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Objective:To explore the therapeutic effect of anti-interleukin (IL)-12/IL-23 p40 antibody on experimental autoimmune uveitis (EAU) and its mechanism.Methods:Sixty-six SPF female C57BL/6N mice aged 6-8 weeks were selected.EAU model was established in 24 mice through immunization with the interphotoreceptor retinoid-binding protein (IRBP) 651-670.The 24 mice were sacrificed before immunization, and on the 3rd, 12th, and 18th day after immunization, with 6 at each time point.Flow cytometry was used to detect the proportion of IL-17A + interferon-γ (IFN-γ) + CD4 + T cells in the spleen, lymph nodes and eyeballs.Another 6 mice were selected to establish EAU model, and fundus images of the mice were taken with a small animal imaging instrument and optical coherence tomography (OCT) 18 days after immunization.The 6 mice were sacrificed after OCT examination and the eyeballs were collected.Hematoxylin-eosin staining was used to observe the retinal inflammation and morphological changes in tissue structure.Flow cytometry was employed to detect the proportion of IL-17A + IFN-γ + CD4 + T cells in lymph nodes.The 6 mice were divided into IL-17A + IFN-γ + highly expressed group and IL-17A + IFN-γ + lowly expressed group according to flow cytometry results, and the retinal injury was compared between the two groups.EAU model was established in another 36 mice, which were divided into anti-IL-12/IL-23 p40 group and IgG group by random number table method, with 18 mice in each group.Anti-IL-12/IL-23 p40 or IgG was injected by tail vein at a 3-day inteval according to grouping.On the 12th and 18th day after immunization, 6 mice were selected from each group to collect lymph nodes and eyeballs, and the proportion of T cell subsets was detected by flow cytometry.Eyeballs of 6 mice in each group were extracted on the 24th day after immunization and retinal damage was observed by hematoxylin-eosin staining.The induced differentiation of CD4 + T cells in vitro was assayed by flow cytometry.The expressions of IL-17 and IFN-γ were detected by enzyme-linked immunosorbent assay (ELISA) after induced differentiation of IL-17A + IFN-γ + CD4 + T cells.The relative expression levels of Th1 transcription factor T-bet and Th17 transcription factor retinoid acid-related orphan nuclear receptor γt (ROR-γt) after induced differentiation of IL-17A + IFN-γ + CD4 + T cells were detected by real-time quantitative PCR.The use and care of animals followed the ARVO statement and this study protocol was approved by an Ethics Committee of Experimental Animals of Tianjin Medical University Eye Hospital (No.TJYY2019111019). Results:There were significant differences in the proportion of IL-17A + IFN-γ + CD4 + T cells in lymph nodes, spleen and eyeballs between wild-type mice and EAU mice at the 3rd, 12th and 18th day after immunization ( H=9.642, 16.531, 10.385; all at P<0.05). Compared with before immunization, the proportion of IL-17A + IFN-γ + CD4 + T cells was significantly increased in lymph nodes of EAU mice on the 12th day following immunization and was significantly increased in spleen and lymph nodes on day 18 after immunization (all at P<0.05). Severe retinal exudation, retinal detachment, severe inflammatory cell infiltration and extensive retinal folds were detected in IL-17A + IFN-γ + highly expressed mice.Mild retinal edema, focal inflammatory cell infiltration and mild retinal folds were found in IL-17A + IFN-γ + lowly expressed mice.The proportion of CD3 and IL-17A + IFN-γ + CD4 + T cells in the eyeballs of anti-IL-12/IL-23 p40 group was lower than that in IgG group at the 18th day after immunization, and the differences were statistically significant ( t=15.304, 8.080; both at P<0.05). On day 12 after immunization, the percentage of IL-17A + IFN-γ + CD4 + T cells in anti-IL-12/IL-23 p40 group was (0.33±0.18)%, which was significantly lower than (4.83±0.45)% in IgG group ( t=15.974, P<0.001). Compared with IgG group, the percentage of Th1, Th17, IL-17A + IFN-γ + CD4 + T cells and the expression levels of IL-17, IFN-γ, T-bet, ROR-γt in anti-IL-12/IL-23 p40 group were significantly decreased, with statistical significances (all at P<0.05). Conclusions:Anti-IL-12/IL-23 p40 has a therapeutic effect on EAU by inhibiting IL-17A + IFN-γ + CD4 + T cells.
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Interleukin-12 (IL-12) is a pro-inflammatory cytokine derived from antigen-presenting cells. IL-12 has a variety of effector mechanisms such as promoting T cells proliferation and cytotoxicity, inducing T cells and NK cells to secrete interferon-γ (IFN-γ), etc. IL-12 shows significant anti-tumor effect in preclinical stage and is quickly used in clinic, but its systemic administration has serious dose-limiting toxicity. At present, most of the research is about the topical use of IL-12 to reduce systemic toxicity, change tumor microenvironment and activate immune system. This article reviews the research progress and clinical trials of IL-12 in the treatment of solid tumors in recent years.
