Abstract
In this study, cytokine release by tumor-draining lymph node cells sensitized in vitro (IVS-TDLN) was examined and correlated with therapeutic efficacy in adoptive immunotherapy. Mice bearing immunologically distinct MCA 207 and MCA 205 sarcoma tumors were utilized in criss-cross experiments. IVS-TDLN obtained from mice bearing 10-day subcutaneous (s. c.) tumors mediated immunologically specific regression of established 3-day pulmonary metastases, but demonstrated non-specific cytolytic reactivity against both tumors in a 4-h51Cr-release assay. By contrast, these IVS-TDLN cells were found specifically to secrete granulocyte/macrophage colony-stimulating factor (GM-CSF) and interferon γ (IFNγ) when restimulated in vitro with irradiated tumor cells. To determine the predictive value of tumor-specific cytokine release with in vivo therapeutic efficacy, a kinetic analysis of antitumor activities of TDLN obtained from animals bearing MCA 207 tumors for increasing lengths of time was performed. IVS-TDLN cells from mice bearing day-7, -10 and-14 s. c. tumors manifested tumor-specific release of GM-CSF and IFNγ, and mediated significant antitumor reactivity in vivo. In contrast IVS-LN cells from day-0 and day-21 tumor-bearing animals did not release significant amounts of GM-CSF and IFNγ, and were not therapeutically efficacious in vivo. Day-4 IVS-TDLN released high levels of GM-CSF and IFNγ non-specifically, and were not therapeutic in adoptive immunotherapy at doses effective for day-7 and day-14 IVS-TDLN cells. In other experiments, IVS cells generated from different lymph node groups in animals bearing 10-day established s. c. tumors were examined and found to have unique profiles of cytokine release. In these studies, the ability of IVS cells to release specifically both cytokines as opposed to one was associated with greater therapeutic efficacy on a per cell basis. Our findings suggest that the tumor-specific releases of GM-CSF and IFNγ are useful parameters to assess the in vivo therapeutic efficacy of immune lymphocytes.
Key words: Sarcoma, Neoplasms, Adoptive immunotherapy, Cytokines
References
- 1.Arca MJ, Krauss JC, Aruga A, Cameron MJ, Shu S, Chang AE (1995) Therapeutic efficacy of T cells derived from lymph nodes draining a poorly immunogenic tumor transduced to secrete GM-CSF. Cancer Gene Ther (in press) [PubMed]
- 2.Barth RJ, Bock SN, Mulé JJ, Rosenberg SA. Unique murine tumor-associated antigens identified by tumor infiltrating lymphocytes. J Immunol. 1990;144:1531–1537. [PubMed] [Google Scholar]
- 3.Barth RJ, Mulé JJ, Spiess PJ, Rosenberg SA. Interferon γ and tumor necrosis factor have a role in tumor regressions mediated by murine CD8+ tumor-infiltrating lymphocytes. J Exp Med. 1991;173:647–658. doi: 10.1084/jem.173.3.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Chen TT, Tao M-H, Levy R. Idiotype-cytokine fusion proteins as cancer vaccines. J Immunol. 1994;153:4775–4787. [PubMed] [Google Scholar]
- 5.Chou T, Chang AE, Shu S. Generation of therapeutic T lymphocytes from tumor-bearing mice by in vitro sensitization. J Immunol. 1988;140:2453–2461. [PubMed] [Google Scholar]
- 6.Chou T, Bertera S, Chang AE, Shu S. Adoptive immunotherapy of microscopic and advanced visceral metastases with in vitro sensitized lymphoid cells from mice bearing progressive tumors. J Immunol. 1988;141:1775–1781. [PubMed] [Google Scholar]
- 7.Dranoff G, Jaffee E, Lazenby A, Golumbek P, Levitsky H, Brose K, Jackson V, Hamada H, Pardoll D, Mulligan RC. Vaccination with irradiated tumor cells engineered to secrete murine granulocyte-macrophage colony-stimulating factor stimulates potent, specific, and long-lasting anti-tumor immunity. Proc Natl Acad Sci USA. 1993;90:3539–3543. doi: 10.1073/pnas.90.8.3539. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8.Geiger JD, Wagner PD, Cameron MJ, Shu S, Chang AE. Generation of T cells reactive to the poorly immunogenic B16-BL6 melanoma with efficacy in the treatment of spontaneous metastases. J Immunother. 1993;13:153–165. doi: 10.1097/00002371-199304000-00002. [DOI] [PubMed] [Google Scholar]
