Analysis of Stage-Specific Protein Expression during Babesia Bovis Development within Female Rhipicephalus Microplus.
Arthropod-borne protozoan pathogens have a complex life cycle that includes asexual reproduction of haploid stages in mammalian hosts and the development of diploid stages in invertebrate hosts. The ability of pathogens to invade, survive, and replicate within distinct cell types is required to maintain their life cycle. In this study, we describe a comparative proteomic analysis of a cattle pathogen, Babesia bovis, during its development within the mammalian and tick hosts with the goal of identifying cell-surface proteins expressed by B. bovis kinetes as potential targets for the development of a transmission blocking vaccine. To determine parasite tick-stage-specific cell-surface proteins, CyDye labeling was performed with B. bovis blood stages from the bovine host and kinetes from the tick vector. Cell-surface kinete-stage-specific proteins were identified using 2D difference in gel electrophoresis and analyzed by mass spectrometry. Ten proteins were identified as kinete-stage-specific, with orthologs found in closely related Apicomplexan pathogens. Transcriptional analysis revealed two genes were highly expressed by kinetes as compared with blood stages. Immunofluorescence using antibodies against the two proteins confirmed kinete-stage-specific expression. The identified cell-surface kinete proteins are potential candidates for the development of a B. bovis transmission blocking vaccine.
Animal Disease Research Unit, USDA-ARS , Pullman, Washington 99164, United States.
Wendell C
WC
Johnson
Animal Disease Research Unit, USDA-ARS , Pullman, Washington 99164, United States.|Program in Vector-borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University , Pullman, Washington 99164, United States.
Naomi S
NS
Taus
Animal Disease Research Unit, USDA-ARS , Pullman, Washington 99164, United States.|Program in Vector-borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University , Pullman, Washington 99164, United States.
Kathryn E
KE
Reif
Program in Vector-borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University , Pullman, Washington 99164, United States.
Gamila A R
GA
Bohaliga
Animal Disease Research Unit, USDA-ARS , Pullman, Washington 99164, United States.
Lowell S
LS
Kappmeyer
Animal Disease Research Unit, USDA-ARS , Pullman, Washington 99164, United States.|Program in Vector-borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University , Pullman, Washington 99164, United States.|Paul G. Allen School for Global Animal Health, Washington State University , Pullman, Washington 99164, United States.
Massaro W
0000-0002-2680-6091
MW
Ueti
D000818:Animals; D016793:Babesia bovis / Q000737:chemistry* / Q000254:growth & development; D002417:Cattle; D005260:Female; D020869:Gene Expression Profiling; D008018:Life Cycle Stages / Q000502:physiology*; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics; D040901:Proteomics / Q000379:methods*; D048494:Rhipicephalus / Q000382:microbiology*
D008565:Membrane Proteins
Babesia bovis; bovine and Rhipicephalus ticks; kinete; stage-specific protein