Serum TGF-beta1 Levels Research Articles

PP037. Relationship of serum leptin levels to circulating cytokines, chemokines, adhesion molecules and angiogenic factors in women with preeclampsia

Serum leptin levels are known to be increased in preeclampsia. To determine circulating levels of leptin along with those of several cytokines, chemokines, adhesion molecules and angiogenic factors in normal pregnancy and preeclampsia, and to investigate whether serum leptin levels are related to the clinical features and measured laboratory parameters of the patients. Serum levels of leptin, IL-1beta, IL-1ra, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p40, IL-12p70, IL-18, IFN-gamma, TNF-alpha, TGF-beta1, IP-10, MCP-1, ICAM-1, VCAM-1, sFlt-1 and PlGF were determined in 60 preeclamptic, 60 healthy pregnant and 59 healthy non-pregnant women. There were significant differences in most of the measured laboratory parameters among the three study groups except for serum IL-1beta and TGF-beta1 levels. Preeclamptic patients had significantly higher leptin levels than healthy pregnant women. A significant positive correlation was observed between serum leptin and IP-10 concentrations of preeclamptic patients. Furthermore, elevated serum leptin level and sFlt-1/PlGF ratio had an additive effect in the risk of preeclampsia, as shown by the substantially higher odds ratios of their combination than of either alone. Increased serum levels of leptin are related to those of IP-10 in preeclampsia, suggesting that circulating leptin might contribute to the development of endothelial dysfunction through the anti-angiogenic effect of IP-10.

Association of cytokine gene polymorphisms and serum concentrations with the outcome of chronic hepatitis B

ObjectiveThe present paper investigated possible correlations between the clinical presentation of hepatitis B and the TNF-α −308G/A, IFN-γ +874A/T, TGF-beta1 −509C/T, and IL-10 −1081A/G polymorphisms and associated serum levels of these cytokines. MethodsFifty-three hepatitis patients were selected and divided into two groups: A – inactive (n=30) and B – chronic hepatitis/cirrhosis (n=23). The control group consisted of 100 subjects who were positive for anti-HBc and anti-HBs. The serum concentrations of the cytokines were determined by immunoenzymatic assays. The polymorphisms of the cytokines genes were assessed by PCR and PCR-SSP. ResultsThe mean serum levels of IFN-γ of the control group were significantly higher than those of groups A and B, whereas the mean levels TGF-beta1 were significantly higher in groups A and B in comparison with the control. In the case of IL-10, the mean serum level recorded in the control group was significantly higher than that of group B. The TNF-α −308AG genotype was considerably more frequent in group B (43.3%) than the control (14.4%). ConclusionHigher serum levels of IFN-γ and TGF-beta1 were associated with chronic hepatitis B, and lower serum levels of IL-10 were found in patients with the active disease. Furthermore the presence of allele A of the TNF-α −308 polymorphism suggest a risk of the progressive disease.

Increased Serum Transforming Growth Factor-beta1 (Tgf-beta1) Is Related to a Better Outcome in MM, WM and CLL Patients

AbstractAbstract 4976 Background and Aims:TGF-beta1 normally down-regulates B-cell proliferation. Resistance to its effects is reported in malignant cells of B-lineage such as chronic lymphocytic leukemia (CLL) and multiple myeloma (MM) cells. There are however very few studies on serum TGF-beta1 levels in patients with B-cell lymphoproliferative diseases and their eventual correlations with parameters of disease activity and survival. The purpose of our study was to assess any eventual relationship between serum TGF-beta1 levels and disease parameters, as well as time to first treatment (TFT) and overall survival (OS) in a series of MM, Waldenstrom's macroglobulinemia (WM) and CLL patients at diagnosis. Patients and Methods:92 MM, 64 WM and 110 CLL patients were studied. In the MM group, paraprotein was IgG in 64, IgA in 18, LC in 8 and IgD in 2 patients while 32, 25 and 43% of patients were in ISS stage 1, 2 and 3 respectively. MM patients’ median follow-up was 51 months. All patients required treatment a some point, 55% immediately. In the WM group, 44, 27 and 29% of patients were in WM – IPSS stage 1, 2 and 3 respectively. WM patients’ median follow-up was 49 months; 75% of them required treatment during follow – up. In the CLL group, 54% and 74%, 25% and 20%, 14% και 6%, were in stage 0 and A, 1 and β, 2 and C according to Rai and Binet respectively. Median follow up was 43 months; 14 patients needed immediate treatment, 30 more required treatment during follow up while the others are still just followed. Patients’ frozen sera, drawn at diagnosis, were retrospectively analyzed while sera from 20 healthy individuals (HI) were tested as controls. Serum TGF-beta1 measurements were done by ELISA according to the manufacturer’s instructions. Statistical analysis was performed by SPSS software, version 15. 0. Results:Median serum TGF-beta1 levels were 36330 pg/ml (range 4. 6 – 77976) in MM patients, 33965 pg/ml (range 1665–615000 pg/ml) in WM, 12207 pg/ml in CLL (range 0 – 42970) and 32902 pg/ml in HI (range 1941 – 58123). CLL patients presented significantly lower TGF-beta1 levels than those with MM and WM (p<0. 00001). Patients with TGF-beta1 levels above median presented a longer TFT than the others in MM and CLL (p=0. 0003, and p=0. 002 respectively) while in addition a longer OS was observed in MM and WM patients with increased serum TGF-beta1 median (p=0. 001, and 0. 02 respectively). Furthermore, serum TGF-beta1 levels stongly correlated with clinical and laboratory parameters of adverse prognosis such as staging, anemia, beta-2 microglobulin levels, serum LDH and serum free light chain concentrations. Conclusions:Our results of longer TFT in MM and CLL and improved OS in MM and WM patients with increased serum TGF-beta1 levels, suggest that a degree of sensitivity to the suppressive effect of TGF-beta1 may remain in neoplastic B-cells in the majority of B-cell lymphoproliferative disorders. Disclosures:No relevant conflicts of interest to declare.

