Follicular lymphoma (FL) is a disease that responds to current treatment regimens; however, patients in general relapse with increasingly refractory disease. Idiotype-based vaccines are currently under trial for the treatment of FL. These vaccines comprise the patient's BCR idiotype (Id) as the tumor antigen conjugated to the protein carrier Keyhole Limpet Hemocyanin (KLH); however, other protein carriers may enhance the immune response to the lymphoma Id. In this study we investigated whether an alternate carrier, Listeriolysin O (LLO), would amplify the immune response to Id protein and provide better protection against challenge by 38C13 murine lymphoma. The Id-LLO vaccine compared favorably against Id-KLH in tumor-protection studies and both vaccines provided systemic immunity against 38C13 lymphoma. However, the immune response to the two conjugates was different in that Id-LLO induced a more powerful Th1 response characterized by high titer IgG2a anti-Id antibodies after one immunization and the presence of CD4 cells secreting IFN-gamma. In vivo studies demonstrated that immune serum contributed to the anti-lymphoma efficacy seen following Id-LLO immunization. Interestingly, Id-LLO immunized mice, when challenged twice with 38C13 lymphoma provided better protection against challenge by the BCR loss variant 38C13-V2, suggesting that Id-LLO immunized mice have more potential to develop epitope spreading than Id-KLH. In conclusion, Id-LLO compared favorably against Id-KLH in its anti-lymphoma efficacy. Furthermore, Id-LLO induced a more potent humoral and cell-mediated immune response and promoted epitope spreading after lymphoma challenge. Thus, anti-Id vaccines incorporating LLO may be a better therapeutic option for treatment of B-cell lymphoma.