Serum B7 homologous protein 2, tumor necrosis factor-related apoptosis-inducing ligand, interleukin-37, interleukin-17A levels in patients with primary immune thrombocytopenia and their clinical significance / 中国医师进修杂志

Objective:To explore the changes of serum B7 homolog 2 (B7-H2), tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), interleukin(IL)-37 and IL-17A levels in patients with primary immune thrombocytopenia (ITP), and to analyze the clinical guiding significance of each index level in the diagnosis and treatment of ITP.Methods:A total of 90 patients with ITP treated in Jining Hospital of Traditional Chinese Medicine from September 2018 to September 2020 were retrospectively selected as the research group, and 90 healthy patients during the same period were selected as the normal control group. The levels of serum B7-H2, TRAIL, IL-37, IL-17A and platelet count (PLT) in the two groups were compared, and the clinical guidance significance of each index level in the diagnosis and treatment of ITP were analyzed by receiver operating characteristic(ROC) curve.Results:The serum levels of B7-H2, TRAIL, IL-37 and IL-17A in the research group were higher than those in the normal control group, and PLT was lower than that in the normal control group: (25.12 ± 4.29) μg/L vs. (12.26 ± 3.10) μg/L, (0.92 ± 0.20) μg/L vs.(0.46 ± 0.13) μg/L, (145.02 ± 43.18) ng/L vs. (50.23 ± 14.07) ng/L, (21.63 ± 5.06) ng/L vs. (7.71 ± 2.04) ng/L, (16.12 ± 4.61) × 10 9/L vs. (200.86 ± 61.4) × 10 9/L, there were statistical differences ( P<0.05). After treatment for 3 months, 73 patients obtained remission, and 13 patients were non-remission. The levels of B7-H2, TRAIL, IL-37, IL-17A in the non-remission group before and after treatment were higher than those in the remission group, and PLT was lower than that in the remission group, there were statistical differences ( P<0.05). Pearson correlation analysis showed that the levels of serum B7-H2, TRAIL, IL-37 and IL-17A were negatively correlated with PLT ( P<0.05). The ROC curve analysis showed that the area under the curve of serum B7-H2, TRAIL, IL-37 and IL-17A in prognostic prediction was 0.916, the sensitivity and the specificity were 94.12% and 86.30%. Conclusions:The serum levels of B7-H2, TRAIL, IL-37 and IL-17A in patients with ITP are significantly elevated, and are closely related to the level of PLT. Combined detection can help clinically predict the direction of disease outcome.

Effect of Hispidulin on Proliferation and Apoptosis of Leukemia K562 Cells by Up-Regulating IL-37 / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and underlying mechanism of hispidulin on the proliferation and apoptosis of leukemia K562 cells.@*METHODS@#K562 cells were cultured in vitro and treated with 0, 5, 25 or 100 μmol/L hispidulin for 24 h. Cell proliferation and apoptosis were detected by CCK-8 and flow cytometry, respectively. Western blot was used to assess the expression of Bax, Bcl-2 and interleukin (IL)-37 proteins. Bone marrow mononuclear cells were extracted from 17 chronic myeloid leukemia patients and 21 healthy individuals by Ficoll-Hypaque density gradient method, and the expression of IL-37 protein was measured by Western blot. K562 cells with IL-37 overexpression or knockdown were constructed, and then treated with 0 or 100 μmol/L hispidulin for 24 h. Cell proliferation, apoptosis and protein expression of Bax and Bcl-2 were determined in the same way as above.@*RESULTS@#After K562 cells were treated with hispidulin, the cell inhibition rate, apoptosis rate, and the protein expression of Bax and IL-37 were significantly increased (P <0.05), but the cell proliferation and expression of Bcl-2 protein were decreased (P <0.05). The expression of IL-37 protein in bone marrow mononuclear cells of the leukemia patient was 0.24±0.03, which was significantly lower than 0.91±0.05 of healthy controls (P <0.05). Overexpression of IL-37 significantly promoted inhibition rate, apoptosis rate, and expression of Bax protein in K562 cells (P <0.05), but suppressed the expression of Bcl-2 protein (P <0.05). In addition, knockdown of IL-37 could reverse the effects of hispidulin on proliferation and apoptosis of K562 cells.@*CONCLUSION@#Hispidulin inhibits the proliferation and induces apoptosis of leukemia K562 cells, which may be related to the up-regulation of IL-37 protein in cells.

