Immune response to an peptide of enolase from Sporothrix spp. administered in mice by intranasal route in formulation with the Gel 01 adjuvant.

Abstract

Sporotrichosis is a subcutaneous mycosis of subacute or chronic evolution caused by pathogenic species of the genus Sporothrix. The disease in Brazil is acquired by zoonotic transmission involving cats infected with S. brasiliensis. As cats with sporotrichosis have severe lesions in the nasal and respiratory mucosa, with a high fungal load in these places, an intranasal administration vaccine may protect these felines from S. brasiliensis infection more efficiently than a parenteral administration vaccine. In this project we propose to evaluate the immunogenic properties of a vaccine formulation composed by a chimeric peptide, containing an immunogenic region (NIAMDVASSEFYKEDV) of enolase from Sporothrix spp. and a peptide (APPHALS) that stimulates the Th1 immune response, which is important in the anti-Sporothrix defense. The chimeric peptide (NIAMDVASSEFYKEDV APPHALS) will be administered in mice intranasal via with the Montanide Gel 01 adjuvant, which has been shown to improve the immunogenicity of antigens after intranasal immunization of mice and cats. The production of antibodies in serum (IgG, IgG1 and IgG2a) and in saliva (IgA) of vaccinated mice will be evaluated by ELISA. In the supernatant of splenocytes from control and vaccinated animals stimulated in vitro with the vaccine antigen will be quantified cytokines of Th1/Th2/Th17 response pattern by flow cytometry. We hope that the formulation can induce the expected immune response to test its protective efficacy in a murine model of intranasal infection by Sporothrix brasiliensis.