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. 1991 Mar;82(3):332–338. doi: 10.1111/j.1349-7006.1991.tb01850.x

Characterization and Purification of an Immunosuppressive Factor Produced by a Small Cell Lung Cancer Cell Line

Toshiyuki Ikeda 1,✉, Tomiya Masuno 1, Takeshi Ogura 2, Masatoshi Watanabe 1, Takuma Shirasaka 1, Hideki Kara 1, Yoshiro Tanio 1, Ichiro Kawase 1, Susumu Kishimoto 1
PMCID: PMC5918405  PMID: 1850726

Abstract

The present study was undertaken to determine whether small cell lung cancer (SCLC) cell lines produce immunosuppressive factors and, if they do, to characterize the factors. The supernatants of SCLG cell lines, H69 and N857, inhibited not only the blastogenic response of human peripheral blood lymphocytes (PBL) to phytohemagglutinin or concanavalin A, but also the cytotoxic activity of lymphokine‐activated killer cells. Neither was inhibited by supernatants from non‐SCLC cell lines PC9, QG56, and A549. The immunosuppressive activity of H69 supernatant was stable upon heating to 56°C for 60 min, but labile when heated to 70°C for 10 min. The activity was abolished after dialysis at pH 2.0 or pH 11.0, but not at pH 4.5 or pH 9.0. Digestion with trypsin or proteinase eliminated the immunosuppressive activity, whereas treatment with neuraminidase, mixed glycosidase, DNase or RNase had no effect, suggesting that the immunosuppressive activity in H69 supernatant is due to a protein factor. This H69‐derived immunosuppressive factor was isolated by ion exchange chromatography using a gradient of 0.04 to 0.08 M NaCl solution. Gel filtration and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis showed the factor to have molecular weights of 98 kD and 102 kD, respectively. These results suggest that SCLC cells produce a potent immunosuppressive factor which may account for the immune deficiency in SCLC patients.

Keywords: Small cell lung cancer, Immunosuppressive factor, Lymphocyte blastogenesis

