Chitosan)-Entrapped ADP-Ribosylation Factor 1 of Haemonchus contortus Enhances the Immune Responses in ICR Mice

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Nanoparticles(PLGA and Chitosan)-Entrapped ADP-Ribosylation ...

2021中国畜牧兽医学会兽医寄生虫学分会第二届青年科学家学术论坛

作者： Muhammad Waqqas Hasan Muhammad Haseeb Muhammad Ehsan Javaid Ali Gadahi Muhammad Ali-Ul-Husnain Naqvi Qiang Qiang Wang Xinchao Liu Shakeel Ahmed Lakho ruofeng yan Lixin Xu Xiaokai Song 李祥瑞 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) against *** infection 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(CD11 c+CD83+,CD11 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)-γ)induced by rHcARFl+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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旋毛虫反式-2-烯酰辅酶A还原酶1对大鼠免疫功能的影响

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南京农业大学学报 2020年第1期43卷 116-124页

作者：孙晓轲储稳徐立新宋小凯李祥瑞严若峰南京农业大学动物医学院

[目的]本文旨在分析旋毛虫反式-2-烯酰辅酶A还原酶1(TER1)对大鼠免疫系统功能的影响。[方法]通过RT-PCR方法克隆旋毛虫TER 1基因,构建表达载体,获得重组蛋白(rTER1)并检测其酶活性,用Western blot分析rTER1的反应原性。将大鼠外周血单... 详细信息

[目的]本文旨在分析旋毛虫反式-2-烯酰辅酶A还原酶1(TER1)对大鼠免疫系统功能的影响。[方法]通过RT-PCR方法克隆旋毛虫TER 1基因,构建表达载体,获得重组蛋白(rTER1)并检测其酶活性,用Western blot分析rTER1的反应原性。将大鼠外周血单个核细胞(PBMC)与rTER1共孵育,用免疫荧光试验(IFA)检验rTER1与大鼠PBMC的结合情况,分析rTER1对细胞增殖、迁移、NO分泌、吞噬功能和细胞凋亡的影响。大鼠腹腔注射rTER1后,测定血清中IFN-γ、IL-4、IL-9、IL-17和TGF-β等细胞因子的变化。[结果]体外试验表明,rTER1能够促进大鼠PBMC的增殖与迁移,增强细胞NO分泌和吞噬功能,提高细胞凋亡比例;体内试验显示,rTER1能促进大鼠IFN-γ分泌,抑制IL-17分泌,但对IL-4、IL-9和TGF-β的分泌无显著影响。[结论]旋毛虫反式-2-烯酰辅酶A还原酶1能通过多种途径影响宿主免疫功能。

关键词：旋毛虫反式-2-烯酰辅酶A还原酶1 外周血单个核细胞免疫功能大鼠

Glyceraldehyde-3-phosphate dehydrogenase from Eimeria acervulina modulates the functions of chicken dendritic cells to boost Th1 type immune response and stimulates autologous CD4+T cells differentiation in-vitro

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Glyceraldehyde-3-phosphate dehydrogenase from Eimeria acervu...

2021中国畜牧兽医学会兽医寄生虫学分会第二届青年科学家学术论坛

作者： Shakeel Ahmed Lakho Muhammad Haseeb Jianmei Huang Zhang yang Muhammad Waqqas Hasan Muhammad Tahir Aleem Muhammad Ali-Ul-Husnain Naqvi Muhammad Ali Memon XiaoKai Song ruofeng yan Lixin Xu 李祥瑞 College of Veterinary Medicine Nanjing Agricultural University

Dendritic cells(DCs) play a pivotal role to amplify antigen-specific immune *** that sensitize T cells via antigen-presentation by DCs could enhance the capacity of host immunity to fight *** this study,we tested the immunogenic profiles of chicken DCs towards Glyceraldehyde-3-phosphate dehydrogenase from Eimeria acervulina(EaGAPDH).Immunoblot analysis showed that recombinant EaGAPDH(rEaGAPDH) protein was successfully recognized by rat sera generated against *** and internalisation of rEaGAPDH by chicken splenic-derived DCs(chSPDCs) was confirmed by immunofluorescence *** cytometry revealed that chSPDCs upregulated MHCII,CD 1.1,CD11 c,CD80,and CD86 cell-surface ***,mRNA expressions of DC maturation biomarkers(CCL5,CCR7,and CD83) and TLR signalling genes(TLR15 and MyD88) were also upregulated whereas those of Wnt signalling were non-significant compared to negative *** treatment induced IL-12 and IFN-γ secretion in chSPDCs but had no effect on IL-10 and TGF-β.Likewise,DC-T cell co-culture promoted IFN-γ secretion and the level of IL-4 was *** of T cells and their differentiation into CD3+/CD4+T cells were triggered in chSPDCs-T cells co-culture *** together,rEaGAPDH could promote Th1 polarization by activating both host DCs and T cells and sheds new light on the role of this important molecule which might contribute to the development of new DCs-based immunotherapeutic strategies against coccidiosis.

