Objective To investigate the relationship between human cytomegalovirus(HCMV)infection and peripheral blood CD14+CD16+monocytes in the pathogenesis of coronary heart disease(CHD),and to elucidate the mechanism of path...Objective To investigate the relationship between human cytomegalovirus(HCMV)infection and peripheral blood CD14+CD16+monocytes in the pathogenesis of coronary heart disease(CHD),and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection,inflammation,and CHD,to provide a basis for the prevention,evaluation,and treatment of the disease.Methods In total,192 patients with CHD were divided into three groups:latent CHD,angina pectoris,and myocardial infarction.HCMV-IgM and-IgG antibodies were assessed using ELISA;CD14+CD16+monocytes were counted using a five-type automated hematology analyzer;mononuclear cells were assessed using fluorescence-activated cell sorting;and an automatic biochemical analyzer was used to measure the levels of triglyceride,cholesterol,high-and low-density lipoprotein cholesterols,lipoprotein,hs-CRp and Hcy.Results The positive rates of HCMV-IgM and-IgG were significantly higher in the CHD groups than in the control group.HCMV infection affects lipid metabolism to promote immune and inflammatory responses.Conclusion HCMV infection has a specific correlation with the occurrence and development of CHD.The expression of CD14+CD16+mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection.Thus,HCMV antibody as well as peripheral blood CD14+CD16+mononuclear cells can be used to monitor the occurrence and development of CHD.展开更多
AIM: Dendritomas formed by fusing cancer cells to dendritic cells have already been applied to clinical treatment trial of several types of cancers. Dendritic cells for the fusion in most trials and experiments were f...AIM: Dendritomas formed by fusing cancer cells to dendritic cells have already been applied to clinical treatment trial of several types of cancers. Dendritic cells for the fusion in most trials and experiments were from blood monocytes in standard 7-d protocol culture, which requires 5-7 d of culture with granulocyte-macrophage-colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4), followed by 2-3 d of activation with a combination of proinflammatory mediators such as tumor necrosis factorα (TNFα), interleukin-1β (IL-1β), interleukin-6 (IL-6) and prostaglandin E2 (PGE2).One study showed that mature monocyte-derived dendritic cells could be obtained within 48 h of in vitro culture with the same protocol as standard 7-d culture and referred to as FastDCs. Here we aimed to fuse human hepatocellular carcinoma cell line HCCLM3 cells with mature monocytederived dendritic cells within 48 h of in vitro culture (FastDC).METHODS: HCCLM3 cells were cultured in RPMI 1640 with 150 mL/L fetal calf serum (FCS). CD14+monocytes from healthy human peripheral blood were purified with MACS CD14 isolation kit and cultured in six-well plates in fresh complete DC medium containing RPMI-1640, 20 mL/L heat inactivated human AB serum, 2 mmol/1 L-glutamine,100 μg/mL gentamicin, 1000 U/mL GM-CSF and 500 U/mL IL-4 for 24 h, then proinflammatory mediators such as TNFα(1000 U/mL), IL-1β (10 ng/mL), IL-6 (10 ng/mL) and PGE2(1 μg/mL) were supplemented for another 24 h, and thus mature FastDCs were generated. HCCLM3 cells and FastDCs were labeled with red fluorescent dye PKH26-GL and green fluorescent dye PKH67-GL respectively. After the red fluorescent-stained HCCLM3 cells were irradiated with 50 Gy, FastDCs and irradiated HCCLM3 cells were fused in 500 mL/1 polyethylene glycol(PEG)+100 mL/L dimethyl sulfoxide (DMSO) to generate novel dendritornas. The FastDCs and novel dendritomas were immunostained with antiCD80, anti-CD86, anti-CD83, anti-HLA-DR mAbs and analyzed by fluorescence-activated cell