scholarly journals Scleroderma-specific autoantibodies embedded in immune complexes mediate endothelial damage: an early event in the pathogenesis of systemic sclerosis

2020 ◽  
Author(s):  
Elena Raschi ◽  
Daniela Privitera ◽  
Caterina Bodio ◽  
Paola Adele Lonati ◽  
Maria Orietta Borghi ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Growth Factor ◽  
Immune Complexes ◽  
Topoisomerase I ◽  
Tumour Growth ◽  
Skin Fibroblasts ◽  
Mrna Levels ◽  
Healthy Skin ◽  
Tumour Growth Factor ◽  
Tgf Β1

Abstract Background: Consistently with their diagnostic and prognostic value, autoantibodies specific for systemic sclerosis (SSc) embedded in immune complexes (ICs) elicited a pro-inflammatory and pro-fibrotic cascade in healthy skin fibroblasts, engaging Toll-like Receptors (TLRs) via their nucleic acid components. The objective of this study was to investigate the pathogenicity of SSc-ICs in endothelial cells (ECs).Methods: ICs were purified from sera of SSc patients bearing different autoantibody specificities (antibodies against DNA topoisomerase I, centromeric proteins, RNA polymerase and Th/To), patients with systemic lupus erythematosus (SLE), primary anti-phospholipid syndrome (PAPS) or healthy controls (NHS) using polyethylen glycol precipitation. Human umbilical vein ECs (HUVECs) were incubated with ICs, IL-1β, Poly I:C, LPS or ODN CpG. mRNA levels of endothelin-1 (ET-1), collagenIα1 (colIα1), interferon (IFN)-α and IFN-β were investigated by Real-Time PCR; ICAM-1 expression was evaluated by cell-ELISA; secretion of IL-6, IL-8 and tumour growth factor (TGF)-β1 in culture supernatants was measured by ELISA. Intracellular signalling pathways culminating with NFkB, p38MAPK, SAPK-JNK and Akt were assessed by Western Blotting. Healthy skin fibroblasts were stimulated with supernatants from HUVEC incubated with ICs, and tumour growth factor (TGF)-β1secretion, mRNA levels of colIα1 and matrix metalloproteinase (mmp)-1 were evaluated.Results: All SSc stimulated IL-6 secretion compared to NHS-ICs; ACA-ICs and anti-Th/To-ICs increased ICAM-1 expression; all SSc-ICs but anti-Th/To-ICs augmented IL-8 levels; all SSc-ICs but ACA and ARA-ICs up-regulated et-1 and all SSc-ICs but ARA-ICs affected TGF-β1 secretion. ColIα1, IFN-α and IFN-β mRNA levels were not affected by any SSc-ICs. A differential modulation of tlr expression was observed: tlr2, tlr3 and tlr4 were upregulated by ATA-ICs and ACA-ICs, while anti-Th/To-ICs resulted in tlr9 up-regulation. All SSc-ICs activated p38MAPK and AKT, all SSc-ICs but ARA-ICs yielded the activation of NFκB; ATA-ICs and ACA-ICs increased the activation rate of both subunits of SAPK-JNK.When healthy skin fibroblasts were stimulated with supernatants from HUVECs incubated with SSc-ICs, TGF-β1 secretion and colIα1 expression levels were significantly modulated.Conclusions: These data provide the first demonstration of the pathogenicity of ICs from scleroderma patients with different autoantibodies in ECs. Endothelial activation induced by SSc-ICs ultimately led to a pro-fibrotic phenotype in healthy skin fibroblasts.

scholarly journals Scleroderma-specific autoantibodies embedded in immune complexes mediate endothelial damage: an early event in the pathogenesis of systemic sclerosis

2020 ◽  
Vol 22 (1) ◽  
Author(s):  
Elena Raschi ◽  
Daniela Privitera ◽  
Caterina Bodio ◽  
Paola Adele Lonati ◽  
Maria Orietta Borghi ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Systemic Sclerosis ◽  
Western Blotting ◽  
Immune Complexes ◽  
Topoisomerase I ◽  
Skin Fibroblasts ◽  
Mrna Levels ◽  
Healthy Skin ◽  
Pre Treatment ◽  
Tgf Β1

