The guidance of urine concentration of chemokine IP-10 in clinical use of rabbit anti-human T-lymphocyte immunoglobulin / 实用医学杂志

Objective To investigate evaluation role of IP-10 level in urine of kidney transplant recipients when using rabbit anti-human T-lymphocyte immunoglobulin to treat acute cellular rejection.Methods A total of 40 patients who underwent renal transplantation and had been diagnosed as acute cellular rejection according to the results of histopathological examination were randomly divided them into IP-10 group (n =20) and serum creatinine group (Scr group,n =20).Urinary IP-10 and Scr levels were measured in time and patients then were treated with ATG,of which the doses and duration were adjusted according to IP-10 or Scr levels.We compared the total and daily ATG dosages,ATG administration period,side effects of ATG such as incidence of severe platelet and neutropenia,acute rejection during first 3 months and infection rates during first 1 year.Result The number of ATG duration is 5.35 ± 1.93 for IP-10 group versus 6.70 ± 1.75 for Scr group.We used a daily dose of 2.50 ± 0.57 mg/(kg·d) for IP-10 group and 2.77 ± 0.74 mg/(kg· d) for Scr group,a total dose of 13.40 ± 6.59 mg/kg for IP-10 group and 18.25 ± 7.35 mg/kg for Scr group.There was significance between the two group in above three outcomes (P ＜ 0.05).There was no significance in incidences of severe thrombocytopenia and neutropenia,incidences of acute rejection during first 3 months,incidences of infection during first 1 year between the two group (P ＞ 0.05).Conclusion Urine IP-10 test is effective and reliable indicators which can guide ATG usage in patients with acute rejection and reduce the ATG cost.

Roles of SAA, IP-10 and PCT in differential diagnosis of AECOPD / 实用医学杂志

Objective To assess the clinical significance of SAA, IP-10 and PCT in the diagnosis of AECOPD. Methods Sixty AECOPD patients, 52 with sCOPD, and 28 healthy subjects were assigned to three groups. Clinical data and serum specimen were obtained from another 19 AECOPD patients at stable stage as AECOPD-sCOPD group. Serum levels of SAA, IP10 and PCT were quantitatively measured by ELISA. Levels of multiple serum markers were statistically compared among different groups. Results The concentration of SAA significantly differed between the AECOPD and sCOPD groups (P 0.05). Conclusions As compared with the sCOPD group, levels of serum SAA and IP-10 in the AECOPD group were significantly elevated, which is helpful in the diagnosis of AECOPD with a sensitivity and specificity of 100% and 54.9%for SAA, and 96.1%and 75.0%for IP-10. However, PCT level failed to identify AECOPD from sCOPD.

Helicobacter pylori induces NOD1/NF-κB activation and IFN-βand IP-10 production in gastrics of mice / 中国免疫学杂志

Objective:To construct the Helicobacter pylori infected C57BL/6 mice model to observe the activation of NOD1/NF-κB signaling pathways in the gastric tissues,and study its roles in inflammatory response during Hp infection.Methods:6-8 week-old C57BL/6 mice were randomly divided into two groups,the Hp infection group and the control group,and mice were given by gavage every 48 h for five times with Hp or PBS,respectively.All the animals were sacrificed at different time point and the gastric tissue were stained with hematoxylin-eosin( HE);The mRNA expression of NOD1 and RIP2 in gastric tissues were examined by RT-PCR;Levels of IFN-βand IP-10 in mice serum were assessed by ELISA;Nuclear translocation of p65 in gastric tissue was detected by Western blot.Results:Hp infection elicits an inflammatory cell response,glands in gastric tissue were reduced or atrophic,as compared with that in the control group.The levels of IP-10 and IFN-βincreased in the model group, and peaked at 16 weeks after Hp infection.Hp infection increased the mRNA expression of NOD1 and the p65 content in nuclear between 24-120 h(P<0.05),and the highest level at 48 h,subsequently the expression levels were began to decrease.The mRNA expression level of RIP2 was up-regulated after Hp was administrated, peaked at 48 h and declined after 72 h.However, the expression levels would rise again at 120 h.Conclusion: Hp infection can activate the NOD1/NF-κB signaling pathways and induce the production of IFN-βand IP-10 in gastrics of mice.

