Identification of immunogenic proteins from ovarian tissue and recognized in larval extracts of Rhipicephalus (Boophilus) microplus, through an immunoproteomic approach

• Published in: Experimental parasitology Vol. 170; pp. 227 - 235
• Main Authors: Ramírez Rodríguez, Patricia Berenice, Rosario Cruz, Rodrigo, Domínguez García, Delia Inés, Hernández Gutiérrez, Rodolfo, Lagunes Quintanilla, Rodolfo Esteban, Ortuño Sahagún, Daniel, González Castillo, Celia, Gutiérrez Ortega, Abel, Herrera Rodríguez, Sara Elisa, Vallejo Cardona, Adriana, Martínez Velázquez, Moisés
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.11.2016
• Subjects: Animals; Arachnid Vectors - immunology; Arthropod Proteins - immunology; Blotting, Western; Boophilus microplus; Cattle; Cattle Diseases - prevention & control; DNA, Complementary - biosynthesis; Electrophoresis - methods; Embryonic Development - immunology; Enzyme-Linked Immunosorbent Assay; Female; Immunogen; Immunoglobulin G - blood; Ixodidae; Larva - immunology; Oogenesis - immunology; Ovary; Ovary - immunology; Polymerase Chain Reaction; Proteomics - methods; Reproduction - immunology; Rhipicephalus; Rhipicephalus - immunology; RNA - genetics; RNA - isolation & purification; Tandem Mass Spectrometry; Tick; Tick Control - methods; Tick Infestations - prevention & control; Tick Infestations - veterinary; Vaccine; Vaccines; Vitellogenin; Vitellogenins - biosynthesis; Vitellogenins - immunology; Ovary; Tick; Boophilus microplus; Vaccine; Vitellogenin; Immunogen
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2016.10.005

Rhipicephalus (Boophilus) microplus ticks are obligatory hematophagous ectoparasites of cattle and act as vectors for disease-causing microorganisms. Conventional tick control is based on the use of chemical acaricides; however, their uncontrolled use has increased tSresistant tick populations, as well as food and environmental contamination. Alternative immunological tick control has shown to be partially effective. The only anti-tick vaccine commercially available at present in the world is based on intestinal Bm86 protein, and shows a variable effectiveness depending on tick strains or geographic isolates. Therefore, there is a need to characterize new antigens in order to improve immunological protection. The aim of this work was to identify immunogenic proteins from ovarian tissue extracts of R. microplus, after cattle immunization. Results showed that ovarian proteins complexed with the adjuvant Montanide ISA 50 V generated a strong humoral response on vaccinated cattle. IgG levels peaked at fourth post-immunization week and remained high until the end of the experiment. 1D and 2D SDS-PAGE-Western blot assays with sera from immunized cattle recognized several ovarian proteins. Reactive bands were cut and analyzed by LC-MS/MS. They were identified as Vitellogenin, Vitellogenin-2 precursor and Yolk Cathepsin. Our findings along with bioinformatic analysis indicate that R. microplus has several Vitellogenin members, which are proteolytically processed to generate multiple polypeptide fragments. This apparent complexity of vitellogenic tick molecular targets gives the opportunity to explore their potential usefulness as vaccine candidates but, at the same time, imposes a challenge on the selection of the appropriate set of antigens. [Display omitted] •A classical approach immunizing cattle with total ovarian proteins was followed.•The detection of ovarian proteins was achieved by ELISA plates coated with proteins from larval homogenates.•Several immunogenic proteins such as Vitellogenins and yolk Cathepsin were identified.•Vitellogenin-1 and Vitellogenin-2 transcripts are synthesized in the ovaries of R. microplus.•Vitellogenins represent attractive candidates for development of anti-tick vaccines.