Abstract Background Consistently with their diagnostic and prognostic value, autoantibodies specific for systemic sclerosis (SSc) embedded in immune complexes (ICs) elicited a pro-inflammatory and pro-fibrotic cascade in healthy skin fibroblasts, engaging Toll-like receptors (TLRs) via their nucleic acid components. The objective of this study was to investigate the pathogenicity of SSc-ICs in endothelial cells. Methods ICs were purified from the sera of SSc patients bearing different autoantibody specificities (antibodies against DNA topoisomerase I, centromeric proteins, RNA polymerase, and Th/To), patients with systemic lupus erythematosus (SLE) and primary anti-phospholipid syndrome (PAPS), or healthy controls (NHS) using polyethylene glycol precipitation. Human umbilical vein endothelial cells (HUVECs) were incubated with ICs, positive and negative controls. mRNA levels of endothelin-1 (et-1), collagenIα1 (colIα1), interferon (IFN)-α, and IFN-β were investigated by real-time PCR; et-1 and il-6 mRNA levels were assessed after pre-treatment with bafilomycin. ICAM-1 expression was evaluated by cell ELISA; secretion of IL-6, IL-8, and transforming growth factor (TGF)-β1 in culture supernatants was measured by ELISA. The expression of Fcγ receptors (CD64, CD32, and CD16) was assessed in endothelial cells at FACS analysis. Intracellular signaling pathways culminating with NFκB, p38MAPK, SAPK-JNK, and Akt were assessed by Western blotting. Healthy skin fibroblasts were stimulated with supernatants from HUVECs incubated with ICs, and TGF-β1 secretion and mRNA levels of colIα1 and matrix metalloproteinase (mmp)-1, protein expression of α smooth muscle actin (α-SMA), and IL-6 were evaluated by Western blotting; et-1 mRNA levels were assessed in fibroblasts pre-treated with IL-6 and TGF-β inhibitors and stimulated with ATA-ICs. Results All SSc stimulated IL-6 secretion; ACA-ICs and anti-Th/To-ICs increased ICAM-1 expression; all SSc-ICs but anti-Th/To-ICs augmented IL-8 levels; all SSc-ICs but ACA-ICs and ARA-ICs upregulated et-1, and all SSc-ICs but ARA-ICs affected TGF-β1 secretion. colIα1, IFN-α, and IFN-β mRNA levels were not affected by any SSc-IC. FcγRII (CD32) and FcγRIII (CD16) were not detectable on HUVECs, while FcγRI (CD64) was minimally expressed. A differential modulation of tlr expression was observed: tlr2, tlr3, and tlr4 were upregulated by ATA-ICs and ACA-ICs, while anti-Th/To-ICs resulted in tlr9 upregulation. Pre-treatment with bafilomycin did not affect the upregulation of et-1 and il-6 induced by ATA-ICs, ACA-ICs, and anti-Th/To-ICs; a 23% reduction in both genes was reported for ARA-ICs. All SSc-ICs activated p38MAPK and Akt, and all SSc-ICs but ARA-ICs yielded the activation of NFκB; ATA-ICs and ACA-ICs increased the activation rate of both subunits of SAPK-JNK. When healthy skin fibroblasts were stimulated with supernatants from HUVECs incubated with SSc-ICs, TGF-β1 secretion, colIα1, α-SMA, and IL-6 expression levels were significantly modulated. Pre-treatment with IL-6 and TGF-β inhibitors prevented et-1 upregulation induced by ATA-ICs by 85% and 77%, respectively. Conclusions These data provide the first demonstration of the pathogenicity of ICs from scleroderma patients with different autoantibodies on the endothelium. Endothelial activation induced by SSc-ICs ultimately led to a pro-fibrotic phenotype in healthy skin fibroblasts.

