Marian Nakada - Academia.edu (original) (raw)
Papers by Marian Nakada
Because the b3-antagonist abciximab (c7E3 Fab) has significantly improved late outcomes after cor... more Because the b3-antagonist abciximab (c7E3 Fab) has significantly improved late outcomes after coronary angioplasty, the b3 integrins have been implicated in the arterial response to injury. However, the mechanisms underlying this benefit are unknown. The observation that c7E3 binds b3 integrins on vascular cells (avb3) with affinity equal to that for the platelet glycoprotein IIb/IIIa integrin has led to the
Cancer Research, May 15, 2002
GPIIb/IIIa and ␣v3. We used chimeric (c) 7E3 Fab (ReoPro) and murine (m) 7E3 F(ab) 2 to elucidat... more GPIIb/IIIa and ␣v3. We used chimeric (c) 7E3 Fab (ReoPro) and murine (m) 7E3 F(ab) 2 to elucidate the role of these integrins in angiogenesis, tumor growth, and metastasis. These antibodies are potent inhibitors of GPIIb/IIIa and ␣v3. c7E3 Fab inhibited ␣v3-mediated human umbilical vein endothelial (HUVEC) and melanoma cell adhesion, migration, invasion, and basic fibroblast growth factor stimulated proliferation of HUVECs (IC 50 values range from 0.15 to 5 g/ml for different assays). In an in vitro angiogenesis assay, c7E3 Fab inhibited basic fibroblast growth factor and platelet-stimulated capillary formation of HUVECs (IC 50 ؍ 10 g/ml and 15 g/ml, respectively), demonstrating that endothelial ␣v3 is important for sprouting, and platelet-stimulated sprouting is mediated by GPIIb/IIIa. In an experimental metastasis assay, a single pretreatment of human melanoma cells with c7E3 Fab (2.5 g/ml) inhibited lung colonization of the tumor cells in severe combined immunodeficient mice. In vivo, m7E3 F(ab) 2 partially inhibited growth of human melanoma tumors in nude mice compared with control-treated animals. These data suggest that tumor cell-expressed integrins are important but not the only component involved in tumor growth. Because c7E3 Fab and m7E3 F(ab) 2 do not cross-react with murine integrins, this inhibition of metastasis and tumor growth is attributable to direct blockade of human tumor ␣v3 integrins. m7E3 F(ab) 2 completely blocked tumor formation and growth of human melanoma tumors growing in nude rats. In this xenograft model, m7E3 F(ab) 2 simultaneously binds to both human tumor and host platelet GPIIb/IIIa and endothelial ␣v3 integrins, thus participating as an antiangiogenic and an antitumor agent. Collectively, these results indicate that combined blockade of GPIIb/IIIa and ␣v3 affords significant antiangiogenic and antitumor benefit.
Upregulation of adhesion proteins plays an important role in mediating inflammation. The inductio... more Upregulation of adhesion proteins plays an important role in mediating inflammation. The induction of adhesive molecules has been well studied, but the reversibility of their expression has not been well characterized. A neutralizing anti-TNF monoclonal antibody (cA2) was used to study the down regulation of TNF-induced E-selectin, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) on cultured human umbilical vein endothelial cells (HUVECs). Addition of cA2 following TNF stimulation of HUVECs enhanced the rate of E-selectin and VCAM-1 down-regulation from the cell surface and also reduced steady state E-selectin and VCAM-1 mRNA levels. The cA2-mediated disappearance of E-selectin, but not VCAM-1 protein was microtubule and not microfilament dependent. Neutralization of TNF only slightly reduced ICAM-1 cell surface levels following initial TNF stimulation, suggesting a slower turnover of ICAM-1 compared to E-selectin and VCAM-1. Microtubule inhibition during TNF stimulation partially inhibited E-selectin, VCAM-1 and ICAM-1 mRNA upregulation. VCAM-1 and ICAM-1 cell surface expression were similarly partially inhibited, however, E-selectin levels were unaffected, presumably due to the dual, opposing effect of inhibiting protein expression and inhibiting internalization. Microfilament inhibition during protein induction specifically inhibited the maximal expression of VCAM-1 protein and mRNA, without affecting E-selectin or ICAM-1. These data support the notion that E-selectin, VCAM-1, and ICAM-1 expression are differentially regulated on HUVECs and suggest that TNF neutralizing therapies may be effective because of their ability to reduce the levels of pre-existing adhesion proteins.
