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. 1989 Feb;83(2):728–732. doi: 10.1172/JCI113939

Cryopreservable neutrophil surrogates. Stored cytoplasts from human polymorphonuclear leukocytes retain chemotactic, phagocytic, and microbicidal function.

S E Malawista 1, G Van Blaricom 1, M G Breitenstein 1
PMCID: PMC303736  PMID: 2536406

Abstract

Cryopreservation of polymorphonuclear leukocytes (PMN) has largely failed, probably because of their rich content of granular (lysosomal) enzymes. We have been developing granule-poor cytoplasts (anucleate fragments) from PMN which retain motile functions of the parent cell. The two types studied here were induced either by brief heating on surfaces (cytokineplasts) or by discontinuous gradient centrifugation (Ficoll) without heat or drugs (U-cytoplasts). Freshly made, these cytoplasts respond chemotactically to formyl peptide (fMet-Leu-Phe), and they take up and kill roughly half as many Staphylococcus aureus as their (larger, granular) parent PMN. Unlike their parent cells, after cryopreservation both cytoplasts remain chemotactic, and in matched experiments they take up and kill staphylococci with undiminished avidity. These findings are the first indications that PMN cytoplasts suitable for clinical use may be feasible.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Babior B. M., Kipnes R. S., Curnutte J. T. Biological defense mechanisms. The production by leukocytes of superoxide, a potential bactericidal agent. J Clin Invest. 1973 Mar;52(3):741–744. doi: 10.1172/JCI107236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dyett D. E., Malawista S. E., Naccache P. H., Sha'afi R. I. Stimulated cytokineplasts from human polymorphonuclear leukocytes mobilize calcium and polymerize actin. Cytoplasts made in cytochalasin B retain a defect in actin polymerization. J Clin Invest. 1986 Jan;77(1):34–37. doi: 10.1172/JCI112297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dyett D. E., Malawista S. E., Van Blaricom G., Melnick D. A., Malech H. L. Functional integrity of cytokineplasts: specific chemotactic and capping responses. J Immunol. 1985 Sep;135(3):2090–2094. [PubMed] [Google Scholar]
  4. Eid N. S., Kravath R. E., Lanks K. W. Heat-shock protein synthesis by human polymorphonuclear cells. J Exp Med. 1987 May 1;165(5):1448–1452. doi: 10.1084/jem.165.5.1448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Frim J., Mazur P. Approaches to the preservation of human granulocytes by freezing. Cryobiology. 1980 Jun;17(3):282–286. doi: 10.1016/0011-2240(80)90035-8. [DOI] [PubMed] [Google Scholar]
  6. Keller H. U., Bessis M. Chemotaxis and phagocytosis in anucleated cytoplasmic fragments of human peripheral blood leucocytes. Nouv Rev Fr Hematol. 1975 Jul-Aug;15(4):439–446. [PubMed] [Google Scholar]
  7. Lehrer R. I., Ganz T., Selsted M. E. Oxygen-independent bactericidal systems. Mechanisms and disorders. Hematol Oncol Clin North Am. 1988 Mar;2(1):159–169. [PubMed] [Google Scholar]
  8. Lionetti F. J., Hunt S. M., Lin P. S., Kurtz S. R., Valeri C. R. Preservation of human granulocytes. II. Characteristics of granulocytes obtained by counterflow centrifugation. Transfusion. 1977 Sep-Oct;17(5):465–472. doi: 10.1046/j.1537-2995.1977.17578014585.x. [DOI] [PubMed] [Google Scholar]
  9. Malawista S. E., Bodel P. T. The dissociation by colchicine of phagocytosis from increased oxygen consumption in human leukocytes. J Clin Invest. 1967 May;46(5):786–796. doi: 10.1172/JCI105579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Malawista S. E., De Boisfleury Chevance A. The cytokineplast: purified, stable, and functional motile machinery from human blood polymorphonuclear leukocytes. J Cell Biol. 1982 Dec;95(3):960–973. doi: 10.1083/jcb.95.3.960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Malawista S. E. Simple screening test on clotted blood for chronic granulomatous disease of childhood. Lancet. 1978 Apr 29;1(8070):943–943. doi: 10.1016/s0140-6736(78)90723-7. [DOI] [PubMed] [Google Scholar]
  12. Malawista S. E., Van Blaricom G., Cretella S. B. Cytokineplasts from human blood polymorphonuclear leukocytes. Lack of oxidase activity and extended functional longevity. Inflammation. 1985 Mar;9(1):99–106. doi: 10.1007/BF00915416. [DOI] [PubMed] [Google Scholar]
  13. Malawista S. E., Van Blaricom G. Cytoplasts made from human blood polymorphonuclear leukocytes with or without heat: preservation of both motile function and respiratory burst oxidase activity. Proc Natl Acad Sci U S A. 1987 Jan;84(2):454–458. doi: 10.1073/pnas.84.2.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Malawista S. E., Van Blaricom G. Phagocytic capacity of cytokineplasts from human blood polymorphonuclear leukocytes. Blood Cells. 1986;12(1):167–177. [PubMed] [Google Scholar]
  15. Malawista S. E., de Boisfleury-Chevance A., Maunoury R., Bessis M. Heat as a probe of centrosomal function: a phase-contrast and immunofluorescent study of human blood monocytes. Blood Cells. 1983;9(3):443–453. [PubMed] [Google Scholar]
  16. Petrequin P. R., Todd R. F., 3rd, Smolen J. E., Boxer L. A. Expression of specific granule markers on the cell surface of neutrophil cytoplasts. Blood. 1986 Apr;67(4):1119–1125. [PubMed] [Google Scholar]
  17. Roos D., Voetman A. A., Meerhof L. J. Functional activity of enucleated human polymorphonuclear leukocytes. J Cell Biol. 1983 Aug;97(2):368–377. doi: 10.1083/jcb.97.2.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rowe A. W., Lenny L. L. Cryopreservation of granulocytes for transfusion: studies on human granulocyte isolation, the effect of glycerol on lysosomes, kinetics of glycerol uptake and cryopreservation with dimethyl sulfoxide and glycerol. Cryobiology. 1980 Jun;17(3):198–212. doi: 10.1016/0011-2240(80)90027-9. [DOI] [PubMed] [Google Scholar]
  19. Voetman A. A., Bot A. A., Roos D. Cryopreservation of enucleated human neutrophils (PMN cytoplasts). Blood. 1984 Jan;63(1):234–237. [PubMed] [Google Scholar]

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