RESUMEN
We determined the prophylactic effect of both the d-mannose-binding lectin ArtinM extracted from the seeds of Artocarpus integrifolia (jackfruit) and its recombinant counterpart during the course of experimental paracoccidioidomycosis induced in BALB/c mice. Four experimental protocols of prophylaxis were employed to evaluate the most protective regimen of ArtinM administration. It was demonstrated that the best effect was obtained by administration of two ArtinM doses on days 10 and 3 before the challenge with Paracoccidioides brasiliensis. By following this protocol, the lungs of mice that received native or recombinant ArtinM exhibited reduced fungal burden and granuloma incidence. In addition, the protocol augmented contents of IL-12, IFN-gamma, TNF-alpha and NO. On the other hand, the control group consisting of untreated infected mice had higher pulmonary levels of IL-4 and IL-10. In conclusion, prophylaxis with ArtinM significantly reproduces the effect of its therapeutic administration, i.e, it confers resistance to P. brasiliensis infection in mouse models by promoting IL-12 production and favours Th1-immunity.
Asunto(s)
Antifúngicos/uso terapéutico , Artocarpus/química , Quimioprevención/métodos , Lectinas/uso terapéutico , Paracoccidioides/efectos de los fármacos , Paracoccidioidomicosis/prevención & control , Animales , Antifúngicos/aislamiento & purificación , Citocinas/análisis , Modelos Animales de Enfermedad , Femenino , Lectinas/aislamiento & purificación , Pulmón/inmunología , Pulmón/patología , Ratones , Ratones Endogámicos BALB C , Óxido Nítrico/análisis , Paracoccidioidomicosis/inmunología , Paracoccidioidomicosis/microbiología , Paracoccidioidomicosis/patología , Proteínas Recombinantes/genética , Proteínas Recombinantes/uso terapéutico , Semillas/químicaRESUMEN
KM(+) is a mannose-binding lectin from Artocarpus integrifolia that induces interleukin (IL)-12 production by macrophages and protective T helper 1 immune response against Leishmania major infection. In this study, we performed experiments to evaluate the therapeutic activity of jackfruit KM(+) (jfKM(+)) and its recombinant counterpart (rKM(+)) in experimental paracoccidioidomycosis. To this end, jfKM(+) or rKM(+) was administered to BALB/c mice 10 days after infection with Paracoccidiodes brasiliensis. Thirty days postinfection, lungs from the KM(+)-treated mice contained significantly fewer colony-forming units and little to no organized granulomas compared to the controls. In addition, lung homogenates from the KM(+)-treated mice presented higher levels of nitric oxide, IL-12, interferon-gamma, and tumor necrosis factor-alpha, whereas higher levels of IL-4 and IL-10 were detected in the control group. With mice deficient in IL-12, Toll-like receptor (TLR) 2, TLR4, or TLR adaptor molecule MyD88, we demonstrated that KM(+) led to protection against P. brasiliensis infection through IL-12 production, which was dependent on TLR2. These results demonstrated a beneficial effect of KM(+) on the severity of P. brasiliensis infection and may expand its potential use as a novel immunotherapeutic molecule.
Asunto(s)
Interleucina-12/biosíntesis , Lectinas Tipo C/metabolismo , Lectinas de Unión a Manosa/metabolismo , Paracoccidioides/efectos de los fármacos , Paracoccidioidomicosis/tratamiento farmacológico , Lectinas de Plantas/uso terapéutico , Receptores de Superficie Celular/metabolismo , Receptor Toll-Like 2/metabolismo , Animales , Artocarpus , Células Cultivadas , Recuento de Colonia Microbiana , Citocinas/biosíntesis , Interleucina-12/genética , Pulmón/metabolismo , Pulmón/microbiología , Pulmón/patología , Masculino , Receptor de Manosa , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Factor 88 de Diferenciación Mieloide/metabolismo , Óxido Nítrico/metabolismo , Paracoccidioidomicosis/metabolismo , Lectinas de Plantas/farmacología , Proteínas Recombinantes/farmacología , Proteínas Recombinantes/uso terapéutico , Receptor Toll-Like 4/metabolismoRESUMEN
Paracoccidioides brasiliensis components interact with host cells and can influence the pathogenesis of paracoccidioidomycosis (PCM). Among the components released by P. brasiliensis, gp 43 and a heavily glycosylated antigen with MM>160 kDa are the most recognized by serum antibodies from patients with PCM. In order to isolate the high MM glycoconjugate, we carried out affinity chromatography of a crude exoantigen preparation on immobilized jacalin. The bound fraction (JBE, jacalin binding exoantigen) consisted of a major antigen of high MM and frequently of an additional 70-kDa minor protein. This protein, designated paracoccin, exhibited selective binding to immobilized GlcNAc, a property that was used for its purification. The structural data of paracoccin obtained by mass spectrometry of tryptic peptides did not match any known protein. Anti-paracoccin serum localized the lectin on the surface of P. brasiliensis yeasts, especially in the budding regions. Paracoccin was able to interact with laminin in a dose-dependent manner. This interaction was inhibited by GlcNAc, followed by D-glucose and D-mannose, but not by D-galactose, N-acetyl-galactosamine or L-fucose. Interestingly, paracoccin induced both resident and elicited mouse peritoneal cavity macrophages to release high and persistent levels of TNF-alpha in vitro, a fact that was associated with high nitric oxide production in elicited cells. Because binding to laminin can favor yeast adhesion and invasion of host tissues, and overproduction of NO has been associated with suppression of cell immunity, paracoccin is suggested to play an important role in PCM pathogenesis.