The polymorphism of MHC class II B gene in 14 Chinese alligators was analyzed, which came from three different areas： a wild population from Xuancheng, Anhui, a captive population from Changxing, Zhejiang, and a capt...The polymorphism of MHC class II B gene in 14 Chinese alligators was analyzed, which came from three different areas： a wild population from Xuancheng, Anhui, a captive population from Changxing, Zhejiang, and a captive population from Anhui Research Center for Reproduction of Chinese Alligators. The gene fragment was amplified using a pair of specific primers designed from the MHC gene sequence of the spectacled caiman. A total of 34 sequence haplotypes of exon 3 were detected in the sampled Chinese alligators. The numbers of haplotypes of the 3 Chinese alligator populations were 15, 10, and 9, respectively. The overall estimation of the MHC polymorphism in the Chinese alligator population was higher than those in mammals and in cypdnid fish, The rates of nonsynonymous substitutions （dN） occurred at a significantly lower frequency than that of synonymous substitutions （ds）, which were not consistent with the common rule. This result might suggest that the polymorphism of exon 3 seemed not to be maintained by the balancing selection. The neutrality test of Tajima excluded the null hypothesis that the polymorphism of exon 3 was generated by a random drift, and the fact that D = -0.401 indicated an excess of rare mutations in the Chinese alligator. The nucleotide diversity of the sequences and the phylogenetic relations were also analyzed, and the results suggested that there was no significant difference in genetic diversity among the 3 populations of Chinese alligator.展开更多
The histological hallmark of autoimmune hepatitis（AIH） is a dense portal mononuclear cell infiltrate that invades the surrounding parenchyma and comprises T and B lymphocytes,macrophages,and plasma cells.An unknown ...The histological hallmark of autoimmune hepatitis（AIH） is a dense portal mononuclear cell infiltrate that invades the surrounding parenchyma and comprises T and B lymphocytes,macrophages,and plasma cells.An unknown but powerful stimulus must be promoting the formation of this massive inflammatory cellular reaction that is likely to initiate and perpetuate liver damage.An autoimmune attack can follow different pathways to inflict damage on hepatocytes.Liver damage is likely to be orchestrated by CD4^＋ T lymphocytes recognizing an autoantigenic liver peptide.To trigger an autoimmune response,the peptide must be embraced by an HLA class Ⅱ molecule and presented to naive CD4^＋ T helper（Th0） cells by professional antigen presenting cells,with the co-stimulation of ligand-ligand fostering interaction between the two cells.Th0 cells become activated,differentiate into functional phenotypes according to the cytokines prevailing in the microenvironment and the nature of the antigen,and initiate a cascade of immune reactions determined by the cytokines produced by the activated T cells.Th1 cells,arising in the presence of the macrophage-derived interleukin（IL） -12,secrete mainly IL-2 and interferon-gamma（IFN-γ）,which activate macrophages,enhance expression of HLA classⅠ（increasing liver cell vulnerability to a CD8^＋ T cell cytotoxic attack）,and induce expression of HLA class Ⅱ molecules on hepatocytes.Th2 cells,which differentiate from Th0 if the microenvironment is rich in IL-4,produce mainly IL-4,IL-10,and IL-13 which favour autoantibody production by B lymphocytes.Physiologically,Th1 and Th2 antagonize each other.Th17 cells,a recently described population,arise in the presence of transforming growth factor beta（TGF-β） and IL-6 and appear to have an important effector role in inflammation and autoimmunity.Theprocess of autoantigen recognition is strictly controlled by regulatory mechanisms,such as those exerted by CD4^＋CD25^＋ regulatory T cells,which derive from Th0 in the presence of TGF-β,but in the absence of IL-6.If regulatory mechanisms fail,the autoimmune attack is perpetuated.Over the past three decades different aspects of the above pathogenic scenario have been investigated.In particular,a defect in immunoregulation affecting CD4^＋CD25^＋ regulatory T cells（T-regs） has been demonstrated in AIH,particularly at diagnosis or during relapse.Advances in the study of autoreactive T cells have occurred mostly in AIH type 2,since the knowledge that CYP2D6 is the main autoantigen has enabled the characterization of both CD4 and CD8 T cells targeting this cytochrome.CD4 T cells from patients with type 2 AIH positive for the predisposing HLA allele DRB10701 recognize seven regions of CYP2D6,five of which are also recognized by CD8 T cells.High numbers of IFN-γ producing CD4 T cells and CD8 T cells are associated with biochemical evidence of liver damage,suggesting a combined cellular immune attack.展开更多