Biochem Biophys Res Commun, 2000
Contortrostatin is a homodimeric disintegrin from snake venom. We have shown that contortrostatin... more Contortrostatin is a homodimeric disintegrin from snake venom. We have shown that contortrostatin binds to integrins ␣IIb3, ␣51, and ␣v3. We now use several criteria to demonstrate the binding of contortrostatin to ␣v5. First, incubation of T24 cells, which express ␣v3 and ␣v5, with antibody against ␣v3 failed to completely inhibit adhesion of cells to vitronectin. However, pretreatment of the cells with contortrostatin or the combination of antibodies against ␣v3 and ␣v5 completely blocked adhesion to vitronectin. By contrast, either anti-␣v5 alone or contortrostatin blocked adhesion of an ␣v3-negative T24 subline. Second, contortrostatin as well as anti-␣v5 inhibits invasion of OVCAR-5, which express only ␣v5. Third, contortrostatin binds to purified ␣v5 in a saturable manner. Finally, radioligand binding assays yielded a K d value of 24 nM for [ 125 I]contortrostatin binding to ␣v5. This investigation identifies ␣v5 as a binding site for contortrostatin. Blockage of ␣v5 by contortrostatin inhibits ␣v5-mediated adhesion and invasion.
Neonatal exposure to Gross murine leukemia virus results in a profound inhibition of the virusspe... more Neonatal exposure to Gross murine leukemia virus results in a profound inhibition of the virusspecific T and B cell responses of adult animals. Animals exposed to virus as neonates exhibit a marked depression in virus-specific T cell function as measured by the virtual absence of in vivo delayed type hypersensitivity responses and in vitro proliferative responses to virally infected stimulator cells. Further, serum obtained from neonatally treated mice failed to either immunoprecipitate viral proteins or neutralize virus in an in vitro plaque assay, suggesting the concurrent induction of a state of B cell hyporesponsiveness . The specificity of this effect at the levels of both T and B cells was demonstrated by the ability ofneonatally treated mice to respond normally after adult challenge with either irrelevant reovirus or influenza virus. The replication of Gross virus within both stromal and lymphocytic compartments of the neonatal thymus suggests that thymic education plays a key role in the induction ofimmunologic nonresponsiveness to viruses. mmunologic nonresponsiveness to autologous molecules and a variety of exogenous antigens can be linked to their presence during neonatal maturation (1-4). The consequences of neonatal exposure to viruses are considerably more variable (5, 6). Mice infected either neonatally or congenitally with lymphocytic choriomeningitis virus (LCMV)' or endogenous ecotropic retroviruses mount potent Ig responses and produce functional CTL precursors (7-9) . However, several recent studies have shown that exposure of neonatal mice to Moloney murine leukemia virus significantly reduced the frequency ofMoloney virus-specific CTL precursors and Ig levels (6, 10) . These contrasting results raise important questions concerning the ability of different viruses to induce tolerance, and the contributions of T and B cell components to virus-specific tolerance . To address these questions, the immune response ofmice exposed as neonates to NB-tropic Gross murine leukemia virus (GMuLV) was investigated . Exposure of neonates to GMuLV induces a state ofimmunologic nonresponsiveness that may be linked to intrathymic replication of this retrovirus .
Angiogenesis, 1999
Angiogenesis plays an essential role in the growth and dissemination of solid tumor cancers. The ... more Angiogenesis plays an essential role in the growth and dissemination of solid tumor cancers. The expression of endothelial cell integrin alpha(v)beta3 has been shown to increase during vascular proliferation associated with human tumors. Selective antagonists of alpha(v)beta3 can block angiogenesis and tumor growth by inducing programmed cell death in proliferating endothelial cells. Monoclonal antibody 7E3, an antagonist of the human, but not murine, integrins alpha(v)beta3 and alphaIIbbeta3 (GPIIb/IIIa), inhibits platelet aggregation. It is the parent antibody of a mouse/human chimeric antibody fragment approved for adjunctive therapy of patients undergoing percutaneous coronary interventions to prevent ischemic complications (c7E3Fab; abciximab; ReoPro). To evaluate the potential of 7E3 to inhibit human angiogenesis and tumor growth independent of its antiplatelet effects, we established integrin alpha(v)beta3-negative human melanoma tumors in full-thickness human skin grafted on...
Angiogenesis, 1999
Contortrostatin, a 13.5 kDa disulfide-linked homodimeric polypeptide possessing an Arg-Gly-Asp se... more Contortrostatin, a 13.5 kDa disulfide-linked homodimeric polypeptide possessing an Arg-Gly-Asp sequence, was isolated from venom of the southern copperhead snake. Daily injection of contortrostatin into the primary tumor of human breast cancer MDA-MB-435 carried in nude mice significantly inhibited tumor growth and neovascularization of the tumor tissue. On the chick embryo chorioallantoic membrane, contortrostatin inhibited angiogenesis induced by MDA-MB-435 cells, basic fibroblast growth factor, and vascular endothelial growth factor. In addition, contortrostatin effectively blocked adhesion of human umbilical vein endothelial cells (HUVEC) to immobilized vitronectin and significantly inhibited invasion of HUVEC through a Matrigel barrier. Competitive binding assays and adhesion assays with different integrin antibodies suggested that integrin alpha(v)beta3 is a binding site for contortrostatin on vascular endothelial cells. Detachment of HUVEC from vitronectin by contortrostatin ...
