Muscarinic receptors participation in angiogenic response induced by macrophages from mammary adenocarcinoma-bearing mice

INTRODUCTION: The role of macrophages in tumor progression has generated contradictory evidence. We had previously demonstrated the ability of peritoneal macrophages from LMM3 murine mammary adenocarcinoma-bearing mice (TMps) to increase the angiogenicity of LMM3 tumor cells, mainly through polyamine synthesis. Here we investigate the ability of the parasympathetic nervous system to modulate angiogenesis induced by TMps through the activation of the muscarinic acetylcholine receptor (mAchR). METHODS: Peritoneal macrophages from female BALB/c mice bearing a 7-day LMM3 tumor were inoculated intradermally (3 × 10(5 )cells per site) into syngeneic mice. Before inoculation, TMps were stimulated with the muscarinic agonist carbachol in the absence or presence of different muscarinic antagonists or enzyme inhibitors. Angiogenesis was evaluated by counting vessels per square millimeter of skin. The expression of mAchR, arginase and cyclo-oxygenase (COX) isoforms was analyzed by Western blotting. Arginase and COX activities were evaluated by urea and prostaglandin E(2 )(PGE(2)) production, respectively. RESULTS: TMps, which stimulate neovascularization, express functional mAchR, because carbachol-treated TMps potently increased new blood vessels formation. This response was completely blocked by preincubating TMps with pirenzepine and 4-diphenylacetoxy-N-methylpiperidine (4-DAMP), M(1 )and M(3 )receptor antagonists, and partly by the M(2 )receptor antagonist methoctramine. M(1 )receptor activation by carbachol in TMps triggers neovascularization through arginase products because N(ω)-hydroxy-L-arginine reversed the agonist action. Preincubation of TMps with methoctramine partly prevented carbachol-stimulated urea formation. In addition, COX-derived liberation of PGE(2 )is responsible for the promotion of TMps angiogenic activity by M(3 )receptor. We also detected a higher expression of vascular endothelial growth factor (VEGF) in TMps than in macrophages from normal mice. Carbachol significantly increased VEGF expression in TMps, and this effect was totally reversed by methoctramine and pirenzepine. Arginase and COX inhibitors partly decreased VEGF derived from TMps. CONCLUSION: TMps themselves induce a potent angiogenic response that is augmented by carbachol action. mAchR activation triggers arginine metabolism, PGE(2 )synthesis and VEGF production, promoting neovascularization

Acción del virus de Coxsackie sobre el músculo de embrión de pollo cultivado "in vitro"

Fil: Sacerdote de Lustig, E. Ministerio de Asistencia Social y Salud Pública; Instituto Nacional de Microbiología; ArgentinaFil: Parodi, A. S. Ministerio de Asistencia Social y Salud Pública; Instituto Nacional de Microbiología; ArgentinaEl virus de Coxsackie aislado durante una epidemia local (1955) agregado a un cultivo de músculo de embrión de pollo de 8-15 días, produce parálisis de la contracción automática paralelamente a vacuolización y degeneración de los mioblastos. Histológicamente se observa edema del sarcoplasma, pérdida de la estriación y desplazamiento de los núcleos hacia el centro de las fibras

Acción del virus de Coxsackie sobre el músculo de embrión de pollo cultivado "in vitro"

Fil: Sacerdote de Lustig, E. Ministerio de Asistencia Social y Salud Pública; Instituto Nacional de Microbiología; ArgentinaFil: Parodi, A. S. Ministerio de Asistencia Social y Salud Pública; Instituto Nacional de Microbiología; ArgentinaEl virus de Coxsackie aislado durante una epidemia local (1955) agregado a un cultivo de músculo de embrión de pollo de 8-15 días, produce parálisis de la contracción automática paralelamente a vacuolización y degeneración de los mioblastos. Histológicamente se observa edema del sarcoplasma, pérdida de la estriación y desplazamiento de los núcleos hacia el centro de las fibras

Acción oncolítica del virus "A" de influenza sobre los tumores de ratón : I. Modificaciones macroscópicas y sobrevida del animal

Fil: Parodi, A. S. Instituto Bacteriológico. Departamento Nacional de Higiene; Argentin

Acción oncolítica del virus "A" de influenza sobre los tumores de ratón : I. Modificaciones macroscópicas y sobrevida del animal

Fil: Parodi, A. S. Instituto Bacteriológico. Departamento Nacional de Higiene; Argentin

Hydrogen peroxide is involved in lymphocyte activation mechanisms to induce angiogenesis

