Serum Concentrations Of TGF-β Research Articles

Change of Th17/Treg ratio in peripheral blood and its clinical significance in patients with traumatic sepsis

Objective To investigate changes of Th17/Treg ratio in peripheral blood of patients with traumatic sepsis and discuss the underlying clinical significance.Methods Thrity-five multi-trauma patients enrolled in the study were divided into trauma group (n =22) and sepsis group (n =13 ).Another nine healthy volunteers were designed as the control group.Peripheral blood was collected from all the groups so as to determine the ratios of Th17 and Treg by flow cytometry,detect the mRNA expressions of RORγt and FoxP3 by real time fluorescence quantitative PCR and measure serum levels of IL-4,IL-6,IL-10,IL-17A,IL-23,transforming growth factor-β (TGF-β),interferon-γ (IFN-γ) and C-reaction protein (CRP) by ELISA.Further,the change of Th17/Treg ratio and its clinical significance were analyzed.Results The proportion of Th17 in the trauma group showed no significant difference with that in the control group, whereas the ratio of Th17/Treg was lower than that in the control group (P ＜ 0.05).The proportion of Th17 and ratio of Th17/Treg in the sepsis group were higher than those in the trauma group and control group (P ＜ 0.01 ).Serum concentrations of TGF-β,IL-6 and IL-23 in the sepsis group were all significantly higher than those in the control group ( P ＜ 0.01 ).Serum levels of TGF-β had no significant difference between the sepsis group and the trauma group,while serum concentrations of IL-6 and IL-23 in the sepsis group were higher than those in the trauma group ( P ＜ 0.01 ).Serum level of IL-17A and mRNA expression of RORγt in the sepsis group were higher than those in the trauma and control groups ( both P ＜ 0.01 ).Conclusion Th17/Treg imbalance is closely correlated with the development of sepsis in patients with severe trauma. Key words: Shock, traumatic; Sepsis; T-lymphocytes,regulatory

DECREASED EXPRESSION OF CD4+CD25+ TREG ON PERIPHERAL BLOOD MONONUCLEAR CELLS IN PATIENTS WITH ACUTE CORONARY SYNDROME

ObjectivesTo investigate the expression and function of CD4+CD25+ Treg on PBMCs in patients with acute coronary syndrome (ACS).MethodsThere were 48 patients with ACS and 12 control subjects. Patients were classified...

Evaluation of balance between regulatory T cells and T helper 17 cells in patients with atopic dermatitis

[Objective] To assess the role of imbalance between regulatory T (Treg) cells and T helper 17 (Th17) cells in the pathogenesis of atopic dermatitis (AD).[Methods] Peripheral blood was obtained from 41 patients with AD and 38 age- and sex-matched healthy controls.Flow cytometry was performed to determine the percentage of Treg cells (CD4+CD25+Foxp3+ T cells) and Thl7 cells (CD4+ILl7+ T cells),real-time quantitative reverse transcription (RT)-PCR to detect the mRNA expressions of Foxp3 and RORγt,which are the specific transcription factors of Treg and Th17 cells respectively.Serum concentrations of transforming growth factor (TGF)-β,IL-17 and IL-23 were measured by enzyme linked immunosorbent assay(ELISA).Data were statistically assessed by independent-samples t test and Pearson correlation analysis.[Results] The patients with AD showed an obvious decrease in Treg cell percentage,transcription factor Foxp3 mRNA level and Treg/Th17 ratio (2.01％ ± 0.57％ vs.5.04％ ± 1.44％,t =12.47,P＜ 0.01; 0.65 ± 0.19 vs.1.71 ± 0.69,t=9.47,P＜0.01; 1.26 ± 0.61 vs.14.53 ± 5.77,t =14.11,P ＜ 0.01),but a significant increase in peripheral Th17 cell percentage and transcription factor RORγt mRNA level (1.77％ ± 0.55％ vs.0.39％ ± 0.15％,t =14.82,P ＜0.01; 5.97 ± 1.45 vs.1.49 ± 0.57,t =17.78,P ＜ 0.01 ) compared with the healthy controls.Further comparison revealed that Treg/Th17 ratio was significantly lower in patients with acute AD than in those with subacute AD (0.88 ± 0.04 vs.1.29 ± 0.11,t =4.02,P ＜ 0.01 ) and those with chronic AD (2.05 ± 0.24,t =4.83,P ＜ 0.01 ),statistically different between patients with subacute AD and chronic AD (t =2.89,P ＜ 0.05).There was no significant difference in the serum concentration of TGF-β between patients with AD and healthy controls ((15.28 ± 2.34) μg/L vs.(16.56 ± 3.27) μg/L,t =1.96,P＞ 0.05).A significant increase was observed in the serum levels of IL-17 and IL-23 in patients with AD compared with those in the healthy controls( (33.24 ± 7.06)ng/L vs.(11.68 ± 2.67) ng/L,t =17.96,P＜ 0.01; (56.35 ± 12.16) ng/L vs.(18.43 ± 3.90) ng/L,t =18.36,P＜ 0.01).In patients with moderate and severe AD,SCORing atopic dermatitis (SCORAD) index was negatively correlated with the percentage of Treg ceils (r =-0.40,P＜ 0.05 ),but positively correlated with that of Th17 cells (r =0.42,P ＜ 0.05 ).[Conclusion]s There exists a change in Treg/Th 17 ratio,mRN A expressions of RORγt and Foxp3,and serum levels of relevant cytokines in patients with AD,which may lead to immune imbalance and subsequently contribute to the development of AD. Key words: Dermatitis; atopic; CD4-positive T-lymphocytes; T-lymphocytes; regulatory; Th17 cells

