Programming of CTL with heat-killed Brucella abortus and antigen allows soluble antigen alone to generate effective secondary CTL.

Optimal generation of cytotoxic T cell (CTL) responses continues to be a challenge in the production of vaccines against pathogens such as HIV-1, in part because it is difficult to introduce soluble protein antigens (Ag) into the MHC class I pathway. Using heat-killed Brucella abortus (HKBA) as an adjuvant and ovalbumin (ova) protein as an Ag, we demonstrated that a high dose of Ag was required for systemic and effective CTL. In an adoptive transfer model, primary and secondary ova-specific OT-1 CD8 cell expansion by HKBA plus high dose of ova were partially CD4 T cell-dependent. Interestingly, primary stimulation with HKBA plus ova allowed effective secondary stimulation with ova alone that was equivalent to HKBA plus ova in terms of IFN-gamma production from Ag-specific CD8 cells. Thus a combination of adequate Ag dose, and selection of appropriate adjuvants can meet the threshold not only for primary effective CTL responses to soluble protein Ags but for secondary CTL responses following stimulation with protein Ag alone.