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旋毛虫二肽基肽酶1的免疫保护性研究

畜牧与兽医 2020年第11期52卷 105-109页

作者：廖书漪徐立新宋小凯李祥瑞严若峰南京农业大学动物医学院江苏南京210095 新疆农业大学动物医学学院新疆乌鲁木齐830052

前期发现旋毛虫二肽基肽酶1(dipeptidyl peptidase 1,Dpp1)在体外试验中表现出对宿主的免疫调节作用,但其在宿主体内的免疫调节作用尚不清楚。本研究将重组Dpp1蛋白(rDpp1)腹腔注射大鼠后,检测宿主血清中IL-4、IL-9、IL-17、转化生长因... 详细信息

前期发现旋毛虫二肽基肽酶1(dipeptidyl peptidase 1,Dpp1)在体外试验中表现出对宿主的免疫调节作用,但其在宿主体内的免疫调节作用尚不清楚。本研究将重组Dpp1蛋白(rDpp1)腹腔注射大鼠后,检测宿主血清中IL-4、IL-9、IL-17、转化生长因子-β(TGF-β)和IFN-γ等细胞因子的变化,结果发现rDpp1能提高大鼠体内IFN-γ的分泌水平,下调IL-17的分泌,而IL-4、IL-9和TGF-β的分泌差异不显著。将旋毛虫肌肉幼虫与抗rDpp1抗体共孵育后感染小鼠,与对照组相比其成虫数减少了39.3%。将rDpp1与弗氏佐剂乳化后免疫小鼠,检测动物血清中特异性IgG、IgG1和IgG2a抗体的变化,攻虫后分析宿主体内成虫和肌肉幼虫的荷虫数的变化,结果表明rDpp1能诱导小鼠特异性IgG、IgG1亚型和IgG2a亚型抗体的产生,成虫荷虫量减少了47.7%,肌肉幼虫减少了64.5%。上述结果说明旋毛虫二肽基肽酶1具有较好的免疫保护作用。

关键词：旋毛虫二肽基肽酶1 免疫保护作用细胞因子

旋毛虫重组二肽基肽酶1在体外对大鼠外周血单个核细胞增殖、吞噬、凋亡等的影响

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畜牧兽医学报 2020年第6期51卷 1362-1370页

作者：廖书漪储稳徐立新宋小凯李祥瑞严若峰南京农业大学动物医学院南京210095

本文旨在分析旋毛虫二肽基肽酶1(dipeptidyl-peptidase 1,Dpp1)在体外对大鼠外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)增殖、迁移、凋亡以及一氧化氮(NO)分泌和吞噬功能的影响。根据GenBank中旋毛虫Dpp1基因序列设... 详细信息

本文旨在分析旋毛虫二肽基肽酶1(dipeptidyl-peptidase 1,Dpp1)在体外对大鼠外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)增殖、迁移、凋亡以及一氧化氮(NO)分泌和吞噬功能的影响。根据GenBank中旋毛虫Dpp1基因序列设计一对特异性引物,RT-PCR获得该基因,将其亚克隆到pET-32a原核表达载体,IPTG诱导后获得重组蛋白rDpp1。将大鼠PBMCs与rDpp1共孵育,用间接免疫荧光试验(IFA)分析rDpp1与大鼠PBMCs的结合情况,分析不同质量浓度(10、20、40μg·mL^-1)的rDpp1对细胞增殖、迁移、NO分泌、吞噬功能和细胞凋亡的影响。结果表明,40μg·mL^-1rDpp1极显著促进PBMCs增殖;10μg·mL^-1rDpp1能显著促进PBMCs迁移,而20及40μg·mL^-1时表现极显著的促进;20与40μg·mL^-1rDpp1极显著促进PBMCs分泌NO;10μg·mL^-1rDpp1能显著促进PBMCs的吞噬功能,而20及40μg·mL^-1时表现极显著的促进;各质量浓度蛋白均极显著地促进PBMCs的凋亡。旋毛虫二肽基肽酶1在体外能通过多种途径影响大鼠外周血单个核细胞,对调节宿主免疫应答有一定的促进作用。

关键词：旋毛虫二肽基肽酶1 外周血单个核细胞吞噬凋亡

旋毛虫果糖-1,6-二磷酸酶基因的克隆表达及生物学特性分析

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畜牧与兽医 2015年第9期47卷 5-9页

作者：张萌萌徐立新宋小凯李祥瑞严若峰南京农业大学动物医学院江苏南京210095

根据Gen Bank公布的旋毛虫果糖-1,6-二磷酸酶(FBP)基因序列,设计1对特异性引物,以旋毛虫总RNA为模板,用RT-PCR技术扩增出1条约为1 kb的DNA片段。测序结果表明该片段包含FBP的完整ORF,长1 026 bp,编码341个氨基酸。BLAST分析发现其核酸... 详细信息

根据Gen Bank公布的旋毛虫果糖-1,6-二磷酸酶(FBP)基因序列,设计1对特异性引物,以旋毛虫总RNA为模板,用RT-PCR技术扩增出1条约为1 kb的DNA片段。测序结果表明该片段包含FBP的完整ORF,长1 026 bp,编码341个氨基酸。BLAST分析发现其核酸和氨基酸序列与已知旋毛虫FBP的基因和氨基酸序列的相似性分别为99%和100%。将该ORF亚克隆到p ET-32a(+)载体中,酶切鉴定正确后转化感受态大肠杆菌BL21(DE3),IPTG诱导表达,SDS-PAGE分析,结果表明该融合蛋白大小为55.4 ku,重组蛋白主要以包涵体的形式表达。Western blot结果显示该重组蛋白能被旋毛虫感染小鼠血清识别。对纯化、复性的重组蛋白进行了酶活性测定,结果发现该重组酶的最适p H为7.0,最适温度为37℃。

关键词：旋毛虫果糖-1,6-二磷酸酶原核表达