Advances in application of regulatory T cells in transplant immune tolerance: from basic to clinical research / 器官移植

Regulatory T cells (Treg) are important inhibitory immune cells to establish immune tolerance, which play a pivotal role in regulating excessive immune response and autoimmune diseases of the host. Previous studies related to transplant immune tolerance have confirmed that increasing the number of Treg in vivo or enhancing the function of Treg serve as a therapeutic strategy to induce transplant immune tolerance. At present, Treg-based induction methods for transplant immune tolerance include adoptive infusion of Treg, in vivo amplification of Treg and utilization of antigen-specific Treg. In this article, the characteristics and mechanism of Treg, the latest research progress on basic experiments and clinical practice of Treg related to transplant immune tolerance at home and abroad were reviewed, and future challenges and development of Treg therapy were prospected, aiming to unravel the significance and application prospect of Treg in transplant immune tolerance, explore the advantages and limitations of Treg therapeutic strategies, and provide reference and evidence for subsequent research in this field.

Predictive value of sCTLA-4 and RAD51C proteins for recurrence after interventional therapy for cervical cancer patients / 中华内分泌外科杂志

Objective:To analyze the predictive value of soluble cytotoxic T lymphocyte-associated antigen 4 (sCTLA-4) and RAD51 paralogous gene C (RAD51C) protein in the recurrence of cervical cancer patients after interventional therapy.Methods:A total of 107 patients with cervical cancer who underwent interventional surgery in our hospital from May. 2015 to Apr. 2019 were selected as the research group. postoperative recurrence were recorded. Another 107 patients with benign cervical disease during the same period were selected as the control group. The protein expressions of sCTLA-4 and RAD51C were compared between the two groups and patients with or without recurrence. Logistic regression was used to analyze the influencing factors of postoperative recurrence of cervical cancer patients, and a nomogram model of postoperative recurrence of cervical cancer patients was constructed and verified by calibration curve. The postoperative recurrence rate of cervical cancer patients with different sCTLA-4 and RAD51C protein expressions was compared.Results:The level of sCTLA-4 and the high expression rate of RAD51C protein in the study group were higher than those in the control group ( P<0.05). High-risk human papillomavirus positive, vascular infiltration, interstitial infiltration ≥1/2, paracterine infiltration, high expression of RAD51C protein and high SCTLA-4 level were independent risk factors for postoperative recurrence of cervical cancer ( P<0.05). High-risk human papillomavirus, vascular invasion, interstitial invasion, parametrial invasion, RAD51C protein and sCTLA-4 levels were used to construct a nomogram prediction model for postoperative recurrence of cervical cancer patients. The consistency indices were 0.610 (95% CI: 0.511-0.702), 0.616 (95% CI: 0.517-0.708), 0.640 (95% CI: 0.541-0.730), 0.609 (95% CI: 0.510-0.702), 0.728 (95% CI ranged from 0.633 to 0.809), 0.817 (95% CI ranged from 0.731 to 0.885), and the calibration curve validation showed high consistency. The net benefit rate of combined detection of sCTLA-4 and RAD51C proteins was greater than that of single detection. Conclusions:sCTLA-4 and RAD51C proteins are highly expressed in cervical cancer patients, and the high expression of both indicates that cervical cancer patients have a higher risk of recurrence after surgery. Clinically, the detection of sCTLA-4 and RAD51C protein expression can be used to screen patients with high recurrence risk.

Current research status on immune-related adverse events / 中华胃肠外科杂志

Immune checkpoint inhibitors have progressed rapidly over the past decade and have become one of the most promising oncology treatments. However, immune checkpoint inhibitors reduce T-cell tolerance and lead to a unique spectrum of immune-related adverse events (IRAE). IRAE can involve multiple systems, including endocrine, gastrointestinal, respiratory and skin systems and there is no predictive marker with high specificity and sensitivity. Mild IRAE can be alleviated by discontinuing immune checkpoint inhibitors while severe IRAEs require active intervention. The first-line treatment is glucocorticoids, and immunosuppressants can be considered in refractory cases. However the optimal choice of immunosuppressants is currently controversial. This review provides an overview of the epidemiology and possible mechanisms of immune-related adverse events, outlines some promising predictive biomarkers, and describes several immunotherapy-related organ toxicity and management.

