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Pre-evaluation of humoral immune response of Bactrian camels by the quantification of th2 cytokines using real-time PCR

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the Journal of Biomedical Research 2020年第5期34卷 387-394页

作者： Xinyu Yu Yuan Wu Jiarong Zhang Jirimutu Azhati Zulipikaer Jin Chen Center for Global Health School of Public HealthNanjing Medical UniversityNanjingJiangsu 211166China Department of Medical Oncology Jiangsu Cancer HospitalJiangsu Institute of Cancer Researchthe Affiliated Cancer Hospital of Nanjing Medical UniversityNanjingJiangsu 210009China Key Laboratory of Dairy Biotechnology and Bioengineering Ministry of EducationCollege of Food Science and EngineeringInner Mongolia Agricultural UniversityHohhotInner Mongolia 010018China Xinjiang Academy of Animal Science UrumqiXinjiang 830011China the Key Laboratory of Modern Toxicology Ministry of EducationSchool of Public HealthNanjing Medical UniversityNanjingJiangsu 211166China Key Laboratory of Antibody Technique of National Health Commission Nanjing Medical UniversityNanjingJiangsu 211166China

With the increasing immunological studies on camels due to the advantage of their single-chain antibodies for humanizations,it is demanding to develop an easy-to-handle evaluation method of their humoral immune response before proceeding with immunization of foreign antigens that may be toxic to *** this study,we quantitatively determined the expression levels of T-helper 2(th2) cytokines in peripheral blood lymphocytes obtained from Bactrian camels by real-time *** recorded kinetic profiles resulting from the immunization of ovalbumin(OVA) indicated that after immunization,th2 cytokines including interleukin(IL) families such as IL-4,IL-10,and IL-13 in the camels were up-regulated by a factor of 1.78,3.15,and 1.22,respectively,which was validated by traditional enzyme-linked immunosorbent assay(ELISA) *** ELISA which requires specific enzyme-labeled antibodies,this established method based on the minimal amount of blood samples holds an advantage in the preliminary evaluation of camel humoral immune response with desirable precision,which is meaningful for biomedical explorations of camel-derived antibodies.

关键词： Bactrian camels th2 cytokines humoral immune response real-time PCR

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Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi

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Asian Pacific Journal of Tropical Biomedicine 2021年第1期11卷 20-28页

作者： Vikas Kushwaha Sukhbir Kaur Leishmania Research Laboratory Department of ZoologyPanjab UniversitySector-14Chandigarh 160014India

Objective:To evaluate the immunostimulatory potential of crossreactive molecule heat shock protein 60(HSP60)of filarial parasite Brugia malayi and Leishmania ***:HSP60 of Brugia malayi(BmHSP60)was amplified using gene-specific primer,cloned in p Tri Ex4 vector,expressed in BL21-DE3 cells,and recombinant HSP60(rHSP60)of~65 k Da was purified by affinity chromatography using Ni-NTA *** recombinant protein was desalted by the dialysis membrane,and the presence of endotoxin level was determined by Limulus amebocyte lysate *** recombinant protein was tested for cell proliferation,nitric oxide release,expression of th1 and th2 cytokines,and transcription factors(STATs)in vitro using murine macrophage cell line(J774 A.1).Results:Higher cell proliferation indicated that BmHSP60 had immunostimulatory ***60 exposure upregulated the expression of iNOS,STAT1,STAT4,th1 cytokines(IFN-γ,TNF-α,IL-12),and nitric oxide *** addition,no remarkable change was observed in the expression of IL-6,IL-10,and STAT3 in macrophage cell line J774 *** ELISA analysis showed the levels of IFN-γ,TNF-α,and IL-12 were upregulated while IL-10 level was downregulated,revealing that BmHSP60 triggered a th1 immune ***:Our study demonstrates that rBmHSP60 has immunogenic properties which effectively enhances the th1 type immune responses,and can be used as an immunoprophylactic agent against ***,in vivo studies are in progress to determine the protective role of rBmHSP60 against Leishmania donovani infection in a mouse model.

