Black-legged ticks (Ixodes scapularis) transmit several disease-causing pathogens to humans and animals. In the northeastern and central U.S., Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti parasites are transmitted to vertebrate hosts by infected I. scapularis. This tick’s multifunctional salivary glands are vital to their biological success and likely also play a critical role in transmission of disease-causing agents to their hosts. It may be that tick salivary proteins can be used in an anti-tick feeding vaccination strategy. To begin testing this hypothesis, we have cloned 42 full-length tick salivary gland secretory genes into a mammalian expression vector and evaluated their ability to induce a cellular immune response in guinea pigs sensitized with tick bites, tick saliva, or tick salivary gland homogenate. Only 17 of these salivary genes induced a cellular immune response, as evidenced by delayed-type hypersensitivity in the skin. This reverse immunology approach may be a useful high-throughput screening method for identifying potential anti-tick vaccines. Experiments are underway to evaluate the pathogen-blocking potential of this vaccination strategy.