Cancer research, Jan 15, 2002
In vivo tumor cells interact with a variety of host cells such as endothelial cells and platelets... more In vivo tumor cells interact with a variety of host cells such as endothelial cells and platelets, and these interactions are mediated by integrins GPIIb/IIIa and alphavbeta3. We used chimeric (c) 7E3 Fab (ReoPro) and murine (m) 7E3 F(ab')(2) to elucidate the role of these integrins in angiogenesis, tumor growth, and metastasis. These antibodies are potent inhibitors of GPIIb/IIIa and alphavbeta3. c7E3 Fab inhibited alphavbeta3-mediated human umbilical vein endothelial (HUVEC) and melanoma cell adhesion, migration, invasion, and basic fibroblast growth factor stimulated proliferation of HUVECs (IC(50) values range from 0.15 to 5 microg/ml for different assays). In an in vitro angiogenesis assay, c7E3 Fab inhibited basic fibroblast growth factor and platelet-stimulated capillary formation of HUVECs (IC(50) = 10 microg/ml and 15 microg/ml, respectively), demonstrating that endothelial alphavbeta3 is important for sprouting, and platelet-stimulated sprouting is mediated by GPIIb/II...
The therapeutic options for anti-neutrophil cytoplasm antibody (ANCA)-associated systemic vasculi... more The therapeutic options for anti-neutrophil cytoplasm antibody (ANCA)-associated systemic vasculitis (AASV) remain limited and hampered by adverse effects. One potential novel therapeutic avenue involves inhibition of TNF-, with encouraging uncontrolled data in humans with one agent (infliximab) but disappointing controlled data from another (etanercept). For investigating the potential role of TNF- as a therapeutic target in AASV, the effect of
Drug Discovery Series, 2007
Clinical & experimental metastasis, 2008
CNTO 95 is a fully human monoclonal antibody that recognizes alphav integrins. Previous studies h... more CNTO 95 is a fully human monoclonal antibody that recognizes alphav integrins. Previous studies have shown that CNTO 95 exhibits both anti-tumor and anti-angiogenic activities (Trikha M et al., Int J Cancer 110:326-335, 2004). In this study we investigated the biological activities of CNTO 95 on breast tumor cells both in vitro and in vivo. In vitro treatment with CNTO 95 decreased the viability of breast tumor cells adhering to vitronectin. CNTO 95 inhibited tumor cell adhesion, migration, and invasion in vitro. CNTO 95 treatment also induced tyrosine dephosphorylation of focal adhesion kinase (FAK), and the docking protein paxillin that recruits both structural and signaling molecules to focal adhesions (Turner CE, Int J Biochem Cell Biol 30:955-959, 1998; O'Neil GM et al., Trends Cell Biol 10:111-119, 2000). These results suggest that CNTO 95 inhibits breast tumor cell growth, migration and invasion by interruption of alphav integrin mediated focal adhesions and cell motility...
Platelet endothelial cell adhesion molecule (PECAM-1), a member of the Ig superfamily, is found o... more Platelet endothelial cell adhesion molecule (PECAM-1), a member of the Ig superfamily, is found on endothelial cells and neu- trophils and has been shown to be involved in the migration of leukocytes across the endothelium. Adhesion is mediated, at least in part, through binding interactions involving its first N-terminal Ig-like domain, but it is still unclear which sequences in this
Journal of the American Society of Nephrology, 2006
The therapeutic options for anti-neutrophil cytoplasm antibody (ANCA)-associated systemic vasculi... more The therapeutic options for anti-neutrophil cytoplasm antibody (ANCA)-associated systemic vasculitis (AASV) remain limited and hampered by adverse effects. One potential novel therapeutic avenue involves inhibition of TNF-␣, with encouraging uncontrolled data in humans with one agent (infliximab) but disappointing controlled data from another (etanercept). For investigating the potential role of TNF-␣ as a therapeutic target in AASV, the effect of an anti-rat TNF-␣ mAb (CNTO 1081) in a rat model of AASV was investigated. For testing the effect of TNF-␣ blockade in this model, starting on day 28 after immunization (a point when glomerulonephritis is established), animals were randomized to treatment with CNTO 1081 or control mouse IgG. Treatment with CNTO 1081 significantly reduced albuminuria (mean 1.1 ؎ 0.3 mg/24 h CNTO 1081 versus 8.0 ؎ 1.9 controls; P < 0.05) and crescent formation (0% CNTO 1081 versus 60% controls; P < 0.05). Lung hemorrhage was also reduced (CNTO 1081: median score 0, range 0 to 2; controls: 2, range 1 to 3; P < 0.05). When analyzed by intravital microscopy, there was a 43% inhibition of leukocyte transmigration in mesenteric venules in response to topical CXCL1 (a neutrophil chemoattractant) in the CNTO 1081 group compared with controls (P < 0.001). Anti-myeloperoxidase antibody titers were similar in both groups throughout the study. In conclusion, these findings indicate that TNF-␣ plays an important role in the pathogenesis of experimental autoimmune vasculitis and suggest that blockade of this cytokine with an mAb is effective in treating established vasculitis. The therapeutic action of anti-TNF-␣ reagents may be mediated, in part, by suppression of the enhanced leukocyte-endothelial interactions in this disorder.