Abstract 3558: A pilot clinical study of limonene in women with early stage breast cancer: Effects of short-term treatment on tissue limonene disposition and serum markers

Abstract Background: Limonene is a highly lipophilic monoterpene found in citrus peel oil that has demonstrated anticancer properties in preclinical studies. The purpose of this study was to evaluate whether limonene and its primary metabolite perillic acid (PA) would distribute extensively to the breast tissue and reach an effective drug concentration. Secondary endpoints included evaluation of changes in cancer-related biomarkers in plasma and tissue. Methods: Participants (N = 40) with newly diagnosed stage 0 - 2 breast cancer took 2 grams daily of oral limonene for 2 - 6 weeks (21.5 ± 8.8 days) prior to planned surgical tumor resection. Blood was drawn pre/post intervention for measurement of serum concentration of limonene, PA, and protein biomarkers, as well as to assess toxicity profiles. A small piece of breast tissue adjacent to the tumor mass was also collected for analysis of limonene and PA levels. Limonene and PA levels in tissue and serum were analyzed using specific chromatography-based assays. Results: Limonene was found to preferentially concentrate in breast tissue (332.3 + 336.1 uM) versus plasma (0.49 + 0.67 uM) with tissue-to-plasma concentration ratio (TPCR) of 1,667 + 312.5 (P&amp;lt;0.001). PA did not concentrate in breast tissue (5.73 + 10.28 uM) versus plasma (3.89 + 6.81 uM) with TPCR of 1.4 + 0.56. Post-intervention serum levels of leptin, adiponectin, TGF-beta1, IGFBP-3 and IL-6 were unchanged from baseline. There was a small but statistically significant post-intervention increase in IGF-1, however the change was no longer significant after adjustment for number of days on agent. Possibly or probably related adverse events were primarily GI-related and were not dose-limiting. No clinically significant changes in complete blood count, renal, hepatic or other blood chemistry studies were noted. Analysis of the intervention effects on tissue biomarkers of proliferation, apoptosis, and cell cycle regulation is ongoing. Discussion: Oral limonene preferentially concentrates in the breast tissue and has a favorable side effect profile. PA does not readily concentrate in breast tissue when administered as oral limonene. Short-term limonene intervention resulted in minimal changes in systemic biomarkers. Further clinical trials with a longer intervention are necessary to establish limonene's potential role as a chemopreventive agent in breast cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3558. doi:1538-7445.AM2012-3558

Rapamycin combined with allogenic immature dendritic cells selectively expands CD4+CD25+Foxp3+ regulatory T cells in rats

Dendritic cells (DCs) can initiate the expansion of regulatory T cells (Tregs), which play an indispensable role in inducing transplantation tolerance. Some studies have investigated the effect of the immunosuppressant rapamycin (Rapa) on Tregs in vitro. However, the in vivo effect of Rapa combined with immature DCs (iDCs) on Tregs is unknown. This study was undertaken to determine whether allogenic iDCs combined with a short course of Rapa have the ability to selectively expand the CD4+CD25+Foxp3+ Tregs in a rat model. Brown Norway rats were injected intravenously with 2X10(6) Lewis iDCs followed by 1 mg/kg per day Rapa intraperitoneally for 7 consecutive days. On day 8, the levels of CD4+CD25+Foxp3+ Treg cells in peripheral blood and spleen cells were analyzed by flow cytometry. IL-2, IL-4, TGF-beta1, and IFN-gamma levels in serum were assessed by ELISA. The experimental animals were divided into four groups: control, Rapa-treated, iDC-treated, and combination-treated. CD4+CD25+Foxp3+ Tregs comprised 7%-8% of CD4+ T cells in control rats. Rapa combined with iDCs enhanced this percentage in the peripheral blood and spleen. However, the levels of Tregs did not significantly change after treatment with Rapa or iDCs alone. The levels of CD4+CD25-Foxp3+ T cells and CD4+CD25+Foxp3- T cells in CD4+ T cells did not significantly change in the combined group. The TGF-beta1 level in serum from the combined group increased significantly compared with the other groups. A significantly higher percentage of CD4+ CD25+ Foxp3+ Tregs was found in rats treated with allogenic iDCs and a short course of Rapa, along with an increase in the TGF-beta1 level in serum. This improved protocol may be a promising therapeutic strategy to increase Tregs, which are beneficial to the induction of peritransplant tolerance.

