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Solid-Phase Anti-CD3 Antibody Activation of Murine Tumor-infiltrating Lymphocytes¹

Department of Surgery, Division of Surgical Oncology, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts 02115

Seventeen consecutive s.c. murine tumors, derived from a sarcoma and a colon adenocarcinoma, were cultured in the presence of recombinant interleukin 2 (rIL-2) for growth of tumor-infiltrating lymphocytes (TIL). Identical cultures were activated by solid-phase monoclonal antibody directed against the murine CD3 -chain, in conjunction with rIL-2. Forty-eight h later, cells were replated in rIL-2 alone. Proliferation of anti-CD3-stimulated cultures was 1- to 17-fold greater than those cultured with rIL-2 alone (P < 0.05). Both culture conditions yielded TIL which stained >80% Thy-1.2⁺/Lyt-2⁺ (P > 0.05), <7% Thy-1.2⁺/L3T4⁺ (P > 0.05). Regardless of culture condition, longitudinal studies of in vitro cytotoxicity generated from 10 TIL preparations revealed no significant differences between the ability of TIL to lyse the murine natural killer-sensitive line YAC or heterologous or autologous tumor (P > 0.05). In vivo antitumor activity of TIL was tested by the adoptive transfer of suboptimal doses of TIL plus systemic rIL-2 to mice with pulmonary micrometastatic disease. No difference in tumor regression was noted between the TIL cultured with anti-CD3 plus rIL-2 or with rIL-2 alone (P > 0.05). Anti-CD3 stimulation of murine TIL cultures significantly increases lymphocyte cell yield without alteration of their phenotype, in vitro tumoricidal activity, or in vivo therapeutic effect.

¹ This work was supported in part by NIH Grant CA-45484. Presented at 42nd Annual Cancer Symposium of The Society of Surgical Oncology, San Francisco, CA, May 22, 1989.

² Recipient of an American Cancer Society Career Development Award. To whom requests for reprints should be addressed.

Received 10/ 3/89. Revised 12/21/89.

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