AIM: To investigate the expression of several important molecules involved in major histocompatibility complex (MHC) class I presentation pathway in primary hepatocellular carcinoma (HCC), and to determine whether cyt...AIM: To investigate the expression of several important molecules involved in major histocompatibility complex (MHC) class I presentation pathway in primary hepatocellular carcinoma (HCC), and to determine whether cytotoxic T lymphocyte (CTL) vaccine therapy was suitable for HCC. METHODS: Labeled streptavidin biotin(LSAB) method of immunohisto-chemistry was used to study 33 HCC tissue specimens. RESULTS: Most HCC tissues and adjacent histological normal hepatocytes expressed HLA-I antigens,TAP,and B7, expression of B7 was especially strong,and there was no significant difference between them (P>0.05). CONCLUSION: The MHC class I presentation pathway in primary hepatocellular carcinoma may not be abnormal or dysfunctional,and CTL could kill these tumor cells. Thus, it is suitable and practicable to design and construct CTL vaccine against HCC.展开更多
Objective To compare the characterization and myocardial differentiation capacity of arnniotic fluid-derived mesenchymal stromal cells （AF MSCs） and umbilical cord Wharton＇s Jelly-derived mesenchymal stromal cells ...Objective To compare the characterization and myocardial differentiation capacity of arnniotic fluid-derived mesenchymal stromal cells （AF MSCs） and umbilical cord Wharton＇s Jelly-derived mesenchymal stromal cells （WJ MSCs）. Methods The human AF MSCs were cultured from amniotic fluid samples obtained by amniocentesis. The umbilical cord WJ MSCs were obtained from Wharton＇s Jelly of umbilical cords of infants delivered full-term by normal labor. The morphology, growth curves, and analyses by flow cytometry of cell surface markers were compared between the two types of cells. Myocardial genes （GATA-4, c-TnT, a-actin, and Cx43） were detected by real-time PCR and the corresponding protein expressions were detected by Western blot analysis after myocardial induced in AF MSCs and WJ MSCs. Results Our findings revealed AF MSCs and WJ MSCs shared similar morphological characteristics of the fibroblastoid shape. The AF MSCs were easily obtained than the WJ MSCs and had a shorter time to reach adherence of 2.7 ± 1.6 days to WJ MSCs of 6.5 ± 1.8 days. The growth curves by MTT cytotoxic assay showed the AF MSCs had a similar proliferative capacity at passage 5 and passage 10. However, the proliferative capacities ofWJ MSCs were decreased at 5 passage relative to 10 passage. Both AF stem cells and WJ stem cells had the characteristics of mesenchymal stromal cells with some characteristics of embryonic stem cells. They express CD29 and CD105, but not CD34. They were positive for Class I major histocompatibility （MHC I） antigens （HLA-ABC）, and were negative, or mildly positive, for MHC Class II （HLA-DR） antigen. Oct-4 was positive in all the two cells types. Both AF MSCs and WJ MSCs could differentiate along myocardium. The differentiation capacities were detected by the expression of GATA-4, c-TnT, a-actin, Cx43 after myocardial induction. Conclusions Both AF MSCs and WJ MSCs have the potential clinical application for myogenesis in cardiac regenerative therapy.展开更多
Platichthys stellatus is an economically important marine bony fish species that is cultured in China on a large scale.However,very little is known about its immune-related genes.In this study,the transcriptome of the...Platichthys stellatus is an economically important marine bony fish species that is cultured in China on a large scale.However,very little is known about its immune-related genes.In this study,the transcriptome of the immune organs ofP.stellatus that were intraperitoneally challenged with the pathogen Edwardsiella ictaluri JCM1680 is analyzed.Total RNA from four tissues(spleen,kidney,liver,and intestine) was mixed equally and then sequenced on an Illumina HiSeq 2000 platform.Overall,28 465 813 quality reads were generated and assembled into 43 061 unigenes.Similarity searches against public protein sequence databases were used to annotate 28 291 unigenes(65.7%of the total),368 of which were associated with immunoregulation,including 188 related to immunity response.Additionally,the transcript levels of immunity response unigenes annotated as related to tumor necrosis factor(TNF),TNF receptor,chemokine,major histocompatibility complex,and interleukin-6 were investigated in the different tissues of normal and infected P.stellatus by real-time quantitative PCR.The results confirmed that the unigenes identified in the transcriptome database were indeed expressed and up-regulated in infected P.stellatus.To our knowledge,this is the first report of the sequencing and analysis of the transcriptome of P.stellatus.These findings provide insights into the transcriptomics and immunogenetics of bony fish.展开更多
基金the National Natural Science Foundation of China (No. 30270213 and 30470244)a grant of Special Scholar Foundation from Anhui Province (No. 04043049)the Key Laboratory of Biotic Environment and Ecological Safety in Anhui Province.