Interaction between smoking and functional polymorphism in the TGFB1 gene is associated with ischaemic heart disease and myocardial infarction in patients with rheumatoid arthritis: a cross-sectional study

IntroductionTransforming growth factor-beta1 (TGF-beta1) is a pleiotropic cytokine that plays important roles in immunity and inflammation. Some studies have suggested that polymorphism in the TGFB1 gene is associated with heart disease in the general population. The purpose of the present study was to determine whether common single-nucleotide polymorphisms (SNP) in the TGFB1 gene are associated with ischaemic heart disease (IHD) and/or myocardial infarction (MI) in patients with rheumatoid arthritis (RA), and to investigate the influence of smoking on any association.MethodsPCR-based assays were used to determine the genotypes of TGFB1 SNPs including TGFB1-509 C/T (rs1800469, in the promoter region), +868 T/C (rs1800470, in exon 1) and +913 G/C (rs1800471, in exon 1) in 414 subjects with established RA. Genotyping for the +868 SNP was also carried out on a second study population of RA patients (n = 259) with early disease. Serum levels of TGF-beta1 were measured using a commercial ELISA kit. Smoking history and IHD/MI status were obtained on each patient. Associations with IHD/MI were assessed using contingency tables and logistic regression analyses.ResultsThe heterozygous genotype of TGFB+868 was associated with an increased risk of IHD (OR 2.14, 95% CI 1.30 - 3.55) and MI (OR 2.42, 95% CI 1.30-4.50), compared to the homozygous genotypes combined. Smoking was an independent risk for IHD and MI, and evidence of interaction between smoking and TGFB+868 was found. Multivariate analyses indicated that the strongest associations with IHD and MI were due to the combined effect of the TGFB1+868 TC genotype and smoking (OR 2.75, 95% CI 1.59-4.75; and OR 2.58 95% CI 1.33-4.99, respectively), independent of other cardiovascular risk factors. The association of the +868 TC genotype and evidence of +868 TC-smoking interaction with IHD were replicated in a second population of RA patients with early disease. Serum TGF-beta1 levels were not associated with TGFB1 genetic variations, smoking or IHD/MI status.ConclusionsInteraction between smoking and polymorphism in the TGFB1 gene may influence the risk of IHD and MI in patients with RA.

Serum leptin levels in relation to circulating cytokines, chemokines, adhesion molecules and angiogenic factors in normal pregnancy and preeclampsia

ObjectiveIn this study, we determined circulating levels of C-reactive protein, several cytokines, chemokines, adhesion molecules and angiogenic factors along with those of leptin in healthy non-pregnant and pregnant women and preeclamptic patients, and investigated whether serum leptin levels were related to the clinical characteristics and measured laboratory parameters of the study participants.MethodsSixty preeclamptic patients, 60 healthy pregnant women and 59 healthy non-pregnant women were involved in this case-control study. Levels of leptin and transforming growth factor (TGF)-beta1 in maternal sera were assessed by ELISA. Serum levels of interleukin (IL)-1beta, IL-1 receptor antagonist (IL-1ra), IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p40, IL-12p70, IL-18, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interferon-gamma-inducible protein (IP)-10, monocyte chemotactic protein (MCP)-1, intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 were determined by multiplex suspension array. Serum C-reactive protein (CRP) concentrations were measured by an autoanalyzer. Serum total soluble fms-like tyrosine kinase-1 (sFlt-1) and biologically active placental growth factor (PlGF) levels were determined by electrochemiluminescence immunoassay. For statistical analyses, non-parametric methods were applied.ResultsThere were significant differences in most of the measured laboratory parameters among the three study groups except for serum IL-1beta and TGF-beta1 levels. Serum leptin levels were significantly higher in preeclamptic patients and healthy pregnant women than in healthy non-pregnant women. Additionally, preeclamptic patients had significantly higher leptin levels as compared to healthy pregnant women. Serum leptin levels were independently associated with BMI in healthy non-pregnant women. In healthy pregnant women, both BMI and serum CRP concentrations showed significant positive linear association with leptin levels. There were significant positive correlations between serum leptin concentrations of healthy pregnant women and systolic blood pressure, as well as serum levels of IP-10, while their serum leptin levels correlated inversely with fetal birth weight. In preeclamptic patients, a significant positive correlation was observed between serum concentrations of leptin and IP-10. Furthermore, elevated serum leptin level and sFlt-1/PlGF ratio had an additive (joint) effect in the risk of preeclampsia, as shown by the substantially higher odds ratios of their combination than of either alone.ConclusionsSimultaneous measurement of leptin with several inflammatory molecules and angiogenic factors in this study enabled us to investigate their relationship, which can help to understand the role of circulating leptin in normal pregnancy and preeclampsia.

