• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
J Clin Invest. Jan 15, 1998; 101(2): 429–441.
PMCID: PMC508583

Importance of cyclophosphamide-induced bystander effect on T cells for a successful tumor eradication in response to adoptive immunotherapy in mice.

Abstract

Cyclophosphamide (CTX) increases the antitumor effectiveness of adoptive immunotherapy in mice, and combined immunotherapy regimens are now used in some clinical trials. However, the mechanisms underlying the synergistic antitumor responses are still unclear. The purpose of this study was (a) to evaluate the antitumor response to CTX and adoptive immunotherapy in mice bearing four different syngeneic tumors (two responsive in vivo to CTX and two resistant); and (b) to define the mechanism(s) of the CTX-immunotherapy synergism. Tumor-bearing DBA/2 mice were treated with a single injection of CTX followed by an intravenous infusion of tumor-immune spleen cells. In all the four tumor models, a single CTX injection resulted in an impressive antitumor response to the subsequent injection of spleen cells from mice immunized with homologous tumor cells independently of the in vivo response to CTX alone. Detailed analysis of the antitumor mechanisms in mice transplanted with metastatic Friend leukemia cells revealed that (a) the effectiveness of this combined therapy was dependent neither on the CTX-induced reduction of tumor burden nor on CTX-induced inhibition of some putative tumor-induced suppressor cells; (b) the CTX/immune cells' regimen strongly protected the mice from subsequent injection of FLC, provided the animals were also preinoculated with inactivated homologous tumor together with the immune spleen cells; (c) CD4(+) T immune lymphocytes were the major cell type responsible for the antitumor activity; (d) the combined therapy was ineffective in mice treated with antiasialo-GM1 or anti-IFN-alpha/beta antibodies; (e) spleen and/ or bone marrow cells from CTX-treated mice produced soluble factors that assisted in proliferation of the spleen cells. Altogether, these results indicate that CTX acts via bystander effects, possibly through production of T cell growth factors occurring during the rebound events after drug administration, which may sustain the proliferation, survival, and activity of the transferred immune T lymphocytes. Thus, our findings indicate the need for reappraisal of the mechanisms underlying the synergistic effects of CTX and adoptive immunotherapy, and may provide new insights into the definition of new and more effective strategies with chemotherapy and adoptive immunotherapy for cancer patients.

Full Text

The Full Text of this article is available as a PDF (316K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Shu S, Chou T, Rosenberg SA. In vitro sensitization and expansion with viable tumor cells and interleukin 2 in the generation of specific therapeutic effector cells. J Immunol. 1986 May 15;136(10):3891–3898. [PubMed]
  • Osband ME, Lavin PT, Babayan RK, Graham S, Lamm DL, Parker B, Sawczuk I, Ross S, Krane RJ. Effect of autolymphocyte therapy on survival and quality of life in patients with metastatic renal-cell carcinoma. Lancet. 1990 Apr 28;335(8696):994–998. [PubMed]
  • Mulé JJ, Shu S, Schwarz SL, Rosenberg SA. Adoptive immunotherapy of established pulmonary metastases with LAK cells and recombinant interleukin-2. Science. 1984 Sep 28;225(4669):1487–1489. [PubMed]
  • Greenberg PD, Kern DE, Cheever MA. Therapy of disseminated murine leukemia with cyclophosphamide and immune Lyt-1+,2- T cells. Tumor eradication does not require participation of cytotoxic T cells. J Exp Med. 1985 May 1;161(5):1122–1134. [PMC free article] [PubMed]
  • Yoshizawa H, Sakai K, Chang AE, Shu SY. Activation by anti-CD3 of tumor-draining lymph node cells for specific adoptive immunotherapy. Cell Immunol. 1991 May;134(2):473–479. [PubMed]
  • Gold JE, Osband ME. Autolymphocyte therapy--I. In vivo tumour-specific adoptive cellular therapy of murine melanoma and carcinoma using ex vivo activated memory T-lymphocytes. Eur J Cancer. 1994;30A(12):1871–1882. [PubMed]
