scholarly journals Transforming growth factor beta inhibits megakaryocyte growth and endomitosis

Blood ◽  
1992 ◽  
Vol 79 (3) ◽  
pp. 619-626 ◽  
Author(s):  
DJ Kuter ◽  
DM Gminski ◽  
RD Rosenberg
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Potent Inhibitor ◽  
Neutralizing Antibody ◽  
Maximal Concentration ◽  
Tgf Beta ◽  
Inhibitory Effects ◽  
Growth Factor Beta

Using a rat bone marrow culture system, the effect of transforming growth factor beta 1 (TGF beta 1) on megakaryocyte growth and endoreduplication has been studied. Purified human platelet TGF beta 1 inhibited the number of megakaryocytes that appeared in culture at a half-maximal concentration of 0.66 +/- 0.21 ng/mL and inhibited megakaryocyte endoreduplication at a half-maximal concentration of 0.14 +/- 0.08 ng/mL. Under identical conditions, growth of erythroid precursors was half-maximally inhibited at a concentration of 0.125 ng/mL while myeloid growth was not inhibited at concentrations of TGF beta 1 up to 25 ng/mL. These profound inhibitory effects on megakaryocyte growth and endomitosis suggested that TGF beta might play a role in megakaryocytopoiesis. Therefore, we explored the effect of TGF beta in three different experimental situations by using a neutralizing antibody to TGF beta: (1) Serum but not plasma was found to inhibit the number and ploidy of megakaryocytes that grew in vitro. This inhibitory activity was completely neutralized by antibody to TGF beta or on treatment with dithiothreitol. (2) Plasma from thrombocytotic rats was observed to decrease megakaryocyte ploidy on culture but this effect was not prevented by the addition of antibody to TGF beta. (3) Plasma from thrombocytopenic but not normal rats increased megakaryocyte ploidy on culture. Addition of antibody to TGF beta did not alter these results. Therefore, TGF beta is a potent inhibitor of the number and ploidy of megakaryocytes and accounts for all the inhibition seen when megakaryocytes are cultured in serum. However, the differences in effect on megakaryocyte growth that we observe between normal, thrombocytopenic, and thrombocytotic plasmas are not due to variations in the amount of TGF beta. Furthermore, our results show that release of TGF beta from megakaryocytes during culture does not act as an autocrine regulator of megakaryocyte ploidy in vitro.

scholarly journals Transforming growth factor beta inhibits megakaryocyte growth and endomitosis

Blood ◽  
1992 ◽  
Vol 79 (3) ◽  
pp. 619-626 ◽  
Author(s):  
DJ Kuter ◽  
DM Gminski ◽  
RD Rosenberg
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Potent Inhibitor ◽  
Neutralizing Antibody ◽  
Maximal Concentration ◽  
Tgf Beta ◽  
Inhibitory Effects ◽  
Growth Factor Beta

Abstract Using a rat bone marrow culture system, the effect of transforming growth factor beta 1 (TGF beta 1) on megakaryocyte growth and endoreduplication has been studied. Purified human platelet TGF beta 1 inhibited the number of megakaryocytes that appeared in culture at a half-maximal concentration of 0.66 +/- 0.21 ng/mL and inhibited megakaryocyte endoreduplication at a half-maximal concentration of 0.14 +/- 0.08 ng/mL. Under identical conditions, growth of erythroid precursors was half-maximally inhibited at a concentration of 0.125 ng/mL while myeloid growth was not inhibited at concentrations of TGF beta 1 up to 25 ng/mL. These profound inhibitory effects on megakaryocyte growth and endomitosis suggested that TGF beta might play a role in megakaryocytopoiesis. Therefore, we explored the effect of TGF beta in three different experimental situations by using a neutralizing antibody to TGF beta: (1) Serum but not plasma was found to inhibit the number and ploidy of megakaryocytes that grew in vitro. This inhibitory activity was completely neutralized by antibody to TGF beta or on treatment with dithiothreitol. (2) Plasma from thrombocytotic rats was observed to decrease megakaryocyte ploidy on culture but this effect was not prevented by the addition of antibody to TGF beta. (3) Plasma from thrombocytopenic but not normal rats increased megakaryocyte ploidy on culture. Addition of antibody to TGF beta did not alter these results. Therefore, TGF beta is a potent inhibitor of the number and ploidy of megakaryocytes and accounts for all the inhibition seen when megakaryocytes are cultured in serum. However, the differences in effect on megakaryocyte growth that we observe between normal, thrombocytopenic, and thrombocytotic plasmas are not due to variations in the amount of TGF beta. Furthermore, our results show that release of TGF beta from megakaryocytes during culture does not act as an autocrine regulator of megakaryocyte ploidy in vitro.

scholarly journals Transforming growth factor beta suppresses human immunodeficiency virus expression and replication in infected cells of the monocyte/macrophage lineage.