Expression of interleukin-37, vascular endothelial growth factor A, and transforming growth factor-β1 and their correlation with T cells in children with primary immune thrombocytopenia / 中国当代儿科杂志

OBJECTIVES@#To investigate the expression of interleukin-37 (IL-37), vascular endothelial growth factor A (VEGFA), and transforming growth factor-β1 (TGF-β1) in children with primary immune thrombocytopenia (ITP) and their correlation with T cells.@*METHODS@#A retrospective analysis was conducted on 45 children with ITP (ITP group) who were admitted to Handan Central Hospital from January 2020 to April 2022, and 30 healthy children who underwent physical examination during the same period were included as the healthy control group. The mRNA expression levels of IL-37, VEGFA, and TGF-β1 and the levels of regulatory T cells (Treg) and helper T cells 17 (Th17) were measured before and after treatment, and the correlation between the mRNA expression levels of IL-37, VEGFA, and TGF-β1 and the levels of Treg, Th17, and Treg/Th17 ratio were analyzed.@*RESULTS@#Compared with the healthy control group, the ITP group had a significantly higher mRNA expression level of IL-37 and a significantly higher level of Th17 before and after treatment, as well as significantly lower mRNA expression levels of VEGFA and TGF-β1 and significantly lower levels of Treg and Treg/Th17 ratio (P<0.05). After treatment, the ITP group had significant reductions in the mRNA expression level of IL-37 and the level of Th17 and significant increases in the mRNA expression levels of VEGFA and TGF-β1 and the levels of Treg and Treg/Th17 ratio (P<0.05). Correlation analysis showed that in the ITP group, the mRNA expression levels of IL-37 and TGF-β1 were negatively correlated with the levels of Treg and Treg/Th17 ratio (P<0.05) and were positively correlated with the level of Th17 (P<0.05) before and after treatment; the mRNA expression level of VEGFA was positively correlated with the levels of Treg and Treg/Th17 ratio (P<0.05) and was negatively correlated with the Th17 level (P<0.05) before and after treatment.@*CONCLUSIONS@#Abnormal expression levels of IL-37, VEGFA, and TGF-β1 may be observed in children with ITP, which is significantly associated with the imbalance of Treg/Th17 ratio. It is speculated that the cytokines such as IL-37, VEGFA, and TGF-β1 may be involved in the development and progression of ITP or may become important potential targets for the treatment of children with ITP. Citation:Chinese Journal of Contemporary Pediatrics, 2023, 25(11): 1131-1136.

Role of IL-37 in childhood immune diseases / 国际儿科学杂志

Interleukin-37(IL-37)is an anti-inflammatory cytokine in the IL-1 family that suppresses both innate and adaptive immune responses in two ways: intracellular complex formation and extracellular binding to membrane receptors.Studies have shown that IL-37 plays an important anti-inflammatory role in childhood immune diseases such as juvenile idiopathic arthritis, Kawasaki disease, inflammatory bowel disease, asthma, hand, foot and mouth disease and autism spectrum disease by regulating gene transcription, cell metabolism, cell proliferation and cytokine expression.This article reviews the biological characteristics, signaling pathway of IL-37 and its anti-inflammatory mechanism in childhood immune diseases.