sorting (FACS).Novel dendritomas were nucleus-stained with Hoechst 33258 and analyzed by confocal laser scanning microscopy.RESULTS: Mature FastDCs with highly expressed surface markers CD80, CD86, CD83 and HLA-DR were generated within 48 h in vitro. Novel dendritomas with dual red-green fluorescence were constructed fast and successfully, and FACS analysis showed that the fusion efficiency was 24.27% and the novel dendritomas expressed the same activation markers as FastDCs. Confocal laser scanning microscopy analysis showed representative images of dendritomas.CONCLUSION: Dendritomas can be formed fast with mature FastDCs from healthy human peripheral blood monocytes (PBMC) by incubation with GM-CSF and IL-4 for 24 h and by activation with proinflammatory mediators for an additional period of 24 h. Owing to shorter time required for in vitro DCs development, the generation of these novel dendritomas reduced labor and cost. This rapid method for formation of dendritomas may represent a new strategy for immunotherapy of hepatocellular carcinoma.展开更多
This study aimed to examine the number of circulating Toll-like receptor 4(TLR4) + CD14+ monocytes in patients with different stages of chronic kidney disease(CKD), their responses to lipopolysaccharide(LPS), and to e...This study aimed to examine the number of circulating Toll-like receptor 4(TLR4) + CD14+ monocytes in patients with different stages of chronic kidney disease(CKD), their responses to lipopolysaccharide(LPS), and to explore the potential association of the number of TLR4+CD14+ monocytes with clinical laboratory measures. The numbers of TLR4+CD14+, LPS-stimulated TNF-?+CD14+ and interleukin(IL)-6+CD14+ monocytes were determined by flow cytometry in 9 patients with stage 3 CKD, 11 with stage 4 CKD, 16 with stage 5 CKD, and 19 healthy controls(HCs). Their laboratory tests were performed by routine methods and the potential association among these measures was analyzed by Pearson's correlation analysis. The numbers of CD14+, CD14+TLR4+, LPSstimulated TNF-?+CD14+ and IL-6+CD14+ monocytes in patients with CKD were significantly less than those of HCs(all P<0.05), and were negatively associated with patient disease severity. The number of CD14+TLR4+ monocytes was positively correlated with estimated glomerular filtration rate(e GFR, P<0.001) and the levels of hematocrit(P<0.01), but negatively correlated with the levels of blood urine nitrogen, serum creatinine, and C-reactive protein(P<0.001 for all), in the CKD patients. Our data indicate that significant reduction in the number of TLR4+ monocytes and their impaired responses to LPS may be associated with the progression of CKD in Chinese patients.展开更多
目的Meta分析CD14基因-159C/T多态性与变应性鼻炎(AR)易感性关系,为AR的发病机制及疾病预防提供理论依据。方法检索PubMed、EMBASE、Cochrane等国外数据库及中国期刊全文数据库、中国科技期刊数据库、中国生物医学文献数据库、万方数字...目的Meta分析CD14基因-159C/T多态性与变应性鼻炎(AR)易感性关系,为AR的发病机制及疾病预防提供理论依据。方法检索PubMed、EMBASE、Cochrane等国外数据库及中国期刊全文数据库、中国科技期刊数据库、中国生物医学文献数据库、万方数字期刊全文数据库等中文数据库中发表的CD14基因-159C/T多态性与变应性鼻炎易感性关系的病例对照研究。根据纳入和排除标准对文献进行筛选,用Jadad quantity对纳入的文献进行偏差评估和敏感性分析,用Revman5.3软件对数据进行处理。结果根据纳入和排除标准,共纳入9篇文献,共2319名受试者,其中病例组1195例,对照组1124例。Meta分析结果显示:优势基因模型下(CC vs CT+TT),组合OR值为0.74,95%CI为[0.61,0.90],P=0.002,差异有统计学意义;超显性基因模型下(CT vs CC+TT),联合OR值为1.12,95%CI为[0.95,1.32],P=0.18,差异无统计学意义;隐性基因模型下(TT vs CC+CT),联合OR值为1.14,95%CI为[0.95,1.38],P=0.15;等位基因(C对T),合并OR值为0.95,95%CI为[0.83,1.10],P=0.50,差异无统计学意义。表明与T等位基因相比,在a位点携带C等位基因不会增加AR的易感性。优势基因模型可能增加基因型CD14基因-159C/T的AR易感性,而隐性基因模型和超显性基因模型可能不会增加基因型CD14基因-159C/T的AR易感性。结论CD14基因-159C/T多态性与AR易感性无显著相关性。基因座中的基因型可能增加AR易感性的基因模型是优势基因模型。展开更多
基金Funded by the National Natural Science Foundation of China[81471048]the Natural Science Foundation of Shandong Province[ZR2019MC059]Shandong Province Government-Sponsored Overseas Study Project.&These authors contributed equally to this work.
