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1.
Sci Rep ; 11(1): 23682, 2021 12 08.
Artigo em Inglês | MEDLINE | ID: mdl-34880361

RESUMO

Human immunodeficiency virus-1 (HIV-1) persistence in the presence of antiretroviral therapy (ART) has halted the development of curative strategies. Measuring HIV persistence is complex due to the low frequency of cells containing virus in vivo. Most of the commercially available assays to date measure nucleic acid. These assays have the advantage of being highly sensitive and allow for the analysis of sequence diversity, intactness of the HIV genome or evaluation of diverse RNA species. However, these assays are limited in evaluating translational competent viral reservoirs. In here, we developed an ultrasensitive p24 ELISA that uses the Simoa planar array technology that can detect HIV-1 virions and HIV-1 infected cell with limit of detection similar to nucleic acid assays. Furthermore, the assay is optimized to measure very low levels of p24 in different biological fluids without a major loss of sensitivity or reproducibility. Our results demonstrate that the 'homebrew' planar p24 ELISA immunoassay is a broadly applicable new tool to evaluate HIV persistence in diverse biological fluids and cells.


Assuntos
Ensaio de Imunoadsorção Enzimática , Proteína do Núcleo p24 do HIV/metabolismo , Infecções por HIV/diagnóstico , Infecções por HIV/virologia , HIV-1/metabolismo , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/normas , Proteína do Núcleo p24 do HIV/imunologia , Humanos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
3.
Dis Model Mech ; 14(7)2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-34308967

RESUMO

People of African ancestry living with the human immunodeficiency virus-1 (HIV-1) are at risk of developing HIV-associated nephropathy (HIVAN). Children with HIVAN frequently show high plasma fibroblast growth factor-2 (FGF-2) levels; however, the role of circulating FGF-2 in the pathogenesis of childhood HIVAN is unclear. Here, we explored how circulating FGF-2 affected the outcome of HIVAN in young HIV-Tg26 mice. Briefly, we demonstrated that FGF-2 was preferentially recruited in the kidneys of mice without pre-existing kidney disease, precipitating HIVAN by activating phosphorylated extracellular signal-regulated kinase (pERK) in renal epithelial cells, without inducing the expression of HIV-1 genes. Wild-type mice injected with recombinant adenoviral FGF-2 (rAd-FGF-2) vectors carrying a secreted form of human FGF-2 developed transient and reversible HIVAN-like lesions, including proteinuria and glomerular enlargement. HIV-Tg26 mice injected with rAd-FGF-2 vectors developed more-significant proliferative and pro-fibrotic inflammatory lesions, similar to those seen in childhood HIVAN. These lesions were partially reversed by treating mice with the FGF/VEGF receptor tyrosine kinase inhibitor PD173074. These findings suggest that high plasma FGF-2 levels may be an independent risk factor for precipitating HIVAN in young children.


Assuntos
Nefropatia Associada a AIDS , HIV-1 , Nefropatia Associada a AIDS/genética , Animais , Pré-Escolar , Modelos Animais de Doenças , Fator 2 de Crescimento de Fibroblastos , HIV-1/genética , Humanos , Camundongos , Camundongos Transgênicos
4.
Pediatr Nephrol ; 36(8): 2189-2201, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33044676

RESUMO

HIV-associated nephropathy (HIVAN) predominantly affects people of African ancestry living with HIV who do not receive appropriate antiretroviral therapy (ART). Childhood HIVAN is characterized by heavy proteinuria and decreased kidney function. Kidney histology shows mesangial expansion, classic or collapsing glomerulosclerosis, and microcystic renal tubular dilatation leading to kidney enlargement. The pathogenesis of HIVAN involves the kidney recruitment of inflammatory cells and the infection of kidney epithelial cells. In addition, both viral and genetic factors play key roles in this disease. Modern ART has improved the outcome and decreased the prevalence of childhood HIVAN. However, physicians have had modest success providing chronic ART to children and adolescents, and we continue to see children with HIVAN all over the world. This article discusses the progress made during the last decade in our understanding of the pathogenesis and treatment of childhood HIVAN, placing particular emphasis on the mechanisms that mediate the infection of kidney epithelial cells, and the roles of cytokines, the HIV-Tat gene, and the Apolipoprotein-1 (APOL1) gene risk variants in this disease. In view of the large number of children living with HIV at risk of developing HIVAN, better prevention and treatment programs are needed to eradicate this disease.