Promoter region polymorphisms in the transforming growth factor beta-1 (TGFβ1) gene and serum TGFβ1 concentration in preeclamptic and control Iranian women

Preeclampsia (PE) is a pregnancy associated disorder characterized by hypertension and proteinuria, which causes neonatal and maternal morbidity and mortality. The Th1/Th2 cytokine paradigm of the immune adaptation in pregnancy is now expanded to include Th1/Th2/Th17 and regulatory T (Treg) cells. Among cytokines, TGFβ1 has properties that justify evaluation of its role in PE etiopathology. In this investigation the polymorphisms of the TGFβ1 gene at promoter region, positions −800G→A and −509C→T, were studied in 142 PE and 140 normal pregnant female subjects using PCR-RFLP. Additionally, serum TGFβ1 was determined by ELISA. At position −800G→A genotypes and allele frequencies showed no significant differences between PE patients (GG 73.9%; GA 21.1%; AA 4.93%) and normal control (GG 70%; GA 28.6%; AA 1.4%) women. However the AA genotype at this position was more frequent in PE patients than in the control group. At −509C→T position, genotypes and allele frequencies showed no significant differences between PE patients and control individuals. The CC genotype at −509C→T position was more prevalent in PE patients than in the control group. The mean serum TGFβ1 level was significantly higher (62.14ng/ml) in PE patients compared with pregnant and non-pregnant control groups (and 47.01 and 40.68ng/ml, respectively). In conclusion, the promoter region polymorphisms of TGFβ1 may not be associated with PE, but serum levels of this cytokine may contribute to the etiopathology of PE.

High serum levels of transforming growth factor β1 are associated with increased cortical thickness in cingulate and right frontal areas in healthy subjects

BackgroundTransforming growth factor β (TGF-β) is a cytokine having multiple functions in the central nervous system such as promoting repair mechanisms in degenerative diseases and stroke. To date, however, its neuroprotective effects in non-disease conditions have not been studiedMethodsWith the aim of exploring the relationship between peripheral TGF-β1 expression and brain structural integrity, 70 healthy participants underwent high-resolution structural T1-weighted magnetic resonance imaging scans and blood sampling. Data were processed to obtain brain cortical thickness and serum concentrations of TGF-β1. We investigated the correlation between TGF-β1 and cortical thickness using both region-of-interest- and vertex-based approaches.FindingsRegion-of-interest-based analysis of the cortical mantle showed a correlation between TGF-β1 serum concentrations and cortical thickness bilaterally in cingulate and right frontal and temporal areas. Similar results emerged in the vertex-based analysis, where significant correlations were found bilaterally in cingulate and right frontal cortices.ConclusionsThese results suggest that TGF-β1, through its role in down-regulating inflammatory processes, might have a beneficial effect on the structural integrity of the brain in physiological states.