Belatacept: a new weapon in anti-rejection battlefield / 器官移植

As a co-stimulatory blocker against CD28 receptor, belatacept has been approved and applied to the treatment of rejection in organ transplantation in Europe and America. Belatacept has been proven to outperform calcineurin inhibitor (CNI) in improving the long-term survival rate of recipients and grafts, and enhancing graft function. Nevertheless, it might cause a high incidence of rejection. To resolve this issue, transplant workers have attempted to optimize belatacept immunosuppressive regimen and achieved good clinical efficacy. Although belatacept has been proven to exert poor effect on memory T cells, it has potential value in exploring new co-stimulatory molecular targets to optimize immunosuppressive regimes due to its specificity for immune cells and mild adverse effects. In this article, the advent of co-stimulatory blocker, clinical efficacy and application of belatacept, and the causes of belatacept-resistant rejection were reviewed.

Applying artificial intelligence for cancer immunotherapy

Artificial intelligence (AI) is a general term that refers to the use of a machine to imitate intelligent behavior for performing complex tasks with minimal human intervention, such as machine learning; this technology is revolutionizing and reshaping medicine. AI has considerable potential to perfect health-care systems in areas such as diagnostics, risk analysis, health information administration, lifestyle supervision, and virtual health assistance. In terms of immunotherapy, AI has been applied to the prediction of immunotherapy responses based on immune signatures, medical imaging and histological analysis. These features could also be highly useful in the management of cancer immunotherapy given their ever-increasing performance in improving diagnostic accuracy, optimizing treatment planning, predicting outcomes of care and reducing human resource costs. In this review, we present the details of AI and the current progression and state of the art in employing AI for cancer immunotherapy. Furthermore, we discuss the challenges, opportunities and corresponding strategies in applying the technology for widespread clinical deployment. Finally, we summarize the impact of AI on cancer immunotherapy and provide our perspectives about underlying applications of AI in the future.

Evaluation of I-JS001 for hPD1 immuno-PET imaging using sarcoma cell homografts in humanized mice

JS001 (toripalimab) is a humanized IgG monoclonal antibody which strongly inhibits programmed cell death protein 1 (PD1). In this study, we used a different iodine isotype (I) to label JS001 probes to target the human PD1 (hPD1) antigen. , the half maximal effective concentration (EC) value of I-JS001 did not significantly differ from that of JS001. The uptake of I-JS001 by activated T cells was 5.63 times higher than that by nonactivated T cells after 2 h of incubation. The binding affinity of I-JS001 to T cells of different lineages after phytohemagglutinin (PHA) stimulation reached 4.26 nmol/L. Humanized C57BL/6 mice bearing mouse sarcoma S180 cell tumors were validated for immuno-positron emission tomography (immuno-PET) imaging. Pathological staining was used to assess the expression of PD1 in tumor tissues. The homologous Ihuman IgG (IhIgG) group or blocking group was used as a control group. Immuno-PET imaging showed that the uptake in the tumor area of the I-JS001 group at different time points was significantly higher than that of the blocking group or the I-hIgG group in the humanized mouse model. Taken together, these results suggest that this radiotracer has potential for noninvasive monitoring and directing tumor-specific personalized immunotherapy in PD1-positive tumors.

Effects of Saikosaponin d on differentially expressed genes CTLA-4, IL-10 and IL-17 in mice with autoimmune hepatitis / 上海交通大学学报（医学版）