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REFERENCES

  • 1. ) Faguet , G. B.Quantitation of immunocompetence in Hodgkin's disease . J. Clin. Invest. , 56 , 951 – 957 ( 1975. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. ) Glasgow , A. H. , Nimberg , R. B. , Menzoian , J. O. , Saporoschetz , I. , Cooperband , S. R. , Schmid , K. and Mannick , J. A.Association of anergy with an immuno‐suppressive peptide fraction in the serum of patients with cancer . N. Engl. J. Med. , 291 , 1263 – 1267 ( 1974. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Takasugi , M. , Ramseyer , A , and Takasugi , J.Decline of natural nonselective cell‐mediated cytotoxicity in patients with tumor progression . Cancer Res. , 37 , 413 – 418 ( 1977. ). [PubMed] [Google Scholar]
  • 4. ) Field , E. J. and Caspary , E. A.Lymphocyte sensitization in advanced malignant disease: a study of serum lymphocyte depressive factor . Br. J. Cancer , 26 , 164 – 173 ( 1972. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5. ) Mannick , J. A. , Constantin , M. , Pardridge , D. , Saporoschetz , I. and Badger , A.Improvement of phytohemagglutinin responsiveness of lymphocytes from cancer patients after washing in vitro . Cancer Res. , 37 , 3066 – 3070 ( 1977. ). [PubMed] [Google Scholar]
  • 6. ) Miller , F.Serum‐derived immunosuppressive substances . Transplantation , 21 , 179 – 187 ( 1976. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Ting , C. C.Humoral regulation of cell‐mediated immunity to syngeneic tumor . Cancer Res. , 36 , 3695 – 3701 ( 1976. ). [PubMed] [Google Scholar]
  • 8. ) Wang , B. S. , Badger , A. M. , Nimberg , R. B. , Cooperband , S. R. , Schmid , K. and Mannick , J. A , Suppression of tumor‐specific cell‐mediated cytotoxicity by immunoregulatory α‐globulin and by immunoregulatory α‐globulin‐like peptide from cancer patients . Cancer Res. , 37 , 3022 – 3025 ( 1977. ). [PubMed] [Google Scholar]
  • 9. ) Nimberg , R. B. , Glasgow , A. H. , Menzoian , J. O. , Con‐stantian , M. B. , Cooperband , S. R. , Mannick , J. A. and Schmid , K.Isolation of an immunosuppressive peptide fraction from the serum of cancer patients . Cancer Res. , 35 , 1489 – 1494 ( 1975. ). [PubMed] [Google Scholar]
  • 10. ) Cornelius , J. G. and Normann , S. J.Isolation of a low molecular weight inhibitor of lymphocyte proliferation from tumorous ascites . J. Biol. 141 , 2175 – 2180 ( 1988. . [PubMed] [Google Scholar]
  • 11. ) Hess , A. D. , Gall , S. A. and Dawson , J. R.Partial purification and characterization of a lymphocyte‐inhibitory factor(s) in ascitic fluids from ovarian cancer patients . Cancer Res. , 40 , 1842 – 1851 ( 1980. ). [PubMed] [Google Scholar]
  • 12. ) Medoff , J. R. , Clack , V. D. and Roche , J. K.Characterization of an immunosuppressive factor from malignant ascites that resembles a factor induced in vitro by carcinoembryonic antigen . J. Immunol , 137 , 2057 – 2064 ( 1986. ). [PubMed] [Google Scholar]
  • 13. ) Tamura , K‐ , Shibata , Y. , Matsuda , Y. and Ishida , N.Isolation and characterization of immunosuppressive ascitic protein from ascitic fluids of cancer patients . Cancer Res. , 41 , 3244 – 3252 ( 1981. ). [PubMed] [Google Scholar]
  • 14. ) Hersey , P. , Bindon , C , Czerniecki , M. , Spurling , A. , Wass , J. and McCarthy , W. H.Inhibition of interleukin 2 production by factors released from tumor cells . J. Immunol , 131 , 2837 – 2842 ( 1983. ). [PubMed] [Google Scholar]
  • 15. ) Putnam , J. B.Jr. and Roth , J. A.Identification and characterization of a tumor‐derived immunosuppressive glycoprotein from murine melanoma K‐1735 . Cancer Immunol Immunother. , 19 , 90 – 100 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16. ) Renk , C. M. , Gupta , R. K. and Morton , D. L.Inhibition of normal allogeneic lymphocyte mitogenesis by a factor released from human tumor cells in culture . Cancer Immunol. Immunother. , 11 , 7 – 16 ( 1981. ). [Google Scholar]
  • 17. ) Werktneister , J. , Zbroja , R. , McCarthy , W. and Hersey , P.Detection of an inhibitor of cell division in cultures of tumour cells with immunosuppressive activity in vitro . Clin. Exp. Immunol , 40 , 168 – 177 ( 1980. ). [PMC free article] [PubMed] [Google Scholar]
  • 18. ) Werkmeister , J. , Zaunders , J. , McCarthy , W. and Hersey , P.Characterization of an inhibitor of cell division released in tumour cell cultures , Clin. Exp. Immunol , 41 , 487 – 496 ( 1980. ). [PMC free article] [PubMed] [Google Scholar]
