Characterization of tick cell lines and their use as a substrate for the growth, maintenance and protein expression profile of Anaplasma marginale

• Grant number: 20/03190-9
• Support Opportunities: Regular Research Grants
• Start date: November 01, 2020
• End date: October 31, 2022
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Darci Moraes Barros-Battesti
• Grantee: Darci Moraes Barros-Battesti
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil
• Associated researchers: Daniel Guariz Pinheiro ; Diogo Cavalcanti Cabral de Mello ; Diva Denelle Spadacci Morena ; João Fabio Soares ; Luís Adriano Anholeto ; Marcos Rogério André ; Maria Izabel Souza Camargo ; Pablo Henrique Nunes ; Rosangela Zacarias Machado ; Tiago Santana Balbuena

Abstract

Ticks are arthropods of great medical and veterinary importance, with many species responsible for transmitting viruses, bacteria and protozoa to their hosts. Some of the tick-borne pathogens are difficult to grow in vitro, and do not grow in artificial media. Tick cell culture has been an excellent tool to isolate and study pathogens, allowing the production of antigenic material for diagnostic tests, production of antibodies and vaccines, and for studies of host-vector-pathogen relationships. In this sense, this study aims to characterize tick cell lines and use them as a substrate for the growth, maintenance and protein expression profile of Anaplasma marginale. For this, we will use primary cultures and established lineages of tick cells from Amblyomma sculptum (Berlese) and Rhipicephalus (Boophilus) microplus (Canestrini) eggs. The primary cultures will be prepared with egg masses and kept at 30 ° C in L-15B medium (Leibovitz) supplemented with Bovine Fetal Serum, subcultured when they reach the necessary confluence and cryopreserved in several passages. For the characterization of cells in culture, morphological characterization with histological techniques and karyotyping will be performed. The cells inoculated with Anaplasma marginale canis will be analyzed by slide preparations and counts of infected cells under optical microscopy, by real-time PCR and by Transmission Electron Microscopy. In addition, a differential proteomic analysis of infected and uninfected cells will be carried out, aiming at the search for proteins that may be used as vaccine antigens in the future. (AU)

Scientific publications

(4)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

LIMA-DUARTE, LEIDIANE; CASTRO-SANTIAGO, ANA CAROLINA; CAMARGO, JAQUELINE VALERIA; FERRETTI, ANA BEATRIZ STEIN MACHADO; ANHOLETO, LUIS ADRIANO; PEREIRA, MELISSA CAROLINA; IKEDA, PRISCILA; PEREZ, CARLOS ALBERTO; SANCHES, GUSTAVO SERON; CAMARGO-MATHIAS, MARIA IZABEL; et al. Original Establishment and multiapproach characterization of Amblyomma sculptum (Acari: Ixodidae) cell line (ASE-14) from Brazil. TICKS AND TICK-BORNE DISEASES, v. 13, n. 4, p. 10-pg., 2022-07-01. (20/03190-9, 15/26209-9)

LUZZI, MAYARA DE CASSIA; LOPES DE CARVALHO, LUCAS AMOROSO; PINHEIRO, DANIEL GUARIZ; LIMA-DUARTE, LEIDIANE; CAMARGO, JAQUELINE VALERIA; KISHI, LUCIANO TAKESHI; FERNANDES, CAMILA CESARIO; MACHADO, ROSANGELA ZACARIAS; SOARES, JOAO FABIO; ANDRE, MARCOS ROGERIO; et al. Analysis on the prokaryotic microbiome in females and embryonic cell cultures of Rhipicephalus sanguineus tropical and temperate lineages from two specific localities in Brazil. REVISTA BRASILEIRA DE PARASITOLOGIA VETERINARIA, v. 30, n. 3, 2021. (15/26209-9, 20/03190-9)

LIMA-DUARTE, LEIDIANE; CAMARGO, JAQUELINE VALERIA; CASTRO-SANTIAGO, ANA CAROLINA; MACHADO, ROSANGELA ZACARIAS; ANDRE, MARCOS ROGERIO; CABRAL-DE-MELLO, DIOGO CAVALCANTI; CAMARGO-MATHIAS, MARIA IZABEL; IKEDA, PRISCILA; ANHOLETO, LUIS ADRIANO; PEREIRA, MELISSA CAROLINA; et al. Establishment and characterization of a cell line (RBME-6) of Rhipicephalus (Boophilus) microplus from Brazil. TICKS AND TICK-BORNE DISEASES, v. 12, n. 5, SEP 2021. (15/26209-9, 20/03190-9)

CASTRO-SANTIAGO, ANA CAROLINA; LIMA-DUARTE, LEIDIANE; CAMARGO, JAQUELINE VALERIA; DE ALMEIDA, BEATRIZ ROCHA; SIMONS, SIMONE MICHAELA; MATHIAS, LUIS ANTONIO; BASSINI-SILVA, RICARDO; MACHADO, ROSANGELA ZACARIAS; ANDRE, MARCOS ROGERIO; BARROS-BATTESTI, DARCI MORAES. Artificial Feeding of Ornithodoros fonsecai and O. brasiliensis (Acari: Argasidae) and Investigation of the Transstadial Perpetuation of Anaplasma marginale. MICROORGANISMS, v. 11, n. 7, p. 11-pg., 2023-07-01. (20/03190-9, 21/06758-9, 19/09915-8, 20/11755-6)