scholarly journals Scleroderma-specific autoantibodies embedded in immune complexes mediate endothelial damage: an early event in the pathogenesis of systemic sclerosis

2020 ◽  
Author(s):  
Elena Raschi ◽  
Daniela Privitera ◽  
Caterina Bodio ◽  
Paola Adele Lonati ◽  
Maria Orietta Borghi ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Systemic Sclerosis ◽  
Western Blotting ◽  
Immune Complexes ◽  
Topoisomerase I ◽  
Transforming Growth Factor ◽  
Skin Fibroblasts ◽  
Mrna Levels ◽  
Healthy Skin ◽  
Pre Treatment

Abstract Background: Consistently with their diagnostic and prognostic value, autoantibodies specific for systemic sclerosis (SSc) embedded in immune complexes (ICs) elicited a pro-inflammatory and pro-fibrotic cascade in healthy skin fibroblasts, engaging Toll-like Receptors (TLRs) via their nucleic acid components. The objective of this study was to investigate the pathogenicity of SSc-ICs in endothelial cells.Methods: ICs were purified from sera of SSc patients bearing different autoantibody specificities (antibodies against DNA topoisomerase I, centromeric proteins, RNA polymerase and Th/To), patients with systemic lupus erythematosus (SLE), primary anti-phospholipid syndrome (PAPS) or healthy controls (NHS) using polyethylen glycol precipitation. Human umbilical vein endothelial cells (HUVECs) were incubated with ICs, positive and negative controls. mRNA levels of endothelin-1 (et-1), collagenIα1 (colIa1), interferon (IFN)-α and IFN-β were investigated by Real-Time PCR; et-1 and il-6 mRNA levels were assessed after pre-treatment with bafilomycin. ICAM-1 expression was evaluated by cell-ELISA; secretion of IL-6, IL-8 and transforming growth factor (TGF)-β1 in culture supernatants was measured by ELISA. The expression of Fcg receptors (CD64, CD32 and CD16) was assessed in endothelial cells at FACS analysis. Intracellular signalling pathways culminating with NFkB, p38MAPK, SAPK-JNK and Akt were assessed by Western Blotting. Healthy skin fibroblasts were stimulated with supernatants from HUVEC incubated with ICs, and TGF-b1 secretion, mRNA levels of colIα1 and matrix metalloproteinase (mmp)-1, protein expression of . a smooth muscle actin (a-SMA) and IL-6 were evaluated by Western Blotting; et-1 mRNA levels were assessed in fibroblasts pre-treated with IL-6 and TGF-b inhibitors and stimulated with ATA-ICs.Results: All SSc stimulated IL-6 secretion; ACA-ICs and anti-Th/To-ICs increased ICAM-1 expression; all SSc-ICs but anti-Th/To-ICs augmented IL-8 levels; all SSc-ICs but ACA and ARA-ICs up-regulated et-1 and all SSc-ICs but ARA-ICs affected TGF-b1 secretion. colIa1, IFN-a and IFN-b mRNA levels were not affected by any SSc-ICs. FcgRII (CD32) and FcgRIII (CD16) were not detectable on HUVECs, while FcgRI (CD64) was minimally expressed. A differential modulation of tlr expression was observed: tlr2, tlr3 and tlr4 were up-regulated by ATA-ICs and ACA-ICs, while anti-Th/To-ICs resulted in tlr9 up-regulation. Pre-treatment with bafilomycin did not affect the up-regulation of et-1 and il-6 induced by ATA-ICs, ACA-ICs and anti-Th/To-ICs; a 23% reduction in both genes was reported for ARA-ICs. All SSc-ICs activated p38MAPK and AKT, all SSc-ICs but ARA-ICs yielded the activation of NFkB; ATA-ICs and ACA-ICs increased the activation rate of both subunits of SAPK-JNK.When healthy skin fibroblasts were stimulated with supernatants from HUVECs incubated with SSc-ICs, TGF-b1 secretion, colIa1, a-SMA and IL-6 expression levels were significantly modulated. Pre-treatment with IL-6 and TGF-b inhibitors prevented et-1 up-regulation induced by ATA-ICs by 85% and 77% respectively.Conclusions: These data provide the first demonstration of the pathogenicity of ICs from scleroderma patients with different autoantibodies on the endothelium. Endothelial activation induced by SSc-ICs ultimately led to a pro-fibrotic phenotype in healthy skin fibroblasts.

