CD134, a member of the tumor necrosis factor receptor superfamily, plays a crucial role in T cell survival. In this study, peripheral blood mononuclear cells （PBMCs） stimulated with phytohemagglutinin （PHA, 50 μg...CD134, a member of the tumor necrosis factor receptor superfamily, plays a crucial role in T cell survival. In this study, peripheral blood mononuclear cells （PBMCs） stimulated with phytohemagglutinin （PHA, 50 μg/ml） were treated by CD134 mAb （1μg/ml, 5 μg/ml, 10 μg/ml） for 6 h, 12 h, 24 h and 48 h. The level of perforin mRNA was measured by reverse transcription-polymerase chain reaction （RT-PCR） technique. Our data showed that the expression of perforin mRNA in PBMCs was down-regulated by CD134 mAb in a dose-dependent manner in range of 1 μg/ml to 5 μg/ml and dropped down to its minimum on 24 h （p 〈 0.05）. The level of perforin mRNA reached a plateau when the concentration of CD134 mAb exceeded 5 μg/ml. In conclusion, CD134 mAb can inhibit perforin expression, which may enhance the ability of T cells for survival. Cellular ＆ Molecular Immunology. 2005;2（6）： 467-471.展开更多
DNA damage response （DDR） in different cell cycle status of human peripheral blood lymphocytes （PBLs） and the role of H2AX in DDR were investigated. The PBLs were stimulated into cell cycle with phytohemagglutinin...DNA damage response （DDR） in different cell cycle status of human peripheral blood lymphocytes （PBLs） and the role of H2AX in DDR were investigated. The PBLs were stimulated into cell cycle with phytohemagglutinin （PHA）. The apoptotic ratio and the phosphorylation H2AX （S139） were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin （CPT） or X-ray. The expressions of γH2AX, Bcl-2, caspase-3 and caspase-9 were detected by Western blotting. DDR in 293T cells was detected after H2AX was silenced by RNAi method. Our results showed that DNA double strand breaks （DSBs） were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment. The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes （P0.05）. The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased. No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls. It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates. γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.展开更多
探究低温对红耳龟(Trachemys scripta elegans)免疫功能和应激反应能力的影响,揭示红耳龟适应低温环境的生理学和免疫学基础,为深入了解其入侵生物学提供基础资料。采用活体背甲下静脉窦采血技术,测定了6℃、17℃和26℃条件下处理1、5和...探究低温对红耳龟(Trachemys scripta elegans)免疫功能和应激反应能力的影响,揭示红耳龟适应低温环境的生理学和免疫学基础,为深入了解其入侵生物学提供基础资料。采用活体背甲下静脉窦采血技术,测定了6℃、17℃和26℃条件下处理1、5和10 d时血液中各型白细胞百分比及嗜异性粒细胞与淋巴细胞的比值(ratio of heterophils to lymphocytes,H/L),以及第10天采血后植物血凝素(phytohemagglutinin-P,PHA-P)反应的变化。结果显示:(1)随着温度下降,第1天只有嗜碱性粒细胞的百分比显著升高(P<0.05),此差异在第5天和第10天消失(P>0.05);第5天,嗜酸性粒细胞的百分比和H/L先升后降,淋巴细胞的百分比先降后升,单核细胞的百分比下降(P<0.05);第10天,嗜酸性粒细胞的百分比上升,淋巴细胞、嗜异性粒细胞和单核细胞的百分比,以及H/L下降(P<0.05);(2)随着处理天数延长,除嗜碱性粒细胞的百分比无组内差异外(P>0.05),其他类型白细胞的百分比及H/L都有组内差异(P<0.05);(3)6℃组、17℃组和26℃组对PHA-P的反应分别在注射后48 h、24 h和48 h达到峰值,但组间差异不显著(P>0.05)。红耳龟通过调整血液中嗜酸性粒细胞等不同类型白细胞的百分比、下降的应激反应能力和提前发生反应的免疫应答来适应低温的短期胁迫。展开更多
文摘CD134, a member of the tumor necrosis factor receptor superfamily, plays a crucial role in T cell survival. In this study, peripheral blood mononuclear cells （PBMCs） stimulated with phytohemagglutinin （PHA, 50 μg/ml） were treated by CD134 mAb （1μg/ml, 5 μg/ml, 10 μg/ml） for 6 h, 12 h, 24 h and 48 h. The level of perforin mRNA was measured by reverse transcription-polymerase chain reaction （RT-PCR） technique. Our data showed that the expression of perforin mRNA in PBMCs was down-regulated by CD134 mAb in a dose-dependent manner in range of 1 μg/ml to 5 μg/ml and dropped down to its minimum on 24 h （p 〈 0.05）. The level of perforin mRNA reached a plateau when the concentration of CD134 mAb exceeded 5 μg/ml. In conclusion, CD134 mAb can inhibit perforin expression, which may enhance the ability of T cells for survival. Cellular ＆ Molecular Immunology. 2005;2（6）： 467-471.
基金supported by grants from Program 973 from Ministry of ScienceTechnology of China (Nos. 2004CB518705, 2009CB5218702)the National Natural Sciences Foundation of China (Nos. 30872472, 30800569)
文摘DNA damage response （DDR） in different cell cycle status of human peripheral blood lymphocytes （PBLs） and the role of H2AX in DDR were investigated. The PBLs were stimulated into cell cycle with phytohemagglutinin （PHA）. The apoptotic ratio and the phosphorylation H2AX （S139） were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin （CPT） or X-ray. The expressions of γH2AX, Bcl-2, caspase-3 and caspase-9 were detected by Western blotting. DDR in 293T cells was detected after H2AX was silenced by RNAi method. Our results showed that DNA double strand breaks （DSBs） were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment. The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes （P0.05）. The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased. No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls. It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates. γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.
文摘探究低温对红耳龟(Trachemys scripta elegans)免疫功能和应激反应能力的影响,揭示红耳龟适应低温环境的生理学和免疫学基础,为深入了解其入侵生物学提供基础资料。采用活体背甲下静脉窦采血技术,测定了6℃、17℃和26℃条件下处理1、5和10 d时血液中各型白细胞百分比及嗜异性粒细胞与淋巴细胞的比值(ratio of heterophils to lymphocytes,H/L),以及第10天采血后植物血凝素(phytohemagglutinin-P,PHA-P)反应的变化。结果显示:(1)随着温度下降,第1天只有嗜碱性粒细胞的百分比显著升高(P<0.05),此差异在第5天和第10天消失(P>0.05);第5天,嗜酸性粒细胞的百分比和H/L先升后降,淋巴细胞的百分比先降后升,单核细胞的百分比下降(P<0.05);第10天,嗜酸性粒细胞的百分比上升,淋巴细胞、嗜异性粒细胞和单核细胞的百分比,以及H/L下降(P<0.05);(2)随着处理天数延长,除嗜碱性粒细胞的百分比无组内差异外(P>0.05),其他类型白细胞的百分比及H/L都有组内差异(P<0.05);(3)6℃组、17℃组和26℃组对PHA-P的反应分别在注射后48 h、24 h和48 h达到峰值,但组间差异不显著(P>0.05)。红耳龟通过调整血液中嗜酸性粒细胞等不同类型白细胞的百分比、下降的应激反应能力和提前发生反应的免疫应答来适应低温的短期胁迫。
