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. 1970 Sep 1;132(3):401–416. doi: 10.1084/jem.132.3.401

THE IN VITRO TRANSFORMATION OF FROZEN-STORED LYMPHOCYTES IN THE MIXED LYMPHOCYTE REACTION AND IN CULTURE WITH PHYTOHEMAGGLUTININ AND SPECIFIC ANTIGENS

Richard J Mangi 1, Michael R Mardiney Jr 1
PMCID: PMC2138801  PMID: 5523965

Abstract

We have investigated the in vitro transformation of lymphocytes that have been frozen and stored at low temperatures. Cells frozen at different times and stored for various times do transform in a reproducible manner when placed in culture with PHA or as one population of the two-way or the one-way MLR. When frozen-stored lymphocytes are cultured with specific antigen or as both partners in the MLR the response is minimal. The freezing process destroys neutrophils, and the remaining population transforms in a manner similar to cultures of purified lymphocytes. Practical aspects of the technique are discussed and we have suggested possible practical applications for future investigation.

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Selected References

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  1. Albertini R. J., Bach F. H. Quantitative assay of antigenic disparity at hl-a-the major histocompatibility locus in man. J Exp Med. 1968 Oct 1;128(4):639–651. doi: 10.1084/jem.128.4.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Astaldi G., Costa G., Airo R., Duarte N. Lymphocytes from leukaemic blood cultured with phytohemagglutinin. Eur J Cancer. 1965 Nov;1(3):259–264. doi: 10.1016/0014-2964(65)90058-7. [DOI] [PubMed] [Google Scholar]
  3. Astaldi G., Massimo L., Airo R., Mori P. G. Phytohaemagglutinin and lymphocytes from acute lymphocytic leukaemia. Lancet. 1966 Jun 4;1(7449):1265–1266. doi: 10.1016/s0140-6736(66)90279-0. [DOI] [PubMed] [Google Scholar]
  4. BERNARD C., GERALDES A., BOIRON M. EFFECTS OF PHYTOHAEMAGGLUTININ ON BLOOD-CULTURES OF CHRONIC LYMPHOCYTIC LEUKAEMIAS. Lancet. 1964 Mar 21;1(7334):667–668. doi: 10.1016/s0140-6736(64)91488-6. [DOI] [PubMed] [Google Scholar]
  5. Bach F. H., Voynow N. K. One-way stimulation in mixed leukocyte cultures. Science. 1966 Jul 29;153(3735):545–547. doi: 10.1126/science.153.3735.545. [DOI] [PubMed] [Google Scholar]
  6. Bouroncle B. A., Clausen K. P., Aschenbrand J. F. Studies of the delayed response of phytohemagglutinin (PHA) stimulated lymphocytes in 25 chronic lymphatic leukemia patients before and during therapy. Blood. 1969 Aug;34(2):166–178. [PubMed] [Google Scholar]
  7. Bouroncle B. A. Preservation of human normal and leukemic cells with dimethyl sulfoxide at -80 degrees C. Cryobiology. 1967 May-Jun;3(6):445–455. doi: 10.1016/s0011-2240(67)80154-8. [DOI] [PubMed] [Google Scholar]
  8. Cohen E., Rowe A. W. Detection of leukoagglutinins with dimethylsulfoxide (DMSO) protected leukocytes frozen with liquid nitrogen. Vox Sang. 1965 Sep-Oct;10(5):543–551. doi: 10.1111/j.1423-0410.1965.tb01407.x. [DOI] [PubMed] [Google Scholar]
  9. EPSTEIN L. B., BRECHER G. DNA AND RNA SYNTHESIS OF CIRCULATING ATYPICAL LYMPHOCYTES IN INFECTIOUS MONONUCLEOSIS. Blood. 1965 Feb;25:197–203. [PubMed] [Google Scholar]
  10. Elves M. W., Israëls M. C., Collinge M. An assessment of the mixed leucocyte reaction in renal failure. Lancet. 1966 Mar 26;1(7439):682–685. doi: 10.1016/s0140-6736(66)91628-x. [DOI] [PubMed] [Google Scholar]
  11. Fridman W. H., Kourilsky F. M. Stimulation of lymphocytes by autologous leukaemic cells in acute leukaemia. Nature. 1969 Oct 18;224(5216):277–279. doi: 10.1038/224277a0. [DOI] [PubMed] [Google Scholar]
  12. HASHEM N., CARR D. H. MITOGENIC STIMULATION OF PERIPHERAL LYMPHOCYTE CULTURES BY AUTOLOGOUS LYMPHOCYTE EXTRACTS IN AUTOIMMUNE DISEASES. Lancet. 1963 Nov 16;2(7316):1030–1031. doi: 10.1016/s0140-6736(63)90005-9. [DOI] [PubMed] [Google Scholar]
  13. HIRSCHHORN K., BACH F., RAPAPORT F. T., CONVERSE J. M., LAWRENCE H. S. THE RELATIONSHIP OF IN VITRO LYMPHOCYTE COMPATIBILITY TO HOMOGRAFT SENSITIVITY IN MAN. Ann N Y Acad Sci. 1964 Nov 30;120:303–306. doi: 10.1111/j.1749-6632.1964.tb34729.x. [DOI] [PubMed] [Google Scholar]