Serum heat shock protein 70 levels in relation to circulating cytokines, chemokines, adhesion molecules and angiogenic factors in women with preeclampsia

Background We have previously reported that serum levels of 70 kDa heat shock protein (Hsp70, HSPA1A) are increased and reflect systemic inflammation, oxidative stress and hepatocellular injury in preeclampsia. The purpose of this study was to determine whether increased serum Hsp70 concentrations in women with preeclampsia are related to circulating levels of cytokines, chemokines, adhesion molecules and angiogenic factors, the key players in the pathogenesis of the disease. Methods Sixty preeclamptic patients and 60 normotensive, healthy pregnant women were involved in this case-control study. Levels of Hsp70 (HSPA1A) and transforming growth factor (TGF)-beta1 in maternal sera were assessed by ELISA. Serum levels of interleukin (IL)-1beta, IL-1 receptor antagonist, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p40, IL-12p70, IL-18, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interferon-gamma-inducible protein (IP)-10, monocyte chemotactic protein (MCP)-1, intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 were determined by multiplex suspension array. Serum total soluble fms-like tyrosine kinase-1 (sFlt-1) and biologically active placental growth factor (PlGF) levels were measured by electrochemiluminescence immunoassay. For statistical analyses, the Mann–Whitney U-test, the Fisher exact and Pearson chi-square tests, the Spearman rank order correlation, multiple linear regression and logistic regression were applied. Results Serum levels of Hsp70 were significantly higher in preeclamptic patients than in healthy pregnant women. Additionally, most of the measured inflammatory variables differed significantly between the two study groups except for serum IL-1beta and TGF-beta1 levels and IL-18/IL-12p70 and IL-12p70/IL-12p40 ratios, indicating a bias toward a pro-inflammatory status in preeclampsia. Preeclamptic patients had significantly higher sFlt-1 levels and sFlt-1/PlGF ratio and significantly lower PlGF concentrations as compared to healthy pregnant women. In the preeclamptic group, serum Hsp70 concentrations showed significant correlations with serum levels of IL-12p40 ( R = 0.59, p < 0.001), MCP-1 ( R = 0.43, p < 0.001), ICAM-1 ( R = 0.39, p = 0.0020) and VCAM-1 ( R = 0.46, p < 0.001). Furthermore, elevated serum Hsp70 level and sFlt-1/PlGF ratio had a synergistic (joint) effect in the risk of preeclampsia, as shown by the substantially higher odds ratios of their combination than of either alone. Conclusions Increased serum Hsp70 concentrations in women with preeclampsia were associated with pro-inflammatory changes in circulating cytokine profile, suggesting that circulating Hsp70 might contribute to the development of the excessive systemic inflammatory response characteristic of the maternal syndrome of the disease.

A Pilot Study Evaluating the Clinical and Immunomodulatory Effects of an Orally Administered Extract ofDendrobium Huoshanensein Children with Moderate to Severe Recalcitrant Atopic Dermatitis

Atopic dermatitis (AD) is a common inflammatory skin disorder for which few safe and effective systemic treatments are available. To test the clinical and immunomodulatory effects of a crude polysaccharide fraction isolated from Dendrobium huoshanense for the treatment of AD, we conducted a pilot, uncontrolled case series study. Twenty-seven patients aged 4-18 years (mean∓SD, 10.82±4.4) with AD that had not responded to topical therapy were treated with polysaccharide derived from D. huoshanense for 4 weeks and followed-up for another 4 weeks. Progression of AD was determined with the Lund-Browder chart for children, the Investigator's Global Atopic Dermatitis Assessment (IGADA), and the Scoring Atopic Dermatitis (SCORAD) at weeks 0, 2, 4, and 8. Serum levels of cytokines were evaluated. Safety was determined with laboratory and clinical tests. The lesion area, IGADA score, total SCORAD result, and score for sleeplessness decreased significantly from weeks 0 to 4, but did not change significantly between weeks 4 and 8. The scores for subjective symptoms and pruritus decreased significantly from week 0 to week 4 and increased significantly from week 4 to week 8. Serum levels of IL-5, IL-13, IFN-gamma, and TGF-beta1 decreased significantly between weeks 0 and 4 and between weeks 0 and 8. No significant difference in the levels of IL-10 was found. The polysaccharide from D. huoshanense reduced the levels of some cytokines associated with AD and had beneficial effects on symptoms. No serious adverse effects occurred when it was administered orally for 4 weeks.

Insulin-Like Growth Factor-1 (IGF-1) Reduces ischemic changes and increases circulating angiogenic factors in experimentally - induced myocardial infarction in rats

BackgroundCoronary artery disease is a global health concern in the present day with limited therapies. Extensive efforts have been devoted to find molecular therapies to enhance perfusion and function of the ischemic myocardium. Aim of the present study was to look into the effects of insulin like growth factor -1 (IGF-1) on circulating angiogenic factors after myocardial ischemia in rats.MethodsAdult male Sprague-Dawley rats were randomly divided into 10-days control, myocardial infarction, IGF-1 alone (2 μg/rat/day) and ISO+IGF-1 groups. Isoproterenol (ISO), a synthetic catecholamine was used to induce myocardial infarction. Serum transforming growth factor-β (TGF-β) and vascular endothelial growth factor (VEGF) levels were checked after 10-days of IGF-1 administration.ResultsThere was a significant increase in heart weight after IGF-1 treatment. A significant increase in cardiac enzyme level (CK-MB and LDH) was seen in isoproterenol treated rats when compared to control group. IGF-1treatment induced a significant increase in serum angiogenic factors, IGF-1, VEGF and TGF beta levels. IGF-1 also reduced the ischemic changes in the myocardium when compared to the isoproterenol alone treated group.ConclusionsIn conclusion, treatment with insulin-like growth factor-1 (IGF-1) in myocardial infarction significantly increased circulating angiogenic growth factors like IGF-1, VEGF and TGF beta thus, protecting against myocardial ischemia.

