Lysis of fresh human solid tumor cells by autologous lymphocytes activated in vitro by allosensitization

Abstract

We have previously demonstrated that cancer patients' peripheral blood lymphocytes (PBL) allosensitized against single or pool normal donor PBL are capable of lysing fresh autologous tumor cells in a 4-h 51Cr-release assay. In this report, we present further investigations into this phenomenon. These alloactivated killer cells (A-AK cells) lysed autologous and allogeneic tumors and allogeneic but not autologous PBL. Furthermore, cold target inhibition studies demonstrated that autologous and allogeneic tumors were lysed by the same effector cells. Multiple metastases from the same patient were equivalently lysed by these A-Ak cells. The presence of adherent cells and proliferation of the precursors were necessary to generate A-AK cells, although the effector cell itself was radioresistant and nonadherent. The effects of allosensitization were enhanced by the addition of lectin-free interleukin-2 preparations to the in vitro culture by partial depletion of adherent cells prior to sensitization. The A-Ak effector cell was OKT3+, OKT8+, OKT4-, OKM1- and could be generated by just 3 days of allosensitization. The precursors for A-Ak cells could be separated from NK cells on percoll gradients and lysis could be generated from thoracic duct lymphocytes, a population devoid of NK cells. The phenotype of the majority of the precursor cells was OKT3+, OKT4-. Theses alloactivated PBL could be expanded in crude or lectin-free interleukin-2 without loss of cytotoxicity for fresh autologous tumor cells. Activated T cells represent a population of on-NK cells with broad lytic specificity for fresh tumor cells. Such cells may be of value in the adoptive immunotherapy of human solid tumors and may play a role in immune surveillance.

Lymphokine-activated killer cell phenomenon. Lysis of natural killer-resistant fresh solid tumor cells by interleukin 2-activated autologous human peripheral blood lymphocytes.

“…Such reports include anomolous cytotoxicities, distinct from CTL during alloactivation (13)(14)(15)(16)(17), lectinactivated killing, distinct from lectin-dependent killing (18), fetal calf serum cultureinduced killers (16,(19)(20)(21), and mixed-lymphocyte tumor interaction (MLTI)-induced killers, found only when proliferation was stimulated (8)(9)(10)(11)22). Though fresh tumor targets were rarely used in those studies, our previous reports of lysis of autologous fresh solid tumor cells after allosensitization (23) or lectin activation (24), and those of others after MLTI (8)(9)(10)(11), support this hypothesis and suggest that lymphokine activated killers (LAK) represent a unique and fundamental cytotoxic effector system that may play a role in immune surveillance against NK resistant solid tumor cells, and may have a possible role in the adoptive immunotherapy of tumors.…”

Generation of stimulated, lymphokine activated T killer (T-LAK) cells from the peripheral blood of normal donors and adult patients with recurrent glioblastoma

“…About 50-80% of the cells in these cultures die during the first 2-4 days in vitro. This is the period in which LAK and PAK cells are usually harvested (Grimm et al, 1982; Mazumder et al, 1982Mazumder et al, , 1983Rosenberg et al, 1985;Jacobs et al, 1986a;Merchant et al, 1988). However, between 4-6 days, the cells in T-LAK cultures begin to proliferate in the presence of IL-2 and continue to divide at a constant rate, doubling every 24-30 h. While we terminated most cultures at 12-20 days, seven were continued for 5 months.…”

“…Ingram (1987b) employed similar induction and culture techniques but did not maintain cells beyond 14 days in vitro. Mazumder ( , 1983 employed PHA and ConA to stimulate human PBL but collected the cells at 2-3 days and did not employ IL-2 to continue cell division. The present culture technique can provide large numbers of effector cells at the end of the culture period, or they can be harvested at intervals during the culture.…”

“…It is surprising that no difference in CD4 or CD8 phenotype was noted between T-LAK cells that were excellent vs. poor killers in vitro. The PAK killers in 2-7 day PHA stimulated human PBL cultures were shown to be predominantly CD3 T cells by Mazumder et al (1983). Because PHA is a polyclonal T cell stimulator, it is presumed that the T-LAK cells represent the progeny of various T cell subsets and should be heterogenous.…”

Treatment with interleukin-2 (IL-2) and interferon (IFNalpha 2b) after autologous bone marrow or peripheral blood stem cell transplantation in onco-hematological malignancies with a high risk of relapse

“…7,8 Later some studies were undertaken using different dosages and different approaches to administration. [9][10][11][12][13][14][15][16] In 1990 the first results of the efficiency and toxicity of IL-2 s.c. and IFN alpha 2b i.m. in metastatic renal cancer were reported.…”