Research progress on the role of costimulatory signaling pathway in xenotransplantation / 器官移植

Organ shortage is a critical factor limiting the development of organ transplantation. Xenotransplantation is expected to resolve the problem of organ shortage, which has become a new research hotspot. Study of costimulatory signaling pathway related to T cell regulation is a hot topic in terms of immunity of xenotransplantation. Since the discovery of costimulatory molecule CD28, multiple costimulatory molecules have been identified, including costimulatory and coinhibitory receptors and their related ligands. Specific T cell activation of donors is the key factor leading to acute immune rejection. The expression and induction of costimulatory molecules on T cells differ during different immune stages, and these costimulatory molecules play a key role in maintaining T cell tolerance and the balance of T cell immune response. At present, increasing attention has been diverted to the role of costimulatory signaling pathway in organ transplantation. In this article, the latest research progress in costimulatory signaling pathway related to xenotransplantation immunity was reviewed, aiming to provide reference for the optimization of xenotransplantation immunosuppression regimen.

Role of inducible costimulatory molecules (ICOS) and related cytokines in immune regulation of Echinococcus granulosus infections in mice / 中国血吸虫病防治杂志

Objective To investigate the roles of inducible costimulatory molecules (ICOS) and related cytokines in the immune regulation of Echinococcus granulosus infections in mice. Methods Eighty BALB/c mice (weight 18–22 g) were divided into the control and infection groups, of 40 animals in each group. E. granulosus infection was modeled in mice by intraperitoneal injection of 10 000 protoscoleces per mouse. Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) and peripheral interleukin-4 (IL-4) and IL-10 levels were measured 2, 8, 30, 60, 180 days post-infection. Mouse liver specimens were excised for hematoxylin-eosin (HE) staining and immunostaining, and ICOS expression was quantified in mouse liver specimens using quantitative real-time PCR (qPCR) assay. Results There were no significant differences in serum ALT (F = 12.082, P < 0.05), AST (F = 6.347, P < 0.05) or ALP levels (F = 52.186, P < 0.05) in mice 2, 8, 30, 60 and 180 days post-infection with E. granulosus. The serum ALT levels were significantly higher in the infection group than in the control group 2 [(61.72 ± 9.89) vs. (50.65 ± 4.67)U/L, P < 0.05] and 30 days post-infection [(80.61 ± 23.71)vs.(67.75 ± 9.79)U/L, P < 0.05], and the serum ALT levels were significantly higher in the infection group than in the control group 2 [(181.06 ± 60.61) vs.(115.58 ± 17.66)U/L, P < 0.05] and 180 days post-infection [(137.84 ± 29.01) vs. (108.05 ± 10.33) U/L, P < 0.05], while greater serum ALP levels were measured in the infection group than in the control group 2 [(162.90 ± 21.04)vs.(64.54 ± 5.99)U/L, P < 0.05], 8[(176.36 ± 24.56) vs. (62.70 ± 9.21)U/L, P < 0.05] and 30 days post-infection [(138.86 ± 13.59) vs. (58.60 ± 5.28) U/L, P < 0.05]. A few inflammatory cells were seen in mouse liver in the infection group 30 days post-infection, and no apparent changes were found in the mouse hepatic structure 60 days post-infection. On day 180 post-infection, a large number of epithelium-like cells presented fibrotic growth in mouse liver in the cyst-infiltrating regions, with cuticula formation seen, and plenty of red cells were present in lesions and hepatocyte space. Positive ICOS expression was detected in mouse liver in the infection group, with ICOS-positive cells predominantly seen in the cytoplasm of the hepatocyte, and the ICOS expression increased over time. The relative ICOS mRNA expression was 2.732 ± 0.094 on day 180 post-infection, which was significantly greater than that on day 2 postinfection (0.746 ± 0.049). There were no significant differences in serum IL-4 or IL-10 levels at different time points after E. granulosus infections, while the serum IL-4 and IL-10 levels peaked in the infection group 180 days and 60 days post-infection, respectively. Higher serum IL-4 levels were measured in the infection group than in the control group 8 [(22.50 ± 3.24) vs. (5.82 ± 0.49) pg/mL, P < 0.05], 30 [(15.49 ± 4.73) vs. (5.10 ± 1.38) pg/mL, P < 0.05], 60 [(36.93 ± 6.14) vs. (4.13 ± 1.19) pg/mL, P < 0.05] and 180 days post-infection [(198.35 ± 0.70) vs. (4.19 ± 0.98) pg/mL, P < 0.05], and higher IL-10 levels were measured in the infection group than in the control group 2 [(4.84 ± 1.91) vs. (2.11 ± 1.03) pg/mL, P < 0.05], 8 [(44.72 ± 14.63) vs. (3.16 ± 0.60) pg/mL, P < 0.05], 30 [(25.47 ± 8.00) vs. (3.83 ± 1.87) pg/mL, P < 0.05], 60 [(187.16 ± 60.44) vs. (3.69 ± 1.05) pg/mL, P < 0.05] and 180 days post-infection [(85.40 ± 7.15) vs. (3.25 ± 0.93) pg/mL, P < 0.05]. Conclusions High ICOS expression is present in the liver of mice with E. granulosus infections. The positive ICOS expression and immune activation levels increase with the time of E. granulosus infections, leading to aggravation of hepatocyte injury caused by inflammation.