文摘The polymorphism of MHC class II B gene in 14 Chinese alligators was analyzed, which came from three different areas： a wild population from Xuancheng, Anhui, a captive population from Changxing, Zhejiang, and a captive population from Anhui Research Center for Reproduction of Chinese Alligators. The gene fragment was amplified using a pair of specific primers designed from the MHC gene sequence of the spectacled caiman. A total of 34 sequence haplotypes of exon 3 were detected in the sampled Chinese alligators. The numbers of haplotypes of the 3 Chinese alligator populations were 15, 10, and 9, respectively. The overall estimation of the MHC polymorphism in the Chinese alligator population was higher than those in mammals and in cypdnid fish, The rates of nonsynonymous substitutions （dN） occurred at a significantly lower frequency than that of synonymous substitutions （ds）, which were not consistent with the common rule. This result might suggest that the polymorphism of exon 3 seemed not to be maintained by the balancing selection. The neutrality test of Tajima excluded the null hypothesis that the polymorphism of exon 3 was generated by a random drift, and the fact that D = -0.401 indicated an excess of rare mutations in the Chinese alligator. The nucleotide diversity of the sequences and the phylogenetic relations were also analyzed, and the results suggested that there was no significant difference in genetic diversity among the 3 populations of Chinese alligator.
文摘The histological hallmark of autoimmune hepatitis（AIH） is a dense portal mononuclear cell infiltrate that invades the surrounding parenchyma and comprises T and B lymphocytes,macrophages,and plasma cells.An unknown but powerful stimulus must be promoting the formation of this massive inflammatory cellular reaction that is likely to initiate and perpetuate liver damage.An autoimmune attack can follow different pathways to inflict damage on hepatocytes.Liver damage is likely to be orchestrated by CD4^＋ T lymphocytes recognizing an autoantigenic liver peptide.To trigger an autoimmune response,the peptide must be embraced by an HLA class Ⅱ molecule and presented to naive CD4^＋ T helper（Th0） cells by professional antigen presenting cells,with the co-stimulation of ligand-ligand fostering interaction between the two cells.Th0 cells become activated,differentiate into functional phenotypes according to the cytokines prevailing in the microenvironment and the nature of the antigen,and initiate a cascade of immune reactions determined by the cytokines produced by the activated T cells.Th1 cells,arising in the presence of the macrophage-derived interleukin（IL） -12,secrete mainly IL-2 and interferon-gamma（IFN-γ）,which activate macrophages,enhance expression of HLA classⅠ（increasing liver cell vulnerability to a CD8^＋ T cell cytotoxic attack）,and induce expression of HLA class Ⅱ molecules on hepatocytes.Th2 cells,which differentiate from Th0 if the microenvironment is rich in IL-4,produce mainly IL-4,IL-10,and IL-13 which favour autoantibody production by B lymphocytes.Physiologically,Th1 and Th2 antagonize each other.Th17 cells,a recently described population,arise in the presence of transforming growth factor beta（TGF-β） and IL-6 and appear to have an important effector role in inflammation and autoimmunity.Theprocess of autoantigen recognition is strictly controlled by regulatory mechanisms,such as those exerted by CD4^＋CD25^＋ regulatory T cells,which derive from Th0 in the presence of TGF-β,but in the absence of IL-6.If regulatory mechanisms fail,the autoimmune attack is perpetuated.Over the past three decades different aspects of the above pathogenic scenario have been investigated.In particular,a defect in immunoregulation affecting CD4^＋CD25^＋ regulatory T cells（T-regs） has been demonstrated in AIH,particularly at diagnosis or during relapse.Advances in the study of autoreactive T cells have occurred mostly in AIH type 2,since the knowledge that CYP2D6 is the main autoantigen has enabled the characterization of both CD4 and CD8 T cells targeting this cytochrome.CD4 T cells from patients with type 2 AIH positive for the predisposing HLA allele DRB10701 recognize seven regions of CYP2D6,five of which are also recognized by CD8 T cells.High numbers of IFN-γ producing CD4 T cells and CD8 T cells are associated with biochemical evidence of liver damage,suggesting a combined cellular immune attack.