Research progress of mechanism of IL-37 in rheumatoid arthritis / 上海交通大学学报（医学版）

Rheumatoid arthritis (RA) is a chronic progressive autoimmune disorder, the mechanism of which is not clear yet. Cytokines play a key role in the pathogenesis of RA. Recent studies on cytokine involvement in autoimmune diseases have found that, as a new member of interleukin-1 (IL-1) family, IL-37 can participate in the regulation of signaling pathway and play its anti-inflammatory role by extracellular binding to membrane receptor and intracellular complex formation. Studies have shown that the expression level of IL-37 in healthy people is extremely low and overexpressed in RA patients, and the serum level of IL-37 in RA patients is positively correlated with the level of pro-inflammation cytokines such as tumor necrosis factor-α (TNF-α), IL-6 and IL-1α, the level of C reactive protein and 28-joint disease activity score (DAS28). However, IL-37 treatment can alleviate the inflammation and pathological symptoms of collagen-induced arthritis (CIA) mice, which indicates that IL-37 has anti-inflammatory effect via negative feedback. The specific anti-inflammatory mechanism may be related to mothers against decapentaplegic homolog 3 (SMAD3) pathway and nuclear factor kappa-B (NF-κB) pathway. This article reviews the structure and expression of IL-37, the receptor and its signal pathway and the function and its mechanism in RA, which provides references for the further study of IL-37 and RA.

Advances in Research on IL-37 in Inflammatory Bowel Disease / 胃肠病学

Inflammatory bowel disease (IBD), including ulcerative colitis (UC) and Crohn's disease (CD), is a chronic and nonspecific intestinal inflammatory disease. Immune response disorder is one of the important links in the pathogenesis of IBD. Cytokines participate in the development and remission of IBD by modulating immune response. Interleukin-37 (IL-37), an anti-inflammatory cytokine, can inhibit the excessive immune response through extracellular and intracellular pathways. Several studies have shown that IL-37 can limit intestinal inflammation damage and promote inflammation remission, and play an important protective role in IBD. This article reviewed the research progress on IL-37 in IBD.

Relationship of IL-37 in the serum and peritoneal fluid to TNF-α and IFN-γ in endometriosis patients / 解放军医学杂志

Objective To investigate the relationship of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) to interleukin-37 (IL-37) in the serum and peritoneal fluid, and investigate the pathogenesis of IL-37 in endometriosis (EMT) patients. Methods Thirty-six EMT patients, admitted in the Department of Obstetrics and Gynecology, the Third Xiangya Hospital of Central South University from Jul, 2018 to Jan. 2019, and confirmed by laparoscopic operation and pathology, were selected as the EMT group, of which 13 cases were in stage I and II, and 23 cases in stage III and IV. The other 28 women with simple ovarian cyst, teratoma or hysteromyoma were selected as the control group. ELISA was performed to detect the levels of IL-37, TNF-α and IFN-γ in serum and abdominal fluid of the two groups. And then the levels of IL-37, TNF-α and IFN-γ were compared between the two groups and their relationship was analyzed. Results The serum levels of IL-37 and TNF-α were significantly higher in EMT group than those in control group [(101.78±31.58) pg/ml vs. (63.50±14.48) pg/ml and (30.68±16.22) pg/ml vs. (18.09±1.27) pg/ml, respectively]. The levels of IL-37 and TNF-α in peritoneal fluid were significantly higher in EMT group than those in control group [(133.94±47.07) pg/ml vs. (68.43±12.60) pg/ml and (24.23±6.12) pg/ml vs. (16.63±3.13) pg/ml, respectively]. The levels of IFN-γ in serum and peritoneal fluid were significantly lower in EMT group than that in control group [(18.81±3.20) pg/ml vs. (20.52±1.38) pg/ml and (15.93±2.58) pg/ml vs. (20.32±7.03) pg/ml, respectively]. The differences were statistically significant (P0.05). Conclusion The levels of IL-37 in serum and peritoneal fluid are increased significantly in EMT patients, implying that IL-37 may play a certain role in occurrence and progression of EMT by promoting secretion of TNF-α and inhibiting secretion of IFN-γ.

Expression of the recombinant precursor and putative mature forms of human Interleukin-37 Isoform B (IL-37b) in E. coli expression system