Assuntos
Nefropatia Associada a AIDS , Infecções por HIV , HIV-1 , Nefropatia Associada a AIDS/diagnóstico , Nefropatia Associada a AIDS/epidemiologia , Nefropatia Associada a AIDS/genética , Adolescente , Apolipoproteína L1 , Infecções por HIV/complicações , Infecções por HIV/tratamento farmacológico , Infecções por HIV/epidemiologia , Humanos , Rim
5.
Dis Model Mech ; 13(10)2020 10 28.
Artigo em Inglês | MEDLINE | ID: mdl-32917744

RESUMO

Modern antiretroviral therapies (ART) have decreased the prevalence of HIV-associated nephropathy (HIVAN). Nonetheless, we continue to see children and adolescents with HIVAN all over the world. Furthermore, once HIVAN is established in children, it is difficult to revert its long-term progression, and we need better animal models of childhood HIVAN to test new treatments. To define whether the HIV-1 trans-activator (Tat) gene precipitates HIVAN in young mice, and to develop an inducible mouse model of childhood HIVAN, an HIV-Tat gene cloned from a child with HIVAN was used to generate recombinant adenoviral vectors (rAd-Tat). rAd-Tat and LacZ control vectors (2×109) were expressed in the kidney of newborn wild-type and HIV-transgenic (Tg26) FVB/N mice without significant proteinuria (n=5; 8 per group). Mice were sacrificed 7 and 35 days later to assess their renal outcome, the expression of HIV-genes and growth factors, and markers of cell growth and differentiation by RT-qPCR, immunohistochemistry and/or western blots. HIV-Tat induced the expression of HIV-1 genes and heparin-binding growth factors in the kidney of HIV-Tg26 mice, and precipitated HIVAN in the first month of life. No significant renal changes were detected in wild-type mice infected with rAd-Tat vectors, suggesting that HIV-Tat alone does not induce renal disease. This new mouse model of childhood HIVAN highlights the critical role that HIV-Tat plays in the pathogenesis of HIVAN, and could be used to study the pathogenesis and treatment of HIVAN in children and adolescents.


Assuntos
Nefropatia Associada a AIDS/patologia , Produtos do Gene tat do Vírus da Imunodeficiência Humana/metabolismo , Nefropatia Associada a AIDS/genética , Albuminúria/complicações , Albuminúria/genética , Sequência de Aminoácidos , Animais , Animais Recém-Nascidos , Apoptose , Desdiferenciação Celular , Proliferação de Células , Criança , Modelos Animais de Doenças , Fator 2 de Crescimento de Fibroblastos/genética , Fator 2 de Crescimento de Fibroblastos/metabolismo , HIV-1/genética , Humanos , Rim/lesões , Rim/patologia , Camundongos Transgênicos , Podócitos/metabolismo , Podócitos/patologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , beta-Galactosidase/metabolismo , Produtos do Gene env do Vírus da Imunodeficiência Humana/genética , Produtos do Gene env do Vírus da Imunodeficiência Humana/metabolismo , Produtos do Gene tat do Vírus da Imunodeficiência Humana/química
6.
J Am Soc Nephrol ; 28(3): 862-875, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27811066