Stage-dependent association of BDNF and TGF-beta1 with lung function in stable COPD

BackgroundChronic Obstructive Pulmonary Disease (COPD) is characterised by complex inflammatory, neuronal and fibrotic changes. Brain-derived Neurotrophic Factor (BDNF) is a key regulator of neuronal plasticity, whereas Transforming Growth Factor-β1 (TGF-β1) plays a crucial role in tissue repair and emphysema pathogenesis. Both mediators are stored in platelets and released from platelets in inflammatory conditions and during serum preparation. In patients with asthma, it was previously shown that elevated serum BDNF concentrations correlate with disease severity, whereas TGF-β1 concentrations were normal.MethodsIn the present study, 63 patients with stable COPD (spirometric GOLD stages 2–4) and 17 age- and comorbidity-matched controls were studied. Lung function, smoking history, medication, platelet concentrations in peripheral blood and serum concentrations of BDNF, TGF-β1 and Serotonin (5-HT) were assessed in all participants.ResultsSerum levels of both BDNF and TGF-β1 (but not concentrations of platelets in peripheral blood) were significantly elevated in all stages of COPD as compared to controls. Highest BDNF concentrations were found in spirometric GOLD stage 3, whereas highest TGF-β1 serum levels were found in spirometric GOLD stage 4. There were specific, stage-dependent correlations of these mediators with lung function parameters of the patients.ConclusionsTaken together, we show that, in contrast to asthma, COPD is characterised by elevated concentrations of both BDNF and TGF-β1 in serum. The stage-dependent association with lung function supports the hypothesis that these platelet mediators may play a role in the pathogenesis of COPD.

The influence of low molecular weight heparin on the expression of osteogenic growth factors in human fracture healing

Anticoagulant therapy with low molecular weight heparins (LMWH) and mechanical compression is considered the gold standard for the prevention of thrombosis. However, evidence exists that LMWHs impair bone metabolism. The aim of this study was therefore to analyse alterations in the expression of M-CSF, VEGF and TGF-ß1 after treatment with enoxaparin in patients with long bone fracture to investigate the effect of LMWH on human fracture healing. A total of 81 patients with long bone fractures were included in the study and divided into two groups. One group comprised patients who received enoxaparin and the other group, patients who did not receive enoxaparin postoperatively. Growth factor levels were analysed in patients' serum and different groups were retrospectively compared. M-CSF serum concentrations were found to be significantly higher only at 48 weeks after surgery in enoxaparin. Mean overall VEGF serum concentration was higher in patients with enoxaparin. TGF-β1 serum concentrations were higher at 48 weeks after surgery in patients with enoxaparin. This is the first comparative systemic measurement of M-CSF, VEGF and TGF-ß1 serum levels in patients with and without enoxaparin after long bone fracture. Significant differences of the expression of the growth factors after enoxaparin therapy were only observed at week 48 after surgery for M-CSF and TGF-ß1.

Coordinated Increase in Serum Platelet-Derived Growth Factor-BB and Transforming Growth Factor-β1 in Patients with Chronic Pancreatitis

Background: In vitro studies suggest that platelet-derived growth factor-BB (PDGF-BB) and transforming growth factor-β1 (TGF-β1) play an important role in pancreatic fibrosis. The aim of this study was to evaluate serum PDGF-BB and TGF-β1 concentrations in patients with chronic pancreatitis (CP). Methods: Forty male patients with a history of alcoholic CP and 35 age-matched healthy subjects were examined. Serum concentrations of PDGF-BB, TGF-β1, laminin and hyaluronic acid were determined by ELISA assay. Additionally, we determined serum concentrations of PDGF-BB and TGF(31 in patients with functional dyspepsia, ulcerative colitis and Crohn's disease. Results: Patients with advanced CP had significantly higher serum PDGF-BB and TGF-β1 concentrations compared to control subjects. A strong positive correlation between serum PDGF-BB and TGF-β1 concentrations was found in patientswith CP. Serum laminin and hyaluronic acid were also elevated in patients with CP. No increase in serum PDGF-BB and TGF-β1 concentrations was found in patients with functional dyspepsia, ulcerative colitis or Crohn's disease. Conclusion: The obtained results indicate for the first time that serum levels of PDGF-BB are elevated in patients with CP. However, ROC curve analysis suggests that PDGF-BB is not superior to laminin as a potential marker of advanced CP.