基金Supported by the National Natural Science Foundation of China, No. 30070855
文摘AIM: To investigate the expression of several important molecules involved in major histocompatibility complex (MHC) class I presentation pathway in primary hepatocellular carcinoma (HCC), and to determine whether cytotoxic T lymphocyte (CTL) vaccine therapy was suitable for HCC. METHODS: Labeled streptavidin biotin(LSAB) method of immunohisto-chemistry was used to study 33 HCC tissue specimens. RESULTS: Most HCC tissues and adjacent histological normal hepatocytes expressed HLA-I antigens,TAP,and B7, expression of B7 was especially strong,and there was no significant difference between them (P>0.05). CONCLUSION: The MHC class I presentation pathway in primary hepatocellular carcinoma may not be abnormal or dysfunctional,and CTL could kill these tumor cells. Thus, it is suitable and practicable to design and construct CTL vaccine against HCC.
文摘Objective To compare the characterization and myocardial differentiation capacity of arnniotic fluid-derived mesenchymal stromal cells （AF MSCs） and umbilical cord Wharton＇s Jelly-derived mesenchymal stromal cells （WJ MSCs）. Methods The human AF MSCs were cultured from amniotic fluid samples obtained by amniocentesis. The umbilical cord WJ MSCs were obtained from Wharton＇s Jelly of umbilical cords of infants delivered full-term by normal labor. The morphology, growth curves, and analyses by flow cytometry of cell surface markers were compared between the two types of cells. Myocardial genes （GATA-4, c-TnT, a-actin, and Cx43） were detected by real-time PCR and the corresponding protein expressions were detected by Western blot analysis after myocardial induced in AF MSCs and WJ MSCs. Results Our findings revealed AF MSCs and WJ MSCs shared similar morphological characteristics of the fibroblastoid shape. The AF MSCs were easily obtained than the WJ MSCs and had a shorter time to reach adherence of 2.7 ± 1.6 days to WJ MSCs of 6.5 ± 1.8 days. The growth curves by MTT cytotoxic assay showed the AF MSCs had a similar proliferative capacity at passage 5 and passage 10. However, the proliferative capacities ofWJ MSCs were decreased at 5 passage relative to 10 passage. Both AF stem cells and WJ stem cells had the characteristics of mesenchymal stromal cells with some characteristics of embryonic stem cells. They express CD29 and CD105, but not CD34. They were positive for Class I major histocompatibility （MHC I） antigens （HLA-ABC）, and were negative, or mildly positive, for MHC Class II （HLA-DR） antigen. Oct-4 was positive in all the two cells types. Both AF MSCs and WJ MSCs could differentiate along myocardium. The differentiation capacities were detected by the expression of GATA-4, c-TnT, a-actin, Cx43 after myocardial induction. Conclusions Both AF MSCs and WJ MSCs have the potential clinical application for myogenesis in cardiac regenerative therapy.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(Nos.2012AA10A413,2012AA10A408)the National Marine Public Welfare Research Project(No.201205025)
文摘Platichthys stellatus is an economically important marine bony fish species that is cultured in China on a large scale.However,very little is known about its immune-related genes.In this study,the transcriptome of the immune organs ofP.stellatus that were intraperitoneally challenged with the pathogen Edwardsiella ictaluri JCM1680 is analyzed.Total RNA from four tissues(spleen,kidney,liver,and intestine) was mixed equally and then sequenced on an Illumina HiSeq 2000 platform.Overall,28 465 813 quality reads were generated and assembled into 43 061 unigenes.Similarity searches against public protein sequence databases were used to annotate 28 291 unigenes(65.7%of the total),368 of which were associated with immunoregulation,including 188 related to immunity response.Additionally,the transcript levels of immunity response unigenes annotated as related to tumor necrosis factor(TNF),TNF receptor,chemokine,major histocompatibility complex,and interleukin-6 were investigated in the different tissues of normal and infected P.stellatus by real-time quantitative PCR.The results confirmed that the unigenes identified in the transcriptome database were indeed expressed and up-regulated in infected P.stellatus.To our knowledge,this is the first report of the sequencing and analysis of the transcriptome of P.stellatus.These findings provide insights into the transcriptomics and immunogenetics of bony fish.