Clinical Benefits of Biochemical Markers of Fibrosis in Egyptian Children With Chronic Liver Diseases.

BackgroundThe need for repetition of liver biopsy, especially in assessing the degree of fibrosis and follow-up of treatment protocols, justifies an intensive search for non-invasive alternatives. We attempted to investigate the clinical usefulness of serum fibrogenesis markers in pediatric chronic liver diseases.MethodsWe measured serum levels of TGF-β1, collagen IV, laminin, MMP-2 and EGF-R, in 50 children with chronic liver disease (HBV, HCV and Bilharziasis) and 30 healthy controls, and determined their relationship to frequently used liver function tests and liver biopsy findings in patients.ResultsTGF-β1, collagen IV, laminin and MMP-2, but not EGF-R, were significantly higher in patients than in controls (P < 0.01). None of these markers correlated with the histological fibrosis stage, whereas laminin correlated with necroinflammatory activity (P < 0.01). TGF-β1, collagen IV, laminin and MMP-2 had the ability to discriminate patients with significant fibrosis, while only collagen IV and laminin were able to discriminate those with cirrhosis. Among these markers, collagen IV had the best predictive accuracy for significant fibrosis (AUROC 0.94; PPV 91.5%) and cirrhosis (AUROC 0.85; PPV 80%).ConclusionsIn conclusion, these markers may be useful in reducing but not replacing the need for liver biopsy in the monitoring of disease progression and treatment effectiveness and might be an inseparable part of assessment of chronic hepatopathies.

Transforming growth factor beta 1 (TGF-beta 1) in atrial fibrillation and acute congestive heart failure

Atrial fibrillation (AF) and acute congestive heart failure (aCHF) are characterized by an adverse cardiac remodeling. Arrhythmogenic or structural remodeling can be caused by interstitial fibrosis. Transforming growth factor beta 1 (TGF-beta 1) represents a central regulator of cardiac fibrosis. This study investigates serum levels of TGF-beta 1 in patients with AF and aCHF. 401 patients presenting with symptoms of dyspnea or peripheral edema were prospectively enrolled. Blood samples for measurement of TGF-beta 1 (R&D Systems, Inc.) and amino-terminal pro-brain natriuretic peptide (NT-proBNP) (DadeBehring ltd.) were collected after the initial clinical evaluation. Median TGF-beta 1 levels were lower in patients with AF (21.0ng/ml, interquartile range (IR) 15.4-27.6ng/ml, n=107) compared to those without (25.0ng/ml, IR 18.5-31.6ng/ml, n=294) (p=0.009). Patients with aCHF had lower TGF-beta 1 levels (median 22.0ng/ml, IR 15.6-27.1ng/ml, n=122) than those without (median 24.9ng/ml, IR 18.1-31.9ng/ml, n=279) (p=0.0005). In logistic regression models TGF-beta 1 was still associated with AF (odds ratio (OR) 3.00, 95% CI 1.37-6.61, p=0.0001) and aCHF (OR 3.98, 95% CI 1.55-10.19, p=0.004). TGF-beta 1 inversely correlated with left atrial diameter (r=-0.30, p=0.007) and NT-proBNP (r=-0.14, p=0.007). Low serum levels of TGF-beta 1 are associated with AF and aCHF. This decrease may result from a higher consumption of TGF-beta 1 within the impaired myocardium or antifibrotic functions of natriuretic peptides.

Experiment study about effect of Yinqiao detoxifcation oral liquid on natural killercells and TNF-α, TGF-β1 of BALB/C nude mouse infected by influenza virus

To explore the effect of Yinqiao detoxifcation oral liquid on activity of natural killer cells (NK) and serum content of TNF-alpha, TGF-beta1 of BALB/C nude mouse infected by influenza virus. To establish infected mice model by FM1 followed by intragastric administration of Yinqiao detoxifcation oral liquid for treatment. LDH method was used to observe NK cells. ELISA method was used to determine the levels of TNF-alpha, TGF-beta1, in serum on 1st, 3rd, 5th, 7th days after infection. Comparing to the normal group, the NK activity of the model group was significantly increased on 1 dpi (day post infection), and significantly decreased on 3, 5, 7 dpi. The NK activity of three dosage groups (5, 10, 20 g x kg(-1)) of Yinqiao detoxifcation oral liquid were respectively higher than that of the model on 3, 5, 7 dpi, especially with high dose (P < 0.01). The serum level of TNF-alpha and TGF-beta1 of model group is higher than that of normal group on 1, 3, 5, 7 d. Compared with model group, the serum level of Yinqiao detoxifcation oral liquid groups (5, 10, 20 g x kg(-1)) were decreased in different degree on every time point, especially the serum level of the higher dose of Yinqiao detoxifcation oral liquid decreased on 3 dpi (P < 0.05), Yinqiao detoxifcation oral liquid inhibit the serum level of TGF-beta1 in a dose-dependent manner. Yinqiao detoxifcation oral liquid could enhance the activity of NK cell and decrease the serum level of TNF-alpha and TGF-beta1 of the mice infected by influenza virus.