Dipyrone & 2,5-dimethylcelecoxib suppress Th2-related chemokine production in monocyte.

Background & objectives: Selective cyclooxygenase-2 (COX-2) inhibitor is a form of non steroidal anti-inflammatory drug (NSAID) and is commonly used in autoimmune and rheumatic diseases to control inflammation and alleviate pain. Tumour necrosis factor-alpha (TNF-α) production and an imbalance of T helper 1 (Th1)/Th2 contribute to the pathogenesis of autoimmune and also anti-tumour activity. Dipyrone is a NSAID used to treat pain worldwide. The celecoxib analogue, 2,5-dimethylcelecoxib (DMC), lacks COX-2 inhibitory activity but exhibits anti-tumour properties. However, the effects and the mechanisms of dipyrone and 2,5-dimethylcelecoxib on tumour necrosis factor (TNF)-α and Th1- and Th2-related chemokines in monocytes remain poorly defined. This study was carried out to investigate the effects of dipyrone and 2,5-dimethylcelecoxib on the expression of Th1 (IP-10) and Th2 (I-309 and MDC) and TNF-α in human monocytes and the associated intracellular mechanism. methods: THP-1 cells and peripheral blood mononuclear cells (PBMCs) were pre-treated with dipyrone (10-9 – 10-4 M) and 2,5-dimethylcelecoxib (10-9 – 10-5 M) 2 h before lipopolysaccharide (LPS) stimulation. Cell supernatant was collected 24 h after LPS stimulation. TNF-α, I-309, MDC and IP-10 concentrations of cell supernatants were determined using ELISA. Intracellular signaling was evaluated by western blot. results: Dipyrone and 2,5-dimethylcelecoxib downregulated LPS-induced Th2-related chemokine I-309 and macrophage derived chemokine (MDC) production. Only high dose of 2,5-dimethylcelecoxib (10-5 M), but not dipyrone downregulated LPS-induced IP-10. Only very high dose of 2,5-dimethylcelecoxib had effect on LPS-induced TNF-α expression in PBMCs. Dipyrone and 2,5-dimethylcelecoxib suppressed LPS-induced p65 and JNK MAPK (C-Jun N-terminal kinase mitogen activated protein kinase). expression. Interpretation & conclusions: Dipyrone and 2,5-dimethylcelecoxib downregulated LPS-induced Th2-related chemokine I-309 and MDC in THP-1 cells. The suppressive effect on Th2-related

Benzylideneacetophenone derivatives attenuate IFN-gamma-induced IP-10/CXCL10 production in orbital fibroblasts of patients with thyroid-associated ophthalmopathy through STAT-1 inhibition

The aim of the present study was to identify a new candidate anti-inflammatory compound for use in the active stage of thyroid-associated ophthalmopathy (TAO). Benzylideneacetophenone compound JC3 [(2E)-3-(4-hydroxy-3-methoxyphenyl)phenylpro-2-en-l-one] was synthesized based on a structural modification of yakuchinone B, a constituent of the seeds of Alpinia oxyphylla, which belongs to the ginger family (Zingiberaceae), has been widely used in folk medicine as an anti-inflammatory phytochemical. Orbital fibroblasts were primarily cultured from patients with TAO, and the potential of JC3 to suppress the interferon (IFN)-gamma-induced protein (IP)-10/CXCL10 production in these cells was determined. IFN-gamma strongly increased the level of IP-10/CXCL10 in orbital fibroblasts from patients with TAO. JC3 exerted a significant inhibitory effect on the IFN-gamma-induced increase in IP-10/CXCL10 in a dose-dependent manner; its potency was greater than that of an identical concentration of yakuchinone B with no toxicity to cells at the concentration range used. Moreover, the constructed dimer and trimer polystructures of JC3, showed greater potency than JC3 in suppressing the IFN-gamma-induced production of IP-10/CXCL10. JC3 significantly attenuated the IP-10/CXCL10 mRNA expression induced by IFN-gamma, and a gel-shift assay showed that JC3 suppressed IFN-gamma-induced DNA binding of signal transducer and activator of transcription-1 (STAT-1) in TAO orbital fibroblasts. Our results provide initial evidence that the JC3 compound reduces the levels of IP-10/CXCL10 protein and mRNA induced by IFN-gamma in orbital fibroblasts of TAO patients. Therefore, JC3 might be considered as a future candidate for therapeutic application in TAO that exerts its effects by modulating the pathogenic mechanisms in orbital fibroblasts.