- 9.Goedegebuure PS, Zuber M, Leonard-Vidal DL, Burger UL, Cusack JC, Chang MP, Douville LM, Eberlein TJ. Reactivation of murine tumour-infiltrating lymphocytes with solid-phase anti-CD3 antibody: in vitro cytokine production is associated with in vivo efficacy. Surg Oncol. 1994;3:79–89. doi: 10.1016/0960-7404(94)90003-5. [DOI] [PubMed] [Google Scholar]
- 10.Greenberg PD, Kern DE, Cheever MA. Therapy of disseminated murine leukemia with cyclophosphamide and immune Lyt-1+, 2− T cells. J Exp Med. 1985;161:1122–1134. doi: 10.1084/jem.161.5.1122. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11.Kurzrock R, Talpaz M, Gutterman JU. Interferons α, β, γ: Basic principles and preclinical studies. In: DeVita VT, Hellman S, Rosenberg SA, editors. Biologic therapy of cancer. Philadelphia: Lippincott; 1991. pp. 247–274. [Google Scholar]
- 12.Miller AR, McBride WH, Hunt K, Economou JS. Cytokine-mediated gene therapy for cancer. Ann Surg Oncol. 1994;5:436–450. doi: 10.1007/BF02303818. [DOI] [PubMed] [Google Scholar]
- 13.Mitra RS, Judge TA, Nestle FO, Turka LA, Nickoloff BJ. Psoriatic skin-derived dendritic cell function is inhibited by exogenous IL-10. J Immunol. 1995;154:2668–2677. [PubMed] [Google Scholar]
- 14.Restifo NP, Spiess PJ, Karp SE, Mulé JJ, Rosenberg SA. A nonimmunogenic sarcoma transduced with the cDNA for interferon γ elicits CD8+ T cells against the wild-type tumor: correlation with antigen presentation capability. J Exp Med. 1992;175:1423–1431. doi: 10.1084/jem.175.6.1423. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15.Rosenberg SA, Spiess P, Lafreniere R. A new approach to the adoptive immunotherapy of cancer with tumor-infiltrating lymphocytes. Science. 1986;233:1318–1321. doi: 10.1126/science.3489291. [DOI] [PubMed] [Google Scholar]
- 16.Rosenstein M, Rosenberg SA. Generation of lytic and proliferative lymphoid clones to syngeneic tumor: In vitro and in vivo studies. J Natl Cancer Inst. 1984;72:1161–1165. [PubMed] [Google Scholar]
- 17.Sallusto F, Lanzavecchia A. Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor α. J Exp Med. 1994;179:1109–1118. doi: 10.1084/jem.179.4.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 18.Schwartzentruber DJ, Hom SS, Dadmarz R, White DE, Yannelli JR, Steinberg SM, Rosenberg SA, Topalian SL. In vitro predictors of therapeutic response in melanoma patients receiving tumor-infiltrating lymphocytes and interleukin-2. J Clin Oncol. 1994;12:1475–1483. doi: 10.1200/JCO.1994.12.7.1475. [DOI] [PubMed] [Google Scholar]
- 19.Shu S, Chou T, Sakai K. Lymphocytes generated by in vivo priming and in vitro sensitization demonstrate therapeutic efficacy against a murine tumor that lacks apparent immunogenicity. J Immunol. 1989;143:740–748. [PubMed] [Google Scholar]
- 20.Shu S, Krinock RA, Matsumura T, Sussman JJ, Fox BA, Chang AE, Terman DS. Stimulation of tumor-draining lymph node cells with superantigenic staphylococcal toxins leads to the generation of tumor-specific effector T cells. J Immunol. 1994;152:1277–1288. [PubMed] [Google Scholar]
- 21.Spiess PJ, Yang JC, Rosenberg SA. In vivo antitumor activity of tumor-infiltrating lymphocytes expanded in recombinant interleukin-2. J Natl Cancer Inst. 1987;79:1067–1075. [PubMed] [Google Scholar]
- 22.Stephenson KR, Perry-Lalley D, Griffith KD, Shu S, Chang AE. Development of antitumor reactivity in regional draining lymph nodes from tumor-immunized and tumor-bearing murine hosts. Surgery. 1989;105:523–528. [PubMed] [Google Scholar]
- 23.Yoshizawa H, Chang AE, Shu S. Specific adoptive immunotherapy mediated by tumor-draining lymph node cells sequentially activated with anti-CD3 and IL-2. J Immunol. 1991;147:729–737. [PubMed] [Google Scholar]