title: Molecular and immunological characterization of three strains of Anaplasma marginale grown in cultured tick cells.
abstract: Anaplasma marginale is an economically important tick-borne pathogen of cattle that causes bovine anaplasmosis. A wide range of geographic strains of A. marginale have been isolated from cattle, several of which have been characterized using genomics and proteomics. While many of these strains have been propagated in tick lines, comparative analyses after propagation in tick cells have not been reported. The overall purpose of this research therefore was to compare the degree of conservation of selected genes after propagation in tick cell culture among A. marginale strains from the U.S. (the Virginia strain) and Brazil (UFMG1 and UFMG2 strains). The genes studied herein included those which encode the proteins HSP70 and SODB involved in heat shock and stress responses, respectively, and two genes that encode major surface proteins MSP4 and MSP5. Strain identities were first confirmed by sequencing the tandem repeats of the msp1a gene which encodes for the adhesin, MSP1a. The results of these studies demonstrated that the genes encoding for both stress response and heat shock proteins were highly conserved among the three A. marginale strains. Antibodies specific for MSP4, MSP5, SODB and HSP70 proteins were used to further characterize the A. marginale strains, and they reacted with all of these strains propagated in tick cell culture, providing further evidence for antigenic conservation. Although antigenic differences were not found among the three A. marginale strains, multi-locus sequence analysis (MLSA) performed with nucleotide sequences of these genes demonstrated that the A. marginale Brazilian and U.S. strains fall in different clades. These results showed that phylogenetically distant strains of A. marginale are antigenically conserved, even after several in vitro passages, supporting the use of some of the above conserved proteins as candidates for universal vaccines.
authors.affiliation: Comparative Tropical Medicine and Parasitology, Ludwig-Maximilians-Universität München, Leopoldstrasse 5, 80802 Munich, Germany. Electronic address: Katarzyna.Lis@tropa.vetmed.uni-muenchen.de.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Lis
authors.affiliation: SaBio, Instituto de Investigación de Recursos Cinegéticos, IREC-CSIC-UCLM-JCCM, 13005 Ciudad Real, Spain. Electronic address: MariaIsabel.Garcia@uclm.es.
authors.forename: Isabel G
authors.initials: IG
authors.lastname: Fernández de Mera
authors.affiliation: SaBio, Instituto de Investigación de Recursos Cinegéticos, IREC-CSIC-UCLM-JCCM, 13005 Ciudad Real, Spain. Electronic address: marina.popara@gmail.com.
authors.forename: Marina
authors.initials: M
authors.lastname: Popara
authors.affiliation: SaBio, Instituto de Investigación de Recursos Cinegéticos, IREC-CSIC-UCLM-JCCM, 13005 Ciudad Real, Spain; Center for Infection and Immunity of Lille (CIIL), INSERM U1019 - CNRS UMR 8204, Université Lille Nord de France, Institut Pasteur de Lille, Lille, France. Electronic address: cabezasalejandrocruz@gmail.com.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Cabezas-Cruz
authors.affiliation: SaBio, Instituto de Investigación de Recursos Cinegéticos, IREC-CSIC-UCLM-JCCM, 13005 Ciudad Real, Spain. Electronic address: Nieves.Ayllon@uclm.es.
authors.forename: Nieves
authors.initials: N
authors.lastname: Ayllón
authors.affiliation: Comparative Tropical Medicine and Parasitology, Ludwig-Maximilians-Universität München, Leopoldstrasse 5, 80802 Munich, Germany; Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Onderstepoort 0110, South Africa. Electronic address: zweygarthe@gmail.com.
authors.forename: Erich
authors.initials: E
authors.lastname: Zweygarth
authors.affiliation: Comparative Tropical Medicine and Parasitology, Ludwig-Maximilians-Universität München, Leopoldstrasse 5, 80802 Munich, Germany; Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Antonio Carlo, 6627, Belo Horizonte 30161-970, Brazil. Electronic address: lygia.passos@yahoo.com.
authors.forename: Lygia M F
authors.initials: LM
authors.lastname: Passos
authors.affiliation: Comparative Tropical Medicine and Parasitology, Ludwig-Maximilians-Universität München, Leopoldstrasse 5, 80802 Munich, Germany. Electronic address: Marzena.Broniszewska@tropa.vetmed.uni-muenchen.de.
authors.forename: Marzena
authors.initials: M
authors.lastname: Broniszewska
authors.affiliation: SaBio, Instituto de Investigación de Recursos Cinegéticos, IREC-CSIC-UCLM-JCCM, 13005 Ciudad Real, Spain. Electronic address: margaritam.villar@uclm.es.
authors.forename: Margarita
authors.initials: M
authors.lastname: Villar
authors.affiliation: Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078, USA. Electronic address: Katherine.Kocan@okstate.edu.
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Kocan
authors.affiliation: Departamento de Parasitologia, ICB-UFMG, Belo Horizonte, Brazil. Electronic address: muciobr@icb.ufmg.br.
authors.forename: Mucio F B
authors.initials: MF
authors.lastname: Ribeiro
authors.affiliation: Comparative Tropical Medicine and Parasitology, Ludwig-Maximilians-Universität München, Leopoldstrasse 5, 80802 Munich, Germany. Electronic address: Kurt.Pfister@tropa.vetmed.uni-muenchen.de.
authors.forename: Kurt
authors.initials: K
authors.lastname: Pfister
authors.affiliation: SaBio, Instituto de Investigación de Recursos Cinegéticos, IREC-CSIC-UCLM-JCCM, 13005 Ciudad Real, Spain; Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078, USA. Electronic address: jose_delafuente@yahoo.com.
authors.forename: José
authors.initials: J
authors.lastname: de la Fuente
mesh_terms: D042322:Anaplasma marginale / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D000712:Anaplasmosis / Q000276:immunology* / Q000382:microbiology; D000818:Animals; D000940:Antigenic Variation; D001096:Arachnid Vectors / Q000382:microbiology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology*; D001938:Brazil; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology* / Q000382:microbiology; D017124:Conserved Sequence; D008969:Molecular Sequence Data; D010802:Phylogeny; D013987:Ticks / Q000382:microbiology*; D014481:United States
chemical_list: D001426:Bacterial Proteins
keywords: Anaplasma marginale; Hsp70; IDE8 tick cell line; Major surface proteins; Sodb; Stress response proteins