  • 19. ) Masuno , T. , Ikeda , T. , Yokota , S. , Komuta , K. , Ogura , T. and Kishimoto , S.Immunoregulatory T‐lymphocyte functions in patients with small cell lung cancer . Cancer Res. , 46 , 4195 – 4199 ( 1986. ). [PubMed] [Google Scholar]
  • 20. ) Yokota , S. , Shirasaka , T. , Nishikawa , H. , Hosoe , S. , Ikeda , T. , Komuta , K. , Kawase , L , Masuno , T. , Ogura , T. and Kishimoto , S.Augmentative effects of Nocardia rubra cell‐wall skeleton (N‐CWS) on lymphokine‐activated killer (LAK) cell induction . Cancer Immunol. Immunother. , 26 , 11 – 17 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. ) Halper , J. and Moses , H. L.Epithelial tissue‐derived growth factor‐like polypeptides . Cancer Res. , 43 , 1972 – 1979 ( 1983. ). [PubMed] [Google Scholar]
  • 22. ) Okada , M. , Yoshimura , N. , Ichimori , Y. , Kishimoto , S. , Nakaj , S. , Nishino , N. and Kishimoto , T.Immunologic and molecular characterization of T cell‐derived T cell activating factor . J. Immunol , 136 , 1288 – 1294 ( 1986. ). [PubMed] [Google Scholar]
  • 23. ) Laemmli , U. K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature , 227 , 680 – 685 ( 1970. ). [DOI] [PubMed] [Google Scholar]
  • 24. ) Dent , R. G. and Cole , P.In vitro monocyte maturation in squamous carcinoma of the lung . Br. J. Cancer , 43 , 486 – 495 ( 1981. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. ) Figarella , E. F. , Morillo , F. , Blanca , I. and Bianco , N. E.Failure of cell‐mediated effector mechanisms in lung cancer . J. Natl Cancer Inst. , 73 , 1 – 6 ( 1984. ). [PubMed] [Google Scholar]
  • 26. ) Ginns , L. C. , Goldenheim , P. D. , Miller , L. G. , Burton , R. C. , Gillick , L. , Colvin , R. B. , Goldstein , G. , Kung , P. C. , Hurwitz , C. and Kazemi , H.T‐lymphocyte subsets in smoking and lung cancer . Am. Rev. Respir. Dis. , 126 , 265 – 269 ( 1982. ). [DOI] [PubMed] [Google Scholar]
  • 27. ) Hadjipetrou‐Kourounakis , L. and Tsougranis , A.Diminished lymphocyte response to phytohemagglutinin in lung cancer . Biochem. Exp. Biol , 16 , 357 – 364 ( 1980. ). [PubMed] [Google Scholar]
  • 28. ) Krant , M. J. , Manskopf , G. , Brandrap , C. S. and Madoff , M. A.Immunologic alterations in bronchogenic cancer . Cancer , 21 , 623 – 631 ( 1968. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) Sibbitt , W. L. , Jr. , Bankhurst , A. D. , Jumonville , A. J. , Saiki , J. H. , Saiers , J. H. and Doberneck , R. C.Defect in natural killer cell activity and interferon response in human lung carcinoma and malignant melanoma . Cancer Res. 44 , 852 – 856 ( 1984. . [PubMed] [Google Scholar]
  • 30. ) Bray , M. A. , Gordon , D. and Morley , J.Prostaglandins as regulators in cellular immunity . Prostaglandins Med. , 1 , 183 – 199 ( 1978. ). [DOI] [PubMed] [Google Scholar]
  • 31. ) Chouaib , S. , Welte , K. , Mertelsmann , R. and Dupont , B.Prostaglandin E2 acts at two distinct pathways of T lymphocyte activation: inhibition of interleukin 2 production and down‐regulation of transferrin receptor expression . J. Immunol , 135 , 1172 – 1179 ( 1985. ). [PubMed] [Google Scholar]
  • 32. ) Gordon , D. A. , Bray , M. A. and Morley , J.Control of lymphokine secretion by prostaglandins . Nature , 262 , 401 – 402 ( 1976. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Kehrl , J. H , Wakefield , L. M. , Roberts , A. B , Jakowlew , S. , Alvarez‐Mon , M. , Derynck , R. , Sporn , M. B. and Fauci , A. S.Production of transforming growth factor β by human T lymphocytes and its potential role in the regulation of T cell growth . J. Exp. Med. , 163 , 1037 – 1050 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34. ) Wrann , M. , Bodmer , S. , Martin , R. , Siepl , C. , Hofer‐Warbinek , R. , Frei , K. , Hofer , E. and Fontana , A.T cell suppressor factor from human glioblastoma cells is a 12.5‐kd protein closely related to transforming growth factor‐β . EMBOJ. , 6 , 1633 – 1636 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35. ) Shirakawa , F. , Tanaka , Y. , Oda , S. , Chiba , S. , Suzuki , H. , Eto , S. and Yamashita , U.Immunosuppressive factors from adult T‐cell leukemia cells . Cancer Res. , 46 , 4458 – 4462 ( 1986. ). [PubMed] [Google Scholar]
  • 36. ) Ebert , E. C. , Roberts , A. L , O'Connell , S. M. , Robertson , F. M. and Nagase , H.Characterization of immunosuppressive factor derived from colon cancer cells . J. Immunol , 138 , 2161 – 2168 ( 1987. ). [PubMed] [Google Scholar]

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