Ingegerd Hellström* ¹, Jeffrey A. Ledbetter*, Nathalie Scholler*, Yi Yang*, Zhengmao Ye*, Gary Goodman ², Janice Pullman*, Martha Hayden-Ledbetter*, and Karl Erik Hellström*

* Pacific Northwest Research Institute, 720 Broadway, and  ² Swedish Hospital and Medical Center, Tumor Institute, 747 Broadway, Seattle, WA 98122

¹ To whom reprint requests should be addressed.     E-mail:    ihellstrom@pnri.org. 

Lymphocytes from blood or tumors of patients with advanced cancer did not proliferate and produced very low levels of tumor necrosis factor and IFN-g when cultured with autologous tumor cells. Proliferation and lymphokine production dramatically increased in the presence of beads conjugated with mAbs to CD3 plus mAbs to CD28 and/or CD40, and the lymphocytes destroyed the tumor cells. Expression density of CD3 concomitantly increased from low to normal levels. Furthermore, beads providing a CD3 signal (in combination with CD28 or CD28 plus CD40) gave partial protection against the inhibitory effect of transforming growth factor type ^b1 on lymphocyte proliferation and production of tumor necrosis factor and IFN-g. MHC class I-restricted cytolytic T cells lysing autologous tumor cells in a 4-h Cr⁵¹ release assay were generated when peripheral blood leukocytes were activated in the presence of autologous tumor cells and anti-CD3/CD28 or anti-CD3/CD28/CD40 beads. Experiments performed in a model system using anti-V-^b1 or anti-V-^b2 mAbs to activate subsets of T cells expressing restricted T cell receptor showed that lymphocytes previously activated by anti-V-^b can respond to CD3 stimulation with vigorous proliferation and lymphokine production while retaining their specificity, also in the presence of transforming growth factor type ^b1. Our results suggest that T lymphocytes from cancer patients can proliferate and form Th1 type lymphokines in the presence of autologous tumor cell when properly activated, and that antigen released from killed tumor cells and presented by antigen-presenting cells in the cultures facilitates the selective expansion of tumor-directed, CD8⁺ cytolytic T cells. 

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