1991 ◽  
Vol 173 (3) ◽  
pp. 589-597 ◽  
Author(s):  
G Poli ◽  
A L Kinter ◽  
J S Justement ◽  
P Bressler ◽  
J H Kehrl ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
T Cell ◽  
Growth Factor ◽  
Cell Line ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Tgf Beta ◽  
Immunodeficiency Virus ◽  
Growth Factor Beta

The pleiotropic immunoregulatory cytokine transforming growth factor beta (TGF-beta) potently suppresses production of the human immunodeficiency virus (HIV), the causative agent of the acquired immunodeficiency syndrome, in the chronically infected promonocytic cell line U1. TGF-beta significantly (50-90%) inhibited HIV reverse transcriptase production and synthesis of viral proteins in U1 cells stimulated with phorbol myristate acetate (PMA) or interleukin 6 (IL-6). Furthermore, TGF-beta suppressed PMA induction of HIV transcription in U1 cells. In contrast, TGF-beta did not significantly affect the expression of HIV induced by tumor necrosis factor alpha (TNF-alpha). These suppressive effects were not mediated via the induction of interferon alpha (IFN-alpha). TGF-beta also suppressed HIV replication in primary monocyte-derived macrophages infected in vitro, both in the absence of exogenous cytokines and in IL-6-stimulated cultures. In contrast, no significant effects of TGF-beta were observed in either a chronically infected T cell line (ACH-2) or in primary T cell blasts infected in vitro. Therefore, TGF-beta may play a potentially important role as a negative regulator of HIV expression in infected monocytes or tissue macrophages in infected individuals.

scholarly journals Transforming growth factor beta 1 (TGF-beta 1) induced neutrophil recruitment to synovial tissues: implications for TGF-beta-driven synovial inflammation and hyperplasia.

1991 ◽  
Vol 173 (5) ◽  
pp. 1121-1132 ◽  
Author(s):  
R A Fava ◽  
N J Olsen ◽  
A E Postlethwaite ◽  
K N Broadley ◽  
J M Davidson ◽  
...  
Keyword(s):  
Growth Factor ◽  
Synovial Tissue ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Biologically Active ◽  
Tgf Beta ◽  
Histological Techniques ◽  
Growth Factor Beta

We have studied the consequences of introducing human recombinant transforming growth factor beta 1 (hrTGF-beta 1) into synovial tissue of the rat, to begin to better understand the significance of the fact that biologically active TGF-beta is found in human arthritic synovial effusions. Within 4-6 h after the intra-articular injection of 1 microgram of hrTGF-beta 1 into rat knee joints, extensive recruitment of polymorphonuclear leukocytes (PMNs) was observed. Cytochemistry and high resolution histological techniques were used to quantitate the influx of PMNs, which peaked 6 h post-injection. In a Boyden chamber assay, hrTGF-beta 1 at 1-10 fg/ml elicited a chemotactic response from PMNs greater in magnitude than that evoked by FMLP, establishing that TGF-beta 1 is an effective chemotactic agent for PMNs in vitro as well as in vivo. That PMNs may represent an important source of TGF-beta in inflammatory infiltrates was strongly suggested by a demonstration that stored TGF-beta 1 was secreted during phorbol myristate acetate-stimulated degranulation in vitro. Acid/ethanol extracts of human PMNs assayed by ELISA contained an average of 355 ng of TGF/beta 1 per 10(9) cells potentially available for secretion during degranulation of PMNs. [3H]Thymidine incorporation in vivo and autoradiography of tissue sections revealed that widespread cell proliferation was triggered by TGF-beta 1 injection. Synovial lining cells and cells located deep within the subsynovial connective tissue were identified as sources of at least some of the new cells that contribute to TGF-beta 1-induced hyperplasia. Our results demonstrate that TGF-beta is capable of exerting pathogenic effects on synovial tissue and that PMNs may represent a significant source of the TGF-beta present in synovial effusions.

