Abstract
Cell suspensions from 69 human tumor biopsies and malignant effusions depleted of infiltrating T cells were incubated for 10–14 days with mitomycin-C-treated cells of the transformed T cell line MOT as feeder cells. B lymphocytes proliferated and differentiated as indicated by immunoglobulin (Ig) seerction in the culture supernatants (B cell expansion). Ig was present in culture supernatants of tumor cell suspensions incubated without MOT feeder cells (non-expanded cells), but the addition of MOT feeder cells to these cultures invariably resulted in a significant increase in Ig concentration. While IgG, IgA. and IgM isotypes were all detected in supernatants of both expanded- and nonexpanded tumor cell suspensions, the increase in total Ig induced by MOT feeder cells was mainly due to an increase in IgG. Peripheral blood B lymphocytes (PBBL) from 15 cancer patients and 4 healthy individuals were also successfully expanded by the same method. In these it was shown that IgA was the predominant Ig isotype. Using a modified enzyme-linked immunosorbent assay, IgG of 25/36 expansions from tumor cell suspensions showed reactivity with autologous tumor targets, and that from 10/13 expansions reacted with allogeneic tumor targets of the same histological diagnosis. No reactivity was found against tumor targets of different histology. IgG of 4/10 expansions of PBBL from cancer patients showed reactivity with allogeneic tumor targets of the same histology, while no reactivity was demonstrated against tumor targets of different histology. IgG of expanded PBBL from healthy individuals showed no reactivity against tumor targets. This method allows detailed study of the specific humoral antitumor immune response of intratumoral and peripheral blood B lymphocytes in cancer.
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References
Topalian SL, Rosenberg SA (1991) Adoptive cellular therapy: basic principles. In: DeVita VT Jr, Hellman S, Rosenberg SA (eds) Biologic therapy of cancer. Lippincott, Philadelphia, p 187
Hurlimann J, Saraga P (1985) Mononuclear cells infiltrating human mammary carcinomas: immunohistochemical analysis with monoclonal antibodies. Int J Cancer 35: 753
Whiteside TL, Miescher S, Hurlimann J, Moretta L, Von Fliedner V (1986) Clonal analysis and in situ characterization of lymphocytes infiltrating human breast carcinomas. Cancer Immunol Immunother 23: 169
Ferguson A, Moore M, Fox H (1985) Expression of MHC products and leucocyte differentiation antigens in gynaecological neoplasms: an immunohistological analysis of the tumour cells and infiltrating leucocytes. Br J Cancer 52: 551
Wolf GT, Hudson JL, Peterson KA, Miller HL, McClatchey KD (1986) Lymphocyte subpopulations infiltrating squamous carcinomas of the head and neck: correlations with extent of tumor and prognosis. Otolaryngol Head Neck Surg 95: 142
Schoorl R, Brutel de la Rivière A, Borne AEGKr von dem, Feltkamp-Vroom TM (1976) Identification of T and B lymphocytes in human breast cancer with immunohistochemical techniques. Am J Pathol 84: 529
Husby G, Hoagland PM, Strickland RG, Williams RC (1976) Tissue T and B cell infiltration of primary and metastatic cancer. J Clin Invest 57: 1471
Stevens A, Klöter I, Roggendorf W (1988) Inflammatory infiltrates and natural killer cell presence in human brain tumors. Cancer 61: 738
Vose BM, Moore M (1979) Suppressor cell activity of lymphocytes infiltrating human lung and breast tumours. Int J Cancer 24: 579
Bröcker EB, Zwadlo G, Holzmann B, Macher E, Sorg C (1988) Inflammatory cell infiltrates in human melanoma at different stages of tumor progression. Int J Cancer 41: 562
Withford P, Mallon EA, George WD, Campbell AM (1990) Flow cytometric analysis of tumor infiltrating lymphocytes in breast cancer. Br J Cancer 62: 971
Sikora K, Alderson T, Philips J, Watson JV (1982) Human hybridomas from malignant gliomas. Lancet 1: 11
Sikora K, Alderson T, Ellis J, Philips J, Watson J, (1983) Human hybridomas from patients with malignant diseases. Br J Cancer 47: 135
Jerne NK, Henry C, Nordin AA, et al (1974) Plaque forming cells: methodology and theory. Transplant Rev 18: 130
Henderson E, Miller G, Robinson J, Heston L (1977) Efficiency of transformation of lymphocytes by Epstein-Barr virus. Virology 76: 152
Aman P, Ehlin-Henriksson B, Klein G (1984) Epstein-Barr virus susceptibility of normal human B lymphocyte populations. J Exp Med 159: 208