Expression of resistin, CXCR3, IP-10, CCR5 and MIP-1α in obese patients with different severity of asthma

Asthma studies suggest that alteration in the inflammation pattern may be associated with the severity of asthma. The aim of this study was to compare in vitro the expression of chemokines, chemokine receptors and cytokine production from CD4+ T human lymphocytes of asthmatic, both obese and non-obese patients with different severity levels of asthma. Lymphocytes were labeled with monoclonal anti-human CXCR3/IP-10, MIP-1a/CCR5 antibodies and were analyzed by flow cytometry. Cell culture supernatants were used to measure production of interleukin IL-6 and resistin by ELISA. CXCR3/IP-10 expression increased in non-obese patients with mild persistent asthma (2.2%, p<0.05), moderate persistent asthma (3%, p<0.003) and severe persistent asthma (4%, p<0.004); this effect was stronger in obese patients with severe persistent asthma (35%, p<0.004). MIP-1 α / CCR5 increased in non-obese patients with intermittent asthma (0.65%, p<0.05) and severe asthma (1.4%, p<0.03); in obese patients, this expression was greater in intermittent asthma (8%, p<0.05) and severe persistent asthma (12%, p<0.04). Resistin production strongly increased in obese patients with intermittent (976 ng/ml) and severe persistent asthma (795 ng/ml). IL-6 increased in both lean and obese persons; however, the highest value was registered in the group of severe persistent obese asthmatics (992 pg/ml). Obesity per se increased the inflammatory profile of chemokines / cytokines secreted by cells of the blood, increasing the inflammatory status in asthmatic patients. Resistin showed characteristics of a pro-inflammatory cytokine mainly in severely obese asthmatics.

Toll-like Receptor3-mediated Induction of Chemokines in Salivary Epithelial Cells

Toll-like receptors (TLRs) functionally expressed in salivary epithelial cells, but their roles remain elusive. Among TLRs family, TLR3 is activated by dsRNA, a byproduct of viral infection. The aim of this study was to investigate the role of TLR3 in the inflammatory immune responses using HSG cells. Reverse transcriptase-polymerase chain reaction (RT-PCR), real-time PCR and ELISA were performed to identify expression of TLRs and TLR3-mediated chemokine inductions. The chemotaxis assay of activated T lymphocytes was also performed. Treatment of HSG cells with polyinosinic: polycytidylic acid (poly(I:C)) significantly increased interferon-gamma-inducible protein 10 (IP-10), interferoninducible T-cell alpha chemoattractant (I-TAC), and regulated on activation, normal T-cells expressed and secreted (RANTES) gene expressions in a concentration-dependent manner. Anti-TLR3 antibody blocked the increases of IP-10 and I-TAC genes. Poly(I:C)-induced increases of IP-10 and I-TAC were also confirmed at protein levels from cell lysates, but their release into extracellular medium was detected only in IP-10. We found that the culture media from HSG cells stimulated with poly(I:C) significantly increases T lymphocyte migration. Our results suggest that TLR3 plays an important role in chemokine induction, particularly IP-10, in salivary epithelial cells.

Relationship of chemokine IP-10 and its soluble CXCR3 in bile with acute graft rejection after liver transplantation / 第二军医大学学报