Elevated transforming growth factor-beta 1 (TGF-β1) levels in human fracture healing

IntroductionTransforming growth factor-beta 1(TGF-β1) is a regulatory protein, involved in bone fracture healing. Circulating TGF-β1 levels have been reported to be a predictor of delayed bone healing and non-union, suggesting active relationship between tissue and circulating TGF-β1 in fracture healing. The purpose of this study was to analyse TGF-β1 local and serum concentrations in fracture healing to further contribute to the understanding of molecular regulation of fracture healing. Patients and methodsSerum samples of 113 patients with long bone fractures were collected over a period of 6 months following a standardised time schedule. TGF-β1 serum concentrations were measured using ELISA. Patients were assigned to 2 groups: Group 1 contained 103 patients with physiological healing. Group 2 contained 10 patients with impaired healing. Patients in both groups were matched. One patient of the group 2 had to be excluded because of missing match partner. In addition, fracture haematoma from 11 patients of group 1 was obtained to analyse local TGF-β1 concentrations. 33 volunteers donated serum which served as control. ResultsTGF-β1 serum concentrations increased during the early healing period and were significantly higher in patients with physiological healing compared to controls (P=0.04). Thereafter, it decreased continuously between weeks 2 and 8 and fell again after week 8. TGF-β1 serum concentrations in patients with physiological healing were significantly higher at week 24 compared to controls (P=0.05). In non-unions, serum concentrations differed significantly from those of controls at week 6 (P=0.01). No significant difference in between patients with physiological and impaired fracture healing was observed. Fracture haematoma contained significantly higher TGF-β1 concentrations than peripheral serum of the patients (P=0.017). ConclusionElevated levels of TGF-β1 in haematoma and in serum after bone fracture especially during the entire healing process indicate its importance for fracture healing.

Effects of preparation method, shear force, and exposure to collagen on release of growth factors from equine platelet-rich plasma

To test the hypotheses that preparation method, exposure to shear force, and exposure to collagen affect the release of growth factors from equine platelet-rich plasma (PRP). PRP obtained from 6 horses. PRP was prepared via 2 preparation methods (tube and automated) and subjected to 6 treatment conditions (resting, detergent, exposure to shear via 21- and 25-gauge needles, and exposure to collagen [10 and 20 μg/mL]). Concentrations of platelet-derived growth factor, isoform BB (PDGF-BB); transforming growth factor β, isoform 1 (TGFβ₁); and insulin-like growth factor, isoform 1 (IGF-1) were quantified by use of ELISAs. Statistical analysis was conducted via repeated-measures ANOVA. Platelet numbers were significantly higher in tube-prepared PRP than in automated-prepared PRP Growth factor concentrations did not differ significantly between preparation methods. Mean PDGF-BB concentration ranged from 134 to 7,157 pg/mL, mean TGFβ₁ concentration ranged from 1,153 to 22,677 pg/mL, and mean IGF-1 concentration ranged from 150 to 280 ng/mL. Shear force did not affect growth factor concentrations. Dose-dependent increases in PDGF-BB and TGFβ₁ were detected in response to collagen, but equalled only 10% of the estimated total platelet content. Concentrations of IGF-1 were not significantly different among treatments and negative or positive control treatments. Serum concentrations of PDGF-BB and TGFβ₁ exceeded concentrations in PRP for most treatment conditions. Release of growth factors from equine PRP was negligible as a result of the injection process alone. Investigation of platelet-activation protocols is warranted to potentially enhance PRP treatment efficacy in horses.