Kozbor D, Croce CH (1985) Fusion partners for production of human monoclonal antibodies. In: Engleman EG, Foung SKH, Larrick J, Raubitschek A (eds) Human hybridomas and monoclonal antibodies, Plenum, New York, p 21
Hirohata S, Jelinek DF, Lipsky PE (1988) T cell-dependent activation and differentiation of B cell proliferation by immobilized monoclonal antibodies to CD3. J Immunol 140: 3736
Amoroso K Lipsky PE (1990) Frequency of human B cells that differentiate in response to anti-CD3-activated T cells. J Immunol 145: 3155
Kelly PJ, Pascual V, Capra JD, Lipsky PE (1992) Anti-CD3-stimulated T cells induce the production of multiple Ig H chain isotypes by individual human peripheral B lymphocytes. J Immunol 148: 1294
Barbuto JAM, Verastegui EL, Hersh EM (1993) The use of transformed T cell lines for clonal expansion of human B cells from peripheral blood, spleen, and tumor-infiltrating lymphocytes. Hybridoma 12: 115
Hersh EM, Mavligit GM, Gutterman JU (1976) Immunodeficiency in cancer and the importance of immune evaluation of the cancer patient. Med Clin N Am 60: 623
Miescher S, Whiteside TL, Carrel S, Von Fliedner V (1986) Functional properties of tumor-infiltrating and blood lymphocytes in patients with solid tumors: effects of tumor cells and their supernatants on proliferative responses of lymphocytes. J Immunol 136: 1899
Lane HC, Fauci AS (1985) Immunologic abnormalities in the acquired immunodeficiency syndrome. Annu Rev Immunol 3: 477
Saxon A, Feldhaus J, Robins RA (1976) Single step separation of human T and B cells using AET treated sheep red cells. J Immunol Methods 90: 221
Thiele DL, Kurosaka M, Lipsky PE (1983) Phenotype of the accessory cell necessary for mitogen-stimulated T and B cell responses in human peripheral blood: delineation by its sensitivity to the lysomotropic agent,l-leucine methyl ester. J. Immunol 131: 2282
Bishop GA, Hwang J (1992) Use of a cellular ELISA for the detection of cell surface antigens. Biotechniques 12: 326
Henry C, Marbrook J, Vann DC, Kondlin D, Wofsy C (1980) Limiting dilution analysis. In: Mishell BB, Shiigi SM (eds) Selected methods in cellular immunology. Freeman, San Francisco, p 138
Chandler HM, Cox JC, Healy K, MacGregor A, Premier RR, Hurrell JGR (1982) An investigation of the use of urease-antibody conjugates in enzyme immunoassays. J Immunol Methods 53: 187
Douillard JY, Hoffman T, Herberman RB (1980) Enzyme-linked immunosorbent assay for screening monoclonal antibody production: use of intact cells as antigen. J Immunol Methods 39: 309
Suter L, Bruggen J, Sorg C (1980) Use of an enzyme-linked immunosorbent assay (ELISA) for screening of hybridoma antibodies against cell surface antigens. J Immunol Methods 39: 407
Hellström KE, Hellström I (1991) Principles of tumor immunity: tumor antigens. In: DeVita VT Jr, Hellman S, Rosenberg SA (eds) Biologic therapy of cancer. Lippincott, Philadelphia, p 42
Kawakami Y, Zakut R, Topalian SL, Stötter H, Rosenberg SA (1992) Shared human melanoma antigens. Recognition by tumor-infiltrating lymphocytes in HLA-A2.1-transfected melanomas. J Immunol 148: 638
Sjögren HD, Hellström I, Bansal SC, Warner GA, Hellström KE (1972) Elution of “blocking factors” from human tumors, capable of abrogating tumor-cell destruction by specific immune lymphocytes. Int J Cancer 9: 274
Manson LA (1991) Does antibody-dependent epitope masking permit progressive tumour growth in the face of cell-mediated cytotoxicity? Immunol Today 12: 352
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Work supported by grants from the Share and Concern Foundations and grant CA MOPP from the National Institutes of Health, C.J.A.P. is a visiting scientist from the University of Nijmegen, Department of Medical Oncology, Nijmegen, The Netherlands, and is supported by a Fulbright Senior Research Grant and grants from the Dutch Cancer Society and the Regional Cancer Center of the East Netherlands (IKO). J.A.M.B. is a visiting scientist from the University of Sao Paolo, Department of Immunology, Brazil, and is supported by grant 90/1844-4 from the FAPESP
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Punt, C.J.A., Barbuto, J.A.M., Zhang, H. et al. Anti-tumor antibody produced by human tumor-infiltrating and peripheral blood B lymphocytes. Cancer Immunol Immunother 38, 225–232 (1994). https://doi.org/10.1007/BF01533513
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DOI: https://doi.org/10.1007/BF01533513