RÉSUMÉ
Yellow fever (YF) is a disease caused by the homonymous flavivirus that can be prevented by a vaccine containing attenuated viruses. Since some individuals cannot receive this vaccine, the development of alternatives is desirable. Here, we developed a recombinant baculovirus (rBV) surface display platform utilizing a chimeric E-NS1 protein as a vaccine candidate. A pBacPAK9 vector containing the baculoviral GP64 signal peptide, the YFV prM, E, NS1 and the ectodomain of VSV-G sequences was synthesized. This transfer plasmid and the bAcGOZA bacmid were cotransfected into Sf9 cells, and an rBV-E-NS1 was obtained, which was characterized by PCR, WB, IFI and FACS analysis. Mice immunized with rBV-E-NS1 elicited a specific humoral and cellular immune response and were protected after YFV infection. In summary, we have developed an rBV that expresses YFV major antigen proteins on its surface, which opens new alternatives that can be tested in a mouse model.
Sujet(s)
Anticorps antiviraux , Baculoviridae , Protéines virales non structurales , Fièvre jaune , Virus de la fièvre jaune , Animaux , Baculoviridae/génétique , Baculoviridae/immunologie , Souris , Anticorps antiviraux/immunologie , Anticorps antiviraux/sang , Virus de la fièvre jaune/immunologie , Virus de la fièvre jaune/génétique , Protéines virales non structurales/immunologie , Protéines virales non structurales/génétique , Fièvre jaune/prévention et contrôle , Fièvre jaune/immunologie , Protéines de l'enveloppe virale/immunologie , Protéines de l'enveloppe virale/génétique , Cellules Sf9 , Vaccins antiviraux/immunologie , Vaccins antiviraux/administration et posologie , Vaccins antiviraux/génétique , Femelle , Protéines de fusion recombinantes/immunologie , Protéines de fusion recombinantes/génétique , Immunité cellulaire , Souris de lignée BALB C , Immunité humorale , Vecteurs génétiques/génétiqueRÉSUMÉ
ETHNOPHARMACOLOGICAL RELEVANCE: Taiuiá or tayuya (Cayaponia tayuya, Cucurbitaceae) is a climbing, lignified plant with a large swollen root that has traditionally been used as an anti-inflammatory and anti-rheumatic agent in the folk medicine of Brazil, Peru, and Colombia. THE AIM OF THE STUDY: We have assayed the pharmacological properties of a flavonoid fraction obtained from the butanol extract of Cayaponia tayuya roots using two models of topical mouse ear oedema, paying special attention to its influence on the induction on pro-inflammatory enzymes and peptidic mediators. MATERIAL AND METHODS: The in vivo experiments involved both the acute oedema induced by a single application of TPA and the subchronic inflammation brought on by repeated applications of TPA. The effects on the induction of pro-inflammatory enzymes and peptidic mediators in RAW 264.7 macrophages were analyzed with the aid of Western blot analysis. RESULTS: The extract was identified as a mixture of flavonoids in which vicenin-2, spinosin, isovitexin, and a mixture of swertisin and isoswertisin were found. In acute TPA-induced oedema in mouse ears, the flavonoid-enriched fraction (at a dose of 0.5mg/ear) inhibited the oedema by 66% (4.2+/-0.6 mg vs. 12.3+/-1.4 mg, P<0.01) while in the subchronic model, the inhibition reached 37% at a dose of 0.5mg/ear x 7 applications (7.5+/-0.6 mg vs. 11.9+/-1.3mg, P<0.05). When assayed in vitro, the flavonoid showed no toxicity at 33.45 microg/mL on RAW 264.7 macrophages. Although the nitric oxide production in these cells was moderately reduced (42%) at 33.45 microg/mL, the flavonoid-enriched fraction had no effect on TNF-alpha production. In addition, at 22.30 microg/mL, the test sample inhibited both iNOS and COX-2 expression by 98% and 49%, respectively. CONCLUSION: These results indicate that the anti-inflammatory activity of flavonoids from tayuya roots most likely stems from their inhibition of the induction of the enzymes COX-2 and iNOS.