In vivo tumor cells interact with a variety of host cells such as endo- thelial cells and platele... more In vivo tumor cells interact with a variety of host cells such as endo- thelial cells and platelets, and these interactions are mediated by integrins GPIIb/IIIa and v3. We used chimeric (c) 7E3 Fab (ReoPro) and murine (m) 7E3 F(ab)2 to elucidate the role of these integrins in angiogenesis, tumor growth, and metastasis. These antibodies are potent inhibitors of GPIIb/IIIa
Matrix metalloproteinases (MMPs) are endopeptidases that play pivotal roles in promoting tumor di... more Matrix metalloproteinases (MMPs) are endopeptidases that play pivotal roles in promoting tumor disease progression, including tumor angiogenesis. In many solid tumors, MMP expression could be attributed to tumor stromal cells and is partially regulated by tumor-stroma interactions via tumor cell-associated extracellular matrix metalloproteinase in- ducer (EMMPRIN). The role of EMMPRIN during tumor angiogenesis and growth was explored by modulating EMMPRIN
The FASEB Journal, 2011
Elevated generation of reactive oxygen species (ROS) by endothelial enzymes, including NADPH-oxid... more Elevated generation of reactive oxygen species (ROS) by endothelial enzymes, including NADPH-oxidase, is implicated in vascular oxidative stress and endothelial proinflammatory activation involving exposure of vascular cell adhesion molecule-1 (VCAM-1). Catalase and superoxide dismutase (SOD) conjugated with antibodies to platelet/endothelial cell adhesion molecule 1 (PECAM-1) bind specifically to endothelium and inhibit effects of corresponding ROS, H(2)O(2), and superoxide anion. In this study, anti-PECAM/SOD, but not anti-PECAM/catalase or nontargeted enzymes, including polyethylene glycol (PEG)-SOD, inhibited 2- to 3-fold VCAM expression caused by tumor necrosis factor (TNF), interleukin-1β, and lipopolysaccharide. Anti- PECAM/SOD, but not nontargeted counterparts, accumulated in vascular endothelium after intravenous injection, localized in endothelial endosomes, and inhibited by 70% lipopolysaccharide-caused VCAM-1 expression in mice. Anti-PECAM/SOD colocalized with EEA-1-positive endothelial vesicles and quenched ROS produced in response to TNF. Inhibitors of NADPH oxidase and anion channel ClC3 blocked TNF-induced VCAM expression, affirming that superoxide produced and transported by these proteins, respectively, mediates inflammatory signaling. Anti-PECAM/SOD abolished VCAM expression caused by poly(I:C)-induced activation of toll-like receptor 3 localized in intracellular vesicles. These results directly implicate endosomal influx of superoxide in endothelial inflammatory response and suggest that site-specific interception of this signal attained by targeted delivery of anti-PECAM/SOD into endothelial endosomes may have anti-inflammatory effects.
Seminars in Thrombosis and Hemostasis, 2002
Thromboembolism is one of the most common causes of death in cancer patients. Among the most freq... more Thromboembolism is one of the most common causes of death in cancer patients. Among the most frequent thrombotic complications in patients with cancer are disseminated intravascular coagulation, thrombotic thrombocytopenic purpura, and thrombocytosis. Clearly, these complications arise as tumor cells interact with almost all components of the hemostatic system including platelets. Platelets participate in tumor progression by contributing to the metastatic cascade, protecting tumor cells from immune surveillance, regulating tumor cell invasion, and angiogenesis. Platelets contain one of the largest stores of angiogenic and mitogenic factors and the tumor vasculature is leaky, which allows platelets to come in contact with the tumor and deposit multiple angiogenic factors including vascular endothelial growth factor (VEGF) and thrombin to tumor cells, which in turn contributes to tumor progression. This article reviews the recent literature on how platelets contribute to tumor growth, angiogenesis, and metastasis.
