Adoptive cellular therapy of human breast and colorectal tumor targets using ex vivo activated memory T lymphocytes with potentiation by cis‐diamminedichloroplatinum(II)

Abstract

Autolymphocyte therapy (ALT) is adoptive cellular therapy of cancer using ex vivo activation of autologous peripheral blood lymphocytes (PBL). Memory T cells are the principal effector population in ALT, with in vivo activity in patients with metastatic renal cell carcinoma (RCC) and melanoma, and ex vivo cytotoxicity against autologous tumor targets. However, the noncytolytic lymphocyte portion of ex vivo-activated memory T cells (ALT cells) may also contribute as antitumor effectors. Pretreatment of murine and human tumor cells ex vivo with chemotherapeutic agents can enhance their susceptibility to antitumor lymphocytes ex vivo and in vivo. To determine whether cis-diamminedichloroplatinum(II) (DDP) could enhance ex vivo antitumor effects of ALT cells by immunomodulation, human breast and colorectal carcinoma target cells were derived from both primary and metastatic surgical specimens and incubated in complete medium (CM) with DDP or in CM alone (control group). Viability of each group was confirmed by trypan blue-dye exclusion test. ALT cells were prepared from autologous PBL at surgery. Primary and metastatic tumor cells from each group were used as targets for ALT cells and levels of interferon-gamma (IFN-gamma) release were measured as a determination of antitumor effect and recognition. Primary tumor target cells incubated in DDP showed enhanced antitumor effects and recognition by autologous ALT cells, as measured by the IFN-gamma assay compared to non-DDP-treated controls. Metastatic autologous tumor target cells demonstrated less IFN-gamma release than did the primary targets, although this was enhanced by pre-treating metastatic tumor targets with DDP. ALT cells demonstrated minimal IFN-gamma release when incubated with allogeneic tumor targets. These data suggest that autotumor recognition of metastatic tumor targets is comparable to that of primary lesions following ex vivo pretreatment of metastatic cells with nonlethal doses of certain chemotherapeutic agents. DDP may somehow alter the physical properties of target cells, rendering them susceptible to immune-mediated attack and the combination of ALT and DDP may lead to increased therapeutic efficacy in patients with metastatic breast and colon cancer.

Effect of Contaminating Red Blood Cells on OKT3-Mediated Polyclonal Activation of Peripheral Blood Mononuclear Cells

“…Although CD25 expression on the T cells in the secondary culture was lower than that in the primary culture, RBC did In secondary cultures, CD3 ϩ CD45RO ϩ T cells were 62% of the T cells in the PBMC-alone group; 35% would be observed for resting T cells (1,2), indicating that the CM could stimulate the T cells. However, RBCs had no effect on the percentages of cells expressing CD45RO in the culture.…”

“…The cells were resuspended with 10 ml of complete AIM-V medium to which was added 50 M cimetidine (Tagamet; Smith Kline Beecham Pharmaceutical, Cidra, Pa.) and 10 nM indomethicin (Indocin; Merck Sharp & Dohme, West Point, Pa.), 1 mM sodium pyruvate (Gibco-BRL, Grandview, N.Y.), and 100 g of gentamicin (Gibco-BRL)/ml. Indomethacin and cimetidine were added to reduce tumor-related suppression (1,2), keeping this protocol in line with the clinical protocol. Cell count was determined with a Coulter Counter, and cell viability was determined by trypan blue exclusion.…”

Anticancer Drug-Induced Immunomodulation and Cancer Therapeutics

“…Indeed at low doses Cyclophosphamide inhibited the growth of murine hepatoma MH129 in immunocompetent but not in immunodepleted mice and the inhibition did not occur in mice given CD4+CD25+ T cells [166]. In this study the interesting observation was made that the administration of low dose Cyclophosphamide was more effective in causing depletion of CD4+CD25+ cells when given after tumor inoculation than when given before tumor inoculation: as the authors stated [166], this finding is of relevance to clinical Phenylalinine mustard + [147,[188][189][190][191] Busulfan + + [148,169,[192][193][194] Nitrosoureas + + [148,[195][196][197][198][199][200][201][202] Cis Platinum + + [170,203,204,205,[206][207][208][209][210][211][212][213][214][215] Difluoromethylornithine + + [280][281][282][283][284][285] trials of chemotherapy and immunotherapy combinations. In this study the interesting observation was made that the administration of low dose Cyclophosphamide was more effective in causing depletion of CD4+CD25+ cells when given after tumor inoculation than when given ...…”

“…It was later confirmed that CDDP increases the expression of MHC class I antigens H-2 Dd and H 2 -K on the tumor surface leading to the regression of MOPC-104E plasmacytoma in immunocompetent BalbC mice but not in nude mice or in immunocompetent mice treated with anti T cells monoclonal antibodies [207]. With peripheral blood lymphocytes from patients with breast and colorectal tumors it was demonstrated that incubation of these cells with autochthonous tumor cells in the presence of CDDP resulted in increased autolymphocytes reactivity as measured through IFN release, again suggesting that the drug induced a change in the properties of the target cells which rendered them more susceptible to lymphocyte responses [209]. With peripheral blood lymphocytes from patients with breast and colorectal tumors it was demonstrated that incubation of these cells with autochthonous tumor cells in the presence of CDDP resulted in increased autolymphocytes reactivity as measured through IFN release, again suggesting that the drug induced a change in the properties of the target cells which rendered them more susceptible to lymphocyte responses [209].…”

“…The augmenting effects of CDDP on the antigenicity of murine and human tumors [207][208][209][210][211] have been mentioned above where the action of this drug was discussed. Very recently [307], it has been reported that DOX, and some related anthracyclines, but not Etoposide, Mitomycin C or other cytotoxic agents, augment the immunogenicity of mouse CT26 colon cancer and that this effect is mediated through the induction by drug of calreticulin or of a calreticulin-interacting chaperon by the anthracycline.…”