TGF‐β1 serum concentration as a complementary diagnostic biomarker of lung cancer: establishment of a cut‐point value

Lung cancer is a malignant disease with increasing mortality rates. Cytokines play a role in normal cell growth regulation and differentiation and are also implicated in malignant disease. Among these cytokines, Transforming Growth Factor β type 1 (TGF-β1) acts as a tumor promoter in malignant cells. Several clinical studies have found high levels of TGF-β1 in various cancer types. The aim of this study was to establish a TGF-β1 cut-off point as a complementary diagnostic tool in lung cancer detection. Therefore, 72 clinically well-characterized individuals were studied, 41 lung cancer patients and 31 healthy subjects. Serum TGF-β1 concentration was measured by an enzyme-linked immunosorbent assay (ELISA). We compared statistically the serum TGF-β1 concentration between both groups with analysis of variance, linear regression and receiver operating curve analysis. We observed that lung cancer patients produced higher TGF-β1 levels than healthy individuals (37,225±9,436 vs. 28,416±9,324 pg/ml, P<0.001). The cut-point diagnostic value was 30,500 pg/ml with 80.5% sensitivity, 64.5% specificity and odds ratio: 7.5, 95% CI: 2.6-21.8. We found significantly higher TGF-β1 levels in lung cancer patients than in healthy individuals. We propose the measurement of serum TGF-β1 levels as a complementary diagnostic test in lung cancer detection.

Change of TGF-β1 concentration in blood serum of patients during systemic treatment of severe psoriasis

Change of TGF-β1 concentration in blood serum of patients during systemic treatment of severe psoriasis TGF-β1 is a growth factor with multiple actions, responsible for many pathological and physiological processes. Radiation of the skin with a narrowband 311 nm UVB light source is the most often used method of therapy in psoriasis treatment. The aim of the study was to compare different therapy methods on the change of TGF-β1 serum concentration during two weeks of treatment. The TGF-β1 concentration in blood serum of patients with psoriasis was statistically significantly lower than in the control group. For psoriasis treatment, narrow band 311 nm UVB phototherapy was applied to 37 patients and systemic pharmacological treatment with metothrexate, to a group of 18 patients. After two weeks, there was a significant reduction of Psoriasis Area and Severity Index (PASI) and increase of TGF-β1 serum concentration. Narrowband UVB caused a lower PASI, and higher TGF-β1 serum concentration than did pharmacological systemically treatment. Growth factor TGF-β1 had an effect on both of these processes. Lack of TGF-β1 in psoriatic skin is responsible for TH1-induced immune reactions and intensive proliferation of keratinocytes, which are typical of psoriasis. UVB radiation results in a higher TGF-β1 tissue concentration. Elevation of TGF-β1 concentration in blood serum and skin after UVB may present a possible mechanism for the positive effect of UVB in psoriasis treatment.

E0219 Study on the Role of CD4+CD25+treg on Atherosclerosis in ApoE-/- Mice

ObjectiveTo investigate the mechanism of CD4+CD25+treg in atherosclerosis in ApoE-/- mice.Methods28 weeks-old male ApoE−/− mice were randomly divided into two groups feeding high-fat diet (AH) or normal diet (AN). Ten...

E0186 Study on the mechanism of inhibitory effect of CTLA-4Ig fusion protein on atherosclerosis in apoe deficient mice

ObjectiveTo investigate the mechanism of inhibitory effect of CTLA-4Ig fusion protein on atherosclerosis in mice with an apolipoprotein-E gene defect fed on cholesterol diet.Methods30 male 10-week-old apoE(-/-) mice were fed...

