BACKGROUND Immunological dysfunction-induced low-grade inflammation is regarded as one of the predominant pathogenetic mechanisms in post-infectious irritable bowel syndrome(PI-IBS).γδT cells play a crucial role in ...BACKGROUND Immunological dysfunction-induced low-grade inflammation is regarded as one of the predominant pathogenetic mechanisms in post-infectious irritable bowel syndrome(PI-IBS).γδT cells play a crucial role in innate and adaptive immunity.Adenosine receptors expressed on the surface ofγδT cells participate in intestinal inflammation and immunity regulation.AIM To investigate the role ofγδT cell regulated by adenosine 2A receptor(A2AR)in PI-IBS.METHODS The PI-IBS mouse model has been established with Trichinella spiralis(T.spiralis)infection.The intestinal A2AR and A2AR inγδT cells were detected by immunohistochemistry,and the inflammatory cytokines were measured by western blot.The role of A2AR on the isolatedγδT cells,including proliferation,apoptosis,and cytokine production,were evaluated in vitro.Their A2AR expression was measured by western blot and reverse transcription polymerase chain reaction(RT-PCR).The animals were administered with A2AR agonist,or A2AR antagonist.Besides,γδT cells were also injected back into the animals,and the parameters described above were examined,as well as the clinical features.Furthermore,the A2AR-associated signaling pathway molecules were assessed by western blot and RT-PCR.RESULTS PI-IBS mice exhibited elevated ATP content and A2AR expression(P<0.05),and suppression of A2AR enhanced PI-IBS clinical characteristics,indicated by the abdominal withdrawal reflex and colon transportation test.PI-IBS was associated with an increase in intestinal T cells,and cytokine levels of interleukin-1(IL-1),IL-6,IL-17A,and interferon-α(IFN-α).Also,γδT cells expressed A2AR in vitro and generated IL-1,IL-6,IL-17A,and IFN-α,which can be controlled by A2AR agonist and antagonist.Mechanistic studies demonstrated that the A2AR antagonist improved the function ofγδT cells through the PKA/CREB/NF-κB signaling pathway.CONCLUSION Our results revealed that A2AR contributes to the facilitation of PI-IBS by regulating the function ofγδT cells via the PKA/CREB/NF-κB signaling pathway.展开更多
AIM：To detect the expression of B cell receptor signaling pathway（BCRSP） in lacrimal gland benign lymphoepithelial lesions（LGBLEL）.METHODS：Gene microarray was used to compare whole-genome expression in lacrimal ...AIM：To detect the expression of B cell receptor signaling pathway（BCRSP） in lacrimal gland benign lymphoepithelial lesions（LGBLEL）.METHODS：Gene microarray was used to compare whole-genome expression in lacrimal gland tissues from LGBLEL patients to tissues from orbital cavernous hemangioma（control tissues）. Expression of BCRSP was confirmed by polymerase chain reaction（PCR） and immunohistochemistry. RESULTS：The expression of 22 genes of the BCRSP increased significantly in LGBLEL patients. PCR analysis showed that CD22, CR2, and BTK were all highly expressed in LGBLEL tissues. Immunohistochemical analysis showed that CR2 protein was present in LGBLEL, but CD22 and BTK proteins were negative. CR2, CD22, and BTK were not observed in the orbital cavernous hemangiomas with either PCR or immunohistochemistry. CONCLUSION：BCRSP might be involved in the pathogenesis of LGBLEL.展开更多
Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-κB and the expression of chemokine MOB-1, MCP-1genes in pancreatic acinar cells was investigated. Rat panc...Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-κB and the expression of chemokine MOB-1, MCP-1genes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-κB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10 -3 mol/L, 10 -4 mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10 -3 mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-κB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10 -3 mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10 -5 mol/L atropine) or NF-κB inhibitor (10 -2 mol/L PDTC) could obviously inhibit the activation of NF-κB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-1genes in pancreatic acinar cells in vitro through the activation of NF-κB.展开更多
