Adoptive transfer of tumor-primed, in vitro-activated, CD4(+) T effector cells (T(E)s) combined with CD8(+) T(E)s provides intraturnoral T-E proliferation and synergistic antitumor response

The importance of CD4(+) Th1 cells during the effector phase of the antitumor response has been overshadowed by emphasis on CD8(+) cytotoxic T lymphocytes (CTLs). To determine their respective functions, we purified antigen-primed T cells from tumor-draining lymph nodes and separately activated CD4+ and CD8(+) subsets in vitro. Adoptive transfer of CD4(+) T effector cells (TES) combined with CD8(+) TES provided synergistic therapy for mice bearing subcutaneous, intracranial, or advanced pulmonary metastases. CD4+ TES augmented IFN-gamma production by CD8(+) TES when cells were stimulated by tumor digest-containing antigen-presenting cells (APCs). CD4(+) TES infiltrated and proliferated extensively in pulmonary tumors, while also stimulating tumor antigen-specific CD8(+) T cells. By contrast, CD8(+) TES showed minimal intratumoral proliferation in the absence of CD4+ cells or when systemically transferred CD4(+) cells were prevented from infiltrating pulmonary tumors by pretreatment with pertussis toxin. Irradiation of CD4+ T cells immediately prior to adoptive transfer abrogated their intratumoral proliferation and direct antitumor efficacy but did not block their capacity to stimulate intratumoral CD8+ T-E proliferation or tumor regression. These results highlight the importance of cross-presentation of tumor antigens during the effector phase of immunotherapy and suggest that approaches to stimulate CD4(+) TE function and boost APC cross-presentation within tumors will augment cancer immunotherapy.