Changes and significance of monocytic myeloid-derived suppressor cells during acute phase of Kawasaki disease / 中华微生物学和免疫学杂志

Objective:To investigate the changes of monocytic myeloid-derived suppressor cells (M-MDSC) in children with acute Kawasaki disease (KD) and its roles in the immunological pathogenesis of KD.Methods:A total of 38 children with acute KD were enrolled in the present study and 32 age-matched healthy children were selected as control group. The proportions of HLA-DR -CD11b + CD33 + CD15 -CD14 + M-MDSC and CD4 + CD25 + CD127 - regulatory T cells (Treg) in peripheral blood, concentrations of reactive oxygen species (ROS) and expression of arginase-1 (Arg-1), CD39, CD73, CD40, CD40L and CCR5 at protein levels were detected by flow cytometry. Quantitative real-time PCR was used to evaluate the transcription levels of inducible nitric oxide synthase (iNOS) in M-MDSC and the transcription levels of cytotoxic T-lymphocyte associated antigen 4 (CTLA4) and lymphocyte-activation gene 3 (LAG3) in Treg. Concentrations of NO, CCL3, CCL4, CCL5, IL-10 and TGF-β in the supernatants of cell culture were measured by ELISA. Results:(1) The proportion of HLA-DR -CD11b + CD33 + CD15 -CD14 + M-MDSC, the concentration of intracellular ROS and the expression of iNOS, CD39 and CD73 in M-MDSC decreased significantly in patients with acute KD as compared with those in the control group ( P<0.05), and the concentrations of NO, IL-10 and TGF-β in culture supernatant of M-MDSC were lower than those in the control group upon lipopolysaccharide (LPS) stimulation for 48 h ( P<0.05). All of the aforementioned indexes restored to some extent after intravenous immunoglobulin (IVIG) therapy ( P<0.05). No statistical differences were found in Arg-1 expression between healthy controls and patients with KD before or after IVIG therapy ( P<0.05). (2) CD40 expression on M-MDSC was significantly lower in the acute KD group than in the control group ( P<0.05). The concentrations of CCL3, CCL4 and CCL5 in the culture supernatants of M-MDSC were lower in the acute KD group than in the control group after LPS stimulation ( P<0.05). With IVIG treatment, all of the indexes were up-regulated significantly ( P<0.05), although CD40 expression was still lower in the acute KD group than in the control group ( P<0.05). (3) The proportion of CD4 + CD25 + CD127 -Treg and the expression of CTLA4, LAG3, CD40L and CCR5 reduced significantly in patients with acute KD as compared those in healthy controls ( P<0.05), and all increased remarkably after IVIG therapy ( P<0.05). Pearson correlation analysis showed a positive correlation between the proportions of M-MDSC and Treg in patients with acute KD ( r=0.58, P<0.05). Conclusions:Insufficiency and impaired function of M-MDSC might be a major cause of immune dysfunction in patients with acute KD.

Correlation between T follicular helper cells and eosinophilic gastroenteritis in children / 中华实用儿科临床杂志

Objective:To investigate the role and clinical significance of T follicular helper (Tfh) cells in the pathogenesis of eosinophilic gastroenteritis (EG) in children.Methods:A total of 17 children diagnosed with EG in the Department of Gastroenterology, Children′s Hospital of Nanjing medical University from October 2018 to January 2020 were recruited as EG group.During the same period, 15 children diagnosed with colon polyps were included as control group.Flow cytometry was used to detect Tfh cells and their functional molecules, including inducible costimulatory molecules (ICOS) and programmed cell death protein 1 (PD-1) in peripheral blood of the 2 groups.Results:The median (interquartile range) of Tfh cell frequency in peripheral blood of children with EG group was 7.3 (2.6)%, which was significantly higher than that of controls 2.8 (1.4)% ( P<0.05). There were significant differences in the median (interquartile range) of ICOS [1.5 (1.3)% vs.0.1 (0.2)%] and PD-1 expressions [1.8 (3.2)% vs.0.7 (0.6)%] on Tfh cells between children with EG group and control group (all P<0.05). The frequency of Tfh cells in the peripheral blood of children with EG was positively correlated with the expressions of ICOS ( r=0.746, P<0.05) and PD-1 ( r=0.893, P<0.05), and immunoglobulin E (IgE) level ( r=0.587, P<0.05). Conclusions:The frequency of Tfh cells in peripheral blood of children with EG significantly increases, which are proliferative and overexpressed with ICOS and PD-1.Moreover, the frequency of Tfh cells of EG patients is positively correlated with the level of IgE.The abnormal expression of Tfh cells may play a promoting role in the mechanism of EG.