RESUMO

Studies have shown that podocytes and renal tubular epithelial cells from patients with HIV-associated nephropathy (HIVAN) express HIV-1 transcripts, suggesting that productive infection of renal epithelial cells precipitates development of HIVAN. However, podocytes and renal tubular epithelial cells do not express CD4 receptors, and it is unclear how these cells become productively infected in vivo We investigated the mechanisms underlying the infection by HIV-1 of podocytes cultured from the urine of children with HIVAN. We observed low-level productive infection on exposure of these cells to primary cell-free HIV-1 supernatants. However, envelope-defective recombinant HIV-1 did not infect the renal epithelial cell lines. Moreover, treatment of podocytes to inhibit endocytic transport or dynamin activity or remove cell surface heparan sulfate proteoglycans reduced infection efficiency. Transfection of CD4- 293T cells with a cDNA expression library developed from a podocyte cell line derived from a child with HIVAN led to the identification of TNF-α as a possible mediator of HIV-1 infection. Overexpression of transmembrane TNF-α in cultured CD4- renal tubular epithelial cells, 293T cells, and HeLa cells enabled the infection of these cells; exposure to soluble TNF-α did not. Immunohistochemistry showed TNF-α expression in podocytes of renal sections from children with HIVAN. Furthermore, we found that TNF-α enhanced NF-κB activation and integration of HIV-1 into the podocyte DNA. Finally, inhibition of dynamin activity blocked TNF-α-mediated infection. These data establish a role for transmembrane TNF-α in facilitating the viral entry and integration of HIV-1 into the DNA of renal epithelial cells.


Assuntos
Nefropatia Associada a AIDS/virologia , HIV-1/fisiologia , Podócitos/virologia , Fator de Necrose Tumoral alfa/fisiologia , Células Cultivadas , Criança , Humanos , Proteínas de Membrana
7.
Am J Physiol Heart Circ Physiol ; 309(8): H1314-25, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26276817

RESUMO

Critically ill children can develop bleeding complications when treated with heparin-like drugs. These events are usually attributed to the anticoagulant activity of these drugs. However, previous studies showed that fibroblast growth factor-2 (FGF-2), a heparin-binding growth factor released in the circulation of these patients, could precipitate intestinal hemorrhages in mice treated with the heparin-like drug pentosan polysulfate (PPS). Yet very little is known about how FGF-2 induces bleeding complications in combination with heparin-like drugs. Here, we examined the mechanisms by which circulating FGF-2 induces intestinal hemorrhages in mice treated with PPS. We used a well-characterized mouse model of intestinal hemorrhages induced by FGF-2 plus PPS. Adult FVB/N mice were infected with adenovirus carrying Lac-Z or a secreted form of recombinant human FGF-2, and injected with PPS, at doses that do not induce bleeding complications per se. Mice treated with FGF-2 in combination with PPS developed an intestinal inflammatory reaction that increased the permeability and disrupted the integrity of submucosal intestinal vessels. These changes, together with the anticoagulant activity of PPS, induced lethal hemorrhages. Moreover, a genetically modified form of the endothelial ligand angiopoietin-1 (Ang-1*), which has powerful antipermeability and anti-inflammatory activity, prevented the lethal bleeding complications without correcting the anticoagulant status of these mice. These findings define new mechanisms through which FGF-2 and Ang-1* modulate the outcome of intestinal bleeding complications induced by PPS in mice and may have wider clinical implications for critically ill children treated with heparin-like drugs.


Assuntos
Angiopoietina-1/biossíntese , Fator 2 de Crescimento de Fibroblastos/biossíntese , Hemorragia Gastrointestinal/prevenção & controle , Terapia Genética/métodos , Intestino Delgado/metabolismo , Adenoviridae/genética , Angiopoietina-1/genética , Animais , Coagulação Sanguínea , Permeabilidade Capilar , Modelos Animais de Doenças , Fator 2 de Crescimento de Fibroblastos/genética , Hemorragia Gastrointestinal/induzido quimicamente , Hemorragia Gastrointestinal/genética , Hemorragia Gastrointestinal/metabolismo , Hemorragia Gastrointestinal/patologia , Técnicas de Transferência de Genes , Vetores Genéticos , Mediadores da Inflamação/metabolismo , Doenças Inflamatórias Intestinais/induzido quimicamente , Doenças Inflamatórias Intestinais/genética , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/prevenção & controle , Intestino Delgado/irrigação sanguínea , Intestino Delgado/patologia , Macrófagos/metabolismo , Masculino , Metaloproteinases da Matriz/metabolismo , Camundongos , Poliéster Sulfúrico de Pentosana
8.
J Am Soc Nephrol ; 25(8): 1800-13, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24578133

