Differential expression of the tick protective antigen subolesin in anaplasma marginale- and A. phagocytophilum-infected host cells.

Subolesin was recently shown in vaccine and RNA interference (RNAi) studies to protect against tick infestations and to affect tick feeding, reproduction, and development as well as infection of host cells by Anaplasma marginale and A. phagocytophilum. Recent experiments provided evidence that infection of both tick and vertebrate host cells with these two pathogens modified gene expression. We therefore hypothesized that infection of host cells with A. marginale and A. phagocytophilum affects expression of subolesin. Subolesin mRNA levels were determined by real-time reverse transcriptase (RT)-PCR in uninfected and A. marginale-infected Dermacentor variabilis guts and salivary glands and IDE8-cultured tick cells and in uninfected and A. phagocytophilum-infected Ixodes scapularis nymphs, ISE6-cultured tick cells, and the human cell line HL-60. In addition, the effect of subolesin on Anaplasma spp. infection/multiplication was characterized by RNAi in tick tissues and/or cultured tick and human cells. These experiments presented evidence of differential expression of subolesin in A. marginale- and A. phagocytophilum-infected cells. Subolesin was differentially expressed in A. marginale-infected ticks in a tissue-specific manner in which mRNA levels increased in response to A. marginale infection in tick salivary gland cells but not in the gut cells. Subolesin knockdown by RNAi reduced Anaplasma infection/multiplication only in cells in which infection increased subolesin expression, i.e., in A. marginale-infected D. variabilis salivary glands and IDE8 cells. The results reported herein further support the role of subolesin in Anaplasma-host interactions and suggest a putative role of subolesin in vaccines for the control of pathogen infection/multiplication in ticks.