scholarly journals Transplantation of Telocytes Attenuates Unilateral Ureter Obstruction-Induced Renal Fibrosis in Rats

2018 ◽  
Vol 46 (5) ◽  
pp. 2056-2071 ◽  
Author(s):  
Long Zheng ◽  
Long Li ◽  
Guisheng Qi ◽  
Mushuang Hu ◽  
Chao Hu ◽  
...  
Keyword(s):  
Growth Factor ◽  
Protective Effect ◽  
Renal Fibrosis ◽  
Transforming Growth Factor ◽  
Kidney Tissue ◽  
Collagen I ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mrna Levels ◽  
Mesenchymal Transition ◽  
Tgf Β1

Background/Aims: Previous studies imply that telocytes may have a protective effect on fibrosis in various organs, including the liver, colon, and heart. The effect of telocytes on renal fibrosis remains unknown. Herein, this study was designed to investigate the effect of telocytes on renal fibrosis and the potential mechanisms involved. Methods: In a unilateral ureteral obstruction (UUO)-induced renal fibrosis model, telocytes were injected via the tail vein every other day for 10 days. The degree of renal damage and fibrosis was determined using histological assessment. The expression of collagen I, fibronectin, epithelial-mesenchymal transition markers, and Smad2/3 phosphorylation was examined by western blot analyses. Real-time PCR and enzyme-linked immunosorbent assay were performed in vivo to detect the levels of transforming growth factor (TGF)-β1 and various growth factors. Results: Telocytes attenuated renal fibrosis, as evidenced by reduced interstitial collagen accumulation, decreased expression of fibronectin and collagen I, upregulation of E-cadherin, and downregulation of α-smooth muscle actin. Furthermore, telocytes decreased serum TGF-β1 levels, suppressed Smad2/3 phosphorylation, and increased the expression of hepatocyte growth factor (HGF) in rat kidney tissue following UUO. Blockage of HGF counteracted the protective effect of telocytes on UUO-treated kidneys. Through the detection of HGF mRNA levels in vitro, we found that telocytes had no effect on HGF expression compared with renal fibroblasts. Conclusion: Telocytes attenuated UUO-induced renal fibrosis in rats, likely through enhancing the expression of HGF in an indirect manner.

Differential regulation and expression of hyaluronan synthases in human articular chondrocytes, synovial cells and osteosarcoma cells

2001 ◽  
Vol 354 (1) ◽  
pp. 17-24 ◽  
Author(s):  
Anneliese D. RECKLIES ◽  
Chantal WHITE ◽  
Lee MELCHING ◽  
Peter J. ROUGHLEY
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Cultured Cells ◽  
Articular Chondrocytes ◽  
Synovial Cells ◽  
Mrna Levels ◽  
Cell Type ◽  
Osteosarcoma Cells ◽  
Predominant Species ◽  
Tgf Β1