  • Radov LA, Haskill JS, Korn JH. Host immune potentiation of drug responses to a murine mammary adenocarcinoma. Int J Cancer. 1976 Jun 15;17(6):773–779. [PubMed]
  • Kolb JP, Poupon MF, Lespinats GM, Sabolovic D, Loisillier F. Splenic modifications induced by cyclophosphamide in C3H/He, nude, and "B" mice. J Immunol. 1977 May;118(5):1595–1599. [PubMed]
  • Schwartz A, Askenase PW, Gershon RK. Regulation of delayed-type hypersensitivity reactions by cyclophosphamide-sensitive T cells. J Immunol. 1978 Oct;121(4):1573–1577. [PubMed]
  • Hengst JC, Mokyr MB, Dray S. Importance of timing in cyclophosphamide therapy of MOPC-315 tumor-bearing mice. Cancer Res. 1980 Jul;40(7):2135–2141. [PubMed]
  • Vidović D, Marusić M, Culo F. Interference of anti-tumor and immunosuppressive effects of cyclophosphamide in tumor-bearing rats. Analysis of factors determining resistance or susceptibility to a subsequent tumor challenge. Cancer Immunol Immunother. 1982;14(1):36–40. [PubMed]
  • Turk JL, Parker D. Effect of cyclophosphamide on immunological control mechanisms. Immunol Rev. 1982;65:99–113. [PubMed]
  • Kedar E, Ben-Aziz R, Epstein E, Leshem B. Chemo-immunotherapy of murine tumors using interleukin-2 (IL-2) and cyclophosphamide. IL-2 can facilitate or inhibit tumor growth depending on the sequence of treatment and the tumor type. Cancer Immunol Immunother. 1989;29(1):74–78. [PubMed]
  • Katsanis E, Bausero MA, Ochoa AC, Loeffler CM, Blazar BR, Leonard AS, Anderson PM. Importance in timing of cyclophosphamide on the enhancement of interleukin-2-induced cytolysis. Cancer Immunol Immunother. 1991;34(2):74–78. [PubMed]
  • Laude M, Russo KL, Mokyr MB, Dray S. Cure of mice bearing a late-stage, highly metastatic, drug-resistant tumor by adoptive chemoimmunotherapy. Cancer Immunol Immunother. 1993;36(4):229–236. [PubMed]
  • Evans R. Combination therapy by using cyclophosphamide and tumor-sensitized lymphocytes: a possible mechanism of action. J Immunol. 1983 Jun;130(6):2511–2513. [PubMed]
  • Mitchell MS. Combining chemotherapy with biological response modifiers in treatment of cancer. J Natl Cancer Inst. 1988 Nov 16;80(18):1445–1450. [PubMed]
  • Ootsu K, Gotoh K, Houkan T. Therapeutic efficacy of human recombinant interleukin-2 (TGP-3) alone or in combination with cyclophosphamide and immunocompetent cells in allogeneic, semi-syngeneic, and syngeneic murine tumors. Cancer Immunol Immunother. 1989;30(2):71–80. [PubMed]
  • Kedar E, Rutkowski Y, Leshem B. Chemo-immunotherapy of murine solid tumors: enhanced therapeutic effects by interleukin-2 combined with interferon alpha and the role of specific T cells. Cancer Immunol Immunother. 1992;35(1):63–68. [PubMed]
  • Gold JE, Zachary DT, Osband ME. Adoptively transferred ex vivo activated memory T cells with cyclophosphamide: effective tumor-specific chemoimmunotherapy of advanced metastatic murine melanoma and carcinoma. Clin Immunol Immunopathol. 1994 Oct;73(1):115–122. [PubMed]
  • Gold JE, Zachary DT, Osband ME. Adoptive transfer of ex vivo-activated memory T-cell subsets with cyclophosphamide provides effective tumor-specific chemoimmunotherapy of advanced metastatic murine melanoma and carcinoma. Int J Cancer. 1995 May 16;61(4):580–586. [PubMed]
  • Glaser M. Regulation of specific cell-mediated cytotoxic response against SV40-induced tumor associated antigens by depletion of suppressor T cells with cyclophosphamide in mice. J Exp Med. 1979 Mar 1;149(3):774–779. [PMC free article] [PubMed]
  • Awwad M, North RJ. Cyclophosphamide-induced immunologically mediated regression of a cyclophosphamide-resistant murine tumor: a consequence of eliminating precursor L3T4+ suppressor T-cells. Cancer Res. 1989 Apr 1;49(7):1649–1654. [PubMed]
  • Reissmann T, Voegeli R, Pohl J, Hilgard P. Augmentation of host antitumor immunity by low doses of cyclophosphamide and mafosfamide in two animal tumor models. Cancer Immunol Immunother. 1989;28(3):179–184. [PubMed]