Expression and secretion of transforming growth factor-beta by bleomycin-stimulated rat alveolar macrophages

1993 ◽  
Vol 264 (1) ◽  
pp. L36-L42 ◽  
Author(s):  
E. M. Denholm ◽  
S. M. Rollins
Keyword(s):  
Growth Factor ◽  
Alveolar Macrophages ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Chemotactic Activity ◽  
Beta Activity ◽  
Dependent Manner ◽  
Tgf Beta ◽  
Growth Factor Beta

Bleomycin-induced fibrosis in rodents has been used extensively as a model of human pulmonary fibrosis. The influx of monocytes observed during the early stages of fibrosis is at least partially regulated by the elaboration of chemotactic factors in the lung. Exposure of alveolar macrophages (AM phi) to bleomycin either in vivo or in vitro stimulated secretion of monocyte chemotactic activity (MCA). This MCA has been previously characterized as being primarily due to fibronectin fragments. The present experiments revealed that bleomycin also induced AM phi to secrete a second chemotactic factor, transforming growth factor-beta (TGF-beta). However, the TGF-beta secreted by macrophages was in latent form, since no TGF-beta activity was detected unless AM phi conditioned medium (CM) was acid-activated. After acidification, chemotactic activity in CM from AM phi stimulated with bleomycin in vitro was increased by 3.6, whereas activity in AM phi CM from fibrotic rats increased by 2 and that of a bleomycin-stimulated AM phi cell line increased by 1.6. This acid-activatable chemotactic activity was inhibited by antibody to TGF-beta. Bleomycin-stimulated AM phi s secreted significantly more TGF-beta than did unstimulated controls. Further, in vitro exposure of AM phi to bleomycin induced TGF-beta mRNA expression in a time- and concentration-dependent manner, with maximal mRNA being detected following a 16-h incubation with 1 microgram/ml bleomycin.

scholarly journals Distribution and modulation of the cellular receptor for transforming growth factor-beta.

1987 ◽  
Vol 105 (2) ◽  
pp. 965-975 ◽  
Author(s):  
L M Wakefield ◽  
D M Smith ◽  
T Masui ◽  
C C Harris ◽  
M B Sporn
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Cell Types ◽  
Tgf Beta ◽  
Cell Type ◽  
Down Regulation ◽  
Growth Factor Beta ◽  
Different Cell Types

Scatchard analyses of the binding of transforming growth factor-beta (TGF-beta) to a wide variety of different cell types in culture revealed the universal presence of high affinity (Kd = 1-60 pM) receptors for TGF-beta on every cell type assayed, indicating a wide potential target range for TGF-beta action. There was a strong (r = +0.85) inverse relationship between the receptor affinity and the number of receptors expressed per cell, such that at low TGF-beta concentrations, essentially all cells bound a similar number of TGF-beta molecules per cell. The binding of TGF-beta to various cell types was not altered by many agents that affect the cellular response to TGF-beta, suggesting that modulation of TGF-beta binding to its receptor may not be a primary control mechanism in TGF-beta action. Similarly, in vitro transformation resulted in only relatively small changes in the cellular binding of TGF-beta, and for those cell types that exhibited ligand-induced down-regulation of the receptor, down-regulation was not extensive. Thus the strong conservation of binding observed between cell types is also seen within a given cell type under a variety of conditions, and receptor expression appears to be essentially constitutive. Finally, the biologically inactive form of TGF-beta, which constitutes greater than 98% of autocrine TGF-beta secreted by all of the twelve different cell types assayed, was shown to be unable to bind to the receptor without prior activation in vitro. It is proposed that this may prevent premature interaction of autocrine ligand and receptor in the Golgi apparatus.

scholarly journals Regulation of Trypanosoma cruzi infections in vitro and in vivo by transforming growth factor beta (TGF-beta).

1991 ◽  
Vol 174 (3) ◽  
pp. 539-545 ◽  
Author(s):  
J S Silva ◽  
D R Twardzik ◽  
S G Reed
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Tgf Beta ◽  
Ifn Gamma ◽  
Human Macrophages ◽  
Growth Factor Beta ◽  
Cruzi Infection