Recently three isoforms of hyaluronan synthase (HAS), the enzyme responsible for hyaluronate/hyaluronan (HA) biosynthesis, have been cloned, allowing us to study their expression pattern. Our objective was to determine which of the HAS isoenzymes were expressed in human articular chondrocytes, synovial fibroblasts and osteosarcoma cells, whether their expression could be modulated by growth factors (insulin-like growth factor-1, basic fibroblast growth factor and transforming growth factor (TGF-β1) and cytokines [interleukin 1β1 (IL-1β)], and whether changes in the rate of HA synthesis by the cells correlated with changes in mRNA levels for one or more of the HAS isoforms. All three HAS isoforms were found to be expressed in the cultured cells analysed in this study, although the relative proportions varied for each cell type. HAS2 mRNA was usually predominant in chondrocytes, whereas synovial cells contained increased amounts of HAS1. HAS3 was always the least abundant message. The rapidly growing osteosarcoma cells contained almost exclusively HAS2 message. HAS usage in uncultured cartilage and synovial tissues was similar to that in the cultured cells, with HAS2 message being the predominant species in cartilage and HAS1 usually being the predominant species in synovium. HA synthesis was stimulated by the growth factors, but the extent of the response was cell-type specific. Synovial cells responded particularly well to IL-1β, and showed a unique synergistic response when IL-1β was used in combination with TGF-β1. This response was much reduced in articular chondrocytes and absent in the osteosarcoma cells. Analysis of changes in HAS message levels indicated that there was often no correlation with the changes in HA secretion following exposure to growth factors. Although HAS-1 mRNA was increased in synovial cells after exposure to TGF-β1/IL-1β, the magnitude of the change was far less than the effect on HA synthesis. Our data thus suggest that HAS gene usage is tissue specific, and the regulation by growth factors is unique for each HAS gene and is further modulated by cell-specific factors. In addition, regulation of HA biosynthesis appears to be multi-faceted, with control of HAS gene expression and mRNA levels being only one aspect of this process.

The expression of transforming growth factor-β by cultured chick growth plate chondrocytes: differential regulation by 1,25-dihydroxyvitamin D3

1996 ◽  
Vol 149 (2) ◽  
pp. 277-285 ◽  
Author(s):  
C Farquharson ◽  
A S Law ◽  
E Seawright ◽  
D W Burt ◽  
C C Whitehead
Keyword(s):  
Growth Factor ◽  
Conditioned Medium ◽  
Growth Plate ◽  
Bone Growth ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Mrna Levels ◽  
Chick Growth ◽  
Growth Plate Chondrocytes ◽  
Tgf Β1

Abstract 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) and transforming growth factor-β (TGF-β) are both important regulators of chondrocyte growth and differentiation. We report here that 1,25(OH)2D3 differentially regulates the expression of the genes for TGF-β1 to -β3 and the secretion of the corresponding proteins in cultured chick chondrocytes. Confluent growth plate chondrocytes were serum-deprived and cultured in varying concentrations of 1,25(OH)2D3. Cells were assayed for TGF-β mRNA and conditioned medium was assayed for TGF-β activity and isoform composition. Active TGF-β was only detected in 10−8m 1,25(OH)2D3-treated cultures (8·37 ng active TGF-β/mg protein). There was a significant decrease in total (latent+active) TGF-β activity in conditioned medium of 10−12 m (23·4%; P<0·05) and 10−10 m (20·7%; P<0·05) 1,25(OH)2D3-treated cultures but 10−8 m 1,25(OH)2D3 significantly increased (30·9%; P<0·01) TGF-β activity. The amounts of TGF-β1, -β2 and -β3 isoforms produced were similar in control, 10−10 or 10−12m 1,25(OH)2D3-treated cultures but the conditioned medium of 10−8 m 1,25(OH)2D3-treated cultures contained significantly higher amounts of all three isoforms. Quantification of TGF-β mRNA demonstrated differential control of TGF-β gene expression with TGF-β1 and -β3 mRNA levels reduced by all concentrations of 1,25(OH)2D3 examined (10−8, 10−10 and 10−12 m) whilst TGF-β2 mRNA concentrations were elevated. Our results indicated that 1,25(OH)2D3 regulates chick growth plate chondrocyte TGF-β secretion and mRNA expression in a concentration-dependent and isoform-specific manner. This interaction may be important in the regulation of chondrocyte metabolism and endochondral bone growth. Journal of Endocrinology (1996) 149, 277–285

scholarly journals Transforming growth factor β1 protein and mRNA levels in inflammatory bowel diseases: towards solving the contradictions by longitudinal assessment of the protein and mRNA amounts.