Objective: To explore the effects of Saikosaponin d (SS-d) on autoimmune hepatitis (AIH) by observing the expression changes of some differentially expressed genes screened with the Agilent-085631 gene chip in the liver of AIH mice. Methods: Forty healthy male SPF C57BL/6 mice were divided into chip group (n=8) and SS-d treatment group (n=32). The mice in the chip group were divided into the normal group and the model group [concanavalin A (Con A) was administered to the tail vein at a dose of 15 mg/kg] (both n=4). The mice were sacrificed after 12 h. The differentially expressed genes of liver were screened, some of which were verified by qRT-PCR. The SS-d treatment group was further divided into the normal group, the model group (treatment was the same with the chip group), SS-d low-dose group and SS-d high-dose group [according to the literature and pre-experiment results, 2.5 and 5.0 mg/kg dose of intraperitoneal injection of SS-d were given respectively, and 15 mg/kg of Con A was administered to the tail vein 8 h later] (all n=8). After 12 h, total venous blood, liver total protein and total RNA of mice were collected. The levels of serum glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), interleukin (IL)-10 and IL-17 were detected by ELISA, and cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) was detected by Western blotting. qRT-PCR technology was used to detect the mRNA levels of IL-10, IL-17 and CTLA-4. Results: A total of 11 512 differentially expressed genes were screened (up 5 189, down 6 323), which were related to 138 signal pathways. The qRT-PCR results of IL-10, IL-17 and CTLA-4 gene were consistent with the results of chip screening. Compared with the normal group, the serum levels of GPT and GOT in the model group increased, IL-17 mRNA level increased, IL-10 mRNA and CTLA-4 mRNA levels decreased, the content of serum IL-17 increased, the content of serum IL-10 decreased, and the level of CTLA-4 protein expression in the liver tissues decreased. Compared with the model group, the serum GPT and GOT levels of SS-d in the low-dose and high-dose groups decreased, IL-17 mRNA level decreased, the levels of IL-10 mRNA and CTLA-4 mRNA increased, the content of serum IL-17 decreased, the content of serum IL-10 increased, and the level of CTLA-4 protein expression in the liver tissue increased. Conclusion: Multiple signaling pathways are involved in the pathogenesis of AIH, and SS-d can alleviate the liver inflammation in AIH mice by regulating the expression of IL-10, CTLA-4, and IL-17.

Cytotoxic T Lymphocyte-associated Antigen 4 Polymorphisms Correlated with Graves' Disease in Patients of Han Ethnicity in Yunnan, China / Polimorfismos del antígeno 4 del linfocito T citotóxico correlacionados con la enfermedad de Graves en pacientes de la etnia Han en Yunnan, China

ABSTRACT Objective: To investigate the correlations between polymorphisms at position 49 in exon 1 and position 318 in the promoter of the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) gene and autoimmune thyroid diseases in a Han Chinese population. Methods: Polymerase chain reaction-restriction fragment length polymorphism was utilized. The MseI and BbvI restriction endonucleases were used to detect and analyse position 49 in exon 1 and position 318 in the promoter as well as the T/C alleles of the CTLA-4 gene in peripheral blood samples from 112 patients with Graves' disease (GD), 101 with Hashimoto's thyroiditis (HT) and 100 healthy individuals. Results: At position 49 of exon 1, the frequencies of the GG genotype and the G allele in the GD group (χ2 = 12.147; p = 0.002) were statistically significantly higher than those in the control group (χ2 = 9.925; p = 0.002), while no statistically significant differences were found between the frequencies of the GG genotype and the G allele in the HT group (χ2 = 1.195; p = 0.550) and those in the control group (χ2 = 0.984; p = 0.321). No statistically significant differences in the promoter (−318) or the T/C alleles were observed among the three groups. Position 49 in the 17th codon of exon 1 of the CTLA-4 gene may be a candidate susceptibility marker in patients of Han ethnicity with GD. Conclusion: This finding helps us to better understand the genetic risks for GD and provides a direction for targeted gene therapy.

RESUMEN Objetivo: Investigar las correlaciones entre los polimorfismos en la posición 49 en el exón 1 y la posición 318 en el promotor del gen del antígeno 4 asociado al linfocito T citotóxico (CTLA-4), con las enfermedades autoinmunes de la tiroides en una población China de Han. Métodos: Se utilizó la reacción en cadena de la polimerasa-polimorfismo de la longitud de los fragmentos de restricción. Las endonucleasas de restricción de MseI y BbvI se utilizaron para detectar y analizar la posición 49 en el exón 1 y la posición 318 en el promotor, así como los alelos T/C del gen CTLA-4 en muestras de sangre periférica de 112 pacientes con enfermedad de Graves (EG), 101 con tiroiditis de Hashimoto (TH) y 100 individuos sanos. Resultados: En la posición 49 de exón 1, las frecuencias del genotipo GG y el alelo G en el grupo de EG (χ2 = 12.147; p = 0.002) fueron estadísticamente significativamente más altas que las del grupo de control (χ2 = 9.925; p = 0.002), pero no se encontraron diferencias estadísticamente significativas entre las frecuencias del genotipo GG y el alelo G en el grupo de TH (χ2 = 1.195; p = 0.550) y las del grupo de control (χ2 = 0.984; p = 0.321). No se observaron diferencias estadísticamente significativas en el promotor (−318) ni en los alelos T/C entre los tres grupos. La posición 49 en el codón17.° del exón 1 del gen CTLA-4 puede ser un marcador de susceptibilidad candidato en pacientes de la etnia Han con EG. Conclusión: Este hallazgo nos ayuda a comprender mejor los riesgos genéticos de la EG y ofrece una dirección para la terapia génica dirigida.