Abnormal expression of Endoglin and its receptor complex (TGF-β1 and TGF-β receptor II) as early angiogenic switch indicator in premalignant lesions of the colon mucosa

The precise timing of the angiogenic switch in colorectal cancer development is still unclear. The simultaneous expression of Endoglin (CD105), transforming growth factor (TGF)-β1 and TGF-β receptor (R) II were quantified in surgical specimens comprising normal human colon, pre-malignant dysplastic tissue, in situ, and invasive colon cancer specimens, at mRNA and protein levels, respectively by real-time PCR and immunohistochemistry. Serum concentrations of soluble Endoglin and TGF-β1 were evaluated. mRNA and CD105+-microvessel density (MVD) increased significantly in dysplastic colon and carcinoma versus normal tissues; values correlated respectively with dysplasia degree and Dukes' stages. TGF-β1 expression was significantly upregulated in most severe dysplastic adenoma specimens, while TGF-β1 transcript and protein signals were intense in carcinoma, positively-correlated with tumor progression. TGF-β1 RII was overexpressed in adenoma and carcinoma versus normal samples, but unrelated with dysplasia or Dukes' stage. Soluble Endoglin serum levels were equivalent in adenoma and normal tissues; in carcinoma the highest levels were in invasive tumor. Circulating TGF-β1 levels were increased in severe dysplasia and progressed with tumor progression. Correlations between adenoma dysplasia degree and TGF-β RII and CD105+-MVD, and between tumor Dukes' staging and TGF-β1 and CD105+-MVD, were significant. TGF-β1 and Endoglin and TGF-β1 serum levels, TGF-β1 staining and CD105+-MVD were significantly and inversely associated with disease-free survival. TGF-β1 levels were an independent and significant prognostic factor of disease-free survival. These findings suggest active angiogenesis occurs in many pre-malignant colon cases and supports more careful evaluation of different chemopreventive agents.

Cigarette smoking decreases TGF-β1 serum concentrations after long bone fracture

TGF-β1 serum concentrations are considered to be one of the most promising markers of fracture healing. Previously, we demonstrated significant differences in the post-traumatic time courses of patients with timely and delayed fracture healing. The aim of this study was to evaluate possible differences in the serum concentrations of TGF-β1 in cigarette-smoking vs. non-smoking patients with timely and delayed fracture healing in order to understand pathophysiological pathways through which smoking impairs fracture healing. Serum samples were collected from 248 patients undergoing surgical treatment for long bone fractures within 1 year of surgery. Samples from 14 patients with atrophic-type delayed fracture healing were compared with 14 matched patients with normal bone healing. Each group included seven smokers and seven non-smokers. Post-operative serum concentrations were analysed at 1, 2, 4, 8, and 12 weeks as well as 1 year after surgery. The patients were monitored both clinically and radiologically for the entire duration of the study. All patients increased TGF-β1 serum concentrations after surgery. In patients with normal fracture healing, significantly higher TGF-β1 levels were observed in non-smokers (70 ng/ml) than in smokers (50 ng/ml) at the 4th week after surgery ( p = 0.007). Also at the 4th week, in patients with delayed healing, significantly lower TGF-β1 levels were observed in smokers than in non-smokers (38 ng/ml vs. 47 ng/ml, p = 0.021). However, no significant differences between non-smokers with delayed healing and smokers with normal healing ( p = 0.151) were observed at the 4th week after surgery. TGF-β1 serum concentrations reached a plateau in all groups from the 6th to the 12th week after surgery, with a slight decrease observed in the final measurement taken 1 year after surgery. This study demonstrates that, after fracture, TGF-β1 serum concentrations are reduced by smoking, and this reduction is statistically significant during the 4th week after surgery. Our findings may help reveal the mechanism by which smoking impairs fracture healing. Furthermore, these results may help to establish a serological marker that predicts impaired fracture healing soon after the injury. Surgeons will not only be able to monitor the bone healing, but they will also be able to monitor the success of additional treatments such as ultrasound and bone morphologic proteins (BMPs).