RESUMO

Podocyte injury has a critical role in the pathogenesis of HIV-associated nephropathy (HIVAN). The HIV-1 transactivator of transcription (Tat), combined with fibroblast growth factor-2 (FGF-2), can induce the dedifferentiation and proliferation of cultured human podocytes. Cellular internalization of Tat requires interactions with heparan sulfate proteoglycans and cholesterol-enriched lipid rafts (LRs). However, the specific distribution of Tat in human podocytes and its ability to associate with LRs have not been documented. Here, we found that Tat is preferentially recruited to LRs in podocytes isolated from children with HIVAN. Furthermore, we identified arginines in the basic domain (RKKRRQRRR) of Tat as essential for (1) targeting Tat to LRs, (2) Tat-mediated increases in the expression of Rho-A and matrix metalloproteinase-9 in LRs, and (3) Tat-mediated enhancement of FGF-2 signaling in human podocytes and HIV-transgenic mouse kidneys and the exacerbation of renal lesions in these mice. Tat carrying alanine substitutions in the basic domain (AKKAAQAAA) remained localized in the cytosol and did not associate with LRs or enhance FGF-2 signaling in cultured podocytes. These results show the specific association of Tat with LRs in podocytes isolated from children with HIVAN, confirm Tat as a regulator of FGF-2 signaling in LRs, and identify the key domain of Tat responsible for promoting these effects and aggravating renal injury in HIV-transgenic mice. Moreover, these results provide a molecular framework for developing novel therapies to improve the clinical outcome of children with HIVAN.


Assuntos
Nefropatia Associada a AIDS/metabolismo , Fator 2 de Crescimento de Fibroblastos/metabolismo , HIV-1 , Microdomínios da Membrana/fisiologia , Podócitos/fisiologia , Transdução de Sinais/fisiologia , Produtos do Gene tat do Vírus da Imunodeficiência Humana/fisiologia , Nefropatia Associada a AIDS/patologia , Animais , Arginina/metabolismo , Técnicas de Cultura de Células , Criança , Humanos , Metaloproteinase 9 da Matriz/metabolismo , Camundongos Transgênicos , Proteína rhoA de Ligação ao GTP/metabolismo
9.
Pediatr Nephrol ; 27(3): 469-83, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21959768

RESUMO

Fibroblast growth factor-2 (FGF-2) is an angiogenic growth factor involved in renal growth and regeneration. Previous studies in rodents revealed that single intrarenal injections of FGF-2 improved the outcome of acute kidney injury (AKI). Septic children usually show elevated plasma levels of FGF-2, and are at risk of developing AKI. However, the role of circulating FGF-2 in the pathogenesis of AKI is not well understood. We have developed a mouse model to determine how FGF-2 released into the circulation modulates the outcome of AKI induced by lipopolysaccharide (LPS). Young FVB/N mice were infected with adenoviruses carrying a secreted form of human FGF-2 or control LacZ vectors. Subsequently, when the circulating levels of FGF-2 were similar to those seen in septic children, mice were injected with a non-lethal dose of LPS or control buffer. All mice injected with LPS developed hypotension and AKI, from which they recovered after 5 days. FGF-2 did not improve the outcome of AKI, and induced more significant renal proliferative and apoptotic changes during the recovery phase. These findings suggest that circulating FGF-2 may not necessarily prevent the development or improve the outcome of AKI. Moreover, the renal accumulation of FGF-2 might cause further renal damage.