The effects of transforming growth factor beta (TGF-beta) on interferon gamma-mediated killing of the intracellular protozoan parasite Trypanosoma cruzi and on the course of T. cruzi infection in mice were investigated. Spleen cells from mice with acute T. cruzi infections were found to produce elevated levels of biologically active TGF-beta in vitro, and the possibility that TGF-beta may mediate certain aspects of T. cruzi infection was then addressed. When mouse peritoneal macrophages were treated with TGF-beta in vitro, the ability of IFN-gamma to activate intracellular inhibition of the parasite was blocked. This occurred whether cells were treated with TGF-beta either before or after IFN-gamma treatment. TGF-beta treatment also blocked the T. cruzi-inhibiting effects of IGN-gamma on human macrophages. Additionally, treatment of human macrophages with TGF-beta alone led to increased parasite replication in these cells. The effects of TGF-beta on T. cruzi infection in vivo were then investigated. Susceptible C57BL/6 mice developed higher parasitemias and died earlier when treated with TGF-beta during the course of infection. Resistant C57BL/6 x DBA/2 F1 mice treated with TGF-beta also had increased parasitemias, and 50% mortality, compared with no mortality in infected, saline-treated controls. A single dose of TGF-beta, given at the time of infection, was sufficient to significantly decrease resistance to infection in F1 mice and to exacerbate infection in susceptible C57BL/6 mice. Furthermore, a single injection of TGF-beta was sufficient to counter the in vivo protective effects of IFN-gamma. We conclude that TGF-beta, produced during acute T. cruzi infection in mice, is a potent inhibitor of the effects of macrophage activating cytokines in vivo and in vitro and may play a role in regulating infection.

scholarly journals Human natural killer cell expansion is regulated by thrombospondin- mediated activation of transforming growth factor-beta 1 and independent accessory cell-derived contact and soluble factors

Blood ◽  
1996 ◽  
Vol 87 (1) ◽  
pp. 180-189 ◽  
Author(s):  
BA Pierson ◽  
K Gupta ◽  
WS Hu ◽  
JS Miller
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Direct Contact ◽  
Transforming Growth Factor ◽  
Neutralizing Antibody ◽  
Accessory Cell ◽  
Tgf Beta ◽  
Soluble Factors ◽  
Marrow Stroma ◽  
Growth Factor Beta

Natural killer cells (NK) were studied to determine factors important in their expansion. Flourescence-activated cell sorter (FACS) purified CD56+/CD3- NK cells cultured alone for 18 days in rIL-2 containing medium (1,000 U/mL) showed enhanced cytotoxicity but only minimal expansion. NK expansion was increased (12.5 +/- 1.6-fold) by coculturing NK with soluble factors produced by irradiated peripheral blood mononuclear cells (PBMNC) in which the two populations were separated by a microporous membrane. However, maximal NK expansion was always observed when NK were cocultured in direct contact with irradiated PBMNC (49.4 +/- 5.9-fold). To determine if marrow stroma, which supports differentiation of primitive NK progenitors, was a better accessory cell population than irradiated PBMNC, NK were cocultured in direct contact with primary marrow stromal layers. NK expansion with marrow stroma was similar to PBMNC. Fibroblast cell lines (M2–10B4, NRK-49F, NIH-3T3) and human umbilical vein endothelial cells (HUVEC), all homogeneous populations and devoid of monocytes, also exhibited a similar contact-dependent increase in NK expansion. Experiments were designed using fixed M2–10B4 stromal cells to separate the contact-induced proliferative stimuli from soluble factors. NK plated directly on ethanol/acetic acid-fixed M2–10B4, which leaves stromal ligands (cell membrane components and ECM) intact, resulted in increased NK expansion compared with medium alone. We further show that the combination of independent contact and soluble factors is responsible for maximal late NK expansion (days 28 through 40) but paradoxically inhibits early NK expansion (day 7). The proliferation inhibitory effects were verified by 3H-thymidine uptake and could be detected at days 2 through 6 but no longer 14 days after the initiation of the culture. We show that both laminin and thrombospondin inhibit early NK proliferation, whereas only thrombospondin was capable of also stimulating late NK expansion. The effect of thrombospondin on early NK proliferation is related to activation of transforming growth factor-beta 1 (TGF-beta) because anti-TGF-beta neutralizing antibody completely abrogated thrombospondin-mediated inhibition of early NK proliferation. Although inhibitory early in culture, active TGF-beta added only at culture initiation increases late NK expansion similar to thrombospondin. TGF-beta was not present in the thrombospondin preparation but latent TGF-beta in serum, or TGF-beta transcripts identified in IL-2-activated NK could explain paracrine or autocrine mechanisms for the regulation of NK proliferation. Finally, anti-TGF-beta neutralizing antibody only minimally affects stroma-mediated inhibition of early NK proliferation suggesting that aside from thrombospondin/TGF-beta, additional contact factors are important for the regulation of NK proliferation.

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