@#<p style="text-align: justify;"><strong>BACKGROUND AND OBJECTIVES: </strong>IL-37b is a cytokine that may exist in several forms, including a full-length precursor protein and its putative mature forms (IL-37b cleaved at E21, 146, and K53, respectively). In recent years, the role of IL-37b has been associated with the regulation of inflammation and inflammatory diseases. Previous studies focused on the intracellular activity of the cytokine, while the bioactivities of its variants when introduced in the extracellular environment has been limited and require further investigation. To enable this, the study produced precursor and truncated forms of IL-37b in an E.coli expression system.</p><p style="text-align: justify;"><strong>METHODOLOGY:</strong> Recombinant proteins of the full-length (FL) and shorter forms (E21, 146, and K53) of IL-37b were produced in IPTG-induced E. coli BL21-CodonPlus(DE3)-RIPL strain and subsequently purified using Ni2+ NTA affinity, ion exchange, and size exclusion chromatography. The identity of the proteins was confirmed through western blotting and LC-MS.</p><p style="text-align: justify;"><strong>RESULTS:</strong> Findings showed that the masses of the expressed proteins correspond to their respective theoretical masses with 24,134.75 +0.04 Da for FL, 21,919.63 +0.80 Da for E21, 19,298.57 +0.04 Da for 146, and 18,551.21 +0.04 Da for K53 at 90-95% purity. This confirms that the correct proteins have been produced and at high purity. Further, the tendency of FL to homodimerize was observed in this study, which may have implications in the extracellular processing and bioactivity of FL.</p><p style="text-align: justify;"><strong>CONCLUSION:</strong> This study describes the successful expression and purification of recombinant precursor and putative mature forms of IL-37b in E.coli, which can be utilized for downstream characterization. </p>

Enhancement of IL-37 in chemosensitivity of cervical cancer HeLa cells to cisplatin / 吉林大学学报(医学版)

Objective:To transfer the interleukin 37 (IL-37) gene to cervical cancer Hela cells,and to explore the killing effect of IL-37 on the HeLa cells and its enhancement in the chemotherapy sensitivity of HeLa cells.Methods:The pIRES2-EGFP (NC group) and pIRES2-EGFP/IL-37 (IL-37 group) plasmids were transfected into the HeLa cells.Q-PCR and Western blotting methods were used to detect the expression levels of IL-37 mRNA and protein.The activities of HeLa cells in NC group,IL-37 group,DDP group and IL-37+DDP group were detected by CCK8 method,and the inhibitory rates of cells were calculated.The gene expressions of signal transducer and activator of transcription 3 (STAT3) and Cyclin D1 were detected by RT-PCR method.Results:Compared with NC group,the expression levels of IL-37 mRNA and protein in IL-37 group were significantly increased (P＜0.01).The activities of HeLa cells in DDP (5-15 mg · L-1) groups were inhibited after administration for 24-72 h (P＜ 0.01);the inhibitory rates in IL-37 + DDP group were higher than those in DDP group within 48 h after administration (P＜0.05).Compared with IL-37 group,the inhibitory rates in IL-37+DDP group was increased with 96 h after administration (P＜0.05).Compared with NC group,the expression levels of STAT3 and Cyclin D1 mRNA in IL-37 group were significantly decreased (P＜0.01).Conclusion:The over-expression of IL-37 can inhibit the proliferation of cervical cancer cells and enhance the effect of DDP on the chemotherapy of cervical cancer cells which may be related to the down-regulation of the expressions of STAT3 and Cyclin D1 by IL-37.

Observation on the effect of long-acting interferon injection and nucleoside drugs on serum interleukin 37 levels and clinical effect in patients with hepatitis B / 中国生化药物杂志

Objective To investigate the effect of long-acting interferon injection and nucleoside drugs on serum interleukin 37 levels and clinical effect in patients with hepatitis B.Methods 172 patients with Chronic hepatitis B from the Department of liver disease in our hospital were selected and divided into 2 groups, 86 cases in the control group treated with Entecavir, 86 cases in the experiment group received Polyethylene glycol interferon α-2a treatment on the basis of the control group, serum levels of inflammatory factors, peripheral blood T lymphocyte level, serum level of hepatic fibrosis index, and the clinical effect were compared after the treatment.Results The effective rate in the control group(63.95%)was lower than the experiment group(81.40%), with significant difference (P<0.05), the DNA HBV negative conversion rate, HBeAg negative conversion rate, HBeAg conversion rate and ALT recovery rate in control group after treatment were lower than the experiment group, with significant difference (P<0.05), compared with the control group, serum levels of IL-37、IL-6、IL-18、IL-12 were lower after treatment in the experiment group, peripheral blood levels of CD4 + T lymphocyte and CD4 +/CD8 +ratio were higher, and peripheral blood levels of CD8 +T lymphocyte weaas lower after treatment, serum levels of HA、PCⅢ、Ⅳ-C and LN were lower after treatment, with significant difference (P<0.05).Conclusion Long-acting interferon injection and nucleoside drugs can significantly reduce the serum levels of IL-37, IL-6, IL-18, IL-12 in patients with hepatitis B, reduce the inflammatory reaction, improve the cellular immune function, inhibit liver fibrosis, the clinical effect was good.

