The Gram-negative bacterium Holospora obtusa is a macronucleus-specific symbiont of the ciliate Paramecium caudatum. The infectious form of this bacterium infects the host macronucleus through digestive vacuoles and differentiates into the reproductive form two days after the infection in the nucleus. The monoclonal antibodies IF-3-1 and IF-3-2 reacted with 39 and 15 kDa periplasmic proteins, respectively, that were specific for the infectious form of H. obtusa. Because the antigens were not detected in the reproductive form of the bacterium, it appears that expression of the proteins decreases during or soon after the infection. Using these antibodies, quantitative changes in the antigens in the early infection process were examined by immunoblotting and immunogold electron microscopy. Immunoblotting showed that the amounts of both antigens were reduced within 1 h after the bacteria were engulfed into the digestive vacuoles of the paramecia, but that the amounts of IF-3-2 antigens declined earlier than the IF-3-1 antigen. Immunogold labeling showed that the level of IF-3-2 antigens became very low in the bacteria in the host digestive vacuoles, whereas there was no similar decrease in amount of IF-3-1 antigens. Possible functions of the antigens are discussed. The IF-3-1 antigens decrease in concentration in parallel with the decrease in the periplasmic region.