T-lymphocytes from tumour-bearing mice are able to trigger the angiogenic cascade. Since it is known that tumour growth produces reactive oxygen species (ROS), the aim of this study was to evaluate the role of hydrogen peroxide (H202) on the activation of lymphocytes and their induction of this vascular response. Studies on lymphocytes, stimulated in vitro by ROS to induce angiogen- esis, showed that only the enzyme catalase (CAT) could block the activation. The incubation of nor- mal lymphocytes with H202 stimulated these cells to induce angiogenesis. The administration of H202 or an oxidative stress-producing drug (doxorubicin) to normal mice activated in viva angio- genesis. In tumour-bearing mice, high levels of lipid peroxidation products were observed in the spleen, but not in the liver or kidney. Moreover, when the ROS scavenger enzyme activities (super- oxide dismutase (SDM) and CAT) were determined, we observed low CAT activity in normal spleens, reflected in a high SDM/CAT ratio, when compared to liver or kidney values. We also showed an increasing value of the SDM/CAT ratio with tumour growth. These results strongty suggest that H202 could be involved in the mechanisms of lymphocyte activation and their induction of angiogenesis during tumour growth.Fil: Monte, M.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Davel, L. E.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Sacerdote de Lustig, Eugenia. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaUnidad documental simpl

Inhibition of lymphocyte-induced angiogenesis by free radical scavengers

Solid tumors induce an angiogenic response by the host blood vessels to form a new vascular network for the supply of fresh nutrients and oxygen responsible for tumor growth. Furthermore, tumor growth and metastatic spread is abrogated or markedly reduced in the absence of neovascularization. Spleen Y lymphocytes from tumor-bearing mice elicit a strong neovascular response. It is well known that certain T cell responses require the presence of active oxygen radicals. Because these metabolites are produced during tumor growth, we studied whether oxygen free radicals play a role in the angionesis induction by lymphocytes. In this study, we demonstrated that the administration of a free radical scavenger (EGb-761) to tumor-bearing mice, blocked the angiogenic response and decrease the lung metastatic incidence. On the other hand, when normal lymphocytes were incubated with the xanthine-xanthine oxidase system (X-XO), a known superoxide anion generator, this elicited a dose-response positive angiogenic reaction in normal recipient mice. No angiogenic response was observed in the absence of X-XO, or when EGb-761 or superoxide dismutase (SOD) plus catalase (CAT) were added to the incubation medium. These results suggest that free radicals are involved in some step of the angiogenic process, and that the EGb-761 treatments block this response due to the free radical scavenging activity of this compound.Fil: Monte, Martin. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Davel, Lilia E.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Sacerdote de Lustig, Eugenia. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaUnidad documental simpl

Evidence that indomethacin inhibits lymphocyte-induced angiogenesis

Fil: Davel, Lilia E.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Miguez, Marta M.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Sacerdote de Lustig, Eugenia. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaUnidad documental simpl

Decrease of N-CAM adhesion molecule in serum of advanced Alzheimer's patients

Fil: Sacerdote de Lustig, Eugenia. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Kohan, S.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Todaro, Laura Beatriz. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Famulari, A.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Dominguez, R.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Farias, E.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaUnidad documental simpl

Differential nitric oxide release and sensitivity to injury in different murine mammary tumor cell lines

The purpose of this study was to determine whether nitric oxide (NO) production by different mammary tumor cell lines correlated with their sensitivity to NO mediated injury. Three mammary tumor cell lines LM2, LM3 and LMM3 syngeneic to BALB/c mice were cultured in vitro with IFNgamma + LPS. Different levels of NO production among the three lines were detected in culture supernatants. The only tumor cell line which did not produce NO (LM2) showed the highest sensitivity to SNP-derived NO cytotoxicity (87%), while LM3 and LMM3 which both produced higher levels of NO than LM2, showed lower cytotoxicity by SNP (39% and 22% respectively). Spleen cells (SC) from M2 tumor bearing mice (TBM) were able to lyse LM2 cells by NO-dependent mechanisms. SC from M3-TBM exerted cytotoxicity against LM3 cells mainly by NO-independent mechanisms. Thus, we postulate an inverse correlation between NO production and NO mediated cytotoxicity in the three mammary tumor cell lines. It is possible that tumor cells producing NO develop mechanisms to resist NO injury.Fil: Eijan, Ana Maria. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Davel, L. E.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Rueda, H. A.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Rozenberg, G.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaFil: Sacerdote de Lustig, Eugenia. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Jasnis, Maria Adela. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; ArgentinaUnidad documental simpl