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Cryopreservation of human mononuclear cells for quality control in clinical immunology. I. Correlations in recovery of K- and NK-cell functions, surface markers, and morphology

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Abstract

Cryopreserved human peripheral blood mononuclear cells (PBMC) were tested for natural killer (NK) and antibody-dependent cellular cytotoxicity (ADCC) and for high-affinity (29°C) and total (4°C) rosette formation with sheep erythrocytes. PBMC produced variable NK activity following freezing and thawing, but consistently reacted well in ADCC. A significant correlation was found between low NK activity and a decreased percentage of low-affinity rosette-forming cells. On the contrary, the number of large granular lymphocytes (LGL), among which NK cells are restricted, and the reactivity with the monoclonal antibody OKT10, which recognizes the majority of LGL in the peripheral blood, were not significantly altered by cryopreservation. Cryopreserved cells proved to be excellent controls for determining the day-to-day variability of the NK assay and for selecting optimum conditions for this test in the clinical immunology laboratory.

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References

  1. Weiner RS, Breard J, O'Brien C: Cryopreserved lymphocytes in sequential studies of immune responsives: Problems and prospects. Preliminary report.In Cryopreservation of Normal and Neoplastic Cells, RS Weiner, RK Oldham, L Schwartzenberg (eds). Paris, INSERM, 1973, pp 117–131

    Google Scholar 

  2. Sears HF, Rosenberg SA: Advantages of cryopreserved lymphocytes for sequential evaluation of human immune competence. I. Mitogen stimulation. J Natl Cancer Inst 58:183–187, 1975

    Google Scholar 

  3. Jewett MAS, Gupta S, Hansen JA, Cunningham-Rundles S, Siegel FP, Good RA, Dupont B: The use of cryopreserved lymphocytes for longitudinal studies of immune function and enumeration of subpopulations. Clin Exp Immunol 25:449–454, 1976

    Google Scholar 

  4. Callery CD, Golightly M, Sidell N, Golub SH: Lymphocyte surface markers and cytotoxicity following cryopreservation. J Immunol Methods 35:213–223, 1980

    Google Scholar 

  5. Schendel DJ, Wank R, Hall M, Deupont B: Standardization of the humanin vitro cell-mediated lympholysis technique. Tissue Antigens 13:112–120, 1979

    Google Scholar 

  6. Factor MA, Strong DM, Miller JL, Sell KW: Cell-mediated lymphocytotoxicity followingin vitro sensitization of frozen stored human mononuclear cells in heterologous serum. Cryobiology 12:521–529, 1975

    Google Scholar 

  7. Strong DM, Woody JN, Factor MA, Ahmed A, Sell KW: Immunological responsiveness of frozen thawed human lymphocytes. Clin Exp Immunol 21:442–435, 1975

    Google Scholar 

  8. Karpovitch XL, Rosenkovitch E, Ben-Bassett H, Izak G: Structure and functional alterations in lymphocytes induced by cryopreservation. Cryobiology 17:12–17, 1980

    Google Scholar 

  9. Oldham RK, Dean JH, Cannon GB, Ortaldo JR, Dunston G, Applebaum F, McCoy JL, Djeu J, Herberman RB: Cryopreservation of human lymphocyte functions as measured byin vitro assays. Int J Cancer 18:145–155, 1976

    Google Scholar 

  10. Miller RA, Bean MA, Kodera V, Herr HW: Cryopreservation of human effector cells active in antibody dependent cell-mediated cytotoxicity. Transplantation 21:517–519, 1976

    Google Scholar 

  11. Strong DM, Sell KW: Functional properties of cryopreserved lymphocytes.In Cryoimmunologic/Cryoimmunology, Paris, INSERM, 1976, pp 81–92

    Google Scholar 

  12. Naor D, O'Toole C: Cryopreservation of immunological memory and other lymphoid cell functions. J Immunol Methods 16:361–370, 1977

    Google Scholar 

  13. West WH, Boozer RB, Herberman RB: Low affinity E-rosette formation by the human K cell. J Immunol 120:90–95, 1978

    Google Scholar 

  14. Kay HD, Bonnard GD, West WH, Herberman RB: A functional comparison of human Fc-receptor-bearing lymphocytes active in natural cytotoxicity and antibody-dependent cellular cytotoxicity. J Immunol 118:2058–2066, 1977

    Google Scholar 

  15. Landazuri MO, Silva A, Alvarez J, Herberman RB: Evidence that natural cytotoxicity and antibody dependent cellular cytotoxicity are mediated in humans by the same effector cell populations. J Immunol 123:252–258, 1979

    Google Scholar 

  16. Timonen T, Ortaldo JR, Herberman RB: Characteristics of human large granular lymphocytes and relationship to natural killer and K cells. J Exp Med 153:569–582, 1981