关键词： Brugia malayi Heat shock protein 60 Leishmania donovani th1 cytokines th2 cytokines

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Treatment with Enterococcus faecalis CECT7121 Is Not Effective as therapy in Mice with an Established Allergy Status

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Advances in Microbiology 2017年第10期7卷 696-706页

作者： Ailén M.Díaz Matías A.Molina Guillermo G.Nunez Ana C.Mourelle Mónica D.Sparo Marcela A.Manghi Marisa S.Castro Facultad de Farmacia y Bioquímica Universidad de Buenos AiresInstituto de Estudios de la Inmunidad Humoral“Prof.Dr.R.A.Margni”(IDEHUCONICET-UBA)Buenos AiresArgentina Cátedra Microbiología y Parasitología(CUDEMyP) Facultad de Ciencias MédicasUniversidad Nacional de La PlataLa PlataArgentina

In allergies, an unbalanced immune response towards a T helper (th) 2 profile with high levels of Immunoglobulin (Ig) E is produced. We have demonstrated that the pre-administration of Enterococcus faecalis CECT7121 prevents the development of allergy in ovalbumin-immunized mice. In this work, we evaluated whether this bacterium can also revert an established allergic status. Mice were immunized with ovalbumin (OVA) and after that, were inoculated with an E. faecalis CECT7121 suspension. In immunized animals, serum specific immune response, proliferative activity of memory splenocytes, and levels of th2 cytokines were assessed. the in vivo active cutaneous anaphylaxis test was also performed. the treatment with E. faecalis CECT7121 only increased anti-OVA IgG2a levels. No differences were observed in other specific immunological parameters. Probiotic-treatment did not prove to have any desensitizing effect on mice. these results, together with those recently published, can be concluded that this bacterium would not be appropriate for the treatment of allergic symptoms.

关键词： Enterococcus faecalis CECT7121 Ovalbumin-Allergy Murine Model Immunoglobulin E th2 cytokines Probiotics

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Signal transducer and activator of transcription 6 directly regulates human ORMDL3 expression

Signal transducer and activator of transcription 6 directly ...

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第十二次全国医学遗传学学术会议

作者：邱熔芳杨阳赵海玲辛倩单珊李江夏龚瑶琴刘奇迹山东大学实验畸形学教育部重点实验室医学院医学遗传学研究所

Objective Orosomucoid-like 3(0RMDL3) has been associated with asthma and a series of autoimmune disorders and is involved in endoplasmic reticulum-mediated inflammatory ***,the clinical significance and molecular mechanism underlying its expression are still largely *** To elucidate the mechanisms in human ORMDL3 transcriptional regulation,we cloned the 1.5-kb genomic DNA fragment containing the putative promoter region and evaluated its transcriptional activity in a luciferase reporter system by deletion analysis. EMSA,CMP,ChIP-Re-ChIP was used to identify the key transcription *** We identified a 68-bp region that functions as a minimal *** analysis predicted that the -64 to -56 bp region contained a signal transducer and activator of transcription 6(STAT6) binding *** mobility shift assay and chromatin immunoprecipitation demonstrated that STAT6 bound to its binding site within the ORMDL3 ***6 overexpression or knockdown could trans-activate or trans-inhibit,respectively,the ORMDL3 promoter containing the STAT6-binding *** 4(IL-4)/IL-13 treatment increased ORMDL3 promoter activity as Well as endogenous ORMDL3 *** and ChIP-Re-ChIP assays revealed that STAT6 and p300 exist in the same protein complex binding to the ORMDL3 *** Our study confirmed that STAT6 plays important roles in regulating the expression of human ORMDL3 by directly binding to the promoter region, which can shed light on a possible role in various human diseases.

关键词： STAT6 ORMDL3 transcription regulation p300 th2 cytokines

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