Aliskiren ameliorates chlorhexidine digluconate‐induced peritoneal fibrosis in rats

Peritoneal fibrosis (PF) is a recognized complication of long-term peritoneal dialysis (PD) and can lead to ultrafiltration failure. The present study was designed to investigate the protective effects of aliskiren on chlorhexidine digluconate-induced PF in rats. The PF was induced in Sprague-Dawley rats by daily administration of 0.5 mL 0.1% chlorhexidine digluconate in normal saline via PD tube for 1 week. Rats received daily intravenous injections of low-dose aliskiren (1 mg kg(-1)) or high-dose aliskiren (10 mg kg(-1)) for 1 week. After 7 days, conventional 4.25% Dianeal (30 mL) was administered via a PD catheter with a dwell time of 4 h and assessed of peritoneal function. At the end of dialysis, rats were sacrificed and the liver peritoneum was harvested for microscopically and immunohistochemistry. There was no significant difference in mean arterial pressure and heart rate between groups. After 4 h of PD, the D(4)/P(4) urea level was reduced, the D(4)/D(0) glucose level, serum and dialysate transforming growth factor-beta1 (TGF-beta1) level was increased, the liver peritoneum was markedly thicker, and the expression of TGF-beta1, alpha-smooth muscle actin (alpha-SMA), fibronectin, collagen, and vascular endothelial growth factor (VEGF) were elevated in the PS group compared with the vehicle group. Aliskiren decreased the serum and dialysate TGF-beta1 level, decreased the thickness of the liver peritoneum, and decreased the expression of TGF-beta1, alpha-SMA, fibronectin, collagen, and VEGF-positive cells in liver peritoneum. Moreover, high-dose aliskiren had better protective effects against PF than low dose in rats. Aliskiren protected against chlorhexidine digluconate-induced PF in rats by decreasing TGF-beta1 production.

TGF-beta1 in serum and induced sputum for predicting radiation pneumonitis in patients with non-small cell lung cancer after radiotherapy

Research has confirmed that transforming growth factor-beta1 (TGF-beta1) is one of the cytokines related to radiation pneumonitis. But the level of TGF-beta1 in serum needed to predict radiation pneumonitis is still not clear. This study assessed the value of TGF-beta1 in both serum and induced sputum in predicting radiation pneumonitis, providing a reference for the radiotherapy of patients with non-small cell lung cancer (NSCLC). A total of 23 patients with NSCLC treated with three-dimensional conformal radiotherapy (3D-CRT) or intensity-modulated radiation therapy (IMRT) in our department between November 2007 and January 2009 were analyzed and evaluated. TGF-beta1 levels in both serum and sputum were detected before and near the end of radiotherapy for all the patients. The TGF-beta1 level in serum was measured with enzyme-linked immunosorbent assay (ELISA). Immunocytochemistry assays were used to detect TGF-beta1 expression in sputum sediment. Radiation pneumonitis was graded according to Radiation Therapy Oncology Group (RTOG) radiation scoring criteria every 3 weeks from the start to 3 months after the end of treatment. Radiation pneumonitis was noted in 9 patients in this cohort. The total incidence of radiation pneumonitis was 39.1% (9/23) and those with Grade II or worse was 30.4% (7/23). The absolute TGF-beta1 level in serum after radiotherapy was higher than before radiotherapy, but there was no statistical difference (P = 0.139). Patients with increased levels of TGF-beta1 had a higher incidence of radiation pneumonitis (45.5%) than those with decreased TGF-beta1 levels post-radiotherapy (40.0%). Though there was a tendency of higher incidence of radiation pneumonitis with increases in TGF-beta1 level, no statistical difference was found (P = 1.000). Patients with tumor response had higher incidence of radiation pneumonitis (50.0%) than patients without when TGF-beta1 levels in serum increased, but there was no statistical difference (P = 0.792). TGF-beta1 was positively expressed (brown yellow) in sputum on immunocytochemistry assays and located in the cytoplasm of either macrophages or epithelial cells. Macrophages were the main cells expressing TGF-beta1. A significantly higher positive expression rate (71.4%) was found in sputum post-radiotherapy than pre-radiotherapy (28.6%) (P = 0.015). The higher incidence of radiation pneumonitis (46.7%) was found in patients with positive TGF-beta1 expression in sputum post-radiotherapy than those with negative expression post-radiotherapy (14.3%) (P = 0.193). It may be more reasonable to predict radiation pneumonitis by combining the change of TGF-beta1 levels in serum with tumor response than just the change of TGF-beta1 levels in serum alone. TGF-beta1 can positively express in the sputum of patients with NSCLC, located in macrophages and epithelial cells, with macrophages as the main areas of expression. Patients with positively expressed TGF-beta1 in sputum after radiotherapy have a higher incidence of radiation pneumonitis than those with negative expressions. The positive expression of TGF-beta1 in sputum is expected to become a factor for predicting radiation pneumonitis.

