Ex vivo activated memory t‐lymphocytes as adoptive cellular therapy of human soft‐tissue sarcoma targets with potentiation by Cis‐diamminedichloroplatinum (II)

Abstract

Autolymphocyte therapy (ALT) is tumor-specific, adoptive cellular therapy of neoplastic disease using nonspecific ex vivo activation of autologous peripheral blood lymphocytes (PBL), which are composed primarily of memory T-cells (ALT-cells) and are active in patients with metastatic renal cell carcinoma and melanoma. Ex vivo pretreatment of tumor target cells with certain chemotherapeutic agents can enhance susceptibility to lysis by antitumor lymphocytes. To determine if cis-diamminedichloroplatinum(II) (CDDP) enhances ex vivo antitumor cytotoxicity of ALT-cells and if this lysis is mediated by T- and/or NK-cells and is human leukocyte antigen (HLA)-restricted, human soft tissue sarcoma (STS) target cells were derived from primary and metastatic surgical specimens and were incubated with and without CDDP. ALT-cells were prepared from autologous PBL obtained prior to surgery. Primary (PSTS) and metastatic (MSTS) target cells from each group were labelled with chromium 51 (51Cr) and used as targets for ALT-cells, CD45-depleted ALT-cells, CD56 (NK)-depleted ALT-cells, and PBL in a standard (4-hour) and delayed (18-hour) 51Cr release assay. Interferon-gamma (IFN-gamma) release was measured as an indication of antitumor effect and recognition by the noncytolytic lymphocytes in ALT-cells. Primary tumor target cells incubated in CDDP showed enhanced lysis as measured by the 51Cr release assay compared to non-CDDP-treated controls. Metastatic tumor target cells showed less lysis than the primary targets, although this was enhanced by pretreating metastatic tumor targets with CDDP. Lysis of all tumor targets was significantly greater when ALT-cells were used as the effector cells rather than PBL. Depletion of memory T-cells abrogated ex vivo lysis. Depletion of NK cells (CD56+) affected ex vivo lysis of autologous targets during the 4-hour but not the 18-hour assay. Ex vivo ALT-cell lysis and IFN-gamma release against only the autologous tumor targets confirmed tumor-specificity in one patient. Restriction of ALT-cell lysis and IFN-gamma release against HLA-A2+ autologous and one allogeneic HLA-A2+ STS tumor target, but not other non-STS targets, was demonstrated in another patient. These data suggest that CDDP may help render STS susceptible to tumor-specific, immune-mediated attack and that the combination of ALT and CDDP may lead to effective tumor-specific chemoimmunotherapy in patients with metastatic STS.

Anticancer Drug-Induced Immunomodulation and Cancer Therapeutics

“…The increase of HLA antigens after incubation of tumor cells with CDDP was also demonstrated measuring the AC-81 adenocarcinoma-associated antigen on gastric cancer cell line KATO-3 [208], increase which resulted in increased susceptibility of the cancer cells to LAK cells and T-lymphocytes cytotoxicity. Similar effects of CDDP had been found by the same group using human soft tissue sarcoma cells [210] or renal cell carcinoma cells [211] as target tumor cells which were modified by CDDP and elicited increased memory T cells responses. Similar effects of CDDP had been found by the same group using human soft tissue sarcoma cells [210] or renal cell carcinoma cells [211] as target tumor cells which were modified by CDDP and elicited increased memory T cells responses.…”

“…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 ...…”

“…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.…”

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.…”