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Objective:To investigate the effects of dexamethasone-assisted ademetionine and ursodeoxycholic acid on pregnancy outcome and serum thyroid peroxidase antibody (TPOAb) and interleukin-12 (IL-12) expressions in patients with intrahepatic cholestasis (ICP) during pregnancy.Methods:A prospective randomized controlled study of 80 patients with ICP in Pingyang Hospital Affiliated of Wenzhou Medical University from April 2017 to April 2020 was selected. The patients were divided into the observation group and the control group using random number table, with 40 cases in each group. On the basis of conventional treatment, the control group was given ademetionine and ursodeoxycholic acid, and the observation group was given dexamethasone-assisted ademetionine and ursodeoxycholic acid. All patients were treated for 1 week. The efficacy, time of disappearance of symptoms, maternal and infant outcomes and liver function indexes aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bile acid (TBA), immune function indexes immunoglobulin M (IgM), immunoglobulin A (IgA), immunoglobulin G (IgG), serum TPOAb and IL-12 levels before and after treatment were compared between the two groups.Results:After treatment, the total effective rate in the observation group was higher than that in the control group: 95.0% (38/40) vs. 80.0%(32/40), the difference was statistically significant ( χ2 = 4.114, P<0.05). The disappearance time of jaundice and itching were shorter than those in the control group ( P<0.05). After treatment, the levels of serum AST, ALT, TBA, TPOAb, and IL-12 in the two groups were lower than before treatment, and the levels of above index in the observation group were lower than those in the control group: (57.49 ± 11.45) U/L vs. (83.70 ± 13.57) U/L, (87.61 ± 29.03) U/L vs. (126.24 ± 33.28) U/L, (13.24 ± 5.48) μmol/L vs. (21.39 ± 7.20) μmol/L, (9.18 ± 2.41) kU/L vs. (12.97 ± 2.73) kU/L, (11.37 ± 2.05) ng/L vs. (18.26 ± 2.54) ng/L; the levels of serum IgM, IgA and IgG in two groups were higher than before treatment, the levels of above index in the observation group were higher than those in the control group: (1.40 ± 0.09) g/L vs. (1.28 ± 0.11) g/L, (1.96 ± 0.14) g/L vs. (1.82 ± 0.12) g/L, (11.53 ± 2.80) g/L vs. (9.37 ± 2.59) g/L, the differences were statistically significant ( P<0.05). The incidence of cesarean section, premature delivery, postpartum hemorrhage, neonatal asphyxia, and intrauterine distress in the observation group were lower than those in the control group. Conclusions:Dexamethasone-assisted ademetionine and ursodeoxycholic acid treatment of ICP patients can improve liver function and immune function, reduce serum TPOAb and IL-12 levels, alleviate clinical symptoms and improve maternal and infant outcomes.
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OBJECTIVE@#To establish a micellar electrokinetic capillary chromatography-based method for identification and quantitative detection of interleukin-12 (IL-12) and analysis of its unfolding process.@*METHODS@#An uncoated fused-silica capillary (inner diameter 50 μm) with a total length of 48.5 cm (40 cm to the detector) was used for the experiment. The factors influencing the separation efficiency of IL-12 were analyzed, and a standard curve of IL-12 concentration was established. The mixture of IL-12 and anti-IL-12 antibody was incubated in a water bath at 38 ℃ for 40 min, and capillary electrophoresis was then performed under the same conditions. The results were compared with those of IL-12 and anti-IL-12 antibody to identify IL-12. IL-12 and dithiothreitol (DTT) were incubated at 60 ℃ in water bath for different lengths of times, and the unfolding process of IL-12 was analyzed based on electrophoresis results of IL-12 in different states.@*RESULTS@#A micellar capillary electrophoresis on-line sweep method was established with 80 mmol/L borate (pH=9.3) containing 30 mmol/L sodium dodecyl sulfate (SDS) as the buffer solution. This system showed a good linear relationship between the peak area and the mass concentration of IL-12 with a linear correlation coefficient of 0.9991 within the linear range of 2 to 120 ng/L. As the incubation time of IL-12 and DTT prolonged, the disulfide bond of IL-12 gradually opened and resulted in distinct changes in the protein peak.@*CONCLUSIONS@#This capillary electrophoresis-based method is simple and sensitive for IL-2 analysis and allows rapid detection of changes in IL-12 content in the setting of tumors and analysis of the possible causes.
