用醋酶染色法、ELISA法、被动保护试验等手段与方法,监测了ANAE阳性T淋巴细胞抗LPS抗体产生的情况及其保护性。试验说明,在LPS免疫后7 d T淋巴细胞明显增多;于14～21 d后抗LPS抗体逐渐产生;抗LPS抗体有较好的保护性,LPS是主要的保护性抗...用醋酶染色法、ELISA法、被动保护试验等手段与方法,监测了ANAE阳性T淋巴细胞抗LPS抗体产生的情况及其保护性。试验说明,在LPS免疫后7 d T淋巴细胞明显增多;于14～21 d后抗LPS抗体逐渐产生;抗LPS抗体有较好的保护性,LPS是主要的保护性抗原,LPS可激活细胞免疫和体液免疫。展开更多
AIM： To investigate the effect of Lactobacillus bulgaricus （LBG） on the Toll-like receptor 4 （TLR4） pathway and interleukin-8 （IL-8） production in SGC-7901 cells treated with Helicobacter pyloriSydney strain 1 ...AIM： To investigate the effect of Lactobacillus bulgaricus （LBG） on the Toll-like receptor 4 （TLR4） pathway and interleukin-8 （IL-8） production in SGC-7901 cells treated with Helicobacter pyloriSydney strain 1 lipopolysaccharide （HpyloriSS1-LPS）. METHODS： SGC-7901 cells were treated with HpyloriSS1-LPS in the presence or absence of pretreatment for 1 h with viable LBG or supernatant recovered from LBG culture MRS broth （LBG-s）. Cellular lysates were prepared for Western blot with anti-TLR4, anti-transforming growth factor β-activated kinase 1 （TAK1）, anti-phospho-TAK1, anti-nuclear factor κB （NF-κB）, anti-p38 mitogen-activated protein kinase （p38MAPK）, and anti-phospho-p38MAPK antibodies. The amount of IL-8 in cell culture medium was measured by ELISA. RESULTS： H pyloriSS1-LPS up-regulated the expression of TLR4, stimulated the phosphorylation of TAKI, subsequently enhanced the activation of NF- κB and the phosphorylation of p38MAPK in a time- dependent manner, leading to augmentation of IL-8 production in SGC-7901 cells. Viable LBG or LBG-s pretreatment attenuated the expression of TLR4, inhibited the phosphorylation of TAK1 and p38MAPK, prevented the activation of NF-κB, and consequently blocked IL-8 production.CONCLUSION： H py/oriSS1-LPS induces IL-8 production through activating TLR4 signaling in SGC-7901 cells and viable LBG or LBG-s prevents H pyloriSS1-LPS-mediated IL-8 production via inhibition of the TLR4 pathway.展开更多
AIM： TO investigate the effect of interaction between enteric epithelial cells and lymphocytes of Peyer＇s patch on the release of nitric oxide （NO） and IL-6 in response to Shigella lipopolysaccharide （LPS）. METH...AIM： TO investigate the effect of interaction between enteric epithelial cells and lymphocytes of Peyer＇s patch on the release of nitric oxide （NO） and IL-6 in response to Shigella lipopolysaccharide （LPS）. METHODS： Human colonic epithelial cells （Caco-2） were mixed cocultured with lymphocytes of Peyer＇s patch from wild-type （C57 mice） and inducible NO synthase knockout mice, and challenged with Shigella F2a-12 LPS. Release of NO and raiL-6 was measured by Griess colorimetric assay and enzyme-linked immunosorbent assay （ELISA）, respectively. RESULTS： In the absence of LPS challenge, NO was detected in the culture medium of Caco-2 epithelial cells but not in lymphocytes of Peyer＇s patch, and the NO release was further up-regulated in both cocultures with lymphocytes from either the wild-type or iNOS knockout mice, with a significantly higher level observed in the coculture with iNOS knockout lymphocytes. After Shigella F2a-12 LPS challenge for 24-h, NO production was significantly increased in both Caco-2 alone and the coculture with lymphocytes of Peyer＇s patch from the wild-type mice but not from iNOS knockout mice. LPS was found to stimulate the release of mIL-6 from lymphocytes, which was suppressed by coculture with Caco-2 epithelial cells. The LPS-induced mIL-6 production in lymphocytes from iNOS knockout mice was significantly greater than that from the wild-type mice. CONCLUSION： Lymphocytes of Peyer＇s patch maintain a constitutive basal level of NO production from the enteric epithelial cell Caco-2. LPS-induced mIL-6 release from lymphocytes of Peyer＇s patch is suppressed by the cocultured epithelial cells. While no changes are detectable in NO production in lymphocytes from both wild-type and iNOS knockout mice before and after LPS challenge, NO from lymphocytes appears to play an inhibitory role in epithelial NO release and their own mIL-6 release in response to LPS.展开更多