Objective: To observe the dynamic changes of chemokine IP-10 and its soluble receptor CXCR3 in bile, and to explore its relationship with acute graft rejection (AR) after liver transplantation. Methods: A total of 28 patients who underwent orthotopic liver transplantation in our hospital between October 2007 and March 2008 were included in the present study. They were divided into 2 groups according to the presence of AR: AR(n = 8) and NAR(n = 20). Another 10 patients who underwent endoscopic nasobiliary drainage (ENBD) for cholelithiasis in our hospital served as controls. The levels of chemokine IP-10 and sCXCR3 in the bile were determined by ELISA assay on 1,3,5,and 7 days after transplantation in all the patients and on 1,3,and 7 days after the anti-rejection therapy with glucocorticoid in patients of AR group; the relationship between their levels and the rejection active index (RAI) obtained by Banff criteria were evaluated; and its diagnostic value for AR was assessed. Results: The levels of IP-10 and sCXCR3 in bile gradually increased after liver transplantation and reached the peak on the 5th day after transplantation, and starting from day 5 after transplantation, their levels were significantly higher in the AR group than in the NAR group and ENBD group (P<0.05). Besides, the levels of IP-10 and sCXCR3 in bile were significantly decreased after treatment with glucocorticoid (P < 0.05). On the day of AR diagnosis, the the levels of IP-10 and sCXCR3 in bile were significantly correlated with RAI (P < 0.05). On the 5th day after transplantation, the diagnostic sensitivity and specificity of IP-10 level for AR were 87.5% and 100%, respectively (cut-off point = 964.45 pg/ml); on the 7th day after transplantation, the diagnostic sensitivity and specificity of sCXCR3 were 87.5% and 80%, respectively (cut-off point=819. 35 pg/ml). Conclusion: The levels of IP-10 and sCXCR3 in bile are closed related with early graft acute rejection, and their levels may serve as a non-invasive diagnostic indicator for AR and outcome of anti-rejection therapy.

IP-10 Decreases TNF-alpha Induced MUC5AC Expression in Human Airway Epithelial Cells: a Possible Relation with Little Sputum Production in Idiopathic Pulmonary Fibrosis / 결핵및호흡기질환

BACKGROUND: IPF is characterized by chronic, fibrosing inflammatory lung disease of unknown etiology. Typical symptoms of IPF are exertional dyspnea with nonproductive cough. Why patients with typical IPF have dry cough rather than productive cough, is unknown. IP-10 plays an important regulatory role in leukocyte trafficking into the lung. The present study investigated the effect of IP-10 in the pathogenesis of dry cough rather than productive cough in IPF patients. METHODS: IP-10 concentration was measured by ELISA from BALF of IPF patients. To evaluate the role of IP-10 in mucin expression, the expression of the MUC5AC mucin gene was measured in NCI-H292 cells, a human pulmonary mucoepidermoid carcinoma cell line, after stimulation by TNF-alpha with or without IP-10 pretreatment. EGFR-MAPK expression was also examined as a possible mechanism. RESULTS: IP-10 levels were significantly higher in the BALF of IPF patients compared to healthy controls. IP-10 pretreatment reduced TNF-alpha induced MUC5AC mucin expression by inhibiting the EGFR-MAPK signal transduction pathway in NCI-H292 cells. CONCLUSION: These findings suggest that little mucus production in IPF patients might be attributable to IP-10 overproduction, which inhibits the EGFR-MAPK signal transduction pathway required for MUC5AC mucin gene expression.

Upregulation of IP-10(CXCL10) mRNA Expression by Interleukin-18 / 영남의대학술지

BACKGROUND: Interleukin-18 (IL-18) is one of the principal inducers of interferon-gamma (IFN-gamma) in lymphocytes. MATERIALS AND METHODS: The effect of IL-18 on the expression of chemokine IP-10(CXCL10) mRNA in C57BL/6 mouse peritoneal macrophages was studied by using Northern blot analysis, enzyme linked immunosobent assay and electrophoretic mobility shift assay. RESULTS: IL-18 was determined to exert no direct effect on the expression of IP-10(CXCL10) mRNA. However, IL-18 pretreatment was determined to play a cooperative role in the synergistic induction of LPS-induced IP-10(CXCL10) mRNA expression. The effect associated with IL-18 pretreatment with regard to the synergistic induction of LPS-induced IP-10 (CXCL10) mRNA expression was detected after 16 hr of IL-18 pretreatment, administered prior to LPS stimulation. The pattern of NF-kB binding activity during IL-18 pretreatment with LPS stimulation was found to coincide with the expression of IP-10(CXCL10) mRNA. CONCLUSION: Although IL-18 alone exerts no direct effect on the expression of chemokine IP-10(CXCL10), a definite period of IL-18 pretreatment induces the synergistic expression of LPS-induced IP-10(CXCL10) mRNA. NF-kB activation is a component of this synergistic effect of IL-18 pretreatment. These results provide useful information, which may facilitate the elucidation of the action mechanisms underlying IL-18 effect on the expression of IP-10(CXCL10) mRNA.