1970 ◽  
Vol 60 (4) ◽  
Author(s):  
Anna Liberek ◽  
Zbigniew Kmieć ◽  
Piotr M Wierzbicki ◽  
Joanna Jakóbkiewicz-Banecka ◽  
Tomasz Liberek ◽  
...  
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Mrna Levels ◽  
Longitudinal Assessment ◽  
Active Stage ◽  
Intestinal Tissue ◽  
Bowel Diseases ◽  
Inflammatory Bowel ◽  
Tgf Β1

Previously published studies on levels of the transforming growth factor-β1 (TGF-β1) protein and mRNA of the corresponding gene in patients suffering from inflammatory bowel diseases (IBD) gave varying results, leading to contradictory conclusions. To solve the contradictions, we aimed to assess longitudinally TGF-β1 protein and mRNA levels at different stages of the disease in children suffering from IBD. The study group consisted of 19 pediatric patients with IBD at the age between 3.5 and 18.4 years. The control group consisted of 42 children aged between 2.0 and 18.0 years. The plasma TGF-β1 concentration was measured with ELISA. mRNA levels of the TGF-β1 gene isolated from samples of the intestinal tissue were assessed by reverse transcription and real-time PCR. Levels of TGF-β1 protein in plasma and corresponding mRNA in intestinal tissue were significantly higher in IBD patients than in controls. TGF-β1 and corresponding transcripts were also more abundant in plasma and intestinal tissue, respectively, in patients at the active stage of the disease than during remission. In every single IBD patient, plasma TGF-β1 level and mRNA level in intestinal tissue was higher at the active stage of the disease than during remission. Levels of TGF-β1 and corresponding mRNA are elevated during the active stage of IBD but not during the remission. Longitudinal assessment of this cytokine in a single patient may help to monitor the clinical course of IBD.

Transforming Growth Factor Betal and Beta2 Induce Down-Modulation of Thrombomodulin in Human Umbilical Vein Endothelial Cells

1995 ◽  
Vol 73 (05) ◽  
pp. 812-818 ◽  
Author(s):  
Taro Ohji ◽  
Hajime Urano ◽  
Akira Shirahata ◽  
Minoru Yamagishi ◽  
Ken Higashi ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Growth Factor ◽  
Time Course ◽  
Transforming Growth Factor ◽  
Umbilical Vein ◽  
Mrna Levels ◽  
Antigen Level ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells ◽  
Tgf Β1

SummaryTo investigate the effects of transforming growth factor-betas (TGF-βs) on endothelial anticoagulant activity, we assayed thrombomodulin (TM) activity and antigen levels of human umbilical vein endothelial cells (HUVECs) incubated with TGF-βs in vitro. TGF-β1 suppressed surface TM activity and surface TM antigen levels maximally 12 h after incubation in dose-dependent manners. TGF-β2 was almost equipotent with TGF-β1 for the suppression of them. Both TGF-βs suppressed total TM antigen level in HUVECs, and the time course of the suppression was similar to that of the cell surface TM antigen level. The maximal reductions of TM mRNA levels by TGF-βs were observed at several hours ahead of those observed in both surface and total TM antigen levels, suggesting that the TGF-β-mediated suppression of TM antigen of HUVECs is primarily regulated at the TM mRNA level. Our present work suggests that the down-modulation of TM level induced by TGF-βs in HUVECs contributes in vivo to promoting the thrombogenesis either at the sites of injury of vessel walls, such as atherosclerotic lesions where TGF-β1 is released from platelets, smooth muscle cells and monocytes, or at neovascular walls in tumors secreting TGF-β2.

scholarly journals Increased Expression of Transforming Growth Factor-β after Cerebral Ischemia in the Baboon: An Endogenous Marker of Neuronal Stress?