The pattern recognition proteins （PRPs） play a major role in immune response of crustacean to resist pathogens. In the present study, as one of PRPs, lipopolysaccharide and 13-1, 3-glucan binding protein （LGBP） ge...The pattern recognition proteins （PRPs） play a major role in immune response of crustacean to resist pathogens. In the present study, as one of PRPs, lipopolysaccharide and 13-1, 3-glucan binding protein （LGBP） gene in the ridge tail white prawn （Exopalaemon carinicauda） （EcLGBP） was isolated. The full-length cDNA of EcLGBP was 1338 bp, encoding a polypeptide of 366 amino acid residules. The deduced amino acid sequence of EcLGBP shared high similarities with LGBP and BGBP from other crus- taceans. Some conservative domains were predicted in EcLGBP sequence. EcLGBP constitutively expressed in most tissues at dif- ferent levels, and the highest expression was observed in hepatopancreas. With infection time, the cumulative mortality increased gradually followed by the proliferation of Vibrio parahaemolyticus and white spot syndrome virus （WSSV）. The expression of EcLGBP in response to E parahaemolyticus infection was up-regulated in hemocytes and hepatopancreas, and the up-regulation in hepatopancreas was earlier than that in hemocytes. EcLGBP expression after WSSV infection increased at 3 h, then significantly decreased in both hemocytes and hepatopancreas. The results indicated that EcLGBP was involved in the immune defense against bacterial and viral infections.展开更多
As a potent pleiotropic cytokine, tumor necrosis factor-alpha （TNF-ct） plays an important role in innate immune responses. The cDNA sequence and genomic structure of the TNF-α gene （AjTNF-c0 in the Japanese eel （...As a potent pleiotropic cytokine, tumor necrosis factor-alpha （TNF-ct） plays an important role in innate immune responses. The cDNA sequence and genomic structure of the TNF-α gene （AjTNF-c0 in the Japanese eel （Anguilla japonica） were identified and characterized. The full-length AjTNF-α cDNA was 1 546 bp, including a 5＇-untranslated region （UTR） of 13 bp, a 3＇-UTR of 879 bp and an open reading frame of 654 bp encoding a protein of 218 amino acids. The full-length genomic sequence of AjTNF-ct was 2 392 bp and included four exons and three introns. The putative AjTNF-α protein contained TNF family signature motifs, including a protease cleavage site, a transmembrane domain and two conserved cysteine residues. Quantitative real-time reverse transcription PCR analysis revealed AjTNF-α expression in a wide range of tissues, with predominant expression in blood and liver. Lower levels of expression were seen in spleen, gills, kidney, intestine, heart, and skin, with very low levels in muscle. The modulation of AjTNF-ct expression after injection of eels with lipopolysaccharide （LPS）, the viral mimic, poly I：C, or Aeromonas hydrophila was assessed in blood, liver, and kidney. In blood, TNF-α mRNA levels increased rapidly and then rapidly decreased after stimulation with LPS, poly I：C or A. hydrophila. However, the response to LPS and A. hydrophila peaked at 6 h while for poly I：C the peak was at 12 h. In liver, after injection with A. hydrophila, an up- and down-regulation of AjTNF-ct expression occurred twice, peaking at 6 h and 24 h, respectively. No remarkable increase of AjTNF-α expression appeared in liver until 72 h after LPS or poly I：C treatment. In kidney, ApiNF-α expression increased significantly only at 72 h post-stimulation with LPS orA. hydrophila. Our results suggest that AjTNF-α plays an important role in fish in the defense against viral and bacterial infection.展开更多
