Skip to main content
SpringerLink
Log in

Phenotypic and functional analysis of lymphocytes infiltrating paediatric tumours, with a characterization of the tumour phenotype

  • Original articles
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Summary

Tumour-infiltrating lymphocytes (TIL) of paediatric tumours obtained from 37 lesions of different histo-type (12 osteosarcomas, 5 Wilms' tumours, 7 soft-tissue sarcomas, 5 neuroblastomas and 8 miscellaneous) were studied to establish their potential for therapy. Fresh isolated TIL were cultured for the first 2 weeks with low doses of interleukin-2 (IL-2) (20 Cetus U/ml) to select for “tumour-specific” lymphocytes potentially present in the neoplastic lesion, followed by culture with high doses of IL-2 (1000 Cetus U/ml) to achieve TIL expansion. TIL were grown with more than 10-fold expansion in only 9 cases (mean expansion: 58-fold, range 13.5–346). In 17 cases no viable cells were obtained. After 30 days of culture with IL-2 the proliferative ability of TIL declined sharply in the majority of cases and TIL became refractory to any further stimulus, including addition of IL-4, tumour necrosis factor α (TNFα) or interferon γ, and activation with OKT3 in solid phase. In 20 out of 37 cases TIL were available for phenotypic and functional analysis. TIL after long-term culture were predominantly CD3+ but 2 cases of osteosarcoma showed a predominance of CD3+TcR γ/δ cells. The CD4/CD8 ratio was more than 1 in 10 cases, without correlation with tumour histology, site of lesion or TIL growth. The number of CD16+ and CD25+ lymphocytes decreased progressively during culture, the latter concomitantly with a reduction of TIL growth rate. The lytic pattern of TIL against allogeneic and autologous tumour (Auto-Tu) cells was variable, but specific lysis of Auto-Tu was seen in only one case (Wilms' tumour) after culture with TNFα and irradiated Auto-Tu cells. The immunohistochemical analysis of tumour lesions revealed a limited lymphocyte infiltrate, a low expression of histocompatibility leukocyte antigens (HLA) class I and of the adhesion molecules ICAM1, LFA3, and a significant production of transforming growth factor β (TGFβ). These data indicate that TIL obtained from paediatric patients are difficult to expand at levels required for immunotherapy and lack a significant number of tumour-specific T lymphocytes. A low expression of immunomodulatory molecules on tumour cells or the production of suppressive factors may prevent activation and expansion of TIL in paediatric tumours.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Anichini A, Mortarini R, Supino R, Parmiani G (1990) Human melanoma cells with high susceptibility to cell-mediated lysis can be identified on the basis of ICAM-1 phenotype, VLA profile and invasive ability. Int J Cancer 46: 508

    Google Scholar 

  2. Balch CM, Riley LB, Bae YJ, Salmeron MA, Platsoucas CD, von Eschenbach A, Itoh K (1990) Patterns of human tumor-infiltrating lymphocytes in 120 human cancers. Arch Surg 125: 200

    Google Scholar 

  3. Belldegrun A, Muul LM, and Rosenberg SA (1988) Interleukin-2 expanded tumor-infiltrating lymphocytes in human renal cancer: isolation, characterization and antitumor activity. Cancer Res 48: 206

    Google Scholar 

  4. Cattoretti G, Berti E, Schiro R, D'Amato L, Valeggio C, Rilke F (1988) Improved avidin-biotin-peroxidase complex (ABC) staining. Histochem J 20: 75

    Google Scholar 

  5. Favrot MC, Floret D, Negrier S, Cochat P, Bouffet E, Dacheng Z, Franks CR, Bijman T, Brunat-Mentigny M, Philip I, Philip T (1989) Systemic interleukin-2 therapy in children with progressive neuroblastoma after high dose chemotherapy and bone marrow transplantation. Bone Marrow Transplant 4: 499

    Google Scholar 

  6. Ferrini S, Biassoni R, Moretta A, Bruzzone M, Nicolin A, Moretta L (1985) Clonal analysis of T lymphocytes isolated from ovarian carcinoma ascitic fluid. Phenotypic and functional characterization of T cell clones capable of lysing autologous carcinoma cells. Int J Cancer 36: 337

