Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1982 Dec 1;95(3):960–973. doi: 10.1083/jcb.95.3.960

The cytokineplast: purified, stable, and functional motile machinery from human blood polymorphonuclear leukocytes

SE Malawista, A De Boisfleury Chevance
PMCID: PMC2112937  PMID: 6891383

Abstract

We examined the formation of motile, chemotactically active, anucleate fragments from human blood polymorphonuclear leukocytes (PMN, granulocytes), induced by the brief application of heat. These granule-poor fragments are former protopods (leading fronts, lamellipodia) that become uncoupled from the main body of the cell and leave it, at first with a connecting filament that breaks and seals itself. The usual random orientation of such filaments can be controlled by preorientation of cells in a gradient of the chemotactic peptide, N-formylmethionylleucylphenylalanine (F-Met-Leu-Phe) (2x10(-9) M- 1x10(-8)). Cytochalsin B, 2.5-5 μg/ml, prevents fragment formation; colchicine, 10(-5) M, does not. In scanning electron micrographs, fragments are ruffled and the cell body rounded up and rather smooth. In transmission electron micrographs, fragments contain microfilaments but lack centrioles and microtubules. Like intact cells, both bound and free fragments can respond chemotactically to an erythrocyte destroyed by laser microirradiation (necrotaxis); the free, anucleate fragments may do so repeatedly, even after having been held overnight at ambient temperatures. We propse the name cytokineplast for the result of this self-purification of motile apparatus. The exodus of the motile machinery from the granulocyte requires anchoring of the bulk of the cell to glass and uncoupling, which may involve heat-induced dysfunction of the centrosome. In ultrastructural studies of the centrosomal region after heat, centriolar structure remains intact, but pericentriolar osmiophilic material appears condensed, and microtubules are sparse. These changes are found in all three blood cell types examined: PMN, eosinophil, and monocyte. Of these, the first two make fragments under our conditions; the more sluggish monocyte does not. Uncoupling is further linked to centrosomal dysfunction by the observation that colchicines-treated granulocytes (10(-5)M, to destroy the centrosome’s efferent arm) make fragments after less heat than controls. If motive force and orientation are specified mainly from the organelle-excluding leading front, then endoplasmic streaming in PMN is a catch-up phenomenon, and microtubules do not provide the vector of locomotion but rather stabilize and orient the “baggage” (nucleus, granuloplasm)—i.e., they prevent fishtailing. Moreover, constraints emanating from the centrosome may now be extended to include, maintenance of the motile machinery as an integral part of the cell.

Full Text

The Full Text of this article is available as a PDF (8.0 MB).

