The objective of this research was to evaluate the effects of early vaccination on the phenotype (i.e., activation marker expression) and functional capacity of B cell populations in neonatal calves. In the first of 2 experiments, 6 calves were vaccinated with ovalbumin at 3 and 5 wk of age. Three of the 6 calves also were vaccinated with Mycobacterium bovis, strain bacillus Calmette-Guerin (BCG) at 3 wk of age. Mycobacterium bovis lipoarabinomannan-reactive IgG1 and IgG2 were detected in calf sera prior to vaccination, indicative of colostral transfer of maternal Ig cross-specific to BCG. Ovalbumin-specific IgG1 and IgG2 were not detected before vaccination. Vaccination of 3-wk-old calves with ovalbumin elicited antigen-specific IgG1 and IgG2 anti-body responses that were amplified by secondary vaccination. Vaccination with BCG did not elicit a measurable antibody response. In the second experiment, 6 calves were vaccinated with ovalbumin at 3 and 5 wk of age in addition to BCG at 3 wk of age. Lymph node cell populations stimulated with ovalbumin had decreased CD5, CD21, and CD40 expression and increased B-B2, CD25, and CD80 expression on IgM+ cells. Stimulation of the same population with purified-protein derivative increased CD25 and CD80 expression on IgM+ cells. Expression of activation molecules on ovalbumin- and purified protein derivative-stimulated CD5+ IgM+ cells was similar to expression on the larger IgM+ cell population. An increased expression of major histocompatibility class II on CD5+ IgM+ cells after stimulation was the only exception. Interestingly, IgM+ cells isolated from the superficial cervical lymph node draining the vaccination site, but not from the opposing cervical lymph node, responded to antigen stimulation in vitro. In conclusion, calves generated B cell responses to ovalbumin and BCG after vaccination. Additional studies are necessary to determine whether maternal immunologic experience transferred via colostral immunoglobulin inhibits production of mycobacteria-specific immunoglobulin production in the calf.