    Google Scholar 

  7. Ferrini S, Pigione I, Bottino C, Ciccone G, Tambussi S, Mannoliti S, Moretta L, Moretta A (1989) Monoclonal antibodies which react with the T cell receptor γ/δ recognize different substes of CD3+ WT31− T lymphocytes. Eur J Immunol 19: 57

    Google Scholar 

  8. Fisch P, Malkovsky M, Klein BS, Morrisey LW, Carper SW, Welch WJ, Sondel PM (1990) Human V γ9/V δ 2T cells recognize a groEL homolog on Daudi Burkitt's lymphoma cells. Science 250: 1269

    Google Scholar 

  9. Haas GP, Solomon D, Rosenberg SA (1990) Tumor-infiltrating lymphocytes from nonrenal urological malignancies. Cancer Immunol immunother 30: 342

    Google Scholar 

  10. Heo DS, Whiteside TL, Johnson JT, Chen KN, Barnes EL, Herberman RB (1987) Long-term interleukin-2-dependent growth and cytotoxic activity of tumor-infiltrating lymphocytes from human squamous cell carcinoma of the head and neck. Cancer Res 47: 6353

    Google Scholar 

  11. Heo DS, Whiteside TL, Kanbour A, Herberman RB (1988) Lymphocytes infiltrating human ovarian tumors: I. Role of Leu19(NKH1)-positive recombinant IL-2 activated cultures of lymphocytes infiltrating human ovarian cancers. J Immunol 140: 4042

    Google Scholar 

  12. Itoh K, Platsoucas CD, Balch CM (1988) Autologous tumor-specific cytotoxic T lymphocytes in the infiltrate of human metastatic melanomas: activation by interleukin-2 and autologous tumor cells, and involvement of the T cell receptor. J Exp Med 168: 1419

    Google Scholar 

  13. Kawakami Y, Rosenberg SA, Lotze MT (1988) Interleukin-4 promotes the growth of tumor-infiltrating lymphocytes cytotoxic for human autologous melanoma. J Exp Med 168: 2183

    Google Scholar 

  14. Kuppner MC, Hamon MF, deTribolet N (1988) Immunohistological and functional analysis of lymphoid infiltrates in human glioblastomas. Cancer Res 48: 6926

    Google Scholar 

  15. Kurnick JT, Kradin RL, Blumberg R, Schneeberger EE, Boyle LA (1986) Functional characterization of T lymphocytes propagated from human lung carcinomas. Clin Immunol Immunopathol 38: 367

    Google Scholar 

  16. Mechtersheimer G, Staudter M, Majdic O, Dorken B, Moldenhauer G, Moller P (1990) Expression of MHC-A,B,C,β2-microglobulin (β2m), MHC-DR,-DP,-DQ and of MHC-D-associated invariant chain (Ii) in soft-tissue tumors. Int J Cancer 46: 813

    Google Scholar 

  17. Miescher S, Stoeck M, Qiao L, Barras L, Barrelet L, Von Fliedner V (1988) Preferential clonogenic deficit of CD8-positive T-lymphocytes infiltrating human solid tumors. Cancer Res 48: 6992–6998

    Google Scholar 

  18. Mulé JJ, Schwarz SL, Roberts AB, Sporn MB, Rosenberg SA (1988) Transforming growth factor-β inhibits the in vitro generation of lymphokine-activated killer cells and cytotoxic T cells. Cancer Immunol Immunother 26: 95

    Google Scholar 

  19. Nasr S, McKolanis J, Pais R, Findley H, Hnath R, Waldrep K, Ragab AH (1989) A phase I study of interleukin-2 in children with cancer and evaluation of clinical and immunological status during therapy — a Pediatric Oncology Group study. Cancer 64: 783

    Google Scholar 

  20. Pais R, Igrim NB, Garcia ML, Abdel-Mageed A, McKolanis J, Igrim ME, Hnath RS, Ziegler K, Ragab AH (1990) Pharmacokinetics of recombinant interleukin-2 in children with malignancies: a Pediatric Oncology Group study. J Biol Response Mod 9: 517

    Google Scholar 

  21. Ranges GE, Figari IS, Espevik T, Palladino MA (1987) Inhibition of cytotoxic T cell development by transforming growth factor-β and reversal by recombinant tumor necrosis factor-α. J Exp Med 166: 991

    Google Scholar 

  22. Rosenberg SA (1988) Immunotherapy of cancer using interleukin 2: current status and future prospects. Immunol Today 9: 58