Production of Plasma Leptin and Expression of Interferon-gamma Inducible Protein-10 (IP-10), Monokine Induced by Interferon-gamma (Mig) and Interleukin-8 (IL-8) mRNA in Kawasaki Disease

BACKGROUND: Kawasaki disease is an acute febrile illness with systemic vasculitis which primarily affects children, We examined the production of leptin in plasma and gene expressions of CXC chemokines in peripheral blood mononuclear cells from patients with Kawasaki disease. METHODS: Consecutive 39 samples from 13 patients according to the different clinical stages (acute, subacute, convalescent) of Kawasaki disease were collected. The plasma leptin levels according to clinical stages of Kawasaki disease were examined by ELISA and the expression of IP-10, Mig and IL-8 mRNAs in 39 samples (13 samples of each stage) from 13 cases were examined by RT-PCR. RESULTS: There were not significant changes of plasma leptin levels according to the clinical stages of Kawasaki disease. The mean values of plasma leptin concentrations during each of the stages (n=13, p>0.05, pg/ml) were 335.8+/-549.0 in acute, 358+/- 347.6 in subacute, and 443.6+/-645.9 in convalescent stage. The mRNAs of IP-10, Mig, and IL-8 were expressed in 13/13 (100%), 2/13 (15%), 9/13 (69%) during acute stage, 13/13 (100%), 6/13 (46%), 13/13 (100%) during subacute stage, and 13/13 (100%), 4/13 (31%), 10/13 (77%) during the convalescent stage, respectively. In three patients, the production of leptin and expression of IP-10 mRNA were dramatically decreased according to the process of the clinical stages. In five patients with prominent cervical lymphadenopathy, the expression of IL-8 mRNA during the subacute stage was more elevated than the acute and convalescent stages. CONCLUSION: This data suggests that the production of leptin and the gene expressions of IP-10, Mig and IL-8 seem to have no significant correlation to the clinical stages of Kawasaki disease. However, expression patterns of IP-10, Mig and IL-8 mRNA may be related to the specific clinical manifestations, and the expression of IP-10 may also be correlated to leptin levels with pericardial involvement.

IFN -r Enhances Induction of Chemokines Mig and IP10 mRNA from THP - 1 Cells Stimulated with Lipoarabinomannan / 대한면역학회지

Lipoarabinomannans (LAM) is believed as a potential virulence factor of Mycobacterium tuberculosis. LAM exhibits marked differences in biological activities depending on the types, arabinofuranosyl-terminated LAM (AraLAM) derived from a rapidly growing Mycobacterium sp. and heavily mannosylated LAM (ManLAM) derived from the Erdman strain. Collaboration between macrophages and T cells, especially macrophage activation by gamma interferon (IFN-r) and chemoattraction of T cells at the very inflammatory foci would be essential in defence against M. tubercu/osis. Chemokines Mig and IP-10 are inducible by IFN-r from macrophages and have been shown to act in vitro as T cell chemoattractants. However, little is known of LAMs capacity to induce chemokines Mig and IP-10 in macrophages. In this experiment, Mig and IP10 mRNA was expressed in the delayed-type hypersensitivity (DTH) against BCG in BCG-immune mice. In some experiments, both Mig and IP-10 mRNA was evidently induced with different time courses in THP-1 cells stimulated with whole live M. tubercu/osis H37Rv (Erdman). To investigate whether Mig and IP-10 genes are differentially induced depending on the type of LAM, PCR amplification was used to detect mRNA of Mig and IP-10 from the THP-1 human monocytic cells stimulated with LAM. AraLAM, but not ManLAM, induced weakly Mig and IP-10 mRNA in the THP-1 cells. The induction of Mig and IP-10 was dependent upon the dose of AraLAM and exhibited different time courses. The mRNA for Mig and IP-10 was induced within 2 hr and 4 hr from the initiation of treatrnent and has disappeared by 8 hr and 24 hr under the experimental conditions used in this study, respectively. IFN-y at 100 U/ml, but not at 10 U/ml, was itself a good stimulus of both Mig and IP- 10 expression, and synergized with either AraLAM or ManLAM for induction of both Mig and IP-10. The expression patterns of MCP-3 were somewhat similar to those of Mig and IP10 in all of the experiments. These data indicate that IFN-r may contribute to effective macrophage function if macrophages are not fully affected by ManLAM, and chemokines Mig and IP-10 may a role in recruitment of T cells at inflammatory foci of tuberculosis.