Sujet(s)
Anti-inflammatoires/pharmacologie , Cucurbitaceae/composition chimique , Flavonoïdes/pharmacologie , Extraits de plantes/pharmacologie , Animaux , Anti-inflammatoires/isolement et purification , Brésil , Lignée cellulaire , Colombie , Cyclooxygenase 2/effets des médicaments et des substances chimiques , Cyclooxygenase 2/métabolisme , Modèles animaux de maladie humaine , Femelle , Flavonoïdes/isolement et purification , Inflammation/traitement médicamenteux , Inflammation/physiopathologie , Macrophages/effets des médicaments et des substances chimiques , Macrophages/métabolisme , Médecine traditionnelle , Souris , Nitric oxide synthase type II/effets des médicaments et des substances chimiques , Nitric oxide synthase type II/métabolisme , Pérou , Extraits de plantes/composition chimique , Racines de planteRÉSUMÉ
The dichloromethane extract and pomolic acid ( 5) obtained from leaves of Cecropia pachystachya both reduced carrageenan-induced paw oedema in mice. Interestingly, while the triterpenoid inhibited the in vivo production of interleukin-1beta by 39 %, it had no effect on tumour necrosis factor-alpha production. We also demonstrated that both the dichloromethane extract and 5 inhibited the viability of human polymorphonuclear (PMN) cells in a time- and dose-dependent fashion. The PMN membrane integrity was determined with the aid of flow cytometry by means of the exclusion of propidium iodide as assay. Although the cell membrane integrity was altered, neither the extract nor 5 produced cellular necrosis. Moreover, the development of hypodiploid nuclei and DNA fragmentation in the PMN cells were both dependent on dose and time. Finally, in the annexin V-FITC binding assay, compound 5 increased the total of apoptotic cells by 42 % at 100 microM and by 71 % at 200 microM with respect to the control group. In conclusion, both the dichloromethane extract of ambay and isolated compound 5 inhibit the viability of PMN cells through apoptosis. Since they can regulate human neutrophil functions in this way, it is likely that these substances can also limit inflammation.
Sujet(s)
Anti-inflammatoires/pharmacologie , Apoptose/effets des médicaments et des substances chimiques , Cecropia/composition chimique , Acide oléanolique/analogues et dérivés , Animaux , Anti-inflammatoires/isolement et purification , Femelle , Souris , Acide oléanolique/isolement et purification , Acide oléanolique/pharmacologieRÉSUMÉ
El virus HTLV-I fue encontrado promotor de la Leucemia T del adulto y de la paraparesia espástica tropical. No es el único virus inculpado con la producción de neoplasias en el hombre. Se trata de un virus muy similar al HTLV-II o HIV, que se propaga en forma muy similar a éste. Tiene como diferencia fundamental, al producción de atipías linfoides y su muy largo período de incubación. Se lo encuentra en casi todo el mundo, pero su origen es el sur del Japón, si bien es endémico en la costa Americana del Pacífico, donde produce el cuadro neurológico mencionado anteriormente
Sujet(s)
Virus T-lymphotrope humain de type 1/pathogénicité , Infections à HTLV-I/épidémiologie , Infections à HTLV-I/microbiologie , Leucémie-lymphome à cellules T de l'adulte/épidémiologie , Leucémie-lymphome à cellules T de l'adulte/microbiologie , Paraparésie spastique tropicale/épidémiologie , Paraparésie spastique tropicale/microbiologie , ArgentineRÉSUMÉ
El virus HTLV-I fue encontrado promotor de la Leucemia T del adulto y de la paraparesia espástica tropical. No es el único virus inculpado con la producción de neoplasias en el hombre. Se trata de un virus muy similar al HTLV-II o HIV, que se propaga en forma muy similar a éste. Tiene como diferencia fundamental, al producción de atipías linfoides y su muy largo período de incubación. Se lo encuentra en casi todo el mundo, pero su origen es el sur del Japón, si bien es endémico en la costa Americana del Pacífico, donde produce el cuadro neurológico mencionado anteriormente (AU)