Because the b3-antagonist abciximab (c7E3 Fab) has significantly improved late outcomes after cor... more Because the b3-antagonist abciximab (c7E3 Fab) has significantly improved late outcomes after coronary angioplasty, the b3 integrins have been implicated in the arterial response to injury. However, the mechanisms underlying this benefit are unknown. The observation that c7E3 binds b3 integrins on vascular cells (avb3) with affinity equal to that for the platelet glycoprotein IIb/IIIa integrin has led to the
Cancer Research, May 15, 2002
GPIIb/IIIa and ␣v3. We used chimeric (c) 7E3 Fab (ReoPro) and murine (m) 7E3 F(ab) 2 to elucidat... more GPIIb/IIIa and ␣v3. We used chimeric (c) 7E3 Fab (ReoPro) and murine (m) 7E3 F(ab) 2 to elucidate the role of these integrins in angiogenesis, tumor growth, and metastasis. These antibodies are potent inhibitors of GPIIb/IIIa and ␣v3. c7E3 Fab inhibited ␣v3-mediated human umbilical vein endothelial (HUVEC) and melanoma cell adhesion, migration, invasion, and basic fibroblast growth factor stimulated proliferation of HUVECs (IC 50 values range from 0.15 to 5 g/ml for different assays). In an in vitro angiogenesis assay, c7E3 Fab inhibited basic fibroblast growth factor and platelet-stimulated capillary formation of HUVECs (IC 50 ؍ 10 g/ml and 15 g/ml, respectively), demonstrating that endothelial ␣v3 is important for sprouting, and platelet-stimulated sprouting is mediated by GPIIb/IIIa. In an experimental metastasis assay, a single pretreatment of human melanoma cells with c7E3 Fab (2.5 g/ml) inhibited lung colonization of the tumor cells in severe combined immunodeficient mice. In vivo, m7E3 F(ab) 2 partially inhibited growth of human melanoma tumors in nude mice compared with control-treated animals. These data suggest that tumor cell-expressed integrins are important but not the only component involved in tumor growth. Because c7E3 Fab and m7E3 F(ab) 2 do not cross-react with murine integrins, this inhibition of metastasis and tumor growth is attributable to direct blockade of human tumor ␣v3 integrins. m7E3 F(ab) 2 completely blocked tumor formation and growth of human melanoma tumors growing in nude rats. In this xenograft model, m7E3 F(ab) 2 simultaneously binds to both human tumor and host platelet GPIIb/IIIa and endothelial ␣v3 integrins, thus participating as an antiangiogenic and an antitumor agent. Collectively, these results indicate that combined blockade of GPIIb/IIIa and ␣v3 affords significant antiangiogenic and antitumor benefit.
Upregulation of adhesion proteins plays an important role in mediating inflammation. The inductio... more Upregulation of adhesion proteins plays an important role in mediating inflammation. The induction of adhesive molecules has been well studied, but the reversibility of their expression has not been well characterized. A neutralizing anti-TNF monoclonal antibody (cA2) was used to study the down regulation of TNF-induced E-selectin, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) on cultured human umbilical vein endothelial cells (HUVECs). Addition of cA2 following TNF stimulation of HUVECs enhanced the rate of E-selectin and VCAM-1 down-regulation from the cell surface and also reduced steady state E-selectin and VCAM-1 mRNA levels. The cA2-mediated disappearance of E-selectin, but not VCAM-1 protein was microtubule and not microfilament dependent. Neutralization of TNF only slightly reduced ICAM-1 cell surface levels following initial TNF stimulation, suggesting a slower turnover of ICAM-1 compared to E-selectin and VCAM-1. Microtubule inhibition during TNF stimulation partially inhibited E-selectin, VCAM-1 and ICAM-1 mRNA upregulation. VCAM-1 and ICAM-1 cell surface expression were similarly partially inhibited, however, E-selectin levels were unaffected, presumably due to the dual, opposing effect of inhibiting protein expression and inhibiting internalization. Microfilament inhibition during protein induction specifically inhibited the maximal expression of VCAM-1 protein and mRNA, without affecting E-selectin or ICAM-1. These data support the notion that E-selectin, VCAM-1, and ICAM-1 expression are differentially regulated on HUVECs and suggest that TNF neutralizing therapies may be effective because of their ability to reduce the levels of pre-existing adhesion proteins.