B7/CD28 costimulatory molecules involved in the immune mechanism of kidney disease / 国际儿科学杂志

The body′s specific immune response is a very complicated process involving a series of immune cells and molecules that regulate and restrict each other.At present, the pathogenesis of most kidney diseases is not clear.B7(CD80)is located on antigen-presenting cells that regulate CD4+ and CD8+ T cells and play a role in enhancing or amplifying immune responses by binding to the cellular glycoprotein CD28.It also binds to cytotoxic T lymphocyte-Antigen4(CTLA-4)to inhibit the immune response.In general, renal tissue has no or low expression of B7.However, some glomerular diseases are associated with increased B7, which reduces the ability of podocytes to attach to the glomerular basement membrane and increases inflammation and renal fibrosis.When the B7-CTLA-4 interaction occurs, the immune response is attenuated.The disease can be prevented by blocking CD28 or enhancing the CTLA-4 signal.This article reviewed the role of costimulatory molecule B7/CD28 in the pathogenesis of kidney diseases, such as pod-cell damage, primary glomerulonephritis, purpura nephritis, lupus nephritis.Furthermore, it discussed the feasibility of B7 blockers were used as targeted therapies for kidney diseases.

Ethanol extracts of Lepidium meyenii Walp (maca) from two different areas in Xinjiang promote the maturation of mouse macrophages and dendritic cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of ethanol extracts of Lepidium meyenii Walp (LMEE) from two different areas in Xinjiang on the maturation of mouse macrophages (RAW264. 7 cells) and dendritic cells (DCs). Methods Ethanol extracts of LMEE from Tashikuergan County (Ta xian) and A La gou of Xinjiang were prepared and named as LMEE-T and LMEE-A, respectively. RAW264. 7 cells and bone marrow-derived DCs from C57BL/6 mice were treated with different concentrations of LMEE-T/A. The viability of RAW264. 7 cells was analyzed by MTT assay. Expression of costimulatory molecules and MHCⅠ on the surface of RAW264. 7 cells and DCs was detected by flow cytometry. Secretion of cytokines and the release of nitrogen oxide (NO) were measured by ELISA and Griess method, respectively. Results LMEE-T/A had no significant influence on the viability of RAW264. 7 cells when the concentration was lower than 1 mg/ml. Treating RAW264. 7 cells with LMEE-T/A promoted surface molecule expression, cytokine secretion and NO release through TLR4 signaling pathway in a dose-dependent manner. Moreover, LMEE-T was more potent than LMEE-A. LMEE-T/A at the concentration of 0. 4 mg/ml promoted the expression of DC surface molecules and the secretion of cytokines. Infrared and ultraviolet spectra showed that LMEE-A and LMEE-T contained polysaccharides, macaenes, macamides and flavanols. Compared with LMEE-A, LMEE-T contained more benzene ring compounds but less polysaccharides. Conclusion Both LMEE-T and LMEE-A could activate RAW264. 7 cells and promote the maturation of DCs. The differences between their effects might be related to the differences in their contents.

Expression and clinical significance of costimulatory molecule B7-H4 and programmed death ligand 1 in bladder cancer / 中华泌尿外科杂志