The relationship between the TGF-β1 gene −509C/T polymorphism and tubulointerstitial damage resulting from primary nephrotic syndrome

Background: The aim of this study was to investigate the correlation between the transforming growth factor (TGF)-β1 gene −509C/T polymorphism and the susceptibility to primary nephrotic syndrome (PNS), and in particular to the severe degree of tubulointerstitial damage (TID) seen in Chinese. Methods: Ninety-eight PNS patients and 128 normal controls were studied. The extent of tubulointerstitial changes was evaluated and patients were divided into two groups according to the severe or mild degree of TID. The TGF-β1gene −509C/T polymorphism was detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, and the serum level of TGF-β1 was determined by enzyme-linked immunosorbent assay (ELISA). Results: No statistical differences in genotype or allele frequency of the TGF-β1 gene −509C/T were found between PNS and normal subjects. However, T allele and CT + T T genotype frequency were higher in the PNS with severe TID than the mild TID and controls. Additionally, the serum concentration of TGF-β1 was significantly higher in the PNS with severe TID group than the other two groups and in the T T genotype individuals than the CC and CT genotype individuals. A logistic regression analysis demonstrated that TGF-β1 gene −509C/T genotype was the risk factor of TID in PNS patients [OR (odd ratio) 2.34, confidence interval (CI) 0.98–3.46, p = 0.012]. Conclusion. TGF-β1 gene −509C/T polymorphism was associated with severe TID. The higher value in serum concentration of TGF-β1 was also associated with severe TID and the T T genotype/T allele. T allele gene might be the important risk factor for susceptibility.

The Kinetics of Transforming Growth Factor beta1 mRNA Expression and Its Serum Concentration in Graft-Versus-Host Disease After Allogeneic Hemopoietic Stem Cell Transplantation for Myeloid Leukemias.

Abstract 4479 IntroductionGraft-versus-host disease (GVHD) remains one of the major complication following allogeneic haematopoietic stem cell transplantation (alloHSCT). Recent data indicates that transforming growth factor beta1 (TGF-β1) may play a role in the development of GVH reaction. Patients and methodsThe study included 40 adult patients who underwent allogeneic bone marrow transplantation for myeloid leukemias between 2001-2004. Twenty four patients were transplanted for acute myeloid leukemia in first complete remission and 16 patients for chronic myeloid leukemia in first chronic phase. 25 patients received stem cells from unrelated donor whereas the donor was sibling for the remaining 15 recipients. Conditioning regimen consisted of busulphan and cyclosphamide (BuCy) and ciclosporin (CyS) and methotrexate (Mtx) were administered for GVHD prophylaxis. In all cases, the expression of TGF-β1 mRNA by real-time quantitive polymerase chain reaction (RT-QPCR) using ABI PRISMTM 7700 (Taqman) and TGF-β1 serum concentration by commercial ELISA were measured fourfold in the peritransplant period (on days: -10, 0, +30 and +100). ResultsAcute GVHD (aGVHD) experienced 20 patients, in 4 patients the symptoms were classified as severe (grades III-IV). Chronic GVHD (cGVHD) occurred in 11 patients (in 4 as a diffuse form). A median TGF-β1 mRNA expression was 7088 copies/μg of total RNA at day -10 before transplant, dropped to 1907 copies/μg of total RNA after conditioning (p<0.005) and then significantly increased on days +30 (p<0.04) and +100 (p<0.003) when compared to the day of transplant (median: 3808 and 7531 copies/μg of total RNA, respectively). A median TGF-β1 serum concentration was 21411 pg/ml at day -10 before transplant, dropped to 8277 pg/ml at the day of transplant (p<0.000063), the low value (median: 12184 pg/ml) was still detected on day +30 (p<0.0026) in comparison to the pretransplant period. A median TGF-β1 serum concentration significantly increased on day +100 (median: 19509 pg/ml) when compared to the day of transplant and to the day +30 (p<0.0025 and p<0.0196, respectively). All patients demonstrated a prompt decrease in TGF-β1 mRNA expression and its serum concentration after conditioning. In patients with acute GVHD, TGF-β1 mRNA expression and its serum concentration remained low until day +30 after transplant if compared to the transplant day (p<0.03 and p<0.006, respectively). TGF-β1 mRNA expression and its serum concentration significantly increased on day +100 if compared to the transplant day (p<0.0009 and p<0.02, respectively) in patients who developed chronic GVHD. ConclusionsWe demonstrate the kinetics of TGF-β1 changes both on molecular and protein level in patients who underwent alloHSCT for myeloid leukemias. These results show that TGF-β1 is likely to be an additional regulator of donor engraftment, probably its low levels contributing to the development of acute GVHD, which may suggest that TGF-β1 exerts beneficial effects in the early post transplant period. In contrast, chronic GVHD symptoms seem to correlate significantly with high TGF-β1 levels, suggesting that TGF-β1 is likely to increase the risk of cGVHD by causing fibrosis of target organs, which remains in coherence with TGF-β1 profibrotic activity. Disclosures:No relevant conflicts of interest to declare.

