To determine if inactivated poliovirus vaccine combined with diphtheria and tetanus toxoids and whole-cell pertussis vaccine interferes with the immunogenicity of pertussis vaccine, we performed a randomized trial of diphtheria and tetanus toxoids and pertussis vaccine combined with inactivated poliovirus vaccine given as a single injection or as two separate injections at the same visit to infants immunized at 2, 4, and 6 months of age. A total of 84 infants were enrolled in the study; 44 received the single injection, and 40 received separate injections. Before immunization, there were no differences in antibody values between the two groups. After two vaccine doses, infants immunized with the single injection had significantly lower values of antibody to filamentous hemagglutinin (8.3 vs. 23.7 ELISA units; P < .001), fimbriae (266.5 vs. 771.8 ELISA units; P < 0.01), and the 69-kD membrane protein (442.2 vs. 1,352 ELISA units; P < .001). After the third dose, these differences persisted, and differences were also detected for antibody to pertussis toxin (10.0 vs. 35.5 ELISA units; P < .001) and a whole-bacteria antigen preparation (2,667 vs. 3,829 ELISA units; P < .05). We conclude that there is a diminished antibody response to the pertussis vaccine when inactivated poliovirus vaccine is combined with the diphtheria and tetanus toxoids and pertussis vaccine.