文摘Objective To investigate the relationship between human cytomegalovirus(HCMV)infection and peripheral blood CD14+CD16+monocytes in the pathogenesis of coronary heart disease(CHD),and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection,inflammation,and CHD,to provide a basis for the prevention,evaluation,and treatment of the disease.Methods In total,192 patients with CHD were divided into three groups:latent CHD,angina pectoris,and myocardial infarction.HCMV-IgM and-IgG antibodies were assessed using ELISA;CD14+CD16+monocytes were counted using a five-type automated hematology analyzer;mononuclear cells were assessed using fluorescence-activated cell sorting;and an automatic biochemical analyzer was used to measure the levels of triglyceride,cholesterol,high-and low-density lipoprotein cholesterols,lipoprotein,hs-CRp and Hcy.Results The positive rates of HCMV-IgM and-IgG were significantly higher in the CHD groups than in the control group.HCMV infection affects lipid metabolism to promote immune and inflammatory responses.Conclusion HCMV infection has a specific correlation with the occurrence and development of CHD.The expression of CD14+CD16+mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection.Thus,HCMV antibody as well as peripheral blood CD14+CD16+mononuclear cells can be used to monitor the occurrence and development of CHD.
基金Supported by the Key Program Foundation for Clinical Subject of the Ministry of Public Health,China (2001)
文摘AIM: Dendritomas formed by fusing cancer cells to dendritic cells have already been applied to clinical treatment trial of several types of cancers. Dendritic cells for the fusion in most trials and experiments were from blood monocytes in standard 7-d protocol culture, which requires 5-7 d of culture with granulocyte-macrophage-colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4), followed by 2-3 d of activation with a combination of proinflammatory mediators such as tumor necrosis factorα (TNFα), interleukin-1β (IL-1β), interleukin-6 (IL-6) and prostaglandin E2 (PGE2).One study showed that mature monocyte-derived dendritic cells could be obtained within 48 h of in vitro culture with the same protocol as standard 7-d culture and referred to as FastDCs. Here we aimed to fuse human hepatocellular carcinoma cell line HCCLM3 cells with mature monocytederived dendritic cells within 48 h of in vitro culture (FastDC).METHODS: HCCLM3 cells were cultured in RPMI 1640 with 150 mL/L fetal calf serum (FCS). CD14+monocytes from healthy human peripheral blood were purified with MACS CD14 isolation kit and cultured in six-well plates in fresh complete DC medium containing RPMI-1640, 20 mL/L heat inactivated human AB serum, 2 mmol/1 L-glutamine,100 μg/mL gentamicin, 1000 U/mL GM-CSF and 500 U/mL IL-4 for 24 h, then proinflammatory mediators such as TNFα(1000 U/mL), IL-1β (10 ng/mL), IL-6 (10 ng/mL) and PGE2(1 μg/mL) were supplemented for another 24 h, and thus mature FastDCs were generated. HCCLM3 cells and FastDCs were labeled with red fluorescent dye PKH26-GL and green fluorescent dye PKH67-GL respectively. After the red fluorescent-stained HCCLM3 cells were irradiated with 50 Gy, FastDCs and irradiated HCCLM3 cells were fused in 500 mL/1 polyethylene glycol(PEG)+100 mL/L dimethyl sulfoxide (DMSO) to generate novel dendritornas. The FastDCs and novel dendritomas were immunostained with antiCD80, anti-CD86, anti-CD83, anti-HLA-DR mAbs and analyzed by fluorescence-activated cell sorting (FACS).Novel dendritomas were nucleus-stained with Hoechst 33258 and analyzed by confocal laser scanning microscopy.RESULTS: Mature FastDCs with highly expressed surface markers CD80, CD86, CD83 and HLA-DR were generated within 48 h in vitro. Novel dendritomas with dual red-green fluorescence were constructed fast and successfully, and FACS analysis showed that the fusion efficiency was 24.27% and the novel dendritomas expressed the same activation markers as FastDCs. Confocal laser scanning microscopy analysis showed representative images of dendritomas.CONCLUSION: Dendritomas can be formed fast with mature FastDCs from healthy human peripheral blood monocytes (PBMC) by incubation with GM-CSF and IL-4 for 24 h and by activation with proinflammatory mediators for an additional period of 24 h. Owing to shorter time required for in vitro DCs development, the generation of these novel dendritomas reduced labor and cost. This rapid method for formation of dendritomas may represent a new strategy for immunotherapy of hepatocellular carcinoma.