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Objective:To study the effect of modified Erchentang on levels of interleukin-12 (IL-12), interferon-γ (IFN-γ), interleukin-9 (IL-9), interleukin-4 (IL-4) and interleukin-13 (IL-13) in plasma and bronchoalveolar lavage fluid (BALF) of all rats, as well as expressions of interleukin-4 (IL-4) receptor (IL-4R1) and interleukin-13 (IL-13) receptor (IL-13RA1) in bronchioles tissue of rats with chronic obstructive pulmonary disease (COPD). Method:Fifty SD rats were randomly divided into 5 groups, namely normal group, model group, and low, middle and high-dose modified Erchentang groups (5, 10, 20 g·kg-1), with 10 rats in each group. COPD in rat was prepared by using cigarette smoke combined with dripping lipopolysaccharide (LPS) in trachea. After the modeling, normal and model groups were given normal saline solution through intragastric (ig) administration, while other groups were given corresponding herbal drugs (5, 10, 20 g·kg-1) intragastrically (ig) for 14 days. The levels of IL-12, IFN-γ, IL-9, IL-4 and IL-13 in plasma and BALF were detected by Enzyme-linked immunosorbent assay (ELISA) method， and immunohistochemistry (IHC) method was used to detect the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue of all of the groups. Result:Compared with the normal group, the levels of IL-12 and IFN-γ were decreased significantly (P<0.01), but the levels of IL-9, IL-4 and IL-13 in plasma and BALF were significantly increased (P<0.01), and the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue were increased significantly (P<0.01) in model group. Compared with the model group, the levels of IL-12 and IFN-γ were increased significantly, while the levels of IL-9, IL-4 and IL-13 in plasma and BALF were decreased significantly (P<0.01), and the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue were decreased significantly (P<0.01) in modified Erchentang groups (10, 20 g·kg-1). Conclusion:Modified Erchentang has effects in resisting inflammatory and protecting tissue structure of bronchioles. Its mechanism may be correlated with increasing the levels of IL-12, IFN-γ and reducing the levels of IL-9, IL-4 and IL-13 in plasma and BALF, and inhibiting the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue.
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OBJECTIVE@#To investigate whether interleukin-12 (IL-12) over-expression in malignant melanoma B16 cells affects the expression level of programmed death-1 (PD-1) on T cells in mice during immune microenvironment reconstruction.@*METHODS@#B16 cells were transfected with an IL-12 expression lentiviral vector, and IL-12 over-expression in the cells was verified qPCR and ELISA. Plate cloning assay was used to compare the cell proliferation activity between B16 cells and B16/IL-12 cells. The expression of IL-12 protein in B16/IL-12 cells-derived tumor tissue were detected by ELISA. C57BL/6 mice were inoculated with B16 cells or B16/IL-12 cells, and 14 days later the proportion of T cells with high expression of PD-1 in the tumor-draining lymph nodes was detected by flow cytometry. Mouse models of immune reconstitution established by 650 cGy X-ray radiation were inoculated with B16 (B16+RT group) or B16/IL-12 (B16/IL-12+RT group) cells, with the mice without X-ray radiation prior to B16 cell inoculation as controls. Tumor growth in the mice was recorded at different time points, and on day 14, flow cytometry was performed to detect the proportion of T cells with high PD-1 expression in the tumor-draining lymph nodes and in the tumor tissue.@*RESULTS@#B16 cells infected with the IL-12-overexpressing lentiviral vector showed significantly increased mRNA and protein levels of IL-12 ( < 0.001) without obvious changes in cell viability (>0.05). B16/IL-12 cells expressed higher levels of IL-12 than B16 cells ( < 0.01). In the tumor-bearing mouse models, the proportion of CD4 PD-1 T cells was significantly lower in B16/IL-12 group than in B16 group ( < 0.01). In the mice with X-ray radiation-induced immune reconstitution, PD-1 expressions on CD4 T cells ( < 0.05) and CD8+ T cells ( < 0.01) were significantly higher in B16+ RT group than in the control mice and in B16/IL-12+RT group ( < 0.01 or 0.001); the tumors grew more slowly in B16/IL-12+RT group than in B16 + RT group ( < 0.001).@*CONCLUSIONS@#During immune microenvironment reconstruction, overexpression IL-12 in the tumor microenvironment can reduce the percentage of PD-1 T cells and suppress the growth of malignant melanoma in mice.
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Animals , Mice , CD8-Positive T-Lymphocytes , Cell Line, Tumor , Immune Reconstitution , Interleukin-12 , Melanoma, Experimental , Mice, Inbred C57BL , Tumor MicroenvironmentABSTRACT
Myeloid derived suppressor cells (MDSCs) are a heterogenous population of immature cells that play a critical role in tumor associated immune suppression. In tumor conditions, the population of MDSCs increases. The main feature of these cells is their ability to suppress the T cell response in antigen specific or nonspecific manners depending on the condition of T cell activation. IL-12 can modulate MDSC in preliminary reports, so we investigated how IL-12 can affect MDSC in a tumor microenvironment. After implanting tumor based cells on syngeneic host, 4T-1/BALB/c or EL4/C57BL6 mice, MDSCs (Gr1+CD11b+) were isolated from splenocytes. Isolated MDSCs were treated with GM-CSF with or without IL-12 and analyzed based on their phenotypes and functions. Treatment of MDSC with IL-12 increased co-stimulatory molecules of CD80, CD86, OX-40L, enhancing the DC phenotype (CD11c) and maturation markers such as p-NF-κB and p-GSK3β. In addition to a change of surface markers, T-cell suppressive function of MDSC after IL-12 treatment was significantly improved compared with the control MDSC. In addition, PD-L1+F4/80+ macrophages, which show aninhibitory effect in phagocytosis, were decreased after IL-12 treatment. The changes of cell surface expression of CD80, CD86, MHC class II were also shown in vivo. Our results showed that the IL-12 can modulate MDSC into APC and recover the macrophage function. These results suggested that IL-12 plays a role in improving the tumor immune microenvironment through MDSC modulation.
Subject(s)
Animals , Mice , Granulocyte-Macrophage Colony-Stimulating Factor , Interleukin-12 , Macrophages , Phagocytosis , Phenotype , T-Lymphocytes , Tumor MicroenvironmentABSTRACT
Background: The incidence of alcoholic liver disease (ALD) is increasing year by year, and is lack of specific diagnostic indicators. Exploring the trend of molecular markers in different stages of ALD is of great significance for its diagnosis and treatment. Aims: To investigate the levels and diagnostic value of serum interleukin-12 (IL-12), carbohydrate-deficient transferrin (CDT), hepatocyte growth factor (HGF) and endotoxin in different stages of ALD. Methods: Nineteen patients with alcoholic cirrhosis (AC), 14 alcoholic hepatitis (AH), 16 alcoholic fatty liver (AFL), 16 subclinical patients from July 2017 to January 2018 at the Second Affiliated Hospital of Baotou Medical College were enrolled, and 15 healthy volunteers were served as healthy controls. Serum IL-12, CDT, HGF levels were detected by ELISA method, and serum endotoxin was detected by limulus test. Diagnostic value of IL-12, CDT for ALD was analyzed by ROC curve. Results: Serum IL-12, endotoxin were significantly increased in AC, AH groups than in AFL, subclinical patients, healthy controls (P<0.05); serum CDT level was significantly increased in AC, AH, AFL, subclinical patients than in healthy controls (P<0.05); serum HGF was significantly increased in AC patients than in AH, AFL, subclinical patients and healthy controls (P<0.05). Serum IL-12, CDT, HGF were positively correlated with serum endotoxin in ALD patients (P<0.05). When the cut-off value of serum IL-12, CDT were 55.06 pg/mL, 354.41 pg/mL, respectively, sensitivity for diagnosing ALD were 0.86, 0.67, respectively, specificity were 0.95, 0.88, respectively. Conclusions: Serum IL-12, CDT, HGF and endotoxin have a trend of change in different stages of ALD, and IL-12 and CDT have high sensitivity and specificity for diagnosing ALD, and can be used as markers for early diagnosis of ALD. Serum HGF and endotoxin are valuable for assessing the severity of ALD.
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Objective: To observe the effect of modified Erchentang on GATA-binding protein-3(GATA3) and T-box expressed in T cells(T-bet) in lung tissue of rats with chronic obstructive pulmonary disease (COPD). Method: Seventy SD rats were randomly divided into seven groups, namely normal group, model group, low, medium and high-dose modified Erchentang group(5,10,20 g ·kg-1), Xiaokechuan group(5 g ·kg-1) and Erchentang group(5 g ·kg-1), with 10 in each group. The rat model of COPD was established by smoking combined with intratracheal dripping of lipopolysaccharide (LPS). After successful modeling, the treatment group was given intragastric administration, and the normal group and the model group were given intragastric administration of equal volume of saline. Enzyme-linked immunosorbent assay (ELISA) was used to determine the concentrations of interleukin-10 (IL-10) and interleukin-12 (IL-12) in rat serum. The expressions of GATA3 and T-bet were detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). The expressions of GATA3 and T-bet in lung tissue were detected by immunohistochemistry (IHC). Result: Compared with the control group, the serum levels of IL-10 in the model group was significantly decreased, while the IL-12 level was significantly increased (PPPPConclusion: Modified Erchentang may reduce the inflammation of lung tissue and improve lung function in COPD rats by reducing IL-12, increasing the content of IL-10, inhibiting the protein and gene expressions of T-bet, and stimulating the protein and gene expressions of GATA3.
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Objective@#To assess the role of T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) in regulation of interleukin (IL) -12 secretion by CD14+ peripheral blood monocytes from patients with psoriasis vulgaris.@*Methods@#From November 2017 to March 2018, a total of 47 patients with psoriasis vulgaris (psoriasis group) and 19 healthy volunteers (control group) were enrolled from Shanxi Provincial People′s Hospital. Peripheral blood mononuclear cells (PBMC) were isolated, and stimulated with Toll-like receptor (TLR) ligands. TIM-3 expression and IL-12 secretion by CD14+ monocytes were measured by flow cytometry. After blockade of TIM-3 pathway by anti-TIM-3 neutralizing antibody, changes in the downstream signaling pathway molecules in and IL-12 secretion by CD14+ monocytes were investigated. Two independent samples t-test was used for comparison between two groups, and Pearson correlation test for correlation analysis.@*Results@#Under the unstimulated condition, the level of IL-12 secreted by CD14+ monocytes was very low, and the proportion of CD14+ TIM-3+ cells was significantly higher in the psoriasis group (12.20% ± 2.83%) than in the control group (9.91% ± 1.77%, t = 3.270, P = 0.001 7) . After the stimulation with TLR ligands, the proportion of CD14+ IL-12+ cells was significantly lower in the psoriasis group (13.49% ± 2.80%) than in the control group (28.97% ± 8.97%, t = 10.71, P < 0.000 1) , but the proportion of CD14+TIM-3+ cells was still higher in the psoriasis group (6.80% ± 1.11%) than in the control group (4.85% ± 1.37%, t = 6.064, P < 0.000 1) . The proportion of CD14+TIM-3+ cells was negatively correlated with that of CD14+ IL-12+ cells in both the control group and psoriasis group (r = -0.473, -0.371 respectively, both P < 0.05) . The TIM-3 pathway blockade could induce the decrease of suppressor of cytokine signaling 1 in CD14+ monocytes, increase the phosphorylation of signaling transducers and activators of transcription 1, and promote IL-12 secretion by CD14+ monocytes induced by keratinocytes isolated from psoriatic skin lesions.@*Conclusion@#TIM-3 plays a crucial role in the negative regulation of CD14+ monocyte-induced innate immune response in psoriasis.
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Objective To investigate the relationship between neonatal umbilical cord blood cytokine interferon-γ (IFN-γ),interleukin-4 (IL-4),interleukin-12 (IL-12),interleukin-18 (IL-18) and hepatitis B virus (HBV) intrauterine infection.Methods Seventy-five newborns delivered by HBsAg-positive pregnant women in the First Hospital of Hebei Medical University and the Fifth Hospital of Shijiazhuang from December 2017 to June 2018 were selected as observation group.According to the results of five items of hepatitis B and HBV DNA test in cord blood of newborns,17 of them were positive as intrauterine infection group,and 58 of them were negative as uninfection intrauterine group.Forty-three newborns delivered by healthy pregnant women with negative HBsAg were taken as control group.The levels of cytokines IFN-γ,IL-4,IL-12 and IL-18 in cord blood of neonates were detected by ELISA,Results The levels of IFN-γ,IL-4,IL-12 and IL-18 in the newborns of intrauterine infection group were (409.51 ±51.77),630.51(612.49,647.33),85.60(56.11,133.99),32.41 (23.04,87.53) ng/L.The levels in the uninfected intrauterine Group were (523.87 ± 38.45),573.33 (531.95,598.38),186.53 (77.77,302.66),125.99(63.32,202.73) ng/L.The levels in the control group were (509.39±73.02),565.83 (443.40,620.82),199.89 (128.92,289.30),152.98 (86.76,188.57) ng/L.There were significant differences in IFN-γ,IL-4,IL-12,IL-18 between the intrauterine infection group and the uninfected intrauterine group and the control group (all P＜0.01).There was no significant difference between the uninfected group and the control group (all P＞0.05).Conclusion The decrease of IFN-γ,IL-12,IL-18 and the increase of IL-4 in cord blood of neonates result in the decrease of viral clearance ability and the failure of HBV clearance,which leads to intrauterine infection of neonates with HBV.
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Objective To assess the role of T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) in regulation of interleukin(IL)-12 secretion by CD14+ peripheral blood monocytes from patients with psoriasis vulgaris.Methods From November 2017 to March 2018,a total of 47 patients with psoriasis vulgaris (psoriasis group) and 19 healthy volunteers (control group) were enrolled from Shanxi Provincial People's Hospital.Peripheral blood mononuclear cells (PBMC) were isolated,and stimulated with Toll-like receptor (TLR) ligands.TIM-3 expression and IL-12 secretion by CD14+ monocytes were measured by flow cytometry.After blockade of TIM-3 pathway by anti-TIM-3 neutralizing antibody,changes in the downstream signaling pathway molecules in and IL-12 secretion by CD14+ monocytes were investigated.Two independent samples t-test was used for comparison between two groups,and Pearson correlation test for correlation analysis.Results Under the unstimulated condition,the level of IL-12 secreted by CD14+ monocytes was very low,and the proportion of CD14+TIM-3+ cells was significantly higher in the psoriasis group (12.20％ ± 2.83％) than in the control group (9.91％ ± 1.77％,t =3.270,P =0.001 7).After the stimulation with TLR ligands,the proportion of CD14+IL-12+ cells was significantly lower in the psoriasis group (13.49％ ± 2.80％) than in the control group (28.97％ ± 8.97％,t =10.71,P ＜0.000 1),but the proportion of CD14+TIM-3+ cells was still higher in the psoriasis group (6.80％ ± 1.11％)than in the control group (4.85％ ± 1.37％,t =6.064,P ＜ 0.000 1).The proportion of CD14+TIM-3+ cells was negatively correlated with that of CD14+IL-12+ cells in both the control group and psoriasis group (r =-0.473,-0.371 respectively,both P ＜ 0.05).The TIM-3 pathway blockade could induce the decrease of suppressor of cytokine signaling 1 in CD14+ monocytes,increase the phosphorylation of signaling transducers and activators of transcription 1,and promote IL-12 secretion by CD 14+ monocytes induced by keratinocytes isolated from psoriatic skin lesions.Conclusion TIM-3 plays a crucial role in the negative regulation of CD 14+ monocyte-induced innate immune response in psoriasis.
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Objective: To evaluate effects of docosahexaenoic acid-enriched phosphatidylcholine (DHA-PC) on cytokine production induced by lipopolysaccharide (LPS). Methods: The culture supernatants of splenocytes exposed to DHA-PC along with LPS were harvested to determine the production of Th 1 (IFN- γ and IL-2) and Th2 [IL-4, IL-5, IL-6 and IL-12/IL-23(p40)] cytokines. Cytokines were measured using ELISA. Results: Co-administration of DHA-PC with LPS resulted in significantly lower IL-2 expression compared to that observed with administration of only LPS (P<0.01). Treatment with DHA-PC and LPS significantly increased IL-5 expression (P<0.01). Moreover, co-administration of DHA-PC with LPS significantly decreased IL-6 and IL-12/IL-23(p40) expressions compared to that observed with administration of only LPS (P<0.01). Conclusions: Our results suggest that DHA-PC inhibits pro-inflammatory cytokines [IL-2, IL-6 and IL-12/IL-23(p40)] expression on induction of inflammation.
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Objective: To investigate the effect of chemotherapy combined with sodium cantharidinate vitamin B6 injection in the patients with non-small cell lung cancer (NSCLC) and its effect on the patients' quality of life, and to clarify its mechanism. Methods: A total of 110 NSCLC patients were divided into chemotherapy group and combined group according to the different treatment methods adopted by the patients. A total of 55 patients were treated with chemotherapy alone (chemotherapy group) and 55 patients treated with chemotherapy combined with sodium cantharidate vitamin B6 injection (combined group). The total effective rate of the patients in two groups were compared; EORTC QLO-C43 scale was used to evaluate the quality of life of the patients in two groups before and after treatment; the serum levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-a), and interleukin-12 (IL-12) of the patients in two groups before and after treatment were detected by double-antibody sandwich enzyme-linked immunoassay method. Results: After treatment, the total effective rate of the patients in combined group was higher than that in chemotherapy group, but the difference was not statistically significant (χ2 = 0.806, P= 0.369). Before treatment, there were no significant differences in the scores of functional scale, overall health status, symptom subscale and lung cancer specific subscale of the patients between combined group and chemotherapy group (P>0. 05). After treatment, the scores of functional scale and overall health status of the patients in combined group were significantly higher than those in chemotherapy group (P0. 05). After treatment, the serum level of VEGF of the patients in combined group was significantly lower than that in chemotherapy group (χ2= 3. 608, P<0. 05), and the serum levels of TNF-a and IL-12 were significantly higher than those in chemotherapy group (t= 3.426, P<0.05; t= 2.849, P < 0. 05). Conclusion: The effect of chemotherapy combined with sodium cantharidate vitamin B6 injection in the treatment of the NSCLC patients is good, and it can significantly improve the quality of life of the patients; its mechanism may be related to decreasing the serum VEGF level and increasing the serum TNF-α and IL-12 levels.
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Objective: To construct the murine chimeric antigen receptor-redirected T lymphocyte (CAR-T) that can target glypican 3 (GPC3) and express interleukin-12 (IL-12) in an activation-dependent manner, and to explore the anti-cancer efficacy of CAR-T cells in immunocompetent mice bearing GPC3 positive (GPC3+) breast cancer. Methods: Gene recombination technology was used to construct the retroviral vector encoding GPC3-specific CAR (m9.28z) or encoding both GPC3-specific CAR and nuclear factor of activated T cells (NFAT)-IL-12 expression system (m9.28z/IL-12). Murine T cells were retrovirally transduced to develop m9.28z CAR-T cells and m9.28z/IL-12 CAR-T cells. In vitro, the cytotoxicity assay and ELISA assay were used to assess the biological functions of m9.28z/IL-12 CAR-T cells. In vivo, the GPC3+ breast cancer E0771-GPC3 cell subcutaneously transplanted model was established in C57BL/6 mice, and the tumor-bearing mice were injected with 5×106 murine untransduced T cells (mUTD, which served as the negative control), 5×106 m9.28z CAR-T cells, 2×106 m9.28z CAR-T cells, 5×106 m9.28z/IL-12 CAR-T cells and 2×106 m9.28z/IL-12 CAR-T cells, respectively. The growth of xenografts in nude mice was observed, and the expression levels of CD8α and forkhead box protein P3 (FOXP3) in tumor tissues were detected by immunohistochemistry. Results: The retroviral vector encoding m9.28z or m9.28z/IL-12 was constructed, and the m9.28z CAR-T cells and m9.28z/IL-12 CAR-T cells were developed subsequently. In vitro, the m9.28z/IL-12 CAR-T cells killed GPC3+ breast cancer cells specifically and effectively, and secreted more tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) than m9.28z CAR-T cells (both P < 0.05). In vivo, as compared with m9.28z CAR-T cells, the m9.28z/ IL-12 CAR-T cells suppressed the growth of xenografts significantly (P < 0.001), and there were more CD8+ T cells and less regulatory T cells (Treg) in tumor tissues treated with m9.28z/IL-12 CAR-T cells (both P < 0.01). Conclusion: The m9.28z/IL-12 CAR-T cells, which can express IL-12 inducibly, can display more potent anti-cancer activity against GPC3+ tumor than m9.28z CAR-T cells in vitro and in immunocompetent breast cancer transplanted mice.
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Objective: To evaluate effects of docosahexaenoic acid-enriched phosphatidylcholine (DHA-PC) on cytokine production induced by lipopolysaccharide (LPS). Methods: The culture supernatants of splenocytes exposed to DHA-PC along with LPS were harvested to determine the production of Th 1 (IFN-γ and IL-2) and Th2 [IL-4, IL-5, IL-6 and IL-12/IL-23(p40)] cytokines. Cytokines were measured using ELISA. Results: Co-administration of DHA-PC with LPS resulted in significantly lower IL-2 expression compared to that observed with administration of only LPS (P<0.01). Treatment with DHA-PC and LPS significantly increased IL-5 expression (P<0.01). Moreover, co-administration of DHA-PC with LPS significantly decreased IL-6 and IL-12/IL-23(p40) expressions compared to that observed with administration of only LPS (P<0.01). Conclusions: Our results suggest that DHA-PC inhibits pro-inflammatory cytokines [IL-2, IL-6 and IL-12/IL-23(p40)] expression on induction of inflammation.
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Objective To investigate the changes of peritoneal macrophage function in sepsis mice after hyperbaric oxygen (HO) therapy.Methods One hundred C57BL/6 mice were randomly(random number) divided into four groups in equal number (n=25):control group (normal saline),sepsis group,hyperbaric oxygen group,sepsis+ hyperbaric oxygen group;the sepstic mice were treated by intra-peritoneal injection with zymosan;The mice of hyperbaric oxygen group and sepsis+ hyperbaric oxygen group received hyperbaric oxygen therapy (2 atm,2 h,100％ O2).The number of surviving mice was obsevred at 72 h after the intra-peritoneal injection with zymosan;The percentage of M1 macrophage was detected by flow cytometry.The expression of mRNA of inducible nitric oxide synthase (iNOS) in peritoneal macrophage was detected with RT-PCR.The levels of IL-12 and IL-10 in peripheral blood were measured by ELISA.Results Compared with sepsis group,the number of surviving mice of sepsis+ hyperbaric oxygen group was significantly higher (9 vs.16,P＜0.01).The percentages of M1 macrophage in sepsis+ hyperbaric oxygen group was significantly lower than that in sepsis group [(4.75+0.14)％ vs.(2.25±0.16)％,F=803.438,P＜0.01].The expression of mRNA of inducible nitric oxide synthase(iNOS) in sepsis+ hyperbaric oxygen group were significantly lower than that in sepsis group [(39.62+1.74) vs.(48.32±3.34),F=31.992,P＜0.01].The level of IL-12 in peripheral blood in sepsis+ hyperbaric oxygen group was significantly lower than that in sepsis group [(242.62±19.10) pg/mL vs.(159.51±6.35)pg/mL,F=102.282,P＜0.01].The level of IL-10 was significantly higher than that in sepsis group [(521.26±6.3) pg/mL vs.(188.83±8.53) pg/mL,F=5 896.006,P＜0.01].Conclusions Hyperbaric oxygen therapy had effective effect on increasing the number of surviving mice of sepsis group,reducing the level of IL-12 in peripheral blood and rising the level of IL-10 in peripheral blood.By inhibiting the expression of M1 macrophage and iNOS,hyperbaric oxygen could reduce the excessive inflammatory response and play a role in the protection of mice.
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Abstract: Approximately 10% of individuals do not respond to hepatitis B virus (HBV) vaccination, i.e. non-responders (NRs). We aimed to investigate the association of interleukin (IL)-4 and IL-12B gene polymorphisms with responsiveness to the HBV vaccine in Korean infants. Among 300 healthy infants (9-12 month), SNPs for the IL-4 gene (rs2243250, rs2070874, and rs2227284) and for the IL-12B gene (rs3213094 and rs17860508) were compared between subgroups in terms of the response to HBV vaccination. The percentages of NRs (< 10 mIU/mL), low-titer responders (LRs, 10-100 mIU/mL), and high-titer responders (HRs, ≥ 100 mIU/mL) were 20.3%, 37.7% and 42.0%, respectively. No SNPs differed in frequency between NRs and responders or between LRs and HRs. We divided the subjects into two groups according to the time interval from the 3rd dose of HBV vaccination to Ab quantification: > 6 months from the 3rd dose (n = 87) and ≤ 6 months from the 3rd dose (n = 213). In the ≤ 6 month subjects, rs2243250C and rs2227284G were significantly frequent in the lower-titer individuals (NRs + LR) than HRs (40.1 vs. 25.9%, p = 0.014 and 45.1 vs. 33.0%, p = 0.018, respectively), and the rs2243250C and rs2227284G frequencies were significantly different among the three subgroups (13.2 vs. 26.9 vs. 25.9%, p = 0.040 and 15.5 vs. 29.6 vs. 33.0%, p = 0.038, respectively). In conclusion, those results suggest that IL-4 gene polymorphisms may play a role in the response to the HBV vaccine in Korean infants.
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Humans , Male , Female , Infant , Interleukin-4/genetics , Hepatitis B Vaccines/administration & dosage , Polymorphism, Single Nucleotide , Interleukin-12 Subunit p40/genetics , Hepatitis B/prevention & control , Pharmacogenetics , Phenotype , Time Factors , Biomarkers/blood , Hepatitis Antibodies/blood , Immunization Schedule , Vaccination , Treatment Outcome , Republic of Korea , Gene Frequency , Hepatitis B/genetics , Hepatitis B/immunology , Hepatitis B Surface Antigens/bloodABSTRACT
Objective To study the influence of isoniazid on lymphocyte factor expression and macrophage function of rats.Methods Healthy male SD rats were randomly divided into three groups,which was treated for one month,three months and withdrawal for one month after treated for three months,and each group was randomly divided into isoniazid group and control group.The isoniazid groups were ig with isoniazid at dose of 120 mg/kg every other day and control groups were fed on normal saline.At the corresponding time points,the level of interleukin-12 (IL-12) and interferon-γ (IFN-γ) was detected with ELISA method,detected serum lysozyme content by agar plate method,and Comori method was used for the detection of acid phosphatase levels in peritoneal fluid.Results At all the time points,levels of IL-12,IFN-γ and lysozyme in isoniazid group were not significantly different compared with control group.There were statistically significant differences in acid phosphatase between isoniazid group and control group after treated for one month (P ＜ 0.05),but the significant differences disappeared at the next two time points.Conclusion Isoniazid of 120 mg/kg may have no obvious influence on the immune function of rat.We don't detect the immune injury.
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Objective To study the influence of isoniazid on lymphocyte factor expression and macrophage function of rats.Methods Healthy male SD rats were randomly divided into three groups,which was treated for one month,three months and withdrawal for one month after treated for three months,and each group was randomly divided into isoniazid group and control group.The isoniazid groups were ig with isoniazid at dose of 120 mg/kg every other day and control groups were fed on normal saline.At the corresponding time points,the level of interleukin-12 (IL-12) and interferon-γ (IFN-γ) was detected with ELISA method,detected serum lysozyme content by agar plate method,and Comori method was used for the detection of acid phosphatase levels in peritoneal fluid.Results At all the time points,levels of IL-12,IFN-γ and lysozyme in isoniazid group were not significantly different compared with control group.There were statistically significant differences in acid phosphatase between isoniazid group and control group after treated for one month (P ＜ 0.05),but the significant differences disappeared at the next two time points.Conclusion Isoniazid of 120 mg/kg may have no obvious influence on the immune function of rat.We don't detect the immune injury.