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Pluripotent stem cells secrete Activin A to improve their epiblast competency after injection into recipient embryos

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Protein & Cell 2018年第8期9卷 717-728页

作者： Jinzhu Xiang Suying Cao Liang Zhong Hanning Wang Yangli Pei Qingqing Wei Bingqiang Wen Haiyuan Mu Shaopeng Zhang Liang Yue Genhua Yue Bing Lim Jianyong Han State Key Laboratory for Agrobiotechnology College of Biological Sciences China Agricultural University Beijing 100193 China Animal Science and Technology College Beijing University of Agriculture Beijing 102206 China Temasek Life Sciences Laboratory National University of Singapore Singapore 117604 Singapore Stem Cell and Developmental Biology Genome Institute of Singapore Singapore 138672 Singapore Beijing Advanced Innovation Center for Food Nutrition and Human Health China Agricultural University Beijing 100193 China State Key Laboratory of Animal Nutrition Institute of Animal Sciences Chinese Academy of Agricultural Sciences Beijing 100193 China

It is not fully clear why there is a higher contribution of pluripotent stem cells （PSCs） to the chimera produced by injection of PSCs into 4-cell or 8-cell stage embryos compared with blastocyst injection. Here, we show that not only embryonic stem cells （ESCs） but also induced pluripotent stem cells （iPSCs） can generate F0 nearly 100% donor cell-derived mice by 4-cell stage embryo injection, and the approach has a ＂dose effect＂. Through an analysis of the PSC-secreted proteins, Activin A was found to impede epiblast （EPI） lineage development while promoting trophectoderm （TE） differentiation, resulting in replacement of the EPI lineage of host embryos with PSCs. Interestingly, the injection of ESCs into blastocysts cultured with Activin A （cultured from 4-cell stage to early blastocyst at E3.5） could increase the contribution of ESCs to the chimera. The results indicated that PSCs secrete protein Activin A to improvetheir EPI competency after injection into recipient embryos through influencing the development of mouse early embryos. This result is useful for optimizing the chimera production system and for a deep understand- ing of PSCs effects on early embryo development.

关键词： pluripotent stem cells 4-cell embryoinjection secreted proteins Activin A chimeric mice

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Human Seroreactivity to secreted Molecules of Corynebacterium pseudotuberculosis

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Advances in Microbiology 2022年第3期12卷 150-158页

作者： Ramon Mendes dos Santos Silvânia Maria Andrade Cerqueira Caio Lopez Borge Andrade Gabriel Saldanha Müller Vitória Costa de Menezes Santos Hévlyn Ribeiro de Araújo Samanta Queiroz Rogério Reis Conceição Luiz Gustavo Freitas Oliveira Marcos Borges Ribeiro Silvana Marchioro Lilia Ferreira de Moura-Costa Fúlvia Soares Campos de Sousa Maria da Conceição Aquino de Sá José Tadeu Raynal Rocha Filho Soraya Castro Trindade Eduardo Martins Netto Roberto Meyer Songelí Menezes Freire Laboratory of Immunology and Molecular Biology of the Health Sciences Institute of the Federal University of Bahia Bahia Brazil Specialized Hospital Otávio Mangabeira Bahia Brazil Microbiology of the Federal University of Bahia Bahia Brazil Laboratory of Infectology of the University Hospital Complex Professor Edgard Santos Salvador Bahia Brazil

Corynebacterium pseudotuberculosis is an infectious agent that occurs in small ruminants causing caseous lymphadenitis, and more rarely in humans causing lymphadenitis and pneumonia. The breeding small ruminants have great economic importance in Brazil. Rural farm workers and veterinary students who acquired this disease suffered from weakening symptoms for weeks, and the identification of the etiological agent was time-consuming and complex. Due to the low prevalence of case records, there is probably no available commercial diagnostic kit for C. pseudotuberculosis infection in humans. This study aimed to describe human seroreactivity to secreted antigens from C. pseudotuberculosis. Reactivity of serum from farm workers (n = 14), individuals who work with the bacillus at laboratory (n = 8) or individuals without contact (n = 25) was tested with secreted proteins from PAT10 strain of C. pseudotuberculosis by Western blotting. Samples of all (100%) farm workers showed reactivity to 31 kDa, 71 kDa and 164 kDa proteins, while laboratory workers showed 87.5%, 62.5 % and 37.5%, and no-contact 20%, 0% and 16%, respectively. All sera recognized the 275 kDa protein. Our data suggest that C. pseudotuberculosis secreted proteins are antigenic in humans and the recognition profiles allowed the identification of individuals with and without prior contact with this bacillus. This is the first paper which describes human reactivity to C. pseudotuberculosis in serum samples of workers in Brazil.

