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
目的:构建携带小鼠MHCII类分子反式激活因子(MHC class II molecule transactivator,CIITA)突变体基因的腺病毒,并观察腺病毒介导该基因的表达情况以及表达产物的体外功能。方法:采用常规分子生物学方法从IFN-γ诱导后的BALB/c小鼠腹腔...目的:构建携带小鼠MHCII类分子反式激活因子(MHC class II molecule transactivator,CIITA)突变体基因的腺病毒,并观察腺病毒介导该基因的表达情况以及表达产物的体外功能。方法:采用常规分子生物学方法从IFN-γ诱导后的BALB/c小鼠腹腔巨噬细胞中获得IV型CIITA cDNA;利用重叠延伸PCR法构建CIITA突变体基因,并克隆入表达载体pIRES;采用pAdEasy-1系统获得具有感染能力的、携带CIITA突变体基因的缺陷型重组腺病毒(Ad-CIITAm)和空载对照病毒(Ad-GFP),并经大量扩增、纯化及滴度测定;将Ad-CIITAm和Ad-GFP分别感染HeLa细胞和Raji细胞,流式细胞术观察对诱导型和组成型HLA-DR分子表达的影响。结果:成功克隆了小鼠CIITA突变体基因,并构建了携带小鼠CIITA突变体基因的重组腺病毒Ad-CIITAm;经流式细胞术证实感染Ad-CIITAm的Hela和Raji细胞较感染Ad-GFP的细胞,其表面HLA-DR分子的表达均受到明显的抑制。结论:本实验证实了重组腺病毒介导表达的小鼠CIITA突变体在体外能够有效地抑制MHCII类分子的表达。展开更多
Background:Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in“carrier”or“pathogenic”states.HLA DQ and HLA DR humanized mice have been used as a small animal model to stu...Background:Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in“carrier”or“pathogenic”states.HLA DQ and HLA DR humanized mice have been used as a small animal model to study the role of two molecules during S.aureus infection.However,the contribution of HLA DP to S.aureus infection is unknown yet.Methods:In this study,we have produced HLA DP401 and HLA DRA0101 humanized mice by microinjection of C57BL/6J zygotes.Neo-floxed IAβ+/-mice were crossbred with Ella-Cre and further crossbred with HLA DP401 or HLA-DRA0101 humanized mice.After several rounds of traditional crossbreeding,we finally obtained HLA DP401-IAβ-/-and HLA DRA-IAβ-/-humanized mice,in which human DP401 or DRA0101 molecule was introduced into IAβ-/-mice deficient in endogenous murine MHC classⅡmolecules.A transnasal infection murine model of S.aureus pneumonia was induced in the humanized mice by administering 2×108CFU of S.aureus Newman dropwise into the nasal cavity.The immune responses and histopathology changes were further assessed in lungs in these infected mice.Results:We evaluated the local and systemic effects of S.aureus delivered intranasally in HLA DP401-IAβ-/-and HLA DRA-IAβ-/-transgenic mice.S.aureus Newman infection significantly increased the m RNA level of IL 12p40 in lungs in humanized mice.An increase in IFN-γand IL-6 protein was observed in HLA DRA-IAβ-/-mice.We observed a declining trend in the percentage of F4/80+macrophages in lungs in HLA DP401-IAβ-/-mice and a decreasing ratio of CD4+to CD8+T cells in lungs in IAβ-/-mice and HLA DP401-IAβ-/-mice.A decreasing ratio of Vβ3+to Vβ8+T cells was also found in the lymph node of IAβ-/-mice and HLA DP401-IAβ-/-mice.S.aureus Newman infection resulted in a weaker pathological injury in lungs in IAβ-/-genetic background mice.Conclusion:These humanized mice will be an invaluable mouse model to resolve the pathological mechanism of S.aureus pneumonia and study what role DP molecule plays in S.aureus infection.展开更多
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.
文摘目的:构建携带小鼠MHCII类分子反式激活因子(MHC class II molecule transactivator,CIITA)突变体基因的腺病毒,并观察腺病毒介导该基因的表达情况以及表达产物的体外功能。方法:采用常规分子生物学方法从IFN-γ诱导后的BALB/c小鼠腹腔巨噬细胞中获得IV型CIITA cDNA;利用重叠延伸PCR法构建CIITA突变体基因,并克隆入表达载体pIRES;采用pAdEasy-1系统获得具有感染能力的、携带CIITA突变体基因的缺陷型重组腺病毒(Ad-CIITAm)和空载对照病毒(Ad-GFP),并经大量扩增、纯化及滴度测定;将Ad-CIITAm和Ad-GFP分别感染HeLa细胞和Raji细胞,流式细胞术观察对诱导型和组成型HLA-DR分子表达的影响。结果:成功克隆了小鼠CIITA突变体基因,并构建了携带小鼠CIITA突变体基因的重组腺病毒Ad-CIITAm;经流式细胞术证实感染Ad-CIITAm的Hela和Raji细胞较感染Ad-GFP的细胞,其表面HLA-DR分子的表达均受到明显的抑制。结论:本实验证实了重组腺病毒介导表达的小鼠CIITA突变体在体外能够有效地抑制MHCII类分子的表达。
基金National Science and Technology Major Project,Grant/Award Number:2016YFD0500208,2017ZX10304402-001-012 and 2017ZX10304402-001-006Shanghai Science and Technology Commission“R&D public service platform and institutional capacity improvement project”,Grant/Award Number:21DZ2291300Shanghai Public Health Clinical Center projects,Grant/Award Number:KY-GW-2021-39,KY-GW-2019-19 and KY-GW-2019-11。
文摘Background:Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in“carrier”or“pathogenic”states.HLA DQ and HLA DR humanized mice have been used as a small animal model to study the role of two molecules during S.aureus infection.However,the contribution of HLA DP to S.aureus infection is unknown yet.Methods:In this study,we have produced HLA DP401 and HLA DRA0101 humanized mice by microinjection of C57BL/6J zygotes.Neo-floxed IAβ+/-mice were crossbred with Ella-Cre and further crossbred with HLA DP401 or HLA-DRA0101 humanized mice.After several rounds of traditional crossbreeding,we finally obtained HLA DP401-IAβ-/-and HLA DRA-IAβ-/-humanized mice,in which human DP401 or DRA0101 molecule was introduced into IAβ-/-mice deficient in endogenous murine MHC classⅡmolecules.A transnasal infection murine model of S.aureus pneumonia was induced in the humanized mice by administering 2×108CFU of S.aureus Newman dropwise into the nasal cavity.The immune responses and histopathology changes were further assessed in lungs in these infected mice.Results:We evaluated the local and systemic effects of S.aureus delivered intranasally in HLA DP401-IAβ-/-and HLA DRA-IAβ-/-transgenic mice.S.aureus Newman infection significantly increased the m RNA level of IL 12p40 in lungs in humanized mice.An increase in IFN-γand IL-6 protein was observed in HLA DRA-IAβ-/-mice.We observed a declining trend in the percentage of F4/80+macrophages in lungs in HLA DP401-IAβ-/-mice and a decreasing ratio of CD4+to CD8+T cells in lungs in IAβ-/-mice and HLA DP401-IAβ-/-mice.A decreasing ratio of Vβ3+to Vβ8+T cells was also found in the lymph node of IAβ-/-mice and HLA DP401-IAβ-/-mice.S.aureus Newman infection resulted in a weaker pathological injury in lungs in IAβ-/-genetic background mice.Conclusion:These humanized mice will be an invaluable mouse model to resolve the pathological mechanism of S.aureus pneumonia and study what role DP molecule plays in S.aureus infection.