    Google Scholar 

  17. Ortaldo JR, Sharrow S, Timonen T, Herberman RB: Determination of surface antigens of highly purified human NK cells by flow cytometry with monoclonal antibodies. J Immunol (in press), 1981

  18. Bolhuis H, Schuit RE: Identification of lymphocyte subpopulations in E-RFC-enriched and E-RFC depleted cell fractions of fresh and cryopreserved lymphocytes. Clin Exp Immunol 35:317–323, 1979

    Google Scholar 

  19. Lozzio CB, Lozzio RB: Cytotoxicity of a factor isolated from human spleen. J Natl Cancer Inst 50:535–538, 1973

    Google Scholar 

  20. Ortaldo JR, Pestka S, Slease RB, Rubinstein M, Herberman RB: Augmentation of human K cell activity with interferon. Scand J Immunol 12:365–369, 1980

    Google Scholar 

  21. Pandolfi F, Kurnick JT, Nilsson K, Forsbeck K, Wigzell H: Rosette format with goat erythrocytes. A marker for human T lymphocytes. Clin Exp Immunol 32:504, 1978

    Google Scholar 

  22. West HW, Cannon GB, Kay HD, Bonnard GD, Herberman RB: Natural cytotoxic reactivity of human lymphocytes against a myeloid cell line: Characterization of effector cells. J Immunol 118:355–361, 1977

    Google Scholar 

  23. Slease RB, Wistar R Jr, Scher I: Surface immunoglobulin density on human peripheral blood mononuclear cells. Blood 54(1):72–87, 1979

    Google Scholar 

  24. Janossy G, Tidmon N, Papageorgiou ES, Kung PC, Goldstein G: Distribution of T lymphocyte subsets in the human bone marrow and thymus: An analysis with monoclonal antibodies. J Immunol 126:1608–1613, 1981

    Google Scholar 

  25. West WH, Payne JM, Weese JL, Herberman RB: Human T lymphocyte subpopulations: Correlation between E-rosette-forming affinity and expression of the Fc receptor. J Immunol 119:548–554, 1977

    Google Scholar 

  26. Strong DM, Hartzman RJ, Light JA: Lymphocyte cryopreservation for histocompatibility testing. Dialy Transplant 7:508–534, 1978

    Google Scholar 

  27. Weiner RS: Cryopreservation of lymphocytes for use inin vitro assays of cellular immunity. J Immunol Methods 10:49–60, 1976

    Google Scholar 

  28. Strong DM, Ahmed A, Sell KW, Greiff D: Differential susceptibility of murine T and B lymphocytes to freeze-thaw and hypotonic shock. Cryobiology 11:127–138, 1974

    Google Scholar 

  29. Strong DM: Cryobiologic approaches to the recovery of immunologic responsiveness of murine and human mononuclear cells. Transplant Proc 8:203–208, 1976

    Google Scholar 

  30. Pandolfi F, Strong DM, Slease RB, Smith ML, Ortaldo JR, Herberman RB: Characterization of a suppressor T-cell chronic lymphocytic leukemia with ADCC but not NK activity. Blood 56:653–660, 1980

    Google Scholar 

  31. Hul ND, Kim YB, Amos DB: Natural killing and antibody-dependent cellular cytotoxicity in specific pathogen-free miniature swine and germ-free piglets. J Immunol 127:2190–2193, 1981

    Google Scholar 

  32. Ford CHJ, Newman CE, Carter AB: The effect of cryopreservation of lymphocytes on E rosetting ability: A study in lung cancer patients and controls. J Immunol Methods 26:113–124, 1979

    Google Scholar 

  33. Persidsky M, Ellit MH: Lysosomes and cell injury. Cryobiology 8:345–353, 1971

    Google Scholar 

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Authors and Affiliations

  1. Naval Medical Research Institute, National Naval Medical Center, 20814, Bethesda, Maryland

    Douglas M. Strong

  2. Uniformed Services University of the Health Sciences, 20814, Bethesda, Maryland

    Douglas M. Strong & Annette Maluish

  3. Laboratory of Immunodiagnosis, National Cancer Institute, National Institutes of Health, 20205, Bethesda, Maryland

    John R. Ortaldo & Ronald B. Herberman

  4. Department of Clinical Immunology, University of Rome, Rome, Italy

    Franco Pandolfi

Authors
  1. Douglas M. Strong
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  2. John R. Ortaldo
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  3. Franco Pandolfi
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  4. Annette Maluish
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  5. Ronald B. Herberman
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Strong, D.M., Ortaldo, J.R., Pandolfi, F. et al. Cryopreservation of human mononuclear cells for quality control in clinical immunology. I. Correlations in recovery of K- and NK-cell functions, surface markers, and morphology. J Clin Immunol 2, 214–221 (1982). https://doi.org/10.1007/BF00915224

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  • DOI: https://doi.org/10.1007/BF00915224

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