Serum HGF and TGF‐β1 levels after right portal vein embolization

The changes in the serum hepatocyte growth factor (HGF) and transforming growth factor (TGF)-beta1 levels after portal vein embolization (PVE), and their clinical significance, remain unclear and we aimed to assess their relationship. The serum HGF and TGF-beta1 levels were prospectively measured in 22 patients before and 1, 3, 5, 7, and 14 day after right PVE. Computed tomographic volumetry was performed before and at a mean of 26 +/- 4 days after right PVE. Three to four weeks after right PVE, the volume of embolized lobe significantly decreased from 704 +/- 157 cm(3) before PVE to 539 +/- 168 cm(3) after PVE (P < 0.001). In contrast, the volume of nonembolized lobe significantly increased from 426 +/- 142 cm(3) to 560 +/- 165 cm(3) (P < 0.001). The serum HGF level significantly increased on day 3 after PVE compared with the pretreatment level (P = 0.005), while the serum TGF-beta1 level significantly decreased and reached its lowest value on day 3 (P = 0.002). Using Pearson's correlation analysis, we found that the serum HGF and TGF-beta1 levels on day 14 negatively associated with the large hypertrophic response in the nonembolized lobe (HGF: r = -0.490, P = 0.021; TGF-beta1: r = -0.473, P = 0.026). PVE induced an increase in the serum HGF level and reduced the serum TGF-beta1 level. Measurement of serum HGF and TGF-beta1 levels on day 14 after right PVE may be useful for assessment of the future liver hypertrophy in nonembolized lobe after right PVE.

Phase II Study of a HER-2/Neu (HER2) Intracellular Domain (ICD) Vaccine Given Concurrently with Trastuzumab in Patients with Newly Diagnosed Advanced Stage Breast Cancer.

Abstract HER2 is a tumor antigen in breast cancer and several trials have demonstrated that breast cancer patients can be immunized against this protein. We have developed HER2 peptide based vaccines that are aimed at eliciting CD4+ Th1 tumor antigen specific T cell responses. Th1 effectors provide immunologic memory, enhance cross priming which will allow the elaboration of tumor specific CD8+ T cells, and stimulate epitope spreading which we have shown to be a potential biomarker of clinical response. 52 patients will be enrolled with the primary objective to determine relapse free survival after active immunization. Eligible patients are newly diagnosed with Stage III (B or C) or Stage IV breast cancer and begin vaccination within 6 months of starting maintenance trastuzumab. This interim report will present data on the first 25 patients enrolled; 21 stage IV and 4 locally advanced patients. The vaccine is well tolerated with all adverse events (AE) being Grade I or 2. The most common AE is injection site reaction. Moreover, the combination of HER2 vaccination with trastuzumab did not result in additive cardiac toxicity in these patients. Immune responses were evaluated by IFN-gamma ELISPOT. To date, 88% of patients immunized developed significant immunity to the components of the ICD vaccine. The majority, 75%, developed robust immunity to the HER2 protein. Our group has recently demonstrated that a broadening of immunity throughout the HER2 protein, to components of the protein that weren't in the vaccine, i.e. epitope spreading, may be associated with improved survival in vaccinated patients. 63% of immunized patients demonstrated evidence of intramolecular epitope spreading. We questioned whether such high frequencies of homing Type 1 T cells might modulate the immunosuppressive tumor microenvironment, so we evaluated whether circulating serum immunosuppressive cytokines were impacted by immunization. TGF-beta is an immunosuppressive cytokine secreted by tumor stroma and regulatory T cells. We found that the levels of serum TGF-beta decreased significantly in the majority of patients after vaccination. We further analyzed the correlation between the change of serum levels of TGF-beta post vaccination and HER2 ICD vaccine-induced T cell responses. We found that the greater the magnitude of the HER2 specific T cell response, as demonstrated by IFN-gamma secretion, the greater the decrease in serum TGF-beta (p=0.0045, r=0.742). The correlation between the increased epitope spreading T cell response and decreased levels of TGF-beta was even more significant (p=0.0003). The median overall survival has not been reached with 100% of patients alive at this time. Relapse free survival data will be presented. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5102.

