Selection of Molecules with Immunological Potential from Excretory and Secretory Products from the Nematode Haemonchus placei by Cell Proliferation and Gene Expression Assays
Por:
Maza-Lopez, Jocelyn, Camas-Pereyra, Rene, Lopez-Arellano, Maria Eugenia, Contreras-Ochoa, Carla O.
Publicada:
20 jun 2023
Resumen:
The nematode Haemonchus placei is a pathogenic parasite, the most
seriously affecting ruminant's health and being responsible for enormous
economic losses all over the world. The present protocol describes
different in vitro techniques to select potential candidate antigens
with immune-protective activity from excretory and secretory products
(ESP) from H. placei transitory infective larvae (xL3). ESP from xL3
were obtained from the in vitro infective larvae (L3) maintained in
Hank's medium at 37 & DEG;C with 5% CO2 for 48 h. Then, the presence
of ESP proteins was confirmed by SDS-PAGE to be used in an in vitro
proliferation assay with bovine peripheral blood mononuclear cells
(PBMCs). The ESP were exposed to the PBMCs during two different periods
(24 and 48 h). Genes associated with immune response against the
nematode were analyzed using relative gene expression and bioinformatic
tools. These are simple, economic, and helpful tools to identify
potential immune-protective molecules under in vitro conditions for
confirming the efficacy of future in vivo assays.
Filiaciones:
Maza-Lopez, Jocelyn:
Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Ciudad Universitaria, Ciudad de México, CP 04510, Mexico
Camas-Pereyra, Rene:
Departamento de Helmintología, Centro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Carretera Federal Cuernavaca-Cuautla 8534, Progreso, Morelos, Jiutepec, CP 62574, Mexico
Lopez-Arellano, Maria Eugenia:
Departamento de Helmintología, Centro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Carretera Federal Cuernavaca-Cuautla 8534, Progreso, Morelos, Jiutepec, CP 62574, Mexico
Contreras-Ochoa, Carla O.:
Centro de Investigación Sobre Enfermedades Infecciosas, Instituto Nacional de Salud Pública, Av. Universidad 655, Col. Santa María Ahuacatitlán, Morelos, Cuernavaca, CP 62100, Mexico
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