Cancer Immunotherapy Related Endocrine Adverse Effects

Cancer immunotherapy has emerged as a promising therapy for a wide variety of tumors. Immune checkpoint inhibitors including anti cytotoxic T lymphocyte antigen 4 (CTLA-4), programmed death 1 (PD-1) and programmed death ligand-1 (PD-L1) monoclonal antibodies have proven to be especially effective in various advanced cancers. However, cancer the immunotherapy disturbs the immune system and may also cause immune related side effects (IRAE) distinguished from cytotoxic chemotherapy toxicity. Among them, endocrine IRAE has been reported with a higher incidence than other organ IRAE. We focus on the most relevant and new aspects related to endocrine IRAE due to cancer immunotherapy in this review.

Expression of costimulators in mice after allogenetic hematopoietic stem cell transplantation and its relationship with acute graft versus host disease / 第二军医大学学报

Objective To explore the expression of costimulators in mice after allogenetic hematopoietic stem cell transplantation and its relationship with acute graft versus host disease (aGVHD). Methods Five C57BL/6J (H2KD－H2KB＋) male mice were selected as donors. Thirty CB6F1 (H2KD＋H2KB＋) female mice were selected as recipients and randomized into three groups: total body irradiation (TBI), bone marrow transplantation (BM) and aGVHD groups, with 10 mice in each group. The mice in the TBI group received radiation only without injecting any cells. The mice in the BM group were injected with 5×106 bone marrow cells without T lymphocytes from donors after radiation. The mice in the aGVHD group were simultaneously injected with 5×106 bone marrow cells without T lymphocytes and 3×107 spleen cells from donors after radiation. The survival rates of mice in the three groups were evaluated using log-rank survival curve. The expression levels of costimulators, including cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), programmed death 1 (PD-1), inducible costimulator (ICOS) and CD28, on CD4＋ and CD8＋ T lymphocytes, were detected using flow cytometry at 7, 14, 21 and 28 d after transplantation. At 21 d after transplantation, the expression levels of interleukin (IL)-17, interferon γ (IFN-γ) and IL-4 in CD4＋ T lymphocytes, and the costimulator ligands in the colon tissues, were detected with flow cytometry and 3,3’-diaminobenzidine (DAB) staining, respectively. Results The mice in the TBI group all died within 19 d after irradiation. The mice in the BM group survived 30 d after irradiation without aGVHD. The median onset time of aGVHD symptoms and survival of mice in the aGVHD group were 14 (11-18) d and 22 (13-30) d, respectively. The expression levels of CTLA-4 and PD-1 on CD4＋ and CD8＋ T lymphocytes were decreased after transplantation, while the expression levels of ICOS and CD28 were on the rise. At 28 d after transplantation, the expression levels of CTLA-4 and PD-1 on CD4＋ and CD8＋ T lymphocytes were significantly lower in the aGVHD group than those in the BM group (all P0.05), and the expression levels of ICOS and CD28 were significantly higher (all P0.05). DAB staining showed that CD80, ICOS ligand (ICOSL), and PD-1 ligand (PD-L1) were negative in the colon tissues of mice in the BM group, and the positive expression rates of CD80, ICOSL, and PD-L1 in the aGVHD group were 40%, 80% and 80%, respectively. Compared with the BM group, the expression levels of IL-17 and IFN-γ in CD4＋ T lymphocytes in the aGVHD group were significantly increased, while the expression level of IL-4 was significantly decreased (all P0.05). Conclusion In aGVHD mice after allogeneic hematopoietic stem cell transplantation, the expression of costimulator CTLA-4 is gradually decreased over time. CTLA-4, ICOS and PD-1 may participate in the development and progression of aGVHD by regulating the distribution of T lymphocyte subgroups such as T-helper cell (Th)1, Th2 and Th17.