Enhancement of IL-37 in chemosensitivity of cervical cancer HeLa cells to cisplatin / 吉林大学学报(医学版)

Objective:To transfer the interleukin 37 (IL-37) gene to cervical cancer Hela cells,and to explore the killing effect of IL-37 on the HeLa cells and its enhancement in the chemotherapy sensitivity of HeLa cells.Methods:The pIRES2-EGFP (NC group) and pIRES2-EGFP/IL-37 (IL-37 group) plasmids were transfected into the HeLa cells.Q-PCR and Western blotting methods were used to detect the expression levels of IL-37 mRNA and protein.The activities of HeLa cells in NC group,IL-37 group,DDP group and IL-37+DDP group were detected by CCK8 method,and the inhibitory rates of cells were calculated.The gene expressions of signal transducer and activator of transcription 3 (STAT3) and Cyclin D1 were detected by RT-PCR method.Results:Compared with NC group,the expression levels of IL-37 mRNA and protein in IL-37 group were significantly increased (P＜0.01).The activities of HeLa cells in DDP (5-15 mg · L-1) groups were inhibited after administration for 24-72 h (P＜ 0.01);the inhibitory rates in IL-37 + DDP group were higher than those in DDP group within 48 h after administration (P＜0.05).Compared with IL-37 group,the inhibitory rates in IL-37+DDP group was increased with 96 h after administration (P＜0.05).Compared with NC group,the expression levels of STAT3 and Cyclin D1 mRNA in IL-37 group were significantly decreased (P＜0.01).Conclusion:The over-expression of IL-37 can inhibit the proliferation of cervical cancer cells and enhance the effect of DDP on the chemotherapy of cervical cancer cells which may be related to the down-regulation of the expressions of STAT3 and Cyclin D1 by IL-37.

Expression and clinical significance of serum IL-37 and soluble PD-1 in patients with rheumatoid arthritis / 中国免疫学杂志

Objective:To investigate the relationship between IL-37 and soluble PD-1 in patients with rheumatoid arthritis ( RA) , and investigate the relationship between IL-37 and soluble PD-1.Methods: Peripheral blood was collected from RA patients and control group.The levels of IL-37 and sPD-1 in 30 RA patients (RA standard score ≥6) and 30 healthy controls were measured by enzyme-linked immunosorbent assay ( ELISA).The expression of IL-18, IL-6 and IL-18BP in the peripheral blood of two groups was analyzed and the correlation was analyzed by Pearson correlation analysis .Results:The results of ELISA detection showed that the ex-pression levels of IL-37 and sPD-1 in the peripheral blood of patients with RA and other inflammatory cytokines such as IL -18, IL-18BP and IL-6 were higher than those of healthy controls (P<0.05).IL-37 was positively correlated with cytokines IL-18, IL-18BP and IL-6;sPD-1 was positively correlated with cytokine IL-6.IL-37 and sPD-1 were positively correlated with the severity of patients.Conclusion:IL-37 and sPD-1 are elevated in RA patients and associated with other inflammatory cytokines and disease severi -ty.Both of them may play an important regulatory role in the progress of RA , and provide a new basis for future RA therapy .

