Ticks are vectors of different pathogens causing human and animal diseases. Particularly, Rickettsia slovaca is zoonotic infectious bacterium transmitted by Dermacentor ticks, agent of tick-borne lymphadenopathy (TIBOLA), common across Europe. Current studies point to extreme complexity of bacterial induced effects in tick host. Systems biology tools, including proteomics, greatly contribute to understanding of molecular details of pathogen-tick-host interactions. Herein we compared laboratory-infected ticks with uninfected control after four weeks of incubation. Propagation of R. slovaca was confirmed by quantitative PCR. Using DNA was confirmed infection with R. slovaca. By proteomic approach, we discovered 33 differentially abundant gel spots, 23 of them accumulated upon artificial infection with R. slovaca. Modest 6.9% of tick proteome was affected. The protein localizations showing that eight proteins spots might be secreted, three cytoplasmic, two mitochondrial, six likely having multiple localizations, one cell membrane and one nucleus. We identified following proteins defensin, serpins, glycine-rich protein, heat shock protein involved in artificially infected tick vector, Dermacentor reticulatus. Discovered differentially abundant proteins should be further evaluated as targets to block the transmission of bacterial pathogen.