Biochem Biophys Res Commun, 2000
Contortrostatin is a homodimeric disintegrin from snake venom. We have shown that contortrostatin... more Contortrostatin is a homodimeric disintegrin from snake venom. We have shown that contortrostatin binds to integrins ␣IIb3, ␣51, and ␣v3. We now use several criteria to demonstrate the binding of contortrostatin to ␣v5. First, incubation of T24 cells, which express ␣v3 and ␣v5, with antibody against ␣v3 failed to completely inhibit adhesion of cells to vitronectin. However, pretreatment of the cells with contortrostatin or the combination of antibodies against ␣v3 and ␣v5 completely blocked adhesion to vitronectin. By contrast, either anti-␣v5 alone or contortrostatin blocked adhesion of an ␣v3-negative T24 subline. Second, contortrostatin as well as anti-␣v5 inhibits invasion of OVCAR-5, which express only ␣v5. Third, contortrostatin binds to purified ␣v5 in a saturable manner. Finally, radioligand binding assays yielded a K d value of 24 nM for [ 125 I]contortrostatin binding to ␣v5. This investigation identifies ␣v5 as a binding site for contortrostatin. Blockage of ␣v5 by contortrostatin inhibits ␣v5-mediated adhesion and invasion.
Neonatal exposure to Gross murine leukemia virus results in a profound inhibition of the virusspe... more Neonatal exposure to Gross murine leukemia virus results in a profound inhibition of the virusspecific T and B cell responses of adult animals. Animals exposed to virus as neonates exhibit a marked depression in virus-specific T cell function as measured by the virtual absence of in vivo delayed type hypersensitivity responses and in vitro proliferative responses to virally infected stimulator cells. Further, serum obtained from neonatally treated mice failed to either immunoprecipitate viral proteins or neutralize virus in an in vitro plaque assay, suggesting the concurrent induction of a state of B cell hyporesponsiveness . The specificity of this effect at the levels of both T and B cells was demonstrated by the ability ofneonatally treated mice to respond normally after adult challenge with either irrelevant reovirus or influenza virus. The replication of Gross virus within both stromal and lymphocytic compartments of the neonatal thymus suggests that thymic education plays a key role in the induction ofimmunologic nonresponsiveness to viruses. mmunologic nonresponsiveness to autologous molecules and a variety of exogenous antigens can be linked to their presence during neonatal maturation (1-4). The consequences of neonatal exposure to viruses are considerably more variable (5, 6). Mice infected either neonatally or congenitally with lymphocytic choriomeningitis virus (LCMV)' or endogenous ecotropic retroviruses mount potent Ig responses and produce functional CTL precursors (7-9) . However, several recent studies have shown that exposure of neonatal mice to Moloney murine leukemia virus significantly reduced the frequency ofMoloney virus-specific CTL precursors and Ig levels (6, 10) . These contrasting results raise important questions concerning the ability of different viruses to induce tolerance, and the contributions of T and B cell components to virus-specific tolerance . To address these questions, the immune response ofmice exposed as neonates to NB-tropic Gross murine leukemia virus (GMuLV) was investigated . Exposure of neonates to GMuLV induces a state ofimmunologic nonresponsiveness that may be linked to intrathymic replication of this retrovirus .
Angiogenesis, 1999
Angiogenesis plays an essential role in the growth and dissemination of solid tumor cancers. The ... more Angiogenesis plays an essential role in the growth and dissemination of solid tumor cancers. The expression of endothelial cell integrin alpha(v)beta3 has been shown to increase during vascular proliferation associated with human tumors. Selective antagonists of alpha(v)beta3 can block angiogenesis and tumor growth by inducing programmed cell death in proliferating endothelial cells. Monoclonal antibody 7E3, an antagonist of the human, but not murine, integrins alpha(v)beta3 and alphaIIbbeta3 (GPIIb/IIIa), inhibits platelet aggregation. It is the parent antibody of a mouse/human chimeric antibody fragment approved for adjunctive therapy of patients undergoing percutaneous coronary interventions to prevent ischemic complications (c7E3Fab; abciximab; ReoPro). To evaluate the potential of 7E3 to inhibit human angiogenesis and tumor growth independent of its antiplatelet effects, we established integrin alpha(v)beta3-negative human melanoma tumors in full-thickness human skin grafted on...
Angiogenesis, 1999
Contortrostatin, a 13.5 kDa disulfide-linked homodimeric polypeptide possessing an Arg-Gly-Asp se... more Contortrostatin, a 13.5 kDa disulfide-linked homodimeric polypeptide possessing an Arg-Gly-Asp sequence, was isolated from venom of the southern copperhead snake. Daily injection of contortrostatin into the primary tumor of human breast cancer MDA-MB-435 carried in nude mice significantly inhibited tumor growth and neovascularization of the tumor tissue. On the chick embryo chorioallantoic membrane, contortrostatin inhibited angiogenesis induced by MDA-MB-435 cells, basic fibroblast growth factor, and vascular endothelial growth factor. In addition, contortrostatin effectively blocked adhesion of human umbilical vein endothelial cells (HUVEC) to immobilized vitronectin and significantly inhibited invasion of HUVEC through a Matrigel barrier. Competitive binding assays and adhesion assays with different integrin antibodies suggested that integrin alpha(v)beta3 is a binding site for contortrostatin on vascular endothelial cells. Detachment of HUVEC from vitronectin by contortrostatin ...
