To evaluate the immunological characteristics of tumor infiltrating lymphocytes(TIL) and autologous peripheral blood lymphocytes(A―PBL) in 20 patients with primary cervical tumors and tumor draining lymph node lymphocytes from, lymphocytes were
cultured in vitro for proliferation, phenotyping and cytotoxicity.
Freshly obtained cervical tumor masses and draining lymph nodes were digested enzymatically. Lymphocytes were separated using ficoll―hypaque density gradient method, and were cultured in Waymouth's MB 752/1 media with 5% human AB serum only or
with,
1.000U/ml recombinant interleukin-2(rIL―2).
@ES The results were as follows:
@EN 1. Lymphocytes from primary cervical tumors constituted from 20% to 90% of single cell tumor suspensions(average lymphocyte cell: tumor cell ratio=0.95) and expanded from 2.6―folds to 130―folds in 12 of 20 cultures. For 10 of 20 patients,
lymohocytes derived from tumor draining lymph nodes proliferated in culture from 2.8―folds to 120―folds.
2. When TIL as well as A―PBL were cultured in 1,000U/ml of rIL―2, TIL significantly proliferated than a―PBL(p<0.05).
3. TIL and tumor involved lymph node lymphocytes were predominantly cytotoxic/suppressor T―lymphocytes with and average of 53.2% CD 8+ in TIL, 54.&% CD 8+ in tumor involved lymph node lymphocytes. The proportion of CD 8+ of TIL and tumor
involved
lymph
node lymphocytes we significantly higher than that of normal draining lymph node lymphocytes and PBL(p<0.01). There were less than 3% CD 56+(natural Killer cell, NK cell) in both of them.
4. Cervical TIL, lymph node lymphocytes. Normal draining lymph node lymphocyte and A―PBL did not demonstrate any significant cytotoxicity against autologous, allogeneic and K 562 target tumor cells, and autologous lymphokine activated
killer(LAK)
cells
demonstrated antitumor cytotoxicity against autologous tumor cells although these lysis were low. But all of 3 rIL―2 cultured cervical TIL demonstrated substantial levels of cytotoxicity against autologous tumor cells(21.8±3.0%) compared to the
cytotoxicity of TIL cultured without rIL―2(9.1±0.1%)(p<0.05).
These results suggest that cervical TIL could be expanded in vitro and reach high levels of antitumor effector function in cultures with rIL―2.
|