Selected References

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

  1. Albrecht-Buehler G. Autonomous movements of cytoplasmic fragments. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6639–6643. doi: 10.1073/pnas.77.11.6639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allan R. B., Wilkinson P. C. A visual analysis of chemotactic and chemokinetic locomotion of human neutrophil leucocytes. Use of a new chemotaxis assay with Candida albicans as gradient source. Exp Cell Res. 1978 Jan;111(1):191–203. doi: 10.1016/0014-4827(78)90249-5. [DOI] [PubMed] [Google Scholar]
  3. Bandmann U., Rydgren L., Norberg B. The difference between random movement and chemotaxis. Effects of antitubulins on neutrophil granulocyte locomotion. Exp Cell Res. 1974 Sep;88(1):63–73. doi: 10.1016/0014-4827(74)90618-1. [DOI] [PubMed] [Google Scholar]
  4. Barrau M. D., Blackburn G. R., Dewey W. C. Effects of heat on the centrosomes of Chinese hamster ovary cells. Cancer Res. 1978 Aug;38(8):2290–2294. [PubMed] [Google Scholar]
  5. Berlin R. D., Oliver J. M. Analogous ultrastructure and surface properties during capping and phagocytosis in leukocytes. J Cell Biol. 1978 Jun;77(3):789–804. doi: 10.1083/jcb.77.3.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bessis M., Breton-Gorius J. Rapports entre noyau et centrioles dans les granulocytes étalés. Rôle des microtubules. Nouv Rev Fr Hematol. 1967 Sep-Oct;7(5):601–620. [PubMed] [Google Scholar]
  7. Bessis M. Editorial: Une forme particuliére de chimiotaxis: le nécrotaxis. Nouv Rev Fr Hematol. 1973 May-Jun;13(3):285–290. [PubMed] [Google Scholar]
  8. Caner J. E. Colchicine inhibition of chemotaxis. Arthritis Rheum. 1965 Oct;8(5):757–764. doi: 10.1002/art.1780080438. [DOI] [PubMed] [Google Scholar]
  9. Chen W. T. Mechanism of retraction of the trailing edge during fibroblast movement. J Cell Biol. 1981 Jul;90(1):187–200. doi: 10.1083/jcb.90.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Craddock P. R., Hammerschmidt D., White J. G., Dalmosso A. P., Jacob H. S. Complement (C5-a)-induced granulocyte aggregation in vitro. A possible mechanism of complement-mediated leukostasis and leukopenia. J Clin Invest. 1977 Jul;60(1):260–264. doi: 10.1172/JCI108763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Davies W. A., Stossel T. P. Peripheral hyaline blebs (podosomes) of macrophages. J Cell Biol. 1977 Dec;75(3):941–955. doi: 10.1083/jcb.75.3.941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gifford R. H., Malawista S. E. A simple rapid micromethod for detecting chronic granulomatous disease of childhood. J Lab Clin Med. 1970 Mar;75(3):511–519. [PubMed] [Google Scholar]
  13. 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]
  14. Lionetti F. J., Lin P. S., Mattaliano R. J., Hunt S. M., Valeri C. R. Temperature effects on shape and function of human granulocytes. Exp Hematol. 1980 Mar;8(3):304–317. [PubMed] [Google Scholar]
  15. Malawista S. E., Bensch K. G. Human polymorphonuclear leukocytes: demonstration of microtubules and effect of colchicine. Science. 1967 Apr 28;156(3774):521–522. doi: 10.1126/science.156.3774.521. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Malawista S. E. Colchicine: a common mechanism for its anti-inflammatory and anti-mitotic effects. Arthritis Rheum. 1968 Apr;11(2):191–197. doi: 10.1002/art.1780110210. [DOI] [PubMed] [Google Scholar]
  18. Malawista S. E. Microtubules and the mobilization of lysosomes in phagocytizing human leukocytes. Ann N Y Acad Sci. 1975 Jun 30;253:738–749. doi: 10.1111/j.1749-6632.1975.tb19242.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Malech H. L., Root R. K., Gallin J. I. Structural analysis of human neutrophil migration. Centriole, microtubule, and microfilament orientation and function during chemotaxis. J Cell Biol. 1977 Dec;75(3):666–693. doi: 10.1083/jcb.75.3.666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maunoury M. R. Localisation immunocytochimique de la centrosphère de cellules tumorales humaines par utilisation d'anticorps naturels de lapin. C R Acad Sci Hebd Seances Acad Sci D. 1978 Feb 13;286(6):503–506. [PubMed] [Google Scholar]
  22. Nahas G. G., Tannieres M. L., Lennon J. F. Direct measurement of leukocyte motility: effects of pH and temperature. Proc Soc Exp Biol Med. 1971 Oct;138(1):350–352. doi: 10.3181/00379727-138-35894. [DOI] [PubMed] [Google Scholar]
  23. Oliver J. M. Cell biology of leukocyte abnormalities--membrane and cytoskeletal function in normal and defective cells. A review. Am J Pathol. 1978 Oct;93(1):221–270. [PMC free article] [PubMed] [Google Scholar]
  24. Oliver J. M., Krawiec J. A., Becker E. L. The distribution of actin during chemotaxis in rabbit neutrophils. J Reticuloendothel Soc. 1978 Dec;24(6):697–704. [PubMed] [Google Scholar]
  25. Penny R., Galton D. A., Scott J. T., Eisen V. Studies on neutrophil function. 1. Physiological and pharmacological aspects. Br J Haematol. 1966 Sep;12(5):623–632. doi: 10.1111/j.1365-2141.1966.tb00145.x. [DOI] [PubMed] [Google Scholar]
  26. Poste G., Lyon N. C. Enucleation of cultured animal cells by cytochalasin B. Front Biol. 1978;46:161–189. [PubMed] [Google Scholar]
  27. Ramsey W. S. Locomotion of human polymorphonuclear leucocytes. Exp Cell Res. 1972 Jun;72(2):489–501. doi: 10.1016/0014-4827(72)90019-5. [DOI] [PubMed] [Google Scholar]
  28. Robbins E., Jentzsch G., Micali A. The centriole cycle in synchronized HeLa cells. J Cell Biol. 1968 Feb;36(2):329–339. doi: 10.1083/jcb.36.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zigmond S. H. Ability of polymorphonuclear leukocytes to orient in gradients of chemotactic factors. J Cell Biol. 1977 Nov;75(2 Pt 1):606–616. doi: 10.1083/jcb.75.2.606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zigmond S. H. Chemotaxis by polymorphonuclear leukocytes. J Cell Biol. 1978 May;77(2):269–287. doi: 10.1083/jcb.77.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES