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Alteration of level of panel reactive antibodies, TGF-βin serum and expression rate of nTreg cells in peripheral blood in women with unexplained recurrent spontaneous abortion after immunotherapy

Xingbin REN ; Wei ZHANG

Chinese Journal of Immunology.2015;(6):831-834. doi:10.3969/j.issn.1000-484X.2015.06.025

Objective: To investigate the effects of immunotherapy on PRA, TGF-β and CD4+CD25+Foxp3+ nTreg cells in women with unexplained recurrent spontaneous abortion. Methods:We detect the level of PRA,TGF-βin serum and the expression of nTreg cells in peripheral blood of pregnant, non pregnant and natural fertility women by enzyme immunoassay and flow cytometry. Results:After immunotherapy,the median level of PRA and TGF-βin non pregnant group was lower than that in pregnant group(27. 5%,107. 7 pg/L vs 68. 75%,189. 9 pg/L,P<0. 01). Compared to pregnant group,the median expression rate of nTreg cells were lower in non pregnant group(5. 05% vs 8. 05%,P<0. 01). Conclusion:The PRA,TGF-βand nTreg cells are critical factors for pregnancy in RSA patients. It is helpful to identify and judge the effect of immunotherapy by detect the level of panel reactive antibodies,TGF-β in serum and the expression rate of nTreg cells in peripheral blood.

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Evaluation of neutrophilic CD64 index and CD32 index as a diagnostic marker of liver cirrhosis with spontaneous bacterial peritonitis in early stage

Jinhai ZOU ; Xiangjun KONG ; Chunying LI ; Xinyi SUN ; Chendi LIU

Chinese Journal of Immunology.2015;(6):827-830. doi:10.3969/j.issn.1000-484X.2015.06.024

Objective:To investigate the early diagnosis value of neutrophilic CD 64 index(nCD64 ID),neutrophilic CD32 index( nCD32 ID) in ascites and CRP in blood of liver cirrhosis patients combined with spontaneous bacterial peritonitis. Methods:The data of 156 cases with liver cirrhosis was analyzed retrospectively, which CD32 index, CD64 index and CRP were detected respectively and ROC curve analysis were performed. Results:The nCD64 ID,nCD32 ID and CRP in bacterial infection group were all significantly higher than that in no infection group(P<0. 001). The sensitivity and specificity of nCD32 ID,nCD64 ID and CRP were 82. 8%,96. 2%,72. 5% and 81. 0%, 95. 8%, 73. 1% respectively. Conclusion: The sensitivity and specificity of nCD64 ID were higher than nCD32 ID and CRP. The nCD64 ID can be used as an effective index for early diagnosis and differential diagnosis of liver cirrhosis combined with spontaneous bacterial peritonitis.

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Expression and biological significance of microRNA-204 in nasopharyngeal carci-noma

Chengyi JIANG ; Hongtao WANG ; Lei ZHOU ; Tao JIANG ; Yajia XU

Chinese Journal of Immunology.2015;(6):822-826. doi:10.3969/j.issn.1000-484X.2015.06.023

Objective:To investigate the expression and biological significance of MicroRNA-204 in nasopharyngeal carcinoma (NPC). Methods: qRT-PCR was applied to detect the relative expression of miR-204 in 43 paired nasopharyngeal carcinoma in comparison to the normal nasal mucosa tissues. Pearson chi-square test was used to analyze the relationship between the miR-204 expression and clinical features. The expressions of Bcl-2 and SIRT1 were measured by immunohistochemistry ( IHC ) , Spearman correlation analysis was used to analyze the relationship between miR-204 and Bcl-2,as well as SIRT1. We then transfected the miR-204 mimics into CNE-2 cells,then the Western blot was used to detect the expression of Bcl-2 and SIRT1,which were considered as the potential targets of miR-204. Results:The relative expressions of miR-204 was significantly downregulated in NPC tissues compared to those of the matched normal tumor-adjacent tissues(P<0. 05). Low expression of miR-204 was significantly associated with lymphatic metastasis(P<0. 05) and advanced TNM stage(Ⅲ+Ⅳ,P<0. 05). The expressions of Bcl-2 and Sirt1 in lower miR-204 level group were both higher than in higher miR-204 level group ( P<0. 05 ) . Both the mRNA and protin expression in CNE-2 cells were down-regulated after transfection. Conclusion: Low expression of miR-204 is related to the malignant clinicopathological features in NPC tissues,and miR-204 may through down-regulate Bcl-2 and SIRT1 to suppress NPC genesis and development.

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Relationship of clinical outcomes and expression of microRNA-199 a/b-3 p in HCC

Lei LIU ; Shouqing LI ; Xuguang MI ; Yanqiu FANG

Chinese Journal of Immunology.2015;(6):806-808. doi:10.3969/j.issn.1000-484X.2015.06.019

Objective:To investigate the expression of microRNA-199a/b-3p (miR-199a/b-3p) in hepatocellular carcinoma ( HCC) tissues,and to explore the relationship with clinical outcomes. Methods: Real time quantitative PCR technique was used to measure the expression of miRNA-199a/b-3p in HCC tissues. The correlation between miR-199a/b-3p expression and the clinic pathological features of patients were analyzed. Results: Comparing with adjacent control, miRNA-199a/b-3p presented lower expressions in HCC tissues (P<0. 05);lower miR-199a/b-3p was found correlated with metastasis and poor survival. Conclusion:MiR-199a/b-3p take a crucial role in HCC metastasis and recurrence.

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Experimental study on inguinal subcutaneous immunotherapy for allergic rhinitis in mice

Zhenan ZHAO ; Ji DAI ; Wanjun ZHAO ; Qingyun WANG ; Zhongsheng CAO

Chinese Journal of Immunology.2015;(6):785-789. doi:10.3969/j.issn.1000-484X.2015.06.014

Objective:To explore the feasibility of inguinal subcutaneous immunotherapy for allergic rhinitis ( AR ) in mice. Methods:36 female BALB/c mice were divided randomly into six groups( n=6 per group) including the control A,the model A, the treatment A groups,and the control B,the model B,the treatment B groups(inguinal subcutaneous immunotherapy for group A, cervical back subcutaneous immunotherapy for for group B). AR model was established with ovalbumin. At 25 to 55 days,ovalbumin im-munotherapy were performed in treatment groups,once two days,15 times totally. After intranasal rechallenge was performed at 56 to 62 days the AR symptom scores were documented. The eosinophils(EOS)in the nasal mucosa were measured by chromotropic acid 2R staining. Ovalbumin-specific IgE( OVA-sIgE) in the serum and expression of interferon-γ and interleukin-4 in the nasal lavage were measured by enzyme-linked immunosorbent assay meanwhile the ratio of interferon-γ and interleukin-4 was calc μlated. SPSS17. 0 software was used to analyze the data. Results:Before treatment ,the AR symptom scores of the model and treatment groups were more than 5. After treatment,the treatment A group were less than 5. The EOS count of the control A,model A,treatment A groups and the control B,model B, treatment B groups was 0. 78 ± 0. 31, 21. 60 ± 2. 90, 10. 43 ± 2. 56, 0. 83 ± 0. 46, 22. 44 ± 3. 39, 23. 40 ± 4. 24, respectively. The EOS count of the treatment A group was significantly lower than those in model A group ( P<0. 05 ) . There was no significant difference between treatment B and model B group ( P>0. 05 ) . OVA-sIgE expressed was negative in control groups and positive in other groups. The ratio of interferon-γ and interleukin-4 was 10. 75 ± 3. 38,10. 38 ± 3. 08,3. 02 ± 0. 69,2. 71 ± 0. 89,2. 52 ± 0. 30,5. 45±1. 41,respectively. The ratio in treatment A group was significantly higher than those in model A group(P<0. 05). There was no significant difference between treatment B and model B group ( P>0. 05 ) . Conclusion: Inguinal subcutaneous immunotherapy has a good effect on this disease. It spends short time ,has simple operation and good feasibility,which is a novel treatment method for AR in mice.

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γδT cells inhibit transdifferentiation of immature dendritic cells into osteoclasts in vitro

Qingjiao CHEN ; Zhiyong ZENG ; Dongbiao QIU ; Junmin CHEN

Chinese Journal of Immunology.2015;(6):778-784. doi:10.3969/j.issn.1000-484X.2015.06.013

Objective:To explore the role of γδ T cells in the transdifferentiation of immature dendritic cells(imDC) into osteoclasts(OC). Methods:(1) Peripheral blood mononuclear cells(PBMNC) were cultured with zoledronate(Zol) and recombinant human interleukin-2(IL-2),and PBMNC from healthy volunteers were cultured with granulocyte macrophage colony-stimulating factor (GM-CSF) and recombinant human interleukin-4(IL-4) to differentiate into imDC,which were then cultured with receptor activator nuclear factor к B ligand(RANKL) and macrophage colony-stimulating factor(M-CSF) to differentiate into OC. The purity of γδ T cells,and phenotype changing of OC transdifferentiated from imDC were investigated by flow cytometry. (2) Co-culture system was es-tablished using millicell inserts.γδT cells isolated with immune magnetic bead were placed in the upper compartment and imDC in the lower compartment in the ratio of 10∶1. To explore the role of γδ T cells during differentiation of imDC into OC,tartrate resistant acid phosphatase( TRAP) staining and bone resorption observation staining were used. Tumor necrosis factor-alpha( TNF-α) of supernatant liquid from different cultures was measured using ELISA(Enzyme linked immunosorbent assay) kit. Results:(1) γδT cells can be ex-panded from PBMNC of MM patients, and the production capacity was similar to that of healthy volunteers ( 68. 87%± 20. 94% vs 69. 33%±16. 84%,P>0. 05 ) . ( 2 ) OC could be transdifferentiated from imDC when cultured with RANKL and M-CSF. ( 3 ) The number of TRAP+ multinuclear cell and the absorption area of dentine were significantly lower in the group of imDC indirectly co-cultured with γδ T cells than in the group of control imDC(5.67±0.58 vs 28.33±2.08,4.97%±4.3% vs 28.47%±12.8%, respectively). (4) Under the circumstance of γδ T cell-imDC indirect coculture,TNF-α got significantly higher. Conclusion: γδ T cells might inhibit the transdifferentiation of imDC into OC.γδ T cells-based immunotherapy is expected to be a new treatment for myeloma bone disease.

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Molecular mechanism of apigenin on inhibition of LPS-induced inflammatory mediators in murine macrophages

Guang WU ; Ping FU ; Yusheng ZHOU ; Runmei ZHOU

Chinese Journal of Immunology.2015;(6):753-757. doi:10.3969/j.issn.1000-484X.2015.06.007

Objective:To investigate the effect and the mechanism of Apigenin on lipopolysaccharides ( LPS )-induced inflammatory mediators production in murine macrophages. Methods:The murine macrophage cell line RAW 264. 7 cells were cultured in vitro,and were treated with different concentration of Apigenin followed by LPS administration. Expression of heme oxygenase-1 ( HO-1),cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS),phosphorylation of p38 and IκB,nuclear translocation of Nrf2 were detected by Western blot. Production of Nitrite and nitrate ( NOx) was analyzed by colorimetric technique. Secretion of prosta-glandin E2 (PGE2) was detected by ELISA. Activation of NF-κB was measured by luciferase assay. Results: Western blot indicated that apigenin could induce RAW 264. 7 cells expression of HO-1, and pretreatment of SB203580, an inhibitor of p38 significantly inhibited apigenin induced HO-1 expression. In addition,Apigenin could also decrease the content of nuclear transcription factor Nrf2 in cytoplasm and increase its level in the nucleus. Silencing of Nrf2 by specific siRNA could inhibit apigenin-induced HO-1 expression. Furthermore,apigenin administration significantly inhibited LPS-induced NOx production and PGE2 secretion, COX-2 and iNOS expression,IκB phosphorylation and NF-κB activation,and transfection of HO-1 siRNA could reverse these actions. Conclusion:Apigenin inhibits LPS-induced inflammatory response through induction of HO-1 and inhibition of NF-κB in macrophages.

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Advances in inflammasome and inflammasome-related diseases

Yan WANG ; Bing SUN

Chinese Journal of Immunology.2015;(6):721-727. doi:10.3969/j.issn.1000-484X.2015.06.001

Inflammasomes are multi-protein complexes that trigger the activation of caspase-1 and the maturation of interleukin-1β ( IL-1β) , which are critical for inflammation and control of pathogen infection. Inflammasomes are involved in and regulate diversified diseases due to its capability to sense multiple danger signals. In this review, we present the activation and regulation mechanisms for different inflammasomes and discuss how genetic mutations in inflammasome-related genes or abnormal activity of in-flammasomes lead to diseases. Insights into the role of inflammasome in various human diseases will provide sound theoretical basis and effective treatment strategy for these disease.

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Immunity mechanism of vertical transmission blocking for HBV immunoglobulin injection to pregnant women

Yuelan LIANG ; Shengmiao FU

Chinese Journal of Immunology.2015;(6):818-821. doi:10.3969/j.issn.1000-484X.2015.06.022

Objective:To discuss the passive immunity effect and mechanism of HBIG injections to matrix to block the mother-to-child vertical transmission of HBV. Methods:94 cases of patients with chronic viral hepatitis were selected and divided into 3 groups randomly. 31 cases of control group were given no HBIG intervention,while 31 cases of baby intervention ( BBI) group were given HBIG injection in 6h of birth,and 32 cases of infant & mom intervention ( IMI) group were given HBIG injection respectively in 28,32,36 weeks of gestation and 6h of birth. Further more,all newborns were vaccinated against hepatitis B in 0,1 and 6 months,after the last vaccination,peripheral blood of the children were extracted and detected for HBV markers,HBV-DNA and immune function. Results:There were significant difference (P<0. 05) in neonatal HBeAg,HBsAg and HBV-DNA positive rate for the three groups,with Control group got the highest while IMI group got the lowest;and there are also significant differences (P<0. 05) HBeAb positive rate,with Control group got the lowest while IMI group got the highest. We also found that the complement (C3,C4) levels and T cell subtypes (CD3+,CD4+,CD8+) count of the three groups of newborns had significant differences too(P<0. 05),with Control group got the lowest while IMI group got the highest;in terms of immunoglobulin,both the IMI and BBI group were higher in IgG and IgM level (P<0. 05), while there was no obvious difference in IgA between groups (P<0. 05). Conclusion:Maternal HBIG injections can effectively activate the maternal humoral immunity and cellular immunity,resulting in the decrease of HBV. It can also improve newborn′s antigen-antibody response and relieve T lymphocytes loss induced by antiviral consumption through placenta,which may play great role in the passive im-munity mechanism of blocking mother-to-fetus transmission.

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Effect of sodium selenite on Treg cells in peripheral blood mononuclear cells in patients with Graves disease

Yu YANG ; Chun LIU

Chinese Journal of Immunology.2015;(6):814-817. doi:10.3969/j.issn.1000-484X.2015.06.021

Objective:Recently,selenium was used for the treatment of Graves disease. However,there has been no reports that selenium affected the immune function of Graves disease at the cellular level so far. It is reported the effect of sodium selenite on Treg cells in peripheral blood mononuclear cells in patients with Graves disease in the present study. Methods:AFS were used to detect the level of serum selenium;PBMC( Peripheral blood mononuclear cells) were extracted,then,cultured with sodium selenite. Foxp3 mRNA expression was tested by Real-time fluorescence PCR,and ELISA was used to detect the secretion of IL-35. Results:The serum sodium selenite levels of GD group are significantly lower than the HC group;The expression of Foxp3 mRNA and secretion of IL-35 are both increased. Conclusion:It shows sodium selenite could improve the state of immune disorders by recovering the function of Treg cells.

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