Expression levels of IL-37 and HBeAg seroconversion in chronic HBV patients / 中国免疫学杂志

Objective:Detecting serum IL-37 concentrations and HBeAg seroconversion in chronic hepatitis B virus(HBV) patients during the treatment of telbivudine( LDT).Methods:Fifty patients with chronic hepatitis B were included in the study;and 20 healthy people were selected as the control group.The expression levels of interleukin ( IL)-37, IL-6 and CRP were measured by enzyme-linked immunosorbent assay.As the same time,the relationship between serum IL-37 concentrations and HBeAg seroconversion was dynamically observed at 0 w,12 w,24 w,36 w,48 w after treatment.Results:The serum levels of IL-37 in chronic HBV patients were higher than the control group at baseline(q=22.716,P0.05).Conclusion:IL-37 may be involved in the process of HBV infection in CHB patients,and can reflect the degree of liver’ s inflammatory damage.IL-37 may play a significant role in the immune response of CHB patients with HBeAg seroconversion.In future,IL-37 may be an effective therapeutic measure to HBV infection.

Construction of eukaryotic expression vector for IL-37 and EGFP fusion protein and its expression in 293T cells / 实用医学杂志

Objective To construct the eukaryotic expression vector for IL-37 fused with the enhanced green fluorescent protein (EGFP) and express the fusion protein IL-37-EGFP in 293T cells. Methods The IL-37 and EGFP gene were amplified by PCR, respectively. Then the gene fragments of IL-37 and EGFP were in-serted into the pcDNA3.1 vector to construct the recombinant eukaryotic expression vector pcDNA3.1/IL-37-EGFP. The pcDNA3.1/IL-37-EGFP recombinant expression vector was identified by enzymatic digestion , DNA sequencing, PCR identification. Then the recombinant plasmid pcDNA3.1/IL-37-EGFP was used to transfect 293T cells , the expression of EGFP in 293T cells was detected by fluorescence microscope , the expression of IL-37 in 293T cells was detected by RT-PCR and Western blot. Results The results of enzymatic digestion and DNA sequencing showed that the fusion gene of IL-37 and EGFP linked with (G4S)3 was cloned correctly into pcDNA3.1 vector. The green fluorescence was observed in 293T cells transfected with pcDNA3.1/IL-37-EGFP by fluorescence microscope. RT-PCR and Western blot showed that the expression with high level of IL-37 mRNA and IL-37 protein in 293T cells transfected with pcDNA3.1/IL-37-EGFP , respectively. Conclusion The recom-binant eukaryotic expression vector for IL-37 fused with EGFP, pcDNA3.1/IL-37-EGFP, was constructed suc-cessfully and expressed effectively in 293T cells.

Effect of telbivudine on serum IL-18,IL-12, IL-37 and TNF-αlevels in patients with HBV-related hepatocirrhosis / 中国生化药物杂志

Objective To explore the effect of telbivudine on serum interleukin-18(IL-18),IL-12, IL-37 and tumor necrosis factor α(TNF-α) levels in patients with HBV-related hepatocirrhosis.Methods 66 cases of patients with HBV-related hepatocirrhosis were collected and divided into treatment group and control group according to the different treatment method , 34 cases in control group were treated with routine treatment , 32 cases in treatment group were treated by telbivudine on the basis of routine treatment.The serum IL-18, IL-12, IL-37, TNF-αlevels and adverse reactions were compared post-treatment.Results Compared with control group, the levels of IL-18, IL-12, IL-37 and TNF-αdecreased significantly post-treatment (P<0.05).The above levels were significantly lower in treatment group post-treatment than that in control group(P<0.05).Conclusion Telbivudine adjuvant therapy could suppress viral replication, and improve liver function and immune response in patients with HBV-related hepatocirrhosis.

The Changes of Serum Interleukin-37 Levels in Patients With Acute Coronary Syndrome / 中国循环杂志

Objective: To study the serum interleukin-37 (IL-37) level changes in patients with acute coronary syndrome (ACS) and to explore the relationship between IL-37 and coronary atherosclerotic plaque. Methods: Our research included 3 groups. ACS group, n=60, SAP (stable angina pectoris) group, n=30 and Control group, the subjects with normal coronary artery, n=15. The peripheral serum levels of IL-37 were examined by ELISA and the differences were compared among different groups. Results: ① The serum levels of IL-37 at admission were as ACS group SAP group>Control group, P77ug/L. Conclusion: The serum IL-37 might be involved in the inlfammatory process in ACS patients, it could be expected as an index for ACS monitor and evaluation in clinical practice.