Polymorphisms in cytotoxic T-lymphocyte associated antigen 4 gene does not affect scytotoxic T-lymphocyte associated antigen 4 levels in human papillomavirus-infected women with or without cervical cancer

Background: Cervical cancer (CaCx) is the second most common cancer in Indian women. Cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) + 49 AA polymorphism is known to be associated with CaCx. Current attempt is to use immunotherapy for the treatment of metastatic melanoma and metastatic castration-resistant prostate cancer, i.e., blocking of CTLA-4 using a fully human monoclonal CTLA-4 antibody to disrupt its inhibitory signal. This allows the CTLs to destroy the cancer cells. There is no information available on the soluble level of CTLA-4 on which the immunotherapy is targeted. This is specifically in Indian population including cases with CaCx. Objective: The aim of this study is to evaluate the levels of soluble CTLA-4 (sCTLA-4) in human papillomavirus (HPV)-infected women with or without CaCx and their association with the polymorphism at CTLA-4 + 49 A/G and CTLA-4 ?318 C/T genotypes. Materials and Methods: This is an exploratory case–control study involving two groups of HPV-infected women, the cases were with invasive CaCx and the control group was women with the healthy cervix. sCTLA-4 levels were measured using ELISA in 92 CaCx cases and 57 HPV-positive women with the healthy cervix. Results: Both cases and controls have similar sCTLA-4 levels. Comparison of CTLA-4 + 49A/G and ?318 C/T genotypes with sCTLA-4 levels among cases and control also did not show any statistically significant difference. Conclusion: The present study suggests sCTLA-4 levels are not affected by a polymorphism at + 49 A>G CTLA-4. Hence, levels of CTLA-4 are similar in both CaCx cases and control group.

Correlation between gene polymorphism of cytotoxic T lymphocyte associated antigen-4 and Graves disease in Qinghai Tibetan / 中华地方病学杂志

Objective To investigate the correlation between gene polymorphism of cytotoxic T lymphocyte associated antigen-4 (CTLA-4) and Graves disease (GD) in Qinghai Tibetan.Methods Using retrospective analysis methods,totally 130 cases of GD were selected randomly from June 2012 to November 2016 in the People's Hospital of Qinghai Province;meanwhile,110 normal control cases were selected randomly from Qinghai Tibetan.Then the genotype and allele of CTLA-4 were detected by the method of polymerase chain reaction restriction fragment length polymorphism (RFLP-PCR).Results The distribution of CTLA-4 genotype frequencies (AA,AG,GG) was different between normal control cases and GD in Qinghai Tibetan [6.2％ (8/130) vs 26.4％ (29/110),50.0％ (65/130)vs 58.2％ (64/110),43.8％ (57/130) vs 15.4％ (17/110),x2 =32.105,P ＜ 0.05].Allele (A,G) frequencies were compared between GD and control,the differences were statistically significant [31.2％(81/260) vs 55.5％ (122/220),68.8％ (179/260) vs 44.5％ (98/220),x2 =28.834,P ＜ 0.05].Conclusion Polymorphisms of CTLA-4 exon 1 (49A/G)genotype and allele are closely correlated with GD in Qinghai Tibetan.

Clinical adverse reactions and treatment strategies of tumor immune checkpoint blockades / 肿瘤

As representatives of immune checkpoint blockades (ICBs), antibodies targeting cytotoxic T-lymphocyte antigen-4 (CTLA-4) and programmed cell death-1 (PD-1)/programmed death-ligand 1 (PD-L1) have significantly improved the treatment efficacies of a variety of malignant tumors. However, ICBs can also produce a wide range of immune-related adverse reactions, forcing patients to stop treatment and even affect the survival of the patients. Therefore, with the wide application of ICBs in clinical practice, clinical oncologists need to fully understand the possible side effects of ICBs therapy and the appropriate treatment strategies to improve the survival rate and therapeutic effect of patients receiving ICBs.

Advances in treatment of melanoma with immunotherapy / 肿瘤

Immunotherapy, primarily headed by immune checkpoint inhibitors, has become a standard, first-line therapeutic methodin treatment for patients with melanoma. Combination immunotherapy, that is the combination of programmed cell death-1 (PD-1) inhibitor and cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) inhibitor, further enhanced the anti-tumor efficacy, improved the objective response rate, and extended both the overall and progressionfree survival of patients in the past years. Therefore, combination immunotherapy has also become a new direction for the treatment of melanoma. Meanwhile, the anti-tumor effects and clinical outcomes of combination immunotherapy in other types of tumors are also encouraging. Combination immunotherapy offers a new treatment option for patients, but there are still many issues that need to be further discussed. In order to maximize the benefit of patients, more large-scale clinical researches are needed to answer the questions which may strongly affect the clinical decisions, such as, how to optimize the regimens of combination therapy, how to identify the appropriate treatment population, and how to balance the risk-benefit ratio of patients.