Objective To detect the expression of costimulatory molecules B7-H4 and programmed death ligand 1 (PD-L1) in bladder cancer,and to explore the correlation between them and clinicopathological features of bladder cancer.Methods Immunohistochemical staining was used to detect the expression of B7-H4 and PD-L1 in 98 cases of bladder urothelial carcinoma,which were confirmed by pathology from August 2014 to December 2015 in our hospital.There were 23 females,aged 45-82 years,with an average age of 67.8 years.Among them,42 cases of adjacent normal tissues were used as controls.The clinical stage,histological grade and recurrence of bladder cancer were collected,and the correlation between them was analyzed.Results The positive rates of B7-H4 and PD-L1 in bladder urothelial carcinoma were 54.1％ (53/98) and 59.2％ (58/98),respectively,and there was no expression in normal bladder tissues (P ＜ 0.05).The positive expression rate of B7-H4 in muscle invasive bladder cancer (MIBC) patients was higher than that in non-muscle invasive bladder cancer (NMIBC) [73.5％ (25/34) vs.43.8％ (28/64),P =0.005].The positive expression rate of B7-H4 in high-grade patients was higher than that of low-grade [70.0％ (21/30) vs.47.1％ (32/68),P =0.036].The expression rate of B7-H4 in high-risk group was higher than that of low-intermediate risk group [57.1％ (20/35) vs.27.6％ (8/29),P =0.018].The positive expression rate of PD-L1 in patients with MIBC was higher than that in NMIBC [79.4％ (27/34) vs.48.4％ (31/64),P =0.003].The PD-L1 expression rate of histological high-level group was higher than that of low-level group [73.3％ (22/30) vs.52.9％ (36/68)],but the difference was not statistically significant (P =0.058).The PD-L1 expression rate in high-risk group was 68.6％ (24/35),and also higher than low-middle group 24.1％ (7/29) (P ＜ 0.05).There was a positive correlation between the expression of B7-H4 and PD-L1 in bladder urothelial carcinoma (r =0.318,P=0.002).The combined recurrence rate of the two groups was significantly higher than that of the negative expression of the two groups [66.7％ (14/21) vs.30.8％ (8/26),P=0.014].Conclusions The expression of B7-H4 and PD-L1 is up-regulated in bladder urothelial carcinoma,which is closely related to the clinical stage,histological grade,risk classification and recurrence of NMIBC.

Effect of human umbilical cord mesenchymal stem cells on CD4+T cells in liver after hepatic ischemia-reperfusion injury in mice / 器官移植

Objective To investigate the effect of human umbilical cord mesenchymal stem cells (HUC-MSCs) on CD4+T cells in liver after hepatic ischemia-reperfusion injury (HIRI) in mice. Methods Two hundred and twenty-five mice were randomly divided into sham group, control group and MSC group, with 75 mice in each group. HIRI model mice were used in MSC group and control group. HUC-MSCs were injected in MSC group through inferior vena cava. Normal saline was injected in control group through inferior vena cava. Only laparotomy and abdominal closure were performed in sham group without blood vessel clipping. At 6, 12 and 24 h after operation, 15 mice of each group were randomly selected to sample eyeball blood and liver tissues, and the 30 mice left in each group were used to extract intrahepatic mononuclear cells. The number of intrahepatic mononuclear cells, percentage, number and positive rate of CD4+T cells in the mice of various groups at different time points were compared. The content of interleukin (IL)-17 in serum and liver tissue as well as expression levels of costimulatory molecules B7-1 and B7-2 messenger RNA (mRNA) in liver tissues of the mice at different time points were compared. Results At 12 and 24 h after operation, the number of intrahepatic mononuclear cells of control group was significantly higher than that of sham group, while the number of intrahepatic mononuclear cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the percentage, number and positive rate of CD4+T cells of control group were significantly higher than those of sham group (all P<0.01), while the percentage of CD4+T cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 12 and 24 h after operation, the number and positive rate of CD4+T cells of MSC group were significantly lower than those of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the IL-17 contents in serum and liver tissues of control group were higher than those of sham group (all P<0.01), while the IL-17 contents in serum and liver tissues of MSC group were lower than those of control group (all P<0.01). At 6 h after operation, the mRNA expression level of B7-2 of control group was higher than that of sham group (P<0.05). At 12 and 24 h after operation, the mRNA expression levels of B7-1 and B7-2 of control group were higher than those of sham group (all P<0.01), while the mRNA expression levels of B7-1 and B7-2 of MSC group were lower than those of control group (all P<0.01). Conclusions HUC-MSCs inhibits the number of CD4+T cells and the secretion of IL-17 in liver after HIRI, as well as decreases the number of intrahepatic mononuclear cells and the mRNA expression of B7-1 and B7-2, thereby alleviating HIRI.