基金Basic Research Project of Science and Technology Offi ce of Sichuan Province, No. 04JY029-090-1
文摘AIM： To investigate the effect of Lactobacillus bulgaricus （LBG） on the Toll-like receptor 4 （TLR4） pathway and interleukin-8 （IL-8） production in SGC-7901 cells treated with Helicobacter pyloriSydney strain 1 lipopolysaccharide （HpyloriSS1-LPS）. METHODS： SGC-7901 cells were treated with HpyloriSS1-LPS in the presence or absence of pretreatment for 1 h with viable LBG or supernatant recovered from LBG culture MRS broth （LBG-s）. Cellular lysates were prepared for Western blot with anti-TLR4, anti-transforming growth factor β-activated kinase 1 （TAK1）, anti-phospho-TAK1, anti-nuclear factor κB （NF-κB）, anti-p38 mitogen-activated protein kinase （p38MAPK）, and anti-phospho-p38MAPK antibodies. The amount of IL-8 in cell culture medium was measured by ELISA. RESULTS： H pyloriSS1-LPS up-regulated the expression of TLR4, stimulated the phosphorylation of TAKI, subsequently enhanced the activation of NF- κB and the phosphorylation of p38MAPK in a time- dependent manner, leading to augmentation of IL-8 production in SGC-7901 cells. Viable LBG or LBG-s pretreatment attenuated the expression of TLR4, inhibited the phosphorylation of TAK1 and p38MAPK, prevented the activation of NF-κB, and consequently blocked IL-8 production.CONCLUSION： H py/oriSS1-LPS induces IL-8 production through activating TLR4 signaling in SGC-7901 cells and viable LBG or LBG-s prevents H pyloriSS1-LPS-mediated IL-8 production via inhibition of the TLR4 pathway.
基金Supported by Strategic Program of Chinese University of Hong KongDistinguished Young Investigator Fund of the National Natural Science Foundation of China, No. 30029002
文摘AIM： TO investigate the effect of interaction between enteric epithelial cells and lymphocytes of Peyer＇s patch on the release of nitric oxide （NO） and IL-6 in response to Shigella lipopolysaccharide （LPS）. METHODS： Human colonic epithelial cells （Caco-2） were mixed cocultured with lymphocytes of Peyer＇s patch from wild-type （C57 mice） and inducible NO synthase knockout mice, and challenged with Shigella F2a-12 LPS. Release of NO and raiL-6 was measured by Griess colorimetric assay and enzyme-linked immunosorbent assay （ELISA）, respectively. RESULTS： In the absence of LPS challenge, NO was detected in the culture medium of Caco-2 epithelial cells but not in lymphocytes of Peyer＇s patch, and the NO release was further up-regulated in both cocultures with lymphocytes from either the wild-type or iNOS knockout mice, with a significantly higher level observed in the coculture with iNOS knockout lymphocytes. After Shigella F2a-12 LPS challenge for 24-h, NO production was significantly increased in both Caco-2 alone and the coculture with lymphocytes of Peyer＇s patch from the wild-type mice but not from iNOS knockout mice. LPS was found to stimulate the release of mIL-6 from lymphocytes, which was suppressed by coculture with Caco-2 epithelial cells. The LPS-induced mIL-6 production in lymphocytes from iNOS knockout mice was significantly greater than that from the wild-type mice. CONCLUSION： Lymphocytes of Peyer＇s patch maintain a constitutive basal level of NO production from the enteric epithelial cell Caco-2. LPS-induced mIL-6 release from lymphocytes of Peyer＇s patch is suppressed by the cocultured epithelial cells. While no changes are detectable in NO production in lymphocytes from both wild-type and iNOS knockout mice before and after LPS challenge, NO from lymphocytes appears to play an inhibitory role in epithelial NO release and their own mIL-6 release in response to LPS.