Expression of CTLA-4 in T lymphocytes of patients with glioma and its clinical significance / 吉林大学学报(医学版)

Objective:To investigate the correlation between the CTLA-4 expression in the peripheral blood CD4+ and CD8+T cells in the patients with glioma and glioma WHO classification and operation,and to clarify the clinical significances of CTLA-4.Methods:60 patients with glioma from our hospital were selected as glioma group, and 46 healthy volunteers were used as control group.Then the CTLA-4 expression levels in the peripheral blood CD4+ and CD8+T cells of the subj ects in glioma group and control group were detected by fluorescence-activated cell sorting (FACS)and the relationship between CTLA-4 and glioma WHO classification and surgery was analyzed. Results:The CTLA-4 expression levels in CD4+ and CD8+T cells in peripheral blood of the patients in glioma group were higher than those in control group (P<0.01).The CTLA-4 expression level in the patients with gradeⅣ glioma was the highest, the expression level of the patients with grade Ⅳ glioma was higher than that of the patients with grade Ⅲ glioma (P<0.01),and the expression level of grade Ⅲ the patients with was higher than that of the patients with grade Ⅱ (P<0.01).The expression level of CTLA 4 in CD4+ and CD8+T cells of the patients with glioma after operation was lower than before operation (P<0.01 ). Conclusion:The CTLA-4 expression levels in the peripheral blood CD4+ and CD8+T cells are increased with the increasing of malignancy degree of the patients with glioma,and it has a promotion role in the occurrence of development of glioma.

Phenotypic Characteristics of PD-1 and CTLA-4 Expression in Symptomatic Acute Hepatitis A

BACKGROUND/AIMS: The immunoregulatory molecules programmed death 1 (PD-1) and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) are associated with the dysfunction of antiviral effector T-cells, which leads to T-cell exhaustion and persistent viral infection in patients with chronic hepatitis C and chronic hepatitis B. Little is known about the role of PD-1 and CTLA-4 in patients with symptomatic acute hepatitis A (AHA). METHODS: Peripheral blood mononuclear cells were isolated from seven patients with AHA and from six patients with nonviral acute toxic hepatitis (ATH) during the symptomatic and convalescent phases of the respective diseases; five healthy subjects acted as controls. The expression of PD-1 and CTLA-4 on T-cells was measured by flow cytometry. RESULTS: PD-1 and CTLA-4 expression during the symptomatic phase was significantly higher in the T-cells of AHA patients than in those of ATH patients or healthy controls (PD-1: 18.3% vs 3.7% vs 1.6%, respectively, p<0.05; CTLA-4: 23.5% vs 6.1% vs 5.9%, respectively, p<0.05). The levels of both molecules decreased dramatically during the convalescent phase of AHA, whereas a similar pattern was not seen in ATH. CONCLUSIONS: Our findings are consistent with a viral-protective effect of PD-1 and CTLA-4 as inhibitory molecules that suppress cytotoxic T-cells and thereby prevent the destruction of virus-infected hepatocytes in AHA.

Association between cytotoxic T lymphocyte-associated antigen-4 gene polymorphism and Graves ophthalmopathy in Qinghai Han population / 中华地方病学杂志

Objective To investigate the association between gene polymorphism of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) and Graves ophthalmopathy (GO) in Qinghai Han population.Methods Ninety cases of Graves disease were selected from June 2011 to February 2014 in The People's Hospital of Qinghai Province,and the 90 patients were divided into two subgroups according to GO (49 cases) and GD without GO(41 cases).Then the genotype and allele of CTLA-4 exon 1 (+ 49A/G) were detected in surum by the method of polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP).Results The distribution of CTLA-4 exon 1 (+ 49 A/G) genotype frequencies (AA,AG,GG) was not different between GO and GD without GO subgroups [4.1％ (2/49) to 7.3％ (3/41),44.9％ (22/49) to 61.0％ (25/41),51.0％ (25/49) to 31.7％ (13/41),Fisher exact probability,P =0.180 ＞ 0.05]; the distribution of CTLA-4 exon 1 (+ 49A/G) allele frequencies (A,G) was not different between GO and GD without GO subgroups [26.5％ (26/98) to 37.8％ (31/82),73.5％ (72/98) to 62.2％ (51/ 82),x2 =2.622,P＞ 0.05].Conclusion CTLA-4 gene exon 1 (+ 49A/G) may not be a candidate susceptibility gene for Qinghai Han GO.