Interleukin-27 inhibits tumor angiogenesis through up-regulating expression of MIG and IP-10 / 中国肿瘤生物治疗杂志

Objective:To study the inhibitory effect of IL-27 against human tumor angiogenesis and the related mechanisms.Methods: Human esophageal carcinoma cells(Eca109/IL-27) stably transfected with IL-27 gene were injected into nude mice to establish tumor-bearing mouse model.The survival time and tumor growth were observed.IFN-? level secreted by splenocytes was measured by ELISA.Expression of VEGF and CD34 was detected by immunohistochemistry method and MVD was calculated according to CD34 level.RT-PCR was used to detect the expression of IP-10 and MIG mRNA in the tumor tissues.Results: The survival time of mice injected with Eca109/IL-27 cells was significantly longer than those of mice injected with wide type Eca109 or Eca109/LXSN(blank vector) cells(P

Expression of IP-10 and its mRNA in liver biopsy tissue of patients with cirrhosis / 中国免疫学杂志

Objective:To study the expression of IP-10 and its mRNA in liver biopsy of patients with cirrhosis.Methods:The typical patients with cirrhosis after HBV infection were selected,and their serum IP-10 concentrations were evaluated with ELISA,the content of IP-10 mRNA in liver biopsy was measured by Real-time PCR.The level of IP-10 mRNA was represented by lg cDNA/lg GAPDH.The expression of IP-10 in liver biopsy tissue was detected by SP immunohistochemistry stain.Results:The levels of IP-10 in serum and its mRNA in liver biopsy of patients with cirrhosis were (299.3?77.2)pg/ml and (0.856 3?0.150 6),lg cDNA/lg GAPDH respectively,which were significantly higher than those of control groups(P

Expression of IP-10 and IFN-? in human cerebral ischemia / 中国免疫学杂志

Objective:To investigate whether interferon-gamma inducible protein-10(IP-10) and interferon-gamma(IFN-?) participated in the human cerebral ischemia injury.Methods:Twenty-one cerebral ischemia specimens, collected from patients died of cerebral infarction, were divided into three groups: less than 7 days, 7-14 days and 15-21 days according to the lasting time of cerebral infarction. The infiltrating of inflammatory cells were observed using HE stain as non-ischemic hemisphere was for controls. Expression of IP-10 and IFN-? in sections both postmortem ischemic hemisphere and non-ischemic hemisphere were detected using immunohistochemistry.Results:In the groups of less than 7 days and 7-14 days large quantity of inflammatory cells were infiltrated in ischemia tissue. Expression of IP-10 in three groups was elevated in the ischemic hemisphere compared with non-ischemic hemisphere(1.74-folds, 1.41-folds and 1.52-folds increases respectively, P0.05).Conclusion:These results showed IP-10 and IFN-? are expressed in human cerebral ischemia. It was suggested that IP-10 and IFN-? involve in cerebral ischemia injury. Moreover, it was also suggested that IP-10 participate in the repair for the later stage of cerebral ischemia injury.

IP10 enhances anti-tumor immunity and its mechanisms / 中国免疫学杂志

Objective:To investigate the effects of IP10(IFN-? inducible protein 10,CXCL10) on anti-tumor immune response and to explore its mechanisms involved in.Methods:A mammary carcinoma cell line 4T1 was transfected with pcDNA3-IP10(IP10-4T1) by electrophoration and positive clones were screened in the presence of G418.Growth kinetics of IP10-4T1 cells was observed in vitro and in vivo.Survival rate among the animals was determined by daily assessment.Proliferation activity of lymphocytes was analyzed with()~3H-TdR incorporation.The phenotypes of lymphocytes isolated from tumors by ficoll density gradient centrifugation were assayed by flow cytometry.Results:The growth rate of IP10-4T1 cells was similar with that of parental 4T1 cells and neo-vector transfected 4T1 cells in vitro.Growth of the tumors formed by IP10-4T1 cells was inhibited in vivo.Compared to those of controls,the size and the weight of the tumors formed by IP10-4T1 cells decreased significantly 35 days post tumor transplantation(P