关键词： Human Immunoreactivity Antigenicity Corynebacterium pseudotuberculosis secreted proteins Western Blot

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Immunogenicity and protective efficacy study using combination of four tuberculosis DNA vaccines

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Science China Life Sciences 2003年第5期46卷 495-502+562页

作者：蔡宏田霞呼西旦潘怡李国利庄玉辉朱玉贤 China Peking University Tuberculosis Research Laboratory 309 Hospital of PLA Institute of Veterinary Xinjiang Academy of Animal Science Beijing 100091 Peking University Beijing 100871 College of Life Science National Laboratory of Protein Engineering and Plant Genetic Engineering Urumqi 830000

Immune response and protective efficacy for the combination of four tuberculosis DNA vaccines were evaluated in this study. We obtained 1︰200 antibody titers against Ag85B 21d after mice were vaccinated for the first time by four recombinant eukaryotic expression vectors containing coding sequences for Ag85B, MPT-64, MPT-63 and ESAT-6. The titers of Ag85B were elevated to 1︰102400 after the second injection and decreased to 1︰12800 after the third injection. Antibody titers for MPT-64 and MPT-63 reached 1︰25600 21 d after the first vaccination, and were then decreased following the second and third injections. No antigen-specific antibody titer against ESAT-6 was detected in sera harvested from immunized mice at any time. These DNA vaccines evoked specific IFN-g responses in the spleens of vaccinated mice as well. When challenged with M. tuberculosis H37Rv, we found that the lungs of the vaccinated mice produced 99.8% less bacterial counts than that of the empty-vector control group and the bacterial counts were also significantly less than that of the BCG group. Histopathological analyses showed that the lungs of vaccinated mice produced no obvious caseation while over 50%—70% of the pulmonary parenchyma tissue produced central caseation in the vector control group. Our results indicated that the combination of four tuberculosis DNA vaccines may generate high levels of immune responses and result in better animal protection.

关键词： Mycobacterium tuberculosis secreted proteins combination DNA vaccines immunogenicity protective efficacy.

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Association Analysis of SP-SNPs and Avirulence Genes in Puccinia striiformis f. sp. tritici, the Wheat Stripe Rust Pathogen

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American Journal of Plant Sciences 2016年第1期7卷 126-137页

作者： Chongjing Xia Meinan Wang Anmin Wan Derick A. Jiwan Deven R. See Xianming Chen Department of Plant Pathology Washington State University Pullman USA United States Department of Agriculture Agricultural Research Service Wheat Health Genetics and Quality Research Unit Pullman USA

Puccinia striiformis f. sp. tritici (Pst) is one of the pathogenic fungi on wheat, caused stripe rust that is a great threat for wheat production all over the world. Intensive efforts have been made to study genetics of wheat resistance to this disease, but few on avirulence of the pathogen due mainly to the nature of obligate biotrophism and the lack of systems for studying its genetics and molecular manipulations. To overcome these limitations, a natural Pst population comprising 352 isolates representative of a diverse virulence spectrum was genotyped using 97 secreted protein-single nucleotide polymorphism (SP-SNP) markers to identify candidate avirulence genes using association analysis. Among avirulence genes corresponding to 19 resistance genes, significantly associated SP-SNP markers were detected for avirulence genes AvYr1, AvYr2, AvYr6, AvYr7, AvYr8, AvYr44, AvYrExp2, AvYrSP, and AvYrTye. These results indicate that association analysis can be used to identify markers for avirulence genes. This study has laid the foundation for developing more SP-SNPs for mapping avirulence genes using segregating populations that can be generated through sexual reproduction on alternate hosts of the pathogen.

关键词： Puccinia striiformis f. sp. tritici Wheat Stripe Rust Avirulence Genes secreted proteins Single Nucleotide Polymorphism Association Analysis

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