  • North RJ, Awwad M. Elimination of cycling CD4+ suppressor T cells with an anti-mitotic drug releases non-cycling CD8+ T cells to cause regression of an advanced lymphoma. Immunology. 1990 Sep;71(1):90–95. [PMC free article] [PubMed]
  • Hoover SK, Barrett SK, Turk TM, Lee TC, Bear HD. Cyclophosphamide and abrogation of tumor-induced suppressor T cell activity. Cancer Immunol Immunother. 1990;31(2):121–127. [PubMed]
  • Rakhmilevich AL, North RJ. Elimination of CD4+ T cells in mice bearing an advanced sarcoma augments the antitumor action of interleukin-2. Cancer Immunol Immunother. 1994 Feb;38(2):107–112. [PubMed]
  • Gold JE, Ross SD, Krellenstein DJ, LaRosa F, Malamud SC, Osband ME. Adoptive transfer of ex vivo activated memory T-cells with or without cyclophosphamide for advanced metastatic melanoma: results in 36 patients. Eur J Cancer. 1995;31A(5):698–708. [PubMed]
  • Gold JE, Malamud SC, LaRosa F, Osband ME. Adoptive chemoimmunotherapy using ex vivo activated memory T-cells and cyclophosphamide: tumor lysis syndrome of a metastatic soft tissue sarcoma. Am J Hematol. 1993 Sep;44(1):42–47. [PubMed]
  • Abrams JS, Eiseman JL, Melink TJ, Sridhara R, Hiponia DJ, Bell MM, Belani CP, Adler WH, Aisner J. Immunomodulation of interleukin-2 by cyclophosphamide: a phase IB trial. J Immunother Emphasis Tumor Immunol. 1993 Jul;14(1):56–64. [PubMed]
  • Contegiacomo A, Pizzi C, De Marchis L, Alimandi M, Delrio P, Di Palma E, Petrella G, Ottini L, French D, Frati L, et al. High cell kinetics is associated with amplification of the int-2, bcl-1, myc and erbB-2 proto-oncogenes and loss of heterozygosity at the DF3 locus in primary breast cancers. Int J Cancer. 1995 Mar 29;61(1):1–6. [PubMed]
  • Puddu P, Locardi C, Sestili P, Varano F, Petrini C, Modesti A, Masuelli L, Gresser I, Belardelli F. Human immunodeficiency virus (HIV)-infected tumor xenografts as an in vivo model for antiviral therapy: role of alpha/beta interferon in restriction of tumor growth in nude mice injected with HIV-infected U937 tumor cells. J Virol. 1991 May;65(5):2245–2253. [PMC free article] [PubMed]
  • Affabris E, Jemma C, Rossi GB. Isolation of interferon-resistant variants of Friend erythroleukemia cells: effects of interferon and ouabain. Virology. 1982 Jul 30;120(2):441–452. [PubMed]
  • Gabriele L, Proietti E, Greco G, Venditti M, Gresser I, Schirrmacher V, Von Hoegen P, Testa U, Modesti A, Cianfriglia M, et al. Isolation and characterization of a metastatic Eb-like tumor variant highly responsive to interleukin (IL)-2 and to combination cytokine therapy with IL-2/IL-1 beta and IL-1 beta/interferon-alpha/beta. Invasion Metastasis. 1993;13(3):147–162. [PubMed]
  • Proietti E, Tritarelli E, Gabriele L, Testa U, Greco G, Pelosi E, Gabbianelli M, Belardelli F, Peschle C. Combined interleukin 1 beta/interleukin 2 treatment in mice: synergistic myelostimulatory activity and protection against cyclophosphamide-induced myelosuppression. Cancer Res. 1993 Feb 1;53(3):569–576. [PubMed]
  • Tritarelli E, Greco G, Testa U, Belardelli F, Peschle C, Proietti E. Combined interleukin-1 beta/interleukin-6 treatment in mice: synergistic myelostimulatory activity and myelorestorative effect after cyclophosphamide-induced myelosuppression. Cancer Res. 1994 Dec 15;54(24):6469–6476. [PubMed]
  • Ferrantini M, Proietti E, Santodonato L, Gabriele L, Peretti M, Plavec I, Meyer F, Kaido T, Gresser I, Belardelli F. Alpha 1-interferon gene transfer into metastatic Friend leukemia cells abrogated tumorigenicity in immunocompetent mice: antitumor therapy by means of interferon-producing cells. Cancer Res. 1993 Mar 1;53(5):1107–1112. [PubMed]
  • Ciolli V, Gabriele L, Sestili P, Varano F, Proietti E, Gresser I, Testa U, Montesoro E, Bulgarini D, Mariani G, et al. Combined interleukin 1/interleukin 2 therapy of mice injected with highly metastatic Friend leukemia cells: host antitumor mechanisms and marked effects on established metastases. J Exp Med. 1991 Feb 1;173(2):313–322. [PMC free article] [PubMed]