Cancer research, Jan 15, 2002
In vivo tumor cells interact with a variety of host cells such as endothelial cells and platelets... more In vivo tumor cells interact with a variety of host cells such as endothelial cells and platelets, and these interactions are mediated by integrins GPIIb/IIIa and alphavbeta3. We used chimeric (c) 7E3 Fab (ReoPro) and murine (m) 7E3 F(ab')(2) to elucidate the role of these integrins in angiogenesis, tumor growth, and metastasis. These antibodies are potent inhibitors of GPIIb/IIIa and alphavbeta3. c7E3 Fab inhibited alphavbeta3-mediated human umbilical vein endothelial (HUVEC) and melanoma cell adhesion, migration, invasion, and basic fibroblast growth factor stimulated proliferation of HUVECs (IC(50) values range from 0.15 to 5 microg/ml for different assays). In an in vitro angiogenesis assay, c7E3 Fab inhibited basic fibroblast growth factor and platelet-stimulated capillary formation of HUVECs (IC(50) = 10 microg/ml and 15 microg/ml, respectively), demonstrating that endothelial alphavbeta3 is important for sprouting, and platelet-stimulated sprouting is mediated by GPIIb/II...
The therapeutic options for anti-neutrophil cytoplasm antibody (ANCA)-associated systemic vasculi... more The therapeutic options for anti-neutrophil cytoplasm antibody (ANCA)-associated systemic vasculitis (AASV) remain limited and hampered by adverse effects. One potential novel therapeutic avenue involves inhibition of TNF-, with encouraging uncontrolled data in humans with one agent (infliximab) but disappointing controlled data from another (etanercept). For investigating the potential role of TNF- as a therapeutic target in AASV, the effect of
Drug Discovery Series, 2007
Clinical & experimental metastasis, 2008
CNTO 95 is a fully human monoclonal antibody that recognizes alphav integrins. Previous studies h... more CNTO 95 is a fully human monoclonal antibody that recognizes alphav integrins. Previous studies have shown that CNTO 95 exhibits both anti-tumor and anti-angiogenic activities (Trikha M et al., Int J Cancer 110:326-335, 2004). In this study we investigated the biological activities of CNTO 95 on breast tumor cells both in vitro and in vivo. In vitro treatment with CNTO 95 decreased the viability of breast tumor cells adhering to vitronectin. CNTO 95 inhibited tumor cell adhesion, migration, and invasion in vitro. CNTO 95 treatment also induced tyrosine dephosphorylation of focal adhesion kinase (FAK), and the docking protein paxillin that recruits both structural and signaling molecules to focal adhesions (Turner CE, Int J Biochem Cell Biol 30:955-959, 1998; O'Neil GM et al., Trends Cell Biol 10:111-119, 2000). These results suggest that CNTO 95 inhibits breast tumor cell growth, migration and invasion by interruption of alphav integrin mediated focal adhesions and cell motility...
Platelet endothelial cell adhesion molecule (PECAM-1), a member of the Ig superfamily, is found o... more Platelet endothelial cell adhesion molecule (PECAM-1), a member of the Ig superfamily, is found on endothelial cells and neu- trophils and has been shown to be involved in the migration of leukocytes across the endothelium. Adhesion is mediated, at least in part, through binding interactions involving its first N-terminal Ig-like domain, but it is still unclear which sequences in this
Journal of the American Society of Nephrology, 2006
The therapeutic options for anti-neutrophil cytoplasm antibody (ANCA)-associated systemic vasculi... more The therapeutic options for anti-neutrophil cytoplasm antibody (ANCA)-associated systemic vasculitis (AASV) remain limited and hampered by adverse effects. One potential novel therapeutic avenue involves inhibition of TNF-␣, with encouraging uncontrolled data in humans with one agent (infliximab) but disappointing controlled data from another (etanercept). For investigating the potential role of TNF-␣ as a therapeutic target in AASV, the effect of an anti-rat TNF-␣ mAb (CNTO 1081) in a rat model of AASV was investigated. For testing the effect of TNF-␣ blockade in this model, starting on day 28 after immunization (a point when glomerulonephritis is established), animals were randomized to treatment with CNTO 1081 or control mouse IgG. Treatment with CNTO 1081 significantly reduced albuminuria (mean 1.1 ؎ 0.3 mg/24 h CNTO 1081 versus 8.0 ؎ 1.9 controls; P < 0.05) and crescent formation (0% CNTO 1081 versus 60% controls; P < 0.05). Lung hemorrhage was also reduced (CNTO 1081: median score 0, range 0 to 2; controls: 2, range 1 to 3; P < 0.05). When analyzed by intravital microscopy, there was a 43% inhibition of leukocyte transmigration in mesenteric venules in response to topical CXCL1 (a neutrophil chemoattractant) in the CNTO 1081 group compared with controls (P < 0.001). Anti-myeloperoxidase antibody titers were similar in both groups throughout the study. In conclusion, these findings indicate that TNF-␣ plays an important role in the pathogenesis of experimental autoimmune vasculitis and suggest that blockade of this cytokine with an mAb is effective in treating established vasculitis. The therapeutic action of anti-TNF-␣ reagents may be mediated, in part, by suppression of the enhanced leukocyte-endothelial interactions in this disorder.