Liver fibrosis secondary to bile duct injury: correlation of Smad7 with TGF-β and extracellular matrix proteins

BackgroundLiver fibrosis is the result of continuous liver injury stemming from different etiological factors. Bile duct injury induces an altered expression of TGF-β, which has an important role in liver fibrosis because this cytokine induces the expression of target genes such as collagens, PAI-1, TIMPs, and others that lead to extracellular matrix deposition. Smad7 is the principal inhibitor that regulates the target gene transcription of the TGF-β signaling. The aim of the study was to determine whether Smad7 mRNA expression correlates with the gene expression of TGF-β, Col I, Col III, Col IV, or PAI-1 in liver fibrosis secondary to bile duct injury (BDI).ResultsSerum TGF-β concentration was higher in BDI patients (39 296 pg/ml) than in liver donors (9008 pg/ml). Morphometric analysis of liver sections showed 41.85% of tissue contained fibrotic deposits in BDI patients. mRNA expression of Smad7, Col I, and PAI-1 was also significantly higher (P < 0.05) in patients with BDI than in controls. Smad7 mRNA expression correlated significantly with TGF-β concentration, Col I and Col III expression, and the amount of fibrosis.ConclusionWe found augmented serum concentration of TGF-β and an increase in the percentage of fibrotic tissue in the liver of BDI patients. Contrary to expected results, the 6-fold increase in Smad7 expression did not inhibit the expression of TGF-β, collagens, and PAI-1. We also observed greater expression of Col I and Col III mRNA in BDI patients and significant correlations between their expression and TGF-β concentration and Smad7 mRNA expression.

Relationship of age-related serum transforming growth factor (TGF)-β1 and TGF-β2 with osteoprotegerin and leptin in women

Objective To investigate the changes of transforming growth factor-β1 (TGF-131)and β2 (TGF-β2) with age in females and their relationships with osteoprotegerin(OPG) and leptin. Methods Serum TGF-β1, TGF-β2, OPG and leptin concentrations in 459 healthy adult women(aged 25-80 yr) were measured by an enzyme-linked immunosorbent assay (ELISA). Results Serum TGF-β1 (r =-0. 335, P < 0.01) was negatively while TGF-β2(r=0.230,P<0.01)positively correlated with the age. The geometric mean values of TGF-β1, OPG and leptin were (29.7±1.69) μg/L, (3.81±1.96) pmol/L and (10.5±2.01) μg/L, respectively. The arithmetic mean value of TGF-β2 was(13.74±3.86) μg/L. Concentrations of TGF-β1 in premenopansal, perimenopausal and postmenopausal women were (37.6±1.64), (33.4±1.69) and (24.3±1.59)μg/L,respectivly. Moreover, the concentration of TGF-β1 in postmenopausal women was significantly lower than that in perimenopausal and premenopausal women. The concentrations of TGF-β2 in premenopausal, perimenopausal and postmenopausal women were (11.7±2.68), (13.5±3.93) and (14.6±3.91) μg/L respectively, being increased in echelon. TGF-β1 was negatively correlated with TGF-β2(r=-0.261 ,P<0.01) and OPG (r =-0. 313, P<0. 01), but positively correlated with leptin (r = 0. 164, P<0.01). TGF-β2 was positively correlated with OPG (r=0. 432,P<0. 01). Conclusions The reference ranges of serum TGF-β1 and TGF-β2 in healthy adult females are established. Both TGF-β1 and TGF-β2 of them are correlated with OPG and ieptin. Key words: Transforming growth factor-β1 ; Transforming growth factor-β2; Osteoprotegerin; Leptin