  • Gresser I, Maury C, Carnaud C, De Maeyer E, Maunoury MT, Belardelli F. Anti-tumor effects of interferon in mice injected with interferon-sensitive and interferon-resistant Friend erythroleukemia cells. VIII. Role of the immune system in the inhibition of visceral metastases. Int J Cancer. 1990 Sep 15;46(3):468–474. [PubMed]
  • Gresser I, Tovey MG, Bandu ME, Maury C, Brouty-Boyé D. Role of interferon in the pathogenesis of virus diseases in mice as demonstrated by the use of anti-interferon serum. I. Rapid evolution of encephalomyocarditis virus infection. J Exp Med. 1976 Nov 2;144(5):1305–1315. [PMC free article] [PubMed]
  • Gresser I, Maury C, Kaido T, Bandu MT, Tovey MG, Maunoury MT, Fantuzzi L, Gessani S, Greco G, Belardelli F. The essential role of endogenous IFN alpha/beta in the anti-metastatic action of sensitized T lymphocytes in mice injected with Friend erythroleukemia cells. Int J Cancer. 1995 Nov 27;63(5):726–731. [PubMed]
  • Gresser I, Kaido T, Maury C, Woodrow D, Moss J, Belardelli F. Interaction of IFN alpha/beta with host cells essential to the early inhibition of Friend erythroleukemia visceral metastases in mice. Int J Cancer. 1994 May 15;57(4):604–611. [PubMed]
  • Belardelli F, Gresser I, Maury C, Maunoury MT. Antitumor effects of interferon in mice injected with interferon-sensitive and interferon-resistant Friend leukemia cells. II. Role of host mechanisms. Int J Cancer. 1982 Dec 15;30(6):821–825. [PubMed]
  • Gresser I, Carnaud C, Maury C, Sala A, Eid P, Woodrow D, Maunoury MT, Belardelli F. Host humoral and cellular immune mechanisms in the continued suppression of Friend erythroleukemia metastases after interferon alpha/beta treatment in mice. J Exp Med. 1991 May 1;173(5):1193–1203. [PMC free article] [PubMed]
  • Gresser I, Maury C, Woodrow D, Moss J, Grütter MG, Vignaux F, Belardelli F, Maunoury MT. Interferon treatment markedly inhibits the development of tumor metastases in the liver and spleen and increases survival time of mice after intravenous inoculation of Friend erythroleukemia cells. Int J Cancer. 1988 Jan 15;41(1):135–142. [PubMed]
  • Belardelli F, Gabriele L, Proietti E, Sestili P, Peretti M, Rozera C, Gresser I. Synergistic anti-tumor effects of combined IL-1/IFN-alpha/beta therapy in mice injected with metastatic Friend erythroleukemia cells. Int J Cancer. 1991 Sep 9;49(2):274–278. [PubMed]
  • Belardelli F, Proietti E, Ciolli V, Sestili P, Carpinelli G, Di Vito M, Ferretti A, Woodrow D, Boraschi D, Podo F. Interleukin-1 beta induces tumor necrosis and early morphologic and metabolic changes in transplantable mouse tumors. Similarities with the anti-tumor effects of tumor necrosis factor alpha or beta. Int J Cancer. 1989 Jul 15;44(1):116–123. [PubMed]
  • Belardelli F, Ciolli V, Testa U, Montesoro E, Bulgarini D, Proietti E, Borghi P, Sestili P, Locardi C, Peschle C, et al. Anti-tumor effects of interleukin-2 and interleukin-1 in mice transplanted with different syngeneic tumors. Int J Cancer. 1989 Dec 15;44(6):1108–1116. [PubMed]
  • Tough DF, Borrow P, Sprent J. Induction of bystander T cell proliferation by viruses and type I interferon in vivo. Science. 1996 Jun 28;272(5270):1947–1950. [PubMed]
  • Testa U, Martucci R, Rutella S, Scambia G, Sica S, Benedetti Panici P, Pierelli L, Menichella G, Leone G, Mancuso S, et al. Autologous stem cell transplantation: release of early and late acting growth factors relates with hematopoietic ablation and recovery. Blood. 1994 Nov 15;84(10):3532–3539. [PubMed]
  • Unutmaz D, Pileri P, Abrignani S. Antigen-independent activation of naive and memory resting T cells by a cytokine combination. J Exp Med. 1994 Sep 1;180(3):1159–1164. [PMC free article] [PubMed]
  • Rocha B, Dautigny N, Pereira P. Peripheral T lymphocytes: expansion potential and homeostatic regulation of pool sizes and CD4/CD8 ratios in vivo. Eur J Immunol. 1989 May;19(5):905–911. [PubMed]
  • Gold JE, Osband ME. Autolymphocyte therapy. II. Dependence of in vivo anti-tumor specificity and long-term immunity against murine melanoma and carcinoma on ex vivo activated donor memory T-cells. Clin Immunol Immunopathol. 1994 Jun;71(3):325–332. [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...