Aim Immunoglobulin D （IgD） is a surface immunoglobulin that is expressed as either membrane IgD（mIgD） or secreted IgD （sIgD）. Researchers have shown that sIgD is often elevated in patients with autoimmune diseas...Aim Immunoglobulin D （IgD） is a surface immunoglobulin that is expressed as either membrane IgD（mIgD） or secreted IgD （sIgD）. Researchers have shown that sIgD is often elevated in patients with autoimmune diseases. The possible roles of sIgD on the function of peripheral blood mononuclear cells （PBMCs） in rheumatoid arthritis （RA） are still unclear and few studies have been performed. The objective of this study was to investigate the abnormal level of immunoglobulin D （IgD） and the effects of it by binding its receptor （IgDR） on peripheral blood mononuclear cells （PBMCs） in rheumatoid arthritis （RA）. Methods Blood samples were obtained from 54 RA patients and 42 healthy controls. The levels of sIgD, human soluble receptor activator of nuclear factor-KB lig- and （sRANKL）, anti-cyclic citrullinated peptide （anti-CCP）, C-reactive protein （CRP） were determined in ser- um samples by ELISA. Rheumatoid factor （RF） was detected by quantitative nephelometry. Erythrocytes sedimen- tation rate （ESR） was tested by Westergren method. IgDR and mIgD were detected by using flow cytometry. After PBMCs were cultured and treated with different concentrations of human IgD. PBMCs proliferation were measured by CCK-8, inflammatory cytokine production were assessed by inflammation antibody array, T-/B- cell subsets and IgDR expression were tested by flow cytometry. Results A significantly higher level of sIgD, mIgD and IgDR were detected in RA patients compared with healthy controls. The concentrations of sIgD were positively correlated with sRANKL, rheumatoid factor and C-reactive protein in RA patients. Strikingly, IgD could enhance the prolifer- ation of PBMCs and induce IL-lα, IL-1β, TNF-α, IL-6 and production from PBMCs. Moreover, the percentage of activated T cell subsets （ CD4 ＋ CD69 ＋ , CD4 ＋ CD154 ＋ ） and activated B cell subsets （ CD19 ＋ CD23 ＋ , CD19 ＋ CD21 ＋ , CD19 ＋ IgD ＋ and CD19- CD138 ＋ ） were increased by IgD. The percentage of unactivated T cell subset （CD4 ＋ CD62L ＋ ） and immature B cell subset （ CD19 ＋ IgM ＋ IgD- ） were decreased by IgD in PBMCs. Further- more, the expressions of IgDR on T and B cells were significantly increased by treatment with IgD. Conclusion IgD enhanced the activation of PBMCs through stimulation of IgDR, which may contribute to RA pathogenesis. IgD represents a potentially novel immunotherapeutic target for the management of RA.展开更多
基金Supported by National Natural Science Foundation of China,No.81160057,No.81860102,and No.82060102Natural Science Foundation of Hainan Province,High-level Personnel Program,No.821RC1116+1 种基金Research Project of Health Industry in Hainan Province,No.20A200066Hainan Provincial Clinical Medical Center.
文摘BACKGROUND Immunological dysfunction-induced low-grade inflammation is regarded as one of the predominant pathogenetic mechanisms in post-infectious irritable bowel syndrome(PI-IBS).γδT cells play a crucial role in innate and adaptive immunity.Adenosine receptors expressed on the surface ofγδT cells participate in intestinal inflammation and immunity regulation.AIM To investigate the role ofγδT cell regulated by adenosine 2A receptor(A2AR)in PI-IBS.METHODS The PI-IBS mouse model has been established with Trichinella spiralis(T.spiralis)infection.The intestinal A2AR and A2AR inγδT cells were detected by immunohistochemistry,and the inflammatory cytokines were measured by western blot.The role of A2AR on the isolatedγδT cells,including proliferation,apoptosis,and cytokine production,were evaluated in vitro.Their A2AR expression was measured by western blot and reverse transcription polymerase chain reaction(RT-PCR).The animals were administered with A2AR agonist,or A2AR antagonist.Besides,γδT cells were also injected back into the animals,and the parameters described above were examined,as well as the clinical features.Furthermore,the A2AR-associated signaling pathway molecules were assessed by western blot and RT-PCR.RESULTS PI-IBS mice exhibited elevated ATP content and A2AR expression(P<0.05),and suppression of A2AR enhanced PI-IBS clinical characteristics,indicated by the abdominal withdrawal reflex and colon transportation test.PI-IBS was associated with an increase in intestinal T cells,and cytokine levels of interleukin-1(IL-1),IL-6,IL-17A,and interferon-α(IFN-α).Also,γδT cells expressed A2AR in vitro and generated IL-1,IL-6,IL-17A,and IFN-α,which can be controlled by A2AR agonist and antagonist.Mechanistic studies demonstrated that the A2AR antagonist improved the function ofγδT cells through the PKA/CREB/NF-κB signaling pathway.CONCLUSION Our results revealed that A2AR contributes to the facilitation of PI-IBS by regulating the function ofγδT cells via the PKA/CREB/NF-κB signaling pathway.
