Successful Induction of Tumor‐specific Cytotoxic T Lymphocytes from Patients with Non‐small Cell Lung Cancer Using CD80‐transfected Autologous Tumor Cells

Abstract

Cytotoxic T lymphocytes (CTL) against human lung cancer cells are difficult to induce by a conventional method using tumor cell stimulation probably due to an insufficiency of tumor antigens (TA) or costimulatory molecules such as CD80. We, therefore, investigated the potential of CD80-transfected tumor cells as stimulators of the in vitro induction of autologous tumor-specific CTL from regional lymph node lymphocytes in patients with lung cancer. Five non-small cell lung cancer cell lines (two adenocarcinomas, 1 squamous cell carcinoma, 1 large cell carcinoma and 1 adenosquamous cell carcinoma) were established from surgical specimens and were successfully transduced with a plasmid constructed with expression vector pBj and human CD80 cDNA, using a lipofection method. CD80-transfected tumor cells (CD80-AT) significantly augmented the proliferation of autologous lymphocytes from all cases as compared with non-transfected tumor cells (AT). AT-stimulated lymphocytes from 4 out of 5 cases did not show any cytotoxicity against AT;however, lymphocytes stimulated with CD80-AT exhibited substantial cytotoxicity against parental AT in all 5 cases tested. AT-stimulated lymphocytes derived from only one out of 5 cases showed major histocompatibility complex (MHC)-class I-restricted cytokine production in response to AT, while the MHC-class I-restricted responses were found in CD80-AT-stimulated lymphocytes from 4 out of 5 cases. These results indicate that CD80 on tumor cells could be a beneficial costimulatory molecule to elicit CTL against lung cancer, and also show that TA recognized by CTL was frequently expressed on lung cancer cells.

Introduction of theCIITAgene into tumor cells produces exosomes with enhanced anti-tumor effects

“…Many strategies in cancer immunotherapy have aimed to trigger anti-tumor immune responses by an MHC class I-restricted tumor-specific CTL response (Takenoyama et al, 2001;Kass et al, 2003). However, to generate efficiently CD8+ T effector and memory cells, CD4+ T helper (Th) cells are essentially required (Baxevanis et al, 2000).…”

A different pattern of cytotoxic T lymphocyte recognition against primary and metastatic tumor cells in a patient with nonsmall cell lung carcinoma

“…One part was sent for a pathologic examination, and the other part was prepared for single cell suspension and was frozen at Ϫ 130°C until use, as described previously. 30 A pathologic examination revealed that the patient's hilar lymph nodes were involved with carcinoma cells; therefore, the it is possible that the RLNLs in this study may have contained lung carcinoma cells. The frozen RLNLs were thawed rapidly, and 3.6 ϫ 10 6 cells were divided into 3 wells of a 24-well plate (1.2 ϫ 10 6 cells per well) (Iwaki Glass).…”

“…30 A stable transfection of CD80 molecules was confirmed by flow cytometry after staining with FITC-conjugated anti-human CD80 (Pharmingen, San Diego, CA).…”

Immunosuppressive effect of regulatory T lymphocytes in lung cancer, with special reference to their effects on the induction of autologous tumor-specific cytotoxic T lymphocytes

“…Each lymph node was divided into two parts; one for the histological diagnosis and one for this study. Lymphocytes from each lymph node were mixed and stored at -80˚C until use, as described previously (17). They were then rapidly thawed and stimulated with an irradiated (100 Gy) CD80-transfected autologous tumor cell line (F1121L and L1023L) weekly at a tumor-to-lymphocyte ratio of 1:10 in culture medium as described previously (17).…”

“…Lymphocytes from each lymph node were mixed and stored at -80˚C until use, as described previously (17). They were then rapidly thawed and stimulated with an irradiated (100 Gy) CD80-transfected autologous tumor cell line (F1121L and L1023L) weekly at a tumor-to-lymphocyte ratio of 1:10 in culture medium as described previously (17). The tumor-specific induction of CTLs was considered to be successful if the CTLs lysed more than 10% of the autologous tumor cells at an effector/target ratio of 30/1 and did not lyse autologous Epstein-Barr virus-transformed B cells during the 4-h standard 51Cr release assay performed on day 28 of the mixed lymphocyte-tumor cell culture (MLTC).…”