Assuntos
Injúria Renal Aguda/etiologia , Fator 2 de Crescimento de Fibroblastos/fisiologia , Lipopolissacarídeos/toxicidade , Actinas/análise , Injúria Renal Aguda/sangue , Injúria Renal Aguda/fisiopatologia , Proteínas de Fase Aguda/urina , Adenoviridae/genética , Animais , Apoptose/efeitos dos fármacos , Nitrogênio da Ureia Sanguínea , Fator 2 de Crescimento de Fibroblastos/sangue , Rim/efeitos dos fármacos , Rim/patologia , Lipocalina-2 , Lipocalinas/urina , Masculino , Camundongos , Proteínas Oncogênicas/urina , Antígeno Nuclear de Célula em Proliferação/análise , Sístole/efeitos dos fármacos
10.
FEBS Lett ; 581(6): 1103-8, 2007 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-17321526

RESUMO

Talin binding of integrins, via its band 4.1, ezrin, radixin, and moesin (FERM)-homologous domain, directly activates the integrin receptor. However, it is not known whether other FERM-containing proteins also possess such an integrin activating capability. We report here that radixin, one of the original FERM-domain proteins, binds to the membrane-proximal region of the integrin beta(2) but not alpha(M) cytoplasmic tail. Importantly, we show that radixin binding significantly enhances the adhesive activity of integrin alpha(M)beta(2). Given the distinct biological activities of radixin and talin, radixin may represent a novel talin-independent pathway for integrin activation under specific settings.


Assuntos
Proteínas do Citoesqueleto/metabolismo , Antígeno de Macrófago 1/metabolismo , Proteínas de Membrana/metabolismo , Animais , Sítios de Ligação , Células CHO , Adesão Celular , Cricetinae , Cricetulus , Citoplasma/química , Ligação Proteica , Transfecção
11.
Am J Physiol Heart Circ Physiol ; 292(2): H743-50, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17071728

RESUMO

Pentosan polysulfate (PPS) is a heparin-like polysaccharide that can affect the binding interactions of fibroblast growth factor (FGF-2) with its high-affinity receptors. Patients with angiogenic tumors frequently show high levels of FGF-2 in the circulation. Since FGF-2 is a heparin-binding angiogenic growth factor, PPS has been used successfully to block its activity in patients with angiogenic tumors. However, because of its heparin-like activity, the major toxic effect of PPS is the development of bleeding disorders. The role that circulating FGF-2 plays in the pathogenesis of bleeding disorders in patients treated with PPS is currently unknown. Here we hypothesized that FGF-2 might play a physiological role in the pathogenesis of intestinal bleeding induced by PPS. This hypothesis is supported by previous studies showing that PPS is accumulated in the intestine and that circulating FGF-2 specifically binds to and modulates the angiogenic activity of intestinal submucosal endothelial cells. We used recombinant adenoviral vectors carrying a secreted form of FGF-2 and LacZ control vectors to determine whether high levels of circulating FGF-2 facilitate the development of intestinal bleeding disorders in FVB/N and C57BL/6J mice treated with PPS. We found that PPS, acting together with FGF-2, induced structural changes in intestinal vessels leading to the development of lethal intestinal hemorrhages. These findings might have wider clinical implications for the systemic use of PPS and other heparinoids in the treatment of patients with angiogenic diseases associated with high levels of circulating FGF-2.