Water extract of Glycyrrhiza uralensis Fisch.enhanced the activation of RAW264.7 cells through TLR4 signaling pathway / 中华微生物学和免疫学杂志

Objective To investigate the effects of water extract of Glycyrrhiza uralensis Fisch.(GUWE) on the activation of RAW264.7 cells and the possible mechanism.Methods RAW264.7 cells were treated with GUWE containing different concentrations of polysaccharide (10, 50, 100, 500 μg/ml).Viability of these cells was analyzed by MTT assay.Phagocytic activity and surface molecules expressed on these cells were detected by flow cytometry.Levels of cytokines were analyzed by ELISA.Western blot assay was performed to analyze the activation of key molecules in TLR4 signaling pathway.Results GUWE at the concentration of 500 μg/ml significantly decreased the viability of RAW264.7 cells, but significantly increased the viability of RAW264.7 cells at concentrations of 50 μg/ml and 100 μg/ml.GUWE significantly enhanced the phagocytic activity of RAW264.7 cells as well as the expression of cytokines and costimulatory molecules in a dose-dependent manner.Further analysis indicated that the activation of RAW264.7 cells induced by GUWE was suppressed by TLR4 inhibitor.Moreover, GUWE enhanced the phosphorylation of NF-kB p65 and TLR4 downstream mitogen-activated protein kinases (MAPKs) including p38, extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK).Conclusion This study indicates that GUWE promotes the activation of RAW264.7 cells through TLR4 signaling pathway.

Comprehensive therapeutic strategies regarding immune checkpoint / 中国肿瘤临床

The rapid development of immunotherapy has exceeded that of standard treatment modes, which include surgery, radio-therapy, chemotherapy, and targeted therapy. Immunotherapy is more durable and less toxic than traditional cancer therapies. More-over, immune checkpoint therapy is an important component of immunotherapy and has been evaluated in preclinical and clinical tri-als and proven to exhibit broad prospects. However, its clinical benefits are limited to a small subset of patients with a subset of tumor types. Therefore, reasonable comprehensive therapeutic strategies are needed to overcome this limitation. Gene targeted therapy, ra-diotherapy, chemotherapy, and tumor vaccine affect the immune system through different mechanisms, and these could provide theo-retical bases for comprehensive treatments. In this review, immune checkpoint therapy and its potential comprehensive therapies with other cancer treatments are introduced.

Dendritic Cell Dysfunction in Patients with End-stage Renal Disease

End-stage renal disease (ESRD) with immune disorder involves complex interactions between the innate and adaptive immune responses. ESRD is associated with various alterations in immune function such as a reduction in polymorphonuclear leukocyte bactericidal activity, a suppression of lymphocyte proliferative response to stimuli, and a malfunction of cell-mediated immunity at the molecular level. ESRD also increases patients' propensity for infections and malignancies as well as causing a diminished response to vaccination. Several factors influence the immunodeficiency in patients with ESRD, including uremic toxins, malnutrition, chronic inflammation, and the therapeutic dialysis modality. The alteration of T-cell function in ESRD has been considered to be a major factor underlying the impaired adaptive cellular immunity in these patients. However, cumulative evidence has suggested that the immune defect in ESRD can be caused by an Ag-presenting dendritic cell (DC) dysfunction in addition to a T-cell defect. It has been reported that ESRD has a deleterious effect on DCs both in terms of their number and function, although the precise mechanism by which DC function becomes altered in these patients is unclear. In this review, we discuss the effects of ESRD on the number and function of DCs and propose a possible molecular mechanism for DC dysfunction. We also address therapeutic approaches to improve immune function by optimally activating DCs in patients with ESRD.

Effects of lung cancer cell-associated B7-H1 on T-cell proliferation in vitro and in vivo

B7 homolog 1 (B7-H1) is the most potent immunoinhibitory molecule in the B7 family. In this study, we examined the effects of tumor-associated B7-H1 on T-cell proliferation in lung cancer. The expression of B7-H1 in human adenocarcinoma A549 and mouse Lewis lung carcinoma (LLC) cells were examined by flow cytometry. To assess the in vitro effect of tumor-associated B7-H1 on T-cell proliferation, we isolated T cells from peripheral blood mononuclear cells (PBMCs) of healthy individuals, labeled them with carboxyfluorescein succinimidyl ester, and co-cultured them with A549 cells in the absence or presence of anti-B7-H1 antibody. For in vivo analysis, LLC cells were subcutaneously injected into mice treated or not with anti-B7-H1 antibody. T-cell proliferation in both in vitro and in vivo assays was analyzed by flow cytometry. In vitro, co-culturing T cells with A549 cells significantly inhibited the proliferation of the former compared with the proliferation of T cells alone (P<0.01), and the addition of B7-H1 blocking antibody dramatically reversed the inhibition of T-cell proliferation by A549 cells. Similarly, in mice bearing LLC-derived xenograft tumors, in vivo administration of anti-B7-H1 antibody significantly increased the total number of spleen and tumor T cells compared to levels in control mice that did not receive anti-B7-H1 antibody. Functionally, in vivo administration of anti-B7-H1 antibody markedly reduced tumor growth. Tumor-associated B7-H1 may facilitate immune evasion by inhibiting T-cell proliferation. Targeting of this mechanism offers a promising therapy for cancer immunotherapy.

