Activation of Colon Cancer-Draining Lymph Node Mononuclear Cells by 30 kDa or Triton X-100 Solubilized Protein Antigen of Mycobacterium tuberculosis

ABSTRACT

Tumor-draining lymph node mononuclear (TDLMN) cells are specifically sensitized to the growing tumor but such cells are deficient for mediating an antitumor response. In this study, we examined the feasibility of using mycobacterial 30 kDa or Triton X-100 solubilized protein (TSP) antigen to stimulate mononuclear cells of colon cancer-draining lymph node for the generation of cell mediated immune effector cells. The proliferative response of TDLMN cells stimulated with mycobacterial 30 kDa or TSP antigen was determined by H-thymidine incorporation assay. The proliferation of TDLMN cells to mycobacterial 30 kDa or TSP antigen was significantly increased in PPD (+) patients, but a poor response to the 30 kDa or TSP antigen was observed in PPD (-). The expression on ro T cells to mycobacterial 30 kDa or TSP antigen was assessed by flow cytometry. The ro T cells from PPD (+) patient responded only to 30 kDa antigen but to TSP antigen. An investigation of cytokine mRNA expression was undertaken using reverse transcription- polymerase chain reaction (RT-PCR) to follow TDLMN cells stimulated with the 30 kDa or TSP antigens for 5 days. The IFN-r and TNF-a mRNA expression was only induced in TDLMN cells of PPD (+) patient in response to the 30 kDa or TSP antigen. The IL-2 mRNA expression was induced in both PPD (+) and PPD (-) in response to the 30 kDa or TSP antigen. But the IL-4 mRNA expression was not induced in response to the 30 kDa or TSP antigen. These results suggest that the 30 kDa and TSP antigens may serve as biologic response modifier for the generation of cell mediated immune effector cells.