Assuntos
Inibidores da Angiogênese/efeitos adversos , Antineoplásicos/efeitos adversos , Fator 2 de Crescimento de Fibroblastos/metabolismo , Hemorragia Gastrointestinal/metabolismo , Enteropatias/metabolismo , Intestinos/efeitos dos fármacos , Poliéster Sulfúrico de Pentosana/efeitos adversos , Adenoviridae/genética , Inibidores da Angiogênese/metabolismo , Animais , Antineoplásicos/metabolismo , Coagulação Sanguínea/efeitos dos fármacos , Vasos Sanguíneos/efeitos dos fármacos , Vasos Sanguíneos/metabolismo , Vasos Sanguíneos/patologia , Permeabilidade Capilar/efeitos dos fármacos , Modelos Animais de Doenças , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Células Endoteliais/ultraestrutura , Feminino , Fator 2 de Crescimento de Fibroblastos/sangue , Fator 2 de Crescimento de Fibroblastos/genética , Fator 2 de Crescimento de Fibroblastos/farmacologia , Hemorragia Gastrointestinal/sangue , Hemorragia Gastrointestinal/induzido quimicamente , Hemorragia Gastrointestinal/patologia , Técnicas de Transferência de Genes , Vetores Genéticos , Enteropatias/sangue , Enteropatias/induzido quimicamente , Enteropatias/patologia , Mucosa Intestinal/metabolismo , Intestinos/irrigação sanguínea , Intestinos/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Poliéster Sulfúrico de Pentosana/metabolismo , Ligação Proteica , Fatores de Tempo
12.
Am J Physiol Renal Physiol ; 291(2): F289-96, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16597610

RESUMO

Basic fibroblast growth factor (bFGF) is a heparin-binding growth factor that is accumulated in human dysplastic and cystic renal diseases. Previous studies have shown that bFGF can modulate the growth of developing renal tubules; however, its role in the pathogenesis of renal cyst formation is not clearly understood. Here, we tested the hypothesis that overexpression of bFGF in developing rodent kidneys induces cyst formation in vivo. We used two different adenoviral-mediated gene-transferring approaches to overexpress bFGF in developing rodent kidneys. Initially, metanephric kidney (MK) explants harvested from embryonic day 15 Sprague-Dawley rats were infected with adenoviral vectors (rAd) encoding human bFGF or LacZ genes and transplanted under the renal capsule of adult female rats. Subsequently, to determine whether bFGF could induce renal cysts in developing kidneys with an intact renal collecting system, we injected rAd-bFGF or LacZ vectors in the retroorbital plexus of newborn mice. Basic FGF induced a more efficient integration of the MK explants into the host kidneys and increased the vascularization and proliferation of developing tubules, leading to tubular dilatation and rapid formation of renal cysts. In addition, we successfully expressed human bFGF in the kidney of newborn mice in vivo and induced tubular dilatation and renal cysts. In contrast, mice injected with rAd-lacZ did not develop tubular dilatation or renal cysts. To the best of our knowledge, these experiments show for the first time that overexpression of bFGF in developing rodent kidneys can induce the formation of renal cysts in vivo.


Assuntos
Fator 2 de Crescimento de Fibroblastos/fisiologia , Doenças Renais Císticas/fisiopatologia , Rim/crescimento & desenvolvimento , Rim/fisiologia , Adenoviridae/genética , Animais , Animais Recém-Nascidos/fisiologia , Proliferação de Células , Feminino , Fator 2 de Crescimento de Fibroblastos/genética , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Técnicas de Transferência de Genes , Vetores Genéticos/genética , Humanos , Rim/química , Rim/citologia , Doenças Renais Císticas/etiologia , Túbulos Renais Coletores/química , Túbulos Renais Coletores/citologia , Túbulos Renais Coletores/crescimento & desenvolvimento , Túbulos Renais Coletores/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Técnicas de Cultura de Órgãos , RNA Mensageiro/análise , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley
13.
Pediatr Nephrol ; 20(12): 1708-16, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16133048