The role of CD19+CD24hiCD27hi regulatory B cells in the severity assessment of acute pancreatitis / 中华微生物学和免疫学杂志

Objective To investigate the prognostic value of regulatory B cells (Bregs) in patients with acute pancreatitis .Methods Flow cytometry analysis was performed to detected the percentages of CD19+IL-10+and CD19+CD24hiCD27hi Bregs in peripheral blood samples collected from patients with acute pancreatitis (36 cases with mild acute pancreatitis and 15 cases with severe acute pancreatitis ) as well as the surface costimulatory molecules including CD80 and CD86 on CD19+CD24hiCD27hi Bregs.Their correlations with lymphocytes and C-reactive protein ( CRP) were further analyzed .Results The numbers of lympho-cytes, CD19+lymphocytes, CD19+IL-10+and CD19+CD24hi CD27hi Bregs in peripheral blood samples col-lected from patients with severe and mild acute pancreatitis as well as the mean fluorescence intensities ( MFI) of CD80 and CD86 were significantly lower than those from healthy subjects .Compared with patients with mild acute pancreatitis , the numbers of lymphocytes and CD 19+lymphocytes , the absolute numbers of CD19+IL-10+and CD19+CD24hiCD27hi Bregs as well as the mean fluorescence intensities (MFI) of CD80 and CD86 in patients with severe acute pancreatitis were significantly decreased .The percentages of CD19+IL-10+and CD19+CD24hiCD27hi Bregs in patients with mild acute pancreatitis were significantly increased af-ter an initial drop , but in patients with severe acute pancreatitis those values were continuously decreased along with the disease progression .The percentage of CD19+IL-10+Bregs was positively correlated with the percentage of CD19+CD24hiCD27hi Bregs and the absolute number of CD19+lymphocytes, but was negatively correlated with CRP .Conclusion The abnormal number and function of CD 19+IL-10+and CD19+CD24 hi CD27hi Bregs might be one of the important reasons causing immune dysfunction in patients with acute pan -creatitis.

Immunomodulatory effects of butyrate on bone marrow derived dendritic cells / 中国免疫学杂志

Objective:To investigate the effect of butyrate produced by bacterial metabolism on immune features of murine bone marrow-derived dendritic cells(BMDCs) and its potential mechanisms.Methods: BMDCs were prepared from bone marrow cells of C57BL/6 mice by being cultured with GM-CSF and IL-4.The expression of CD80,CD86,B7-DC and MHCⅡ on BMDCs and its pri-ming ability on the proliferation of OVA257-264 antigen specific CD8+T cell were analyzed by using Flow cytometry.The mRNA levels of IL-6 and IL-12 in BMDCs were detected by real-time fluorescence quantitative PCR(q-PCR).Simultaneously,Griess reaction and Western blot was used for analyzing the levels of NO2-in BMDCs culture supernatant and the ERK phosphortylation in BMDCs respectivly.Results:Butyrate could decrease the levels of CD80,CD86,MHCⅡand B7-DC,and downregulate the capability of BMDCs in priming the proliferation of CD8+T cells.Furthermore,the secretions of IL-6,IL-12,NO2-and the phosphorylation of ERK were sup-pressed.Conclusion:Butyrate down-regulats the immune functions of BMDCs via inhibition of ERK phosphorylation in TLR 4 signaling pathway.

Inducible costimulatory molecule and its roles in tumor microenvironment / 国际肿瘤学杂志

Inducible costimulatory molecule(ICOS)is a member of the CD28 family,which can be expressed on the tumor tissues and immune cells in the tumor microenvironment. ICOS enhances its anti-tumor activity through participating in CD4 + T and CD8 + T cell immune response and enhancing the secretion of cyto-kines on the activated T cells and NK cells. While the curative effect of cytotoxic T lymphocyte-associated anti-gen-4(CTLA-4)monoclonal antibody is relevant with CD4 + T cells expressing ICOS,which suggesting ICOS may become a novel anti-tumor therapeutic target in the future.