  14. Hersh E. M., Oppenheim J. J. Impaired in vitro lymphocyte transformation in Hodgkin's disease. N Engl J Med. 1965 Nov 4;273(19):1006–1012. doi: 10.1056/NEJM196511042731903. [DOI] [PubMed] [Google Scholar]
  15. Holm G., Perlmann P., Johansson B. Impaired phytohaemagglutinin-induced cytotoxicity in vitro of lymphocytes from patients with Hodgkin's disease or chronic lymphatic leukaemia. Clin Exp Immunol. 1967 May;2(3):351–360. [PMC free article] [PubMed] [Google Scholar]
  16. Houseley J., Oppenheim J. J. Lymphocyte transformation in thymectomized and nonthymectomized patients with myasthenia gravis. Br Med J. 1967 Jun 10;2(5553):679–681. doi: 10.1136/bmj.2.5553.679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. LaPolla J. J., Thurmon T. F., 3rd Lymphocyte transformation in autoimmune disorders: a preliminary study. J Pediatr. 1966 Dec;69(6):1129–1131. doi: 10.1016/s0022-3476(66)80306-2. [DOI] [PubMed] [Google Scholar]
  18. Leikin S. L., Bazelon M., Park K. H. In vitro lymphocyte transformation in ataxia telangiectasia. J Pediatr. 1966 Mar;68(3):477–479. doi: 10.1016/s0022-3476(66)80255-x. [DOI] [PubMed] [Google Scholar]
  19. Moorhead J. F., Connolly J. J., McFarland W. Factors affecting the reactivity of human lymphocytes in vitro. I. Cell number, duration of culture and surface area. J Immunol. 1967 Aug;99(2):413–419. [PubMed] [Google Scholar]
  20. OPPENHEIM J. J., WHANG J., FREI E., 3rd IMMUNOLOGIC AND CYTOGENETIC STUDIES OF CHRONIC LYMPHOCYTIC LEUKEMIC CELLS. Blood. 1965 Aug;26:121–132. [PubMed] [Google Scholar]
  21. Oppenheim J. J., Barlow M., Waldmann T. A., Block J. B. Impaired in vitro lymphocyte transformation in patients with ataxia-telangiectasis. Br Med J. 1966 Aug 6;2(5509):330–333. doi: 10.1136/bmj.2.5509.330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Oppenheim J. J., Leventhal B. G., Hersh E. M. The transformation of column-purified lymphocytes with nonspecific and specific antigenic stimuli. J Immunol. 1968 Aug;101(2):262–267. [PubMed] [Google Scholar]
  23. Oppenheim J. J., Whang J., Frei E., 3rd The effect of skin homograft rejection on recipient and donor mixed leukocyte cultures. J Exp Med. 1965 Oct 1;122(4):651–664. doi: 10.1084/jem.122.4.651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pike M. C. Chemotherapy in Burkitt's Tumour. Lancet. 1966 Oct 15;2(7468):856–856. doi: 10.1016/s0140-6736(66)92280-x. [DOI] [PubMed] [Google Scholar]
  25. ROWE A. W., COHEN E. PHAGOCYTIC ACTIVITY AND ANTIGENIC INTEGRITY OF LEUKOCYTES PRESERVED WITH DIMETHYL SULFOXIDE AT A CRYOGENIC TEMPERATURE (--196 DEGREES C). Vox Sang. 1965 May-Jun;10:382–384. [PubMed] [Google Scholar]
  26. SCHREK R., RABINOWITZ Y. Effects of phytohemagglutinin on rat and normal and leukemic human blood cells. Proc Soc Exp Biol Med. 1963 May;113:191–194. doi: 10.3181/00379727-113-28317. [DOI] [PubMed] [Google Scholar]
  27. Sharman C., Crossen P. E., Fitzgerald P. H. Lymphocyte number and response to phytohae- magglutinin in chronic lymphocytic leukaemia. Scand J Haematol. 1966;3(5):375–382. doi: 10.1111/j.1600-0609.1966.tb02380.x. [DOI] [PubMed] [Google Scholar]
  28. Thomson A. E., Robinson M. A., Wetherley-Mein G. Heterogeneity of lymphocytes in chronic lymphocytic leukaemia. Lancet. 1966 Jul 23;2(7456):200–202. doi: 10.1016/s0140-6736(66)92480-9. [DOI] [PubMed] [Google Scholar]
  29. WINTER G. C., OSMOND D. G., YOFFEY J. M., MAHY D. J. LEUCOCYTE CULTURES WITH PHYTOHAEMAGGLUTININ IN CHRONIC LYMPHATIC LEUKAEMIA. Lancet. 1964 Sep 12;2(7359):563–565. doi: 10.1016/s0140-6736(64)90624-5. [DOI] [PubMed] [Google Scholar]
  30. Wilson D. B., Silvers W. K., Nowell P. C. Quantitative studies on the mixed lymphocyte interaction in rats. II. Relationship of the proliferative response to the immunologic status of the donors. J Exp Med. 1967 Oct 1;126(4):655–665. doi: 10.1084/jem.126.4.655. [DOI] [PMC free article] [PubMed] [Google Scholar]

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