基金Supported by National Natural Science Fund(No.81170875No.81371052)+1 种基金Key Discipline Leading Plan in Beijing Eye Institution(No.201512)Capital of Clinical Characteristics and the Applied Research(No.Z151100004015115)
文摘AIM：To detect the expression of B cell receptor signaling pathway（BCRSP） in lacrimal gland benign lymphoepithelial lesions（LGBLEL）.METHODS：Gene microarray was used to compare whole-genome expression in lacrimal gland tissues from LGBLEL patients to tissues from orbital cavernous hemangioma（control tissues）. Expression of BCRSP was confirmed by polymerase chain reaction（PCR） and immunohistochemistry. RESULTS：The expression of 22 genes of the BCRSP increased significantly in LGBLEL patients. PCR analysis showed that CD22, CR2, and BTK were all highly expressed in LGBLEL tissues. Immunohistochemical analysis showed that CR2 protein was present in LGBLEL, but CD22 and BTK proteins were negative. CR2, CD22, and BTK were not observed in the orbital cavernous hemangiomas with either PCR or immunohistochemistry. CONCLUSION：BCRSP might be involved in the pathogenesis of LGBLEL.
文摘Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-κB and the expression of chemokine MOB-1, MCP-1genes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-κB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10 -3 mol/L, 10 -4 mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10 -3 mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-κB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10 -3 mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10 -5 mol/L atropine) or NF-κB inhibitor (10 -2 mol/L PDTC) could obviously inhibit the activation of NF-κB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-1genes in pancreatic acinar cells in vitro through the activation of NF-κB.
文摘Aim Immunoglobulin D （IgD） is a surface immunoglobulin that is expressed as either membrane IgD（mIgD） or secreted IgD （sIgD）. Researchers have shown that sIgD is often elevated in patients with autoimmune diseases. The possible roles of sIgD on the function of peripheral blood mononuclear cells （PBMCs） in rheumatoid arthritis （RA） are still unclear and few studies have been performed. The objective of this study was to investigate the abnormal level of immunoglobulin D （IgD） and the effects of it by binding its receptor （IgDR） on peripheral blood mononuclear cells （PBMCs） in rheumatoid arthritis （RA）. Methods Blood samples were obtained from 54 RA patients and 42 healthy controls. The levels of sIgD, human soluble receptor activator of nuclear factor-KB lig- and （sRANKL）, anti-cyclic citrullinated peptide （anti-CCP）, C-reactive protein （CRP） were determined in ser- um samples by ELISA. Rheumatoid factor （RF） was detected by quantitative nephelometry. Erythrocytes sedimen- tation rate （ESR） was tested by Westergren method. IgDR and mIgD were detected by using flow cytometry. After PBMCs were cultured and treated with different concentrations of human IgD. PBMCs proliferation were measured by CCK-8, inflammatory cytokine production were assessed by inflammation antibody array, T-/B- cell subsets and IgDR expression were tested by flow cytometry. Results A significantly higher level of sIgD, mIgD and IgDR were detected in RA patients compared with healthy controls. The concentrations of sIgD were positively correlated with sRANKL, rheumatoid factor and C-reactive protein in RA patients. Strikingly, IgD could enhance the prolifer- ation of PBMCs and induce IL-lα, IL-1β, TNF-α, IL-6 and production from PBMCs. Moreover, the percentage of activated T cell subsets （ CD4 ＋ CD69 ＋ , CD4 ＋ CD154 ＋ ） and activated B cell subsets （ CD19 ＋ CD23 ＋ , CD19 ＋ CD21 ＋ , CD19 ＋ IgD ＋ and CD19- CD138 ＋ ） were increased by IgD. The percentage of unactivated T cell subset （CD4 ＋ CD62L ＋ ） and immature B cell subset （ CD19 ＋ IgM ＋ IgD- ） were decreased by IgD in PBMCs. Further- more, the expressions of IgDR on T and B cells were significantly increased by treatment with IgD. Conclusion IgD enhanced the activation of PBMCs through stimulation of IgDR, which may contribute to RA pathogenesis. IgD represents a potentially novel immunotherapeutic target for the management of RA.