Correlation research between CTLA-4Ig impacting on Th17 and Treg cells expressed in spleen of lupus mice and its intervention role in lupus-like symptoms of B6.MRL-Faslpr/J lupus mice / 中国免疫学杂志

Objective:To observe the correlation between the B7 blocker,CD28 and cytotoxic T lymphocyte-associated antigen 4 immunoglobulin (CTLA-4Ig) impacting on Th17 and Treg cells expressed in the spleen of lupus mice and its intervention role in lupus-like symptoms of B6.MRL-Faslpr/J lupus mice.Methods:Sixteen 4-month-old female B6.MRL-Faslpr/J mice were selected and randomly divided into treatment group ( groupⅠ) and control group ( groupⅡ);injected the same amount of CTLA-4Ig and PBS intra-venously,checked their 24 hour urine protein ,ANA antibody,ds-DNA antibodies before and after the intervention.Two weeks after the intervention ,detected serum IL-17A,and the percentage of Th17 and Treg cells in their spleen.Results:Two weeks after the last inter-vention,24-hour urine protein,serum ANA and ds-DNA in groupⅠdecreased,and all the differences were statistically significant (P<0.05) compared with groupⅡ.Two weeks after the last intervention,serum IL-17A and the proportion of Th17 cells in the spleen in groupⅠwere lower than those in groupⅡ, but Treg cells in CD4+T lymphocytes was higher than that in groupⅡ,all the differences were statistically significant ( P<0.05).Conclusion:CTLA4-Ig can relieve the lupus-like symptoms in lupus mice;raising the number of Treg cells and decreasing the number of Th17 cells may be one of the important mechanisms for CTLA-4Ig to alleviate lupus-like symptoms in B6.MRL-Faslpr/J mice.

Analysis of cytotoxic T-lymphocyte-associated antigen-4 and MMP-9 genes’ methylation and their expression profiles with risk of non-alcoholic fatty liver disease.

OBJECTIVE: To investigate the effect of promoter methylation of cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) gene and matrix metalloproteinases (MMPs) on the risk of non-alcoholic fatty liver disease (NAFLD). MATERIALS AND METHODS: CTLA-4 and MMP-9 promoter methylation were investigated using a methylation-specific polymerase chain reaction (MS-PCR) in blood samples taken from 80 NAFLD individuals and 95 healthy controls. The expression levels of CTLA-4 and MMP-9 were also assessed in 10 blood and 9 liver tissues mRNA samples from NAFLD patients. These cases were compared to the blood (n = 10) samples of healthy controls with real-time quantitative reverse transcriptase PCR. RESULTS: No significant relationship was found for methylation of CTLA-4 and MMP-9 between cases and controls. The relative expression of CTLA-4 and MMP-9 mRNA in NAFLD was not significantly different compared to healthy control samples. CONCLUSION: For the first time, our outcomes indicate that the methylation status of CTLA-4 and MMP-9 genes has no significant function on the process of NAFLD.

Meta-analysis of CTLA-4 promoter-49 A/G polymorphism and Graves' disease in China / 中华内分泌外科杂志

Objective To summarize the correlation between cytotoxic T lymphocyte associated antigen-4 (CTLA-4) promoter-49 A/G polymorphism and Graves' disease (GD)in China.Methods The publications before Sep.30,2011 about the correlation between CTLA-4 promoter-49 A/G polymorphism and GD in China were collected by searching CNKI,CBM,and Wanfang database.Meta-analysis was performed to check the difference of genotypes such as AG,GG,GG + AG,and G allele between GD group and the control group.Results A total of 6 studies involving 570 cases in GD group and 486 cases in the control group were included in the meta-analysis.According to the heterogeneity of these studies,meta-analysis was performed to analyze the correlation between AG,GG,GG + AG genotype,G allele and GD by fixed effects model.The pooled OR value(95％ CI) of various studies was 1.97 (1.37-2.82),3.66 (2.42-5.53),2.55 (1.82-3.58),and 1.99 (1.49-2.65).Conclusion The meta-analysis demonstrates that CTLA-4 promoter-49 A/G polymorphism increases the possibility of getting GD in Chinese.