In vivo tumor cells interact with a variety of host cells such as endo- thelial cells and platele... more In vivo tumor cells interact with a variety of host cells such as endo- thelial cells and platelets, and these interactions are mediated by integrins GPIIb/IIIa and v3. We used chimeric (c) 7E3 Fab (ReoPro) and murine (m) 7E3 F(ab)2 to elucidate the role of these integrins in angiogenesis, tumor growth, and metastasis. These antibodies are potent inhibitors of GPIIb/IIIa
Matrix metalloproteinases (MMPs) are endopeptidases that play pivotal roles in promoting tumor di... more Matrix metalloproteinases (MMPs) are endopeptidases that play pivotal roles in promoting tumor disease progression, including tumor angiogenesis. In many solid tumors, MMP expression could be attributed to tumor stromal cells and is partially regulated by tumor-stroma interactions via tumor cell-associated extracellular matrix metalloproteinase in- ducer (EMMPRIN). The role of EMMPRIN during tumor angiogenesis and growth was explored by modulating EMMPRIN
The FASEB Journal, 2011
Elevated generation of reactive oxygen species (ROS) by endothelial enzymes, including NADPH-oxid... more Elevated generation of reactive oxygen species (ROS) by endothelial enzymes, including NADPH-oxidase, is implicated in vascular oxidative stress and endothelial proinflammatory activation involving exposure of vascular cell adhesion molecule-1 (VCAM-1). Catalase and superoxide dismutase (SOD) conjugated with antibodies to platelet/endothelial cell adhesion molecule 1 (PECAM-1) bind specifically to endothelium and inhibit effects of corresponding ROS, H(2)O(2), and superoxide anion. In this study, anti-PECAM/SOD, but not anti-PECAM/catalase or nontargeted enzymes, including polyethylene glycol (PEG)-SOD, inhibited 2- to 3-fold VCAM expression caused by tumor necrosis factor (TNF), interleukin-1β, and lipopolysaccharide. Anti- PECAM/SOD, but not nontargeted counterparts, accumulated in vascular endothelium after intravenous injection, localized in endothelial endosomes, and inhibited by 70% lipopolysaccharide-caused VCAM-1 expression in mice. Anti-PECAM/SOD colocalized with EEA-1-positive endothelial vesicles and quenched ROS produced in response to TNF. Inhibitors of NADPH oxidase and anion channel ClC3 blocked TNF-induced VCAM expression, affirming that superoxide produced and transported by these proteins, respectively, mediates inflammatory signaling. Anti-PECAM/SOD abolished VCAM expression caused by poly(I:C)-induced activation of toll-like receptor 3 localized in intracellular vesicles. These results directly implicate endosomal influx of superoxide in endothelial inflammatory response and suggest that site-specific interception of this signal attained by targeted delivery of anti-PECAM/SOD into endothelial endosomes may have anti-inflammatory effects.
Seminars in Thrombosis and Hemostasis, 2002
Thromboembolism is one of the most common causes of death in cancer patients. Among the most freq... more Thromboembolism is one of the most common causes of death in cancer patients. Among the most frequent thrombotic complications in patients with cancer are disseminated intravascular coagulation, thrombotic thrombocytopenic purpura, and thrombocytosis. Clearly, these complications arise as tumor cells interact with almost all components of the hemostatic system including platelets. Platelets participate in tumor progression by contributing to the metastatic cascade, protecting tumor cells from immune surveillance, regulating tumor cell invasion, and angiogenesis. Platelets contain one of the largest stores of angiogenic and mitogenic factors and the tumor vasculature is leaky, which allows platelets to come in contact with the tumor and deposit multiple angiogenic factors including vascular endothelial growth factor (VEGF) and thrombin to tumor cells, which in turn contributes to tumor progression. This article reviews the recent literature on how platelets contribute to tumor growth, angiogenesis, and metastasis.