The −509C/T polymorphism of transforming growth factor‐β1 is associated with increased risk for development of chronic idiopathic neutropenia

Impaired granulopoiesis in chronic idiopathic neutropenia (CIN) has been associated with an inflammatory bone marrow (BM) microenvironment consisting of pro-inflammatory and pro-apoptotic mediators, such as tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1, and Fas-Ligand (Fas-L). In this study, we evaluated the frequency of TNF-alpha, TGF-beta1 and Fas-L gene polymorphisms in CIN patients and explored their role in excessive cytokine production and their association with CIN development. The TNF-alpha-308G/A, TGF-beta1 -509C/T, +869T/C, +915G/C, and Fas-L -844T/C polymorphisms were studied in 57 CIN patients, and 100 healthy controls from Crete, a well-defined area with genetically homogeneous population, using a polymerase chain reaction-based restriction fragment length polymorphism assay. The mutant genotype C/T or T/T of TGF-beta1 -509C/T polymorphism was more common in CIN patients than in controls (P = 0.033). Compared to wild-type genotype, the TT genotype was associated with increased risk for CIN development (OR: 5.7; 95% CI: 1.18-27.26; P = 0.033). Compared to controls, patients with CT and TT genotypes displayed increased TGF-beta1 levels in serum (P < 0.0001 and P = 0.0002, respectively) and BM (P < 0.0001 and P = 0.0002, respectively). No significant difference was found between patients and controls in the frequency of TNF-alpha-308G/A, TGF-beta1 +869T/C and +915G/C and Fas-L -844T/C polymorphisms. The TGF-beta1 -509C/T polymorphism is associated with increased risk for CIN and contributes to the pathophysiology of the disorder by inducing TGF-beta1 overproduction. This is the first study providing evidence that genetic factors may predispose to CIN and may have a role in the pathophysiology of the disorder.

No predictive value of serum interleukin-6 and transforming growth factor-β1 in identifying patients with a first restenosis, recurrent restenosis or a history of restenosis

The efficacy of percutaneous coronary intervention (PCI) is limited by the need for repeat revascularization resulting from restenosis. The restenosis rate after treatment for in-stent restenosis (recurrent restenosis) is high (> 30%). Numerous studies have suggested the predictive value of interleukin 6 (IL-6) and transforming growth factor beta1 (TGF-beta1). We sought to determine whether serum levels of IL-6 and TGF-beta1 could help identify individuals with recurrent restenosis. Thirty seven patients with a history of stent implantation were enrolled and divided into three groups: (1) patients with a current, first restenosis (n = 9); (2) patients with current restenosis and at least one prior restenosis (recurrent restenosis) (n = 11), and (3) patients with a history of restenosis, but without current restenosis (n = 17). The baseline profile was similar in all three groups. The median (25th-75th percentile) concentrations of IL-6 were: group 1 - 2.8 (1.4-5.5); group 2 - 2.6 (0.6-8.6); group 3 - 2.4 (0.9-4.7) p = 0.69 and TGF-beta1: group 1 - 3.6 (0.2-14.4); group 2 - 4.2 (1.8-57.6); group 3 - 6.6 (2.8-30.0) p = 0.57. Moreover we found no correlation, either between diameter stenosis and IL-6 (R = 0.10; p = 0.38) or TGF-beta1 (R = 0.10; p = 0.57). Both IL-6 (AUC 0.59 p = 0.4 and AUC 0.51 p = 0.9) and TGF-beta1 (AUC 0.64 p = 0.2 and AUC 0.50 p = 0.9) failed to provide significant results in receiver-operating characteristic analysis. We report that there is no association between the severity of diameter stenosis (restenosis) and IL-6 or TGF-beta1 concentrations. Our findings might suggest that levels of IL-6 and TGF-beta1 have no predictive value for identifying patients with recurrent restenosis.

Endothelial to Mesenchymal Transition via Transforming Growth Factor-β1/Smad Activation Is Associated with Portal Venous Stenosis in Idiopathic Portal Hypertension

Idiopathic portal hypertension (IPH) represents noncirrhotic portal hypertension of unknown etiology, mainly due to stenosis of peripheral portal veins. This study was performed to clarify the mechanism of portal venous stenosis in IPH from the viewpoint of the contribution of the endothelial to mesenchymal transition of the portal vein endothelium via transforming growth factor-beta1 (TGF-beta1)/Smad activation. In vitro experiments using human dermal microvascular endothelial cells demonstrated that TGF-beta1 induced myofibroblastic features in human dermal microvascular endothelial cells, including spindle cell morphology, reduction of CD34 expression, and induction of S100A4, alpha-smooth muscle actin, and COL1A1 expression, as well as the increased nuclear expression of phospho-Smad2. Bone morphogenic protein-7 preserved the endothelial phenotype of human dermal microvascular endothelial cells. Immunohistochemical analysis showed that endothelial cells of the peripheral portal veins in IPH were characterized by the decreased expression of CD34 and the enhanced nuclear expression of phospho-Smad2; these results also confirmed the expression of S100A4 and COL1A1 in the portal vein endothelium. Serum TGF-beta1 levels in patients with IPH were significantly higher than those of healthy volunteers and patients with chronic viral hepatitis/liver cirrhosis, while an elevation of serum bone morphogenic protein-7 levels was not observed. These results suggest that the endothelial to mesenchymal transition of the portal venous endothelium via TGF-beta1/Smad activation is associated with portal venous stenosis in IPH, and bone morphogenic protein-7 may therefore be a suitable therapeutic candidate for IPH.