RESUMO

The role of circulating growth factors in the pathogenesis of childhood HIV-1-associated nephropathy (HIVAN) is not clearly understood. In previous studies, we found a significant accumulation of fibroblast growth factor-2 (FGF-2) in the circulation and kidneys of children with HIVAN. The purpose of this study was to determine whether circulating FGF-2 may play a role in the pathogenesis of HIVAN by increasing the renal recruitment and attachment of HIV-infected mononuclear cells to renal epithelial cells. Using in vitro cell adhesion assays, we showed that FGF-2 increased the attachment of peripheral blood mononuclear cells (PBMCs) to fibronectin-coated tissue culture dishes by approximately threefold through a mechanism that involved the alpha5 integrin subunit. In addition, we found that FGF-2 induces a similar increase in the attachment of HIV-infected PBMCs and monocytes/macrophages to plastic tissue culture dishes and to monolayers of primary renal tubular epithelial cells harvested from the urine of HIV-infected children with renal disease. Finally, we injected 16 adult C57Bl6/J male mice with recombinant adenoviral vectors carrying either the LacZ gene or a secreted form of human FGF-2 (5 x 10(8)pfu/mouse) and demonstrated that high levels of circulating FGF-2 can increase the renal recruitment of circulating inflammatory cells and induce transient tubulointerstitial injury in vivo. These data suggest that FGF-2 may have an immunomodulatory role in the pathogenesis of HIVAN by recruiting HIV-infected cells in the kidney.


Assuntos
Nefropatia Associada a AIDS/fisiopatologia , Células Epiteliais/efeitos dos fármacos , Fator 2 de Crescimento de Fibroblastos/farmacologia , Infecções por HIV , HIV-1 , Túbulos Renais/citologia , Monócitos/efeitos dos fármacos , Nefropatia Associada a AIDS/patologia , Nefropatia Associada a AIDS/virologia , Adenoviridae/genética , Animais , Adesão Celular , Técnicas de Cultura de Células , Células Cultivadas , Criança , Técnicas de Cocultura , DNA Viral/genética , DNA Viral/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/virologia , Fator 2 de Crescimento de Fibroblastos/genética , Fibronectinas/metabolismo , Vetores Genéticos , Infecções por HIV/sangue , Infecções por HIV/urina , Humanos , Imuno-Histoquímica , Túbulos Renais/virologia , Macrófagos/efeitos dos fármacos , Macrófagos/virologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Monócitos/virologia , Proteínas Recombinantes/farmacologia
14.
Proc Natl Acad Sci U S A ; 99(5): 2625-9, 2002 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-11867758

RESUMO

Cytokines regulate lymphocyte development and differentiation, but precisely how they control these processes is still poorly understood. By using microarray technology to detect cytokine-induced genes, we identified a cDNA encoding Cybr, which was increased markedly in cells incubated with IL-2 and IL-12. The mRNA was most abundant in hematopoietic cells and tissues. The predicted amino acid sequence is similar to that of GRP-1-associated protein (GRASP), a recently identified retinoic acid-induced cytohesin-binding protein. Physical interaction, dependent on the coiled-coil domains of Cybr and cytohesin-1, was demonstrated by coimmunoprecipitation of the overexpressed proteins from 293T cells. Cytohesin-1, in addition to its role in cell adhesion, is a guanine nucleotide-exchange protein activator of ARF GTPases. Acceleration of guanosine 5prime prime or minute-O-(thiotriphosphate) binding to ARF by cytohesin-1 in vitro was enhanced by Cybr. Because the binding protein modified activation of ADP ribosylation factor by cytohesin-1, we designate this cytokine-inducible protein Cybr (cytohesin binder and regulator).


Assuntos
Proteínas de Transporte/metabolismo , Moléculas de Adesão Celular/metabolismo , Interleucina-12/farmacologia , Interleucina-2/farmacologia , Proteínas de Membrana/metabolismo , Proteínas/metabolismo , Sequência de Aminoácidos , Animais , Linhagem Celular , Clonagem Molecular , Expressão Gênica , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Humanos , Camundongos , Dados de Sequência Molecular , Proteínas/genética , Homologia de Sequência de Aminoácidos , Fatores de Transcrição
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