RESUMO
Capripoxviruses (CaPVs) have been shown to be ideal viral vectors for the development of recombinant multivalent vaccines to enable delivery of immunogenic genes from ruminant pathogens. So far, the viral thymidine kinase (TK) gene is the only gene used to generate recombinants. A putative non-essential gene encoding a G-protein-coupled chemokine receptor subfamily homologue (GPCR) was targeted as an additional insertion site. Peste des petits ruminants (PPR) was chosen as a disease model. A new recombinant CaPV expressing the viral attachment hemagglutinin (H) of the PPR virus (PPRV) in the GPCR insertion site (rKS1-HPPR-GPCR) was generated in the backbone North African isolate KS1 strain of lumpy skin disease virus (LSDV). Comparison with the recombinant CaPV expressing the H of PPRV in the TK gene (rKS1-HPPR-TK) shown to induce protection against both PPR and LSD in both sheep and goats was assessed. The suitability of the GPCR gene to be a putative additional insertion site in the CaPV genome is evaluated and discussed.
Assuntos
Capripoxvirus/genética , Vetores Genéticos , Doença Nodular Cutânea/prevenção & controle , Vírus da Doença Nodular Cutânea/genética , Mutagênese Insercional , Peste dos Pequenos Ruminantes/prevenção & controle , Vírus da Peste dos Pequenos Ruminantes/genética , Receptores de Quimiocinas/genética , Receptores Acoplados a Proteínas G/genética , Vacinas Virais/genética , Animais , Anticorpos Antivirais/sangue , Bovinos , Chlorocebus aethiops , Cabras , Hemaglutininas Virais/administração & dosagem , Hemaglutininas Virais/genética , Hemaglutininas Virais/imunologia , Injeções Subcutâneas , Doença Nodular Cutânea/genética , Doença Nodular Cutânea/imunologia , Doença Nodular Cutânea/virologia , Vírus da Doença Nodular Cutânea/imunologia , Peste dos Pequenos Ruminantes/genética , Peste dos Pequenos Ruminantes/imunologia , Peste dos Pequenos Ruminantes/virologia , Vírus da Peste dos Pequenos Ruminantes/imunologia , Receptores de Quimiocinas/administração & dosagem , Receptores de Quimiocinas/imunologia , Receptores Acoplados a Proteínas G/administração & dosagem , Receptores Acoplados a Proteínas G/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Células Vero , Vacinas Virais/administração & dosagem , Vacinas Virais/imunologia , Cultura de VírusRESUMO
Chemokines trigger leukocyte trafficking and are implicated in cardiovascular disease pathophysiology. Chemokine-binding proteins, called "Evasins" have been shown to inhibit both CC and CXC chemokine-mediated bioactivities. Here, we investigated whether treatment with Evasin-3 (CXC chemokine inhibitor) and Evasin-4 (CC chemokine inhibitor) could influence post-infarction myocardial injury and remodelling. C57Bl/6 mice were submitted in vivo to left coronary artery permanent ligature and followed up for different times (up to 21 days). After coronary occlusion, three intraperitoneal injections of 10 µg Evasin-3, 1 µg Evasin-4 or equal volume of vehicle (PBS) were performed at 5 minutes, 24 hours (h) and 48 h after ischaemia onset. Both anti-chemokine treatments were associated with the beneficial reduction in infarct size as compared to controls. This effect was accompanied by a decrease in post-infarction myocardial leukocyte infiltration, reactive oxygen species release, and circulating levels of CXCL1 and CCL2. Treatment with Evasin-4 induced a more potent effect, abrogating the inflammation already at one day after ischaemia onset. At days 1 and 21 after ischaemia onset, both anti-chemokine treatments failed to significantly improve cardiac function, remodelling and scar formation. At 21-day follow-up, mouse survival was exclusively improved by Evasin-4 treatment when compared to control vehicle. In conclusion, we showed that the selective inhibition of CC chemokines (i.e. CCL5) with Evasin-4 reduced cardiac injury/inflammation and improved survival. Despite the inhibition of CXC chemokine bioactivities, Evasin-3 did not affect mouse survival. Therefore, early inhibition of CC chemokines might represent a promising therapeutic approach to reduce the development of post-infarction heart failure in mice.
Assuntos
Anti-Inflamatórios/administração & dosagem , Infarto do Miocárdio/tratamento farmacológico , Miocárdio/metabolismo , Receptores CXCR/administração & dosagem , Receptores de Quimiocinas/administração & dosagem , Animais , Proteínas de Artrópodes , Movimento Celular/efeitos dos fármacos , Quimiocina CXCL1/sangue , Vasos Coronários/cirurgia , Modelos Animais de Doenças , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/diagnóstico , Miocárdio/patologia , Estresse Oxidativo/efeitos dos fármacos , Proteínas e Peptídeos SalivaresAssuntos
Antígeno CD11b/administração & dosagem , Células Mieloides/transplante , Infarto do Miocárdio/cirurgia , Neovascularização Fisiológica/fisiologia , Receptores de Quimiocinas/administração & dosagem , Administração Intravenosa , Animais , Transplante de Medula Óssea/métodos , Camundongos , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/patologia , Neovascularização Fisiológica/efeitos dos fármacosRESUMO
Depletion of mouse Kupffer cells and splenic macrophages following intravenous administration of liposome-entrapped clodronate severely reduced host resistance to primary infection with Listeria monocytogenes. Infection of clodronate-treated mice with a sublethal dose of L. monocytogenes resulted in death of the mice within 3 days. The macrophage depletion resulted in marked increases in bacterial growth in the liver and spleen, but not in other tissues. The proliferation of L. monocytogenes was observed in a large number of hepatocytes that underwent apoptosis. Infiltration of neutrophils in the liver and rapid formation of microabscesses were observed in the control mice after L. monocytogenes infection. However, there was less accumulation of neutrophils in the liver of Kupffer cell-depleted mice than in the control mice. Expression of macrophage inflammatory protein-2 (MIP-2) was enhanced in the livers of both the control and Kupffer cell-depleted mice after L. monocytogenes infection. MIP-2 was also induced in a murine hepatocyte cell line following L. monocytogenes infection. The administration of neutralizing anti-interleukin-8 receptor homolog antibody severely abrogated neutrophil infiltration into the Listeria-infected mouse liver. Anti-MIP-2 antibody moderately reduced neutrophil infiltration and microabscess formation in the liver. These findings indicate that Kupffer cells protect hepatocytes from L. monocytogenes infection and the resultant apoptosis. Moreover, MIP-2 and its related molecules produced by the infected hepatocytes regulate neutrophil infiltration and microabscess formation in primary listeriosis.