Relationship between Co-expression Levels of ICOS and CD28 on Peripheral Blood CD4+ and CD8+ T Cells and Behcet’s Disease / 华中科技大学学报(医学版)

Objective To study the relationship between the co‐expression levels of inducible costimulatory molecule (ICOS)and CD28 on peripheral blood CD4+ and CD8+ T cells and Behcet’s disease(BD).Methods Flow cytometry was used to detect the expression levels of ICOS and CD28 on peripheral blood CD4+ and CD8+ T cells of BD patients(n=22) ,including those with active BD(ABD ,n= 14)and stable BD(SBD ,n= 8) ,and of normal subjects(n= 22).Results The proportion of CD28+ ICOS+ CD4+ T cells was significantly increased in ABD patients when compared with normal subjects [(34.72 ± 12.87)% vs.(25.36 ± 8.24)% ,P0.05]between the two groups.The proportions of CD28-ICOS+ CD4+ T cells and CD28-ICOS+CD8+ T cells were both obviously increased in ABD patients when compared with those in normal subjects[(2.54 ± 1.63)% vs.(0.76 ± 0.25)% for CD28- ICOS+ CD4+ T cells ,P< 0.05;(8.79 ± 3.41)% vs.(3.04 ± 1.25)% for CD28-ICOS+CD8+ T cells ,P<0.05].Conclusion The increased expression level of ICOS on peripheral blood CD4+ and CD8+ T cells is not closely associated with the expression of CD28 to some extent in BD patients ,suggesting that cells expressing only ICOS play an important role in the development of BD.

Role of costimulatory molecule Tim-1 in the immune reaction of corneal transplantation in rats / 中华实验眼科杂志

Background Corneal transplantation is the most reliable and effective means to treat the corneal blindness in the clinical,immune rejection is a major cause of corneal graft failure after the keratoplasty.Objective This study aimed to investigate the role of Tim-1 in the immune reaction following corneal transplantation in rats.Methods Forty clean female Wistar rats were randomized into normal control group,autologous corneal transplantation group and allogeneic corneal transplantation group.Penetrating corneal transplantation was performed with the Wistar rat donors and Wistar rat receipts in the autologous corneal transplantation group,while with the SD rat donors and Wistar rat receipts in the allogeneic corneal transplantation group.The corneal graft diameter was 3.5 mm and the plant bed diameter was 3.0 mm.The inflammatory response of the grafts was examined under the slit lamp microscope 7 days and 14 days after operation and scored based on the criteria of Larkin.Rejection index (RI),mean survival time and survival rate were calculated.The histopathological examination was performed 7 days and 14 days after surgery to evaluate the inflammatory manifestation,and the expressions of Tim-1 protein and mRNA were assayed by immnunochemistry and real-time fluorescence quantitative PCR (RT-qPCR)in the time points mentioned above.Results Mild edema of the grafts were found 7 days after operation in both the autologous corneal transplantation group and the allogeneic corneal transplantation group.In postoperative 14 days,the grafts were clear in the autologous corneal transplantation group,but the thickening,neovacularization and cloudy of the grafts were exhibited in the allogeneic corneal transplantation group.The survival rate of the grafts was 100％ in the autologous corneal transplantation group and that of the allogeneic corneal transplantation group was 0 with the survival time of (9.8±1.2) days.Histopathological examination revealed the stromal infiltration of inflammatory cells in both the autologous and allogeneic corneal transplantation groups in the seventh day,however,the inflammatory cells were obvious decreased in the autologous group but increased in the allogeneic corneal transplantation group in the fourteenth day.Immunochemistry showed a gradually declined positive cells for Tim-1 protein in the autologous corneal transplantation group,but the positive cells were exactly elevated in the allogeneic corneal transplantation group from 7 days through 14 days after operation;While only few positive cells were seen in the normal control group.The expression levels of Tim-1 mRNA in the grafts were 1.24 ± 0.03,5.85 ± 0.08 and 6.54 ± 0.20 in the normal control group,autologous corneal transplantation group and that of the allogeneic corneal transplantation group,respectively,in the seventh day,and in the fourteen day after operation,the expression level declined to 1.54 ±0.10 in the autologous corneal transplantation group and elevated to 8.62±0.24 in the allogeneic corneal transplantation group,showing significant differences among the different groups and various time points (Fgroup =3 277.590,P =0.000 ; Ftime =136.000,P =0.000).Conclusions Tim-1 may play an important role not only in the inflammatory response but also in the rejection reaction of the corneal transplantation.