    Google Scholar 

  23. Rosenberg SA, Spiess P, Lafreniere R (1986) A new approach to the adoptive immunotherapy of cancer with tumor-infiltrating lymphocytes. Science 233: 1318

    Google Scholar 

  24. Rosenberg SA, Packard BS, Aebersold PM, Solomon D, Topalian SL, Toy ST, Simon P, Lotze MT, Yang JC, Seipp CA, Simpson C, Carter C, Bock S, Schwartzentruber D, Wei J, White D (1988) Use of tumor infiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. N Engl J Med 319: 1676

    Google Scholar 

  25. Schoof DD, Selleck CM, Massaro AF, Jung S-F, Eberlein TJ (1990) Activation of tumor infiltrating lymphocyte by monoclonal antibodies directed to the CD3 complex. Cancer Res 50: 1138

    Google Scholar 

  26. Shimizu Y, Iwatsuki S, Herberman RB, Whiteside TL (1991) Effects of cytokines on in vitro growth of tumor-infiltrating lymphocytes obtained from human primary and metastatic liver tumors. Cancer Immunol Immunother 32: 280

    Google Scholar 

  27. Skornick Y, Topalian S, Rosenberg SA (1990) Comparative studies of the long-term growth of lymphocytes from tumor infiltrates, tumor draining lymph nodes and peripheral blood by repeated in vitro stimulation with autologous tumor. J Biol Response Mod 9: 431

    Google Scholar 

  28. Sporn MB, Roberts AB, Wakefield LM, Assoian RK (1986) Transforming growth factor-β: biological function and chemical structure. Science 233: 532

    Google Scholar 

  29. Takagi S, Chen K, Schwarz R, Iwatsuki S, Herberman RB, Whiteseide TL (1989) Functional and phenotypic analysis of tumor-infiltrating lymphocytes isolated from human primary and metastatic liver tumors and cultured in recombinant interleukin-2. Cancer 63: 102

    Google Scholar 

  30. Topalian SL, Muul LM, Rosenberg SA (1987) Growth and immunological characteristics of lymphocytes infiltrating human tumors. Surg Forum 37: 390

    Google Scholar 

  31. Triebel F, Faure F, Mami-Chouaib F, Jistukawa S, Griscelli AL, Genevee C, Roman-Roman S, Hercend T (1988) A novel human V/δ gene expressed predominantly in the Ti/γ/A+ fraction of γ/δ+ peripheral lymphocytes

  32. Vaccarello L, Wang YL, Whiteside TL (1990) Substained outgrowth of autotumor-reactive T lymphocytes from human solid tumors in the presence of tumor necrosis factor-α and interleukin-2. Hum Immunol 28: 216

    Google Scholar 

  33. Vanky F, Wang P, Patarroyo M, Klein E (1990) Expression of the adhesion molecule ICAM1 and major histocompatibility complex class I antigens on human tumor cells is required for their interaction with autologous lymphocytes in vitro. Cancer Immunol Immunother 31: 19

    Google Scholar 

  34. Weber JS, Rosenberg SA (1990) Effects of murine tumor class I major histocompatibility complex expression on anti-tumor activity of tumor-infiltrating lymphocytes. J Natl Cancer Inst 82: 755

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Experimental Oncology D, Istituto Nazionale Tumori, Via Venezian 1, I-20 133, Milano, Italy

    Licia Rivoltini, Flavio Arienti, Carlo Gambacorti-Passerini & Giorgio Parmiani

  2. Division of Pathology, Istituto Nazionale Tumori, I-20 133, Milan, Italy

    Attilio Orazi

  3. Division of Paediatric Oncology, Istituto Nazionale Tumori, I-20 133, Milan, Italy

    Graziella Cefalo, Marco Gasparini & Franca Fossati-Bellani

Authors
  1. Licia Rivoltini
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Flavio Arienti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Attilio Orazi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Graziella Cefalo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marco Gasparini
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Carlo Gambacorti-Passerini
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Franca Fossati-Bellani
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Giorgio Parmiani
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rivoltini, L., Arienti, F., Orazi, A. et al. Phenotypic and functional analysis of lymphocytes infiltrating paediatric tumours, with a characterization of the tumour phenotype. Cancer Immunol Immunother 34, 241–251 (1992). https://doi.org/10.1007/BF01741792

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01741792

Key words

Search

Navigation