Identification of Anaplasma marginale Proteins Specifically Upregulated during Colonization of the Tick Vector

• Published in: Infection and Immunity 2010-07, Vol.78 (7), p.3047-3052
• Main Authors: Ramabu, Solomon S, Ueti, Massaro W, Brayton, Kelly A, Baszler, Timothy V, Palmer, Guy H
• Format: Article
• Language: English
• Subjects: Anaplasma marginale; Anaplasma marginale - physiology; Anaplasmosis - microbiology; Animals; Arachnid Vectors - microbiology; Arthropoda; Bacterial Infections; Bacterial Proteins - biosynthesis; Bacterial Proteins - isolation & purification; Bacterial Proteins - physiology; Bacteriology; Biological and medical sciences; Blotting, Western; Dermacentor - microbiology; Ehrlichia; Electrophoresis, Gel, Two-Dimensional; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Bacterial - physiology; Ixodidae; Life Cycle Stages - physiology; Microbiology; Miscellaneous; Proteomics; Up-Regulation
• ISSN: 0019-9567; 1098-5522
• DOI: 10.1128/IAI.00300-10

The transition between infection of the mammalian host and colonization of an arthropod vector is required for the ongoing transmission of a broad array of pathogens, from viruses to protozoa. Understanding how this transition is mediated provides opportunities to disrupt transmission through either chemotherapy or immunization. We used an unbiased proteomic screen to identify Anaplasma marginale proteins specifically upregulated in the tick compared to the mammalian host. Comparative mass spectrometric analysis of proteins separated by two-dimensional gel electrophoresis of uninfected and infected ISE6 cells and infected mammalian cells identified 15 proteins exclusively expressed or upregulated in tick cells. All 15 had originally been annotated as hypothetical proteins. We confirmed quantitative upregulation and expression in situ within the midgut epithelial and salivary gland acinar cells of vector ticks during successful transmission. The results support the hypothesis that A. marginale gene expression is regulated by the specific host environment and, in a broader context, that the core genome evolved in the arthropod vector with differential regulation, allowing adaptation to mammalian hosts. Furthermore, the confirmation of the in situ expression of candidates identified in ISE6 cell lines indicates that this approach may be widely applicable to bacteria in the genera Anaplasma and Ehrlichia, removing a major technical impediment to the identification of new targets for vaccine and chemotherapeutic blocking of transmission.