Effect of costimulatory molecule B7-H3 on the biological characteristics of esophageal cancer Eca-109 cell line / 中国癌症杂志

Background and purpose:Esophageal cancer is a serious disease threatening human health, and it is very difficult to understand the development mechanism and find the therapeutic methods for esophageal cancer. In recent years, B7-H3, as a new member of B7 immunoregulatory superfamily, overexpressed in multiple tumor types, is considered to be a new tumor marker and potential therapeutic target. This study aimed to detect the expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13, Eca-109 and exploring the effect of B7-H3 siRNA on cell proliferation, migration and invasionin vitro in human esophageal cancer Eca-109 cell line. Methods:The expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13 and Eca-109 were detected by reverse transcription polymerase chain reaction (RT-PCR). B7-H3 siRNA and control siRNA were transfectedin vitro into human esophageal cancer Eca-109 cells using LipofectamineTM 2000. The expressions of B7-H3 mRNA and protein in Eca-109 cells were analyzed by RT-PCR and Western blot. The proliferation, migration and invasion abilities of Eca-109 cells were measured by MTT assay, wound scrape assay and transwell invasion assayin vitro,respectively.Results:All tested cultured esophageal cancer cell lines constitutively expressed B7-H3 mRNA under normal conditions (TE-1 0.382±0.008, TE-13 0.399±0.008, Eca-109 0.428±0.012). After transfection, the expression of B7-H3 mRNA levels decreased in B7-H3 siRNA transfected group, compared with control siRNA transfected group (0.128 5±0.000 2vs 0.532 4±0.000 7,P0.05).Conclusion:All tested esophageal cancer cell lines constitutively express B7-H3 mRNA. B7-H3 siRNA interference inhibits Eca-109 cell migration and invasion abilities. B7-H3 may have a critical role in regulating Eca-109 cell progression.

Activation state of and expressions of surface co-stimulatory molecules on peripheral CD4 + T cells from patients with pemphigus / 中华皮肤科杂志

Objective To investigate the activation state of and expressions of surface co-stimulatory molecules on peripheral CD4+ T cells from patients with pemphigus and healthy human controls.Methods Ninety patients with pemphigus including 24 patients with first-onset pemphigus,51 with quiescent pemphigus and 15 with recurrent pemphigus,as well as 30 healthy human controls were enrolled in this study.Peripheral blood samples were obtained from these subjects followed by lymphocyte isolation.Flow cytometry was performed to detect the expressions of CD69,intercellular adhesion molecule-1 (ICAM-1),inducible co-stimulatory molecule (ICOS),CD40 ligand (CD40L) and OX40 on CD4+ T cells.Statistical analysis was done by Mann-Whitney test using Graphpad 5.0 software.Results The expression rate of CD69 on peripheral CD4+ T cells from the healthy human controls was significantly lower than that from patients with pemphigus,patients with first-onset pemphigus,patients with quiescent pemphigus,and patients with recurrent pemphigus ((1.26 ± 0.19)％ vs.(2.46 ± 0.19)％,(2.77 ± 0.40)％,(2.15 ± 0.25)％ and (2.36 ± 0.35)％,all P ＜ 0.05).The patients with pemphigus also showed a significant increase in the expression rates of ICAM-1,CD40L and OX40 compared with the healthy human controls ((55.88 ± 1.67)％ vs.(47.75 ± 2.52)％,P＜ 0.05; (2.23 ± 0.22)％ vs.(0.73 ± 0.07)％,P＜ 0.01; (2.55 ± 0.29)％vs.(0.62 ± 0.17)％,P ＜ 0.01).No significant differences were observed between patients with different stages of pemphigus in the expression rates of CD69,ICAM-1,CD40L or OX40 (all P ＞ 0.05).The percentage of ICOS-expressing CD4+ T cells was significantly up-regulated in only patients with first-onset pemphigus as compared to the healthy controls ((3.73 ± 0.60)％ vs.(2.39 ± 0.16)％,P ＜ 0.05).Conclusions The peripheral blood CD4+ T cells from patients with pemphigus are in a relatively active state with up-regulated surface expressions of many costimulatory molecules,suggesting that CD4+ T cells are involved in the initiation and progression of pemphigus by interacting with B cells through co-stimulatory molecules.

Inducible costimulatory molecule and autoimmune uveitis / 中华实验眼科杂志

Inducible costimulatory molecule (ICOS) is one of the costimulatory molecules CD28 superfamily and is upregulated on the surface of T cells following T cell activation.ICOS initiates costimulatory signals after binds with its ligand (ICOSL) and ICOS/ICOSL promote activation of T cells proliferation,differentiation and cytokine production.Clinical and laboratory researches documented that ICOS/ICOSL plays significant roles in various autoimmune diseases,similarly,it plays critical effects on the outcome and evolution of autoimmune uveitis (AIU) and Behcet disease.To further study the influence and mechanism of ICOS/ICOSL pathway on AIU is of helpful for finding out a new therapeutic approach of AIU.This paper reviewed the basic concept of ICOS,the relationship between ICOS/ICOSL pathway and effector cell,the outlook of ICOS/ICOSL in the treatment of AIU.