2001 ◽  
Vol 21 (7) ◽  
pp. 820-827 ◽  
Author(s):  
Carine Ali ◽  
Fabian Docagne ◽  
Olivier Nicole ◽  
Sylvain Lesné ◽  
Jérôme Toutain ◽  
...  
Keyword(s):  
Growth Factor ◽  
Nonhuman Primate ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Oxygen Metabolism ◽  
Artery Occlusion ◽  
Experimental Stroke ◽  
Mrna Levels ◽  
Primate Model ◽  
Tgf Β1

There has been an increasing interest in recent years in the evaluation of the neuronal and glial responses to ischemic insult. Some cytokines, including transforming growth factor-β (TGF-β), that are overexpressed after experimental stroke in rodents are thought to be implicated in the neuronal processes that lead to necrosis. Thus, such cytokines could predict tissue fate after stroke in humans, although data are currently sparse for gyrencephalic species. The current study addressed the expression pattern of TGF-β1 in a nonhuman primate model of middle cerebral artery occlusion. Focal permanent ischemia was induced for 1 or 7 days in 6 baboons and the following investigations were undertaken: cerebral oxygen metabolism (CMRO2) positron emission tomography studies, magnetic resonance imaging, postmortem histology, and reverse transcription-polymerase chain reaction. The aim of the current study was to correlate the expression of TGF-β1 to the underlying metabolic and histologic state of the threatened cerebral parenchyma. The authors evidenced increased TGF-β1 mRNA levels (up to 25-fold) in those regions displaying a moderate (20% to 49%) reduction in CMRO2. The current findings suggest that the greatly enhanced expression of TGF-β1 in the penumbral zones that surround tissue destined to infarction may represent a robust index of potentially salvageable brain. The current investigation, in the nonhuman primate, strengthens the authors' hypothesis, derived from rodent models, that TGF-β1 may be involved in the physiopathology of human stroke.

Dissecting Growth Factors Production in Patients with Agnogenic Myeloid Metaplasia (AMM).

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4749-4749
Author(s):  
Jen-Chin Wang ◽  
Kirugaval Hemavathy ◽  
Amit Goldberg ◽  
Tsong S. Chang ◽  
Allan D. Novetsky ◽  
...  
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Transforming Growth Factor ◽  
Prior History ◽  
Mrna Levels ◽  
Myeloid Metaplasia ◽  
Number Of Patients ◽  
History Of ◽  
Tgf Β1 ◽  
Marrow Fibrosis

Abstract Growth factors including Transforming Growth Factor-β 1 ( TGF- β1), Platelet Derived Growth Factor (PDGF) and Fibroblast Growth Factor (FGF) have been implicated as responsible for bone marrow fibrosis in AMM. Although TGF-β1 and FGF have been demonstrated to be increased in blood hematopoietic progenitor cells, a direct measurement of the production of these growth factors by megakaryocytes has not been performed. The current study was devised to study the production of these growth factors production directly in megakaryocytes and monocytes of AMM patients and correlate these with the clinical features. Twelve patients with AMM and 11 normal healthy volunteers used as controls, were studied. CD 34 + cells and CD 14+ cells were obtained from blood mononuclear cells by MACS Progenitor cell isolation kits ( Miltenyi Biotec, CA). The megakaryocytic (CD41+) cells were then obtained by growing the isolated blood CD34 + for 10 days in Iscove Modified Dulbecco Medium with Stem Cell Growth Factor and Thrombopoietin. The mRNA levels of TGF-β1, FGF and PDGF in the isolated megakaryocytes and blood monocytes were assayed by Real-Time RT-PCR. The results were as shown in Table 1. Among the AMM patients, a patient with prior history of Polycythemia Vera (PV) and a patient with Essential Throbocythemia (ET) were found to have elevated PDGF and FGF expression in their monocytes but the expression of growth factors was not elevated in their megakaryocytes.. These results demonstrate that megakaryocytes are the main source of growth factors responsible for marrow fibrosis. The study also suggests that growth factors produced by monocytes may be responsible for fibrosis in AMM patients with a prior history of PV or ET. Table-1 Megakaryocytes Monocytes * Denotes number of patients with elevated growth factor levels as compared with controls. ** Denotes numbers of patients tested. TGF- β 1 8*/10* 0/10 PDGF 8/10 3/7 FGF 6/10 2/7

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