文摘This study aimed to examine the number of circulating Toll-like receptor 4(TLR4) + CD14+ monocytes in patients with different stages of chronic kidney disease(CKD), their responses to lipopolysaccharide(LPS), and to explore the potential association of the number of TLR4+CD14+ monocytes with clinical laboratory measures. The numbers of TLR4+CD14+, LPS-stimulated TNF-?+CD14+ and interleukin(IL)-6+CD14+ monocytes were determined by flow cytometry in 9 patients with stage 3 CKD, 11 with stage 4 CKD, 16 with stage 5 CKD, and 19 healthy controls(HCs). Their laboratory tests were performed by routine methods and the potential association among these measures was analyzed by Pearson's correlation analysis. The numbers of CD14+, CD14+TLR4+, LPSstimulated TNF-?+CD14+ and IL-6+CD14+ monocytes in patients with CKD were significantly less than those of HCs(all P<0.05), and were negatively associated with patient disease severity. The number of CD14+TLR4+ monocytes was positively correlated with estimated glomerular filtration rate(e GFR, P<0.001) and the levels of hematocrit(P<0.01), but negatively correlated with the levels of blood urine nitrogen, serum creatinine, and C-reactive protein(P<0.001 for all), in the CKD patients. Our data indicate that significant reduction in the number of TLR4+ monocytes and their impaired responses to LPS may be associated with the progression of CKD in Chinese patients.
文摘目的Meta分析CD14基因-159C/T多态性与变应性鼻炎(AR)易感性关系,为AR的发病机制及疾病预防提供理论依据。方法检索PubMed、EMBASE、Cochrane等国外数据库及中国期刊全文数据库、中国科技期刊数据库、中国生物医学文献数据库、万方数字期刊全文数据库等中文数据库中发表的CD14基因-159C/T多态性与变应性鼻炎易感性关系的病例对照研究。根据纳入和排除标准对文献进行筛选,用Jadad quantity对纳入的文献进行偏差评估和敏感性分析,用Revman5.3软件对数据进行处理。结果根据纳入和排除标准,共纳入9篇文献,共2319名受试者,其中病例组1195例,对照组1124例。Meta分析结果显示:优势基因模型下(CC vs CT+TT),组合OR值为0.74,95%CI为[0.61,0.90],P=0.002,差异有统计学意义;超显性基因模型下(CT vs CC+TT),联合OR值为1.12,95%CI为[0.95,1.32],P=0.18,差异无统计学意义;隐性基因模型下(TT vs CC+CT),联合OR值为1.14,95%CI为[0.95,1.38],P=0.15;等位基因(C对T),合并OR值为0.95,95%CI为[0.83,1.10],P=0.50,差异无统计学意义。表明与T等位基因相比,在a位点携带C等位基因不会增加AR的易感性。优势基因模型可能增加基因型CD14基因-159C/T的AR易感性,而隐性基因模型和超显性基因模型可能不会增加基因型CD14基因-159C/T的AR易感性。结论CD14基因-159C/T多态性与AR易感性无显著相关性。基因座中的基因型可能增加AR易感性的基因模型是优势基因模型。