基金supported by the earmarked fund for National ‘863’ Project of China (No.2012AA10A 409)Modern Agro-industry Technology Research System (No.CARS-47)+1 种基金Special Fund for Agro-scientific Research in the Public Interest (No.201103034)Independent Innovation Foundation of Shandong Province (No.2013CXC80202)
文摘The pattern recognition proteins （PRPs） play a major role in immune response of crustacean to resist pathogens. In the present study, as one of PRPs, lipopolysaccharide and 13-1, 3-glucan binding protein （LGBP） gene in the ridge tail white prawn （Exopalaemon carinicauda） （EcLGBP） was isolated. The full-length cDNA of EcLGBP was 1338 bp, encoding a polypeptide of 366 amino acid residules. The deduced amino acid sequence of EcLGBP shared high similarities with LGBP and BGBP from other crus- taceans. Some conservative domains were predicted in EcLGBP sequence. EcLGBP constitutively expressed in most tissues at dif- ferent levels, and the highest expression was observed in hepatopancreas. With infection time, the cumulative mortality increased gradually followed by the proliferation of Vibrio parahaemolyticus and white spot syndrome virus （WSSV）. The expression of EcLGBP in response to E parahaemolyticus infection was up-regulated in hemocytes and hepatopancreas, and the up-regulation in hepatopancreas was earlier than that in hemocytes. EcLGBP expression after WSSV infection increased at 3 h, then significantly decreased in both hemocytes and hepatopancreas. The results indicated that EcLGBP was involved in the immune defense against bacterial and viral infections.
基金Supported by the Ocean and Fishery Bureau of Fujian Province(No.201212140006)the Scientific Research Foundation of Jimei University,China(No.ZQ2008013)+1 种基金the National Natural Science Foundation of China(No.31272685)the Program of the Education Department of Fujian Province(No.JA11150)
文摘As a potent pleiotropic cytokine, tumor necrosis factor-alpha （TNF-ct） plays an important role in innate immune responses. The cDNA sequence and genomic structure of the TNF-α gene （AjTNF-c0 in the Japanese eel （Anguilla japonica） were identified and characterized. The full-length AjTNF-α cDNA was 1 546 bp, including a 5＇-untranslated region （UTR） of 13 bp, a 3＇-UTR of 879 bp and an open reading frame of 654 bp encoding a protein of 218 amino acids. The full-length genomic sequence of AjTNF-ct was 2 392 bp and included four exons and three introns. The putative AjTNF-α protein contained TNF family signature motifs, including a protease cleavage site, a transmembrane domain and two conserved cysteine residues. Quantitative real-time reverse transcription PCR analysis revealed AjTNF-α expression in a wide range of tissues, with predominant expression in blood and liver. Lower levels of expression were seen in spleen, gills, kidney, intestine, heart, and skin, with very low levels in muscle. The modulation of AjTNF-ct expression after injection of eels with lipopolysaccharide （LPS）, the viral mimic, poly I：C, or Aeromonas hydrophila was assessed in blood, liver, and kidney. In blood, TNF-α mRNA levels increased rapidly and then rapidly decreased after stimulation with LPS, poly I：C or A. hydrophila. However, the response to LPS and A. hydrophila peaked at 6 h while for poly I：C the peak was at 12 h. In liver, after injection with A. hydrophila, an up- and down-regulation of AjTNF-ct expression occurred twice, peaking at 6 h and 24 h, respectively. No remarkable increase of AjTNF-α expression appeared in liver until 72 h after LPS or poly I：C treatment. In kidney, ApiNF-α expression increased significantly only at 72 h post-stimulation with LPS orA. hydrophila. Our results suggest that AjTNF-α plays an important role in fish in the defense against viral and bacterial infection.
