Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1984 May;73(5):1408–1417. doi: 10.1172/JCI111345

Rapid changes in light scattering from human polymorphonuclear leukocytes exposed to chemoattractants. Discrete responses correlated with chemotactic and secretory functions.

I Yuli, R Snyderman
PMCID: PMC425164  PMID: 6715544

Abstract

A platelet aggregometer was adapted for the simultaneous measurement of perpendicular light scattering in addition to light transmission. The addition of chemoattractants to polymorphonuclear leukocyte suspensions evoked a single wave of increased light transmission, whereas the perpendicular scattering measurement demonstrated a previously unrecognized biphasic response. The first perpendicular scattering response had no detectable latency and peaked at 10 +/- 1 s, then decayed rapidly. The second response peaked at 40 +/- 5 s, and decayed over several minutes. The dose-response curve of chemoattractants for inducing the rapid (10 +/- 1 s) perpendicular scattering peak corresponded to that which initiated chemotaxis. Initiation of the slow (40 +/- 5 s) peak required 10-fold higher amounts of chemoattractants, and the dose-response curve correlated with the induction of lysosomal enzyme secretion and superoxide anion production. Low doses of aliphatic alcohols, which have been shown to enhance chemotaxis but to inhibit secretion and superoxide anion production, abolished the slow perpendicular light-scattering response but left the fast response intact. Stimulants of secretion induced only slow and prolonged responses that were best observed in transmission measurements. In an attempt to resolve the origin of the light-scattering responses, the morphological changes of polymorphonuclear leukocytes were examined microscopically. Neither aggregation nor morphological whole cell polarization could be correlated with changes in light transmission or perpendicular scattering, which suggested that the source of scattering is of subcellular dimensions. The rapid perpendicular light-scattering response of polymorphonuclear leukocytes to chemoattractants appears to record an initial event in the stimulus-response coupling, and its measurement should provide a useful new tool for the study of leukocyte function. The biphasic nature of the light-scattering responses to chemoattractants, moreover, correlates with the dual regulation of the chemotactic and secretory responses of leukocytes.

Full text

PDF
1408

Images in this article

1414Image
on p.1414

Selected References

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

  1. Alobaidi T., Naccache P. H., Sha'afi R. I. Calmodulin antagonists modulate rabbit neutrophil degranulation, aggregation and stimulated oxygen consumption. Biochim Biophys Acta. 1981 Jul 17;675(3-4):316–321. doi: 10.1016/0304-4165(81)90020-9. [DOI] [PubMed] [Google Scholar]
  2. Antin P. B., Forry-Schaudies S., Friedman T. M., Tapscott S. J., Holtzer H. Taxol induces postmitotic myoblasts to assemble interdigitating microtubule-myosin arrays that exclude actin filaments. J Cell Biol. 1981 Aug;90(2):300–308. doi: 10.1083/jcb.90.2.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aswanikumar S., Corcoran B., Schiffmann E., Day A. R., Freer R. J., Showell H. J., Becker E. L. Demonstration of a receptor on rabbit neutrophils for chemotactic peptides. Biochem Biophys Res Commun. 1977 Jan 24;74(2):810–817. doi: 10.1016/0006-291x(77)90375-8. [DOI] [PubMed] [Google Scholar]
  4. Becker E. L., Showell H. J. The ability of chemotactic factors to induce lysosomal enzyme release. II. The mechanism of release. J Immunol. 1974 Jun;112(6):2055–2062. [PubMed] [Google Scholar]
  5. Bennett J. P., Cockcroft S., Gomperts B. D. Use of cytochalasin B to distinguish between early and late events in neutrophil activation. Biochim Biophys Acta. 1980 Oct 2;601(3):584–591. doi: 10.1016/0005-2736(80)90560-x. [DOI] [PubMed] [Google Scholar]
  6. Bentwood B. J., Henson P. M. The sequential release of granule constitutents from human neutrophils. J Immunol. 1980 Feb;124(2):855–862. [PubMed] [Google Scholar]
  7. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  8. Chenoweth D. E., Hugli T. E. Demonstration of specific C5a receptor on intact human polymorphonuclear leukocytes. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3943–3947. doi: 10.1073/pnas.75.8.3943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cianciolo G. J., Snyderman R. Monocyte responsiveness to chemotactic stimuli is a property of a subpopulation of cells that can respond to multiple chemoattractants. J Clin Invest. 1981 Jan;67(1):60–68. doi: 10.1172/JCI110033. [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. Goldman D. W., Goetzl E. J. Specific binding of leukotriene B4 to receptors on human polymorphonuclear leukocytes. J Immunol. 1982 Oct;129(4):1600–1604. [PubMed] [Google Scholar]
  12. Goldstein I., Hoffstein S., Gallin J., Weissmann G. Mechanisms of lysosomal enzyme release from human leukocytes: microtubule assembly and membrane fusion induced by a component of complement. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2916–2920. doi: 10.1073/pnas.70.10.2916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hammerschmidt D. E., Bowers T. K., Lammi-Keefe C. J., Jacob H. S., Craddock P. R. Granulocyte aggregometry: a sensitive technique for the detection of C5a and complement activation. Blood. 1980 Jun;55(6):898–902. [PubMed] [Google Scholar]
  14. Hammerschmidt D. E., White J. G., Craddock P. R., Jacob H. S. Corticosteroids inhibit complement-induced granulocyte aggregation. A possible mechanism for their efficacy in shock states. J Clin Invest. 1979 Apr;63(4):798–803. doi: 10.1172/JCI109365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hoffstein S. T., Friedman R. S., Weissmann G. Degranulation, membrane addition, and shape change during chemotactic factor-induced aggregation of human neutrophils. J Cell Biol. 1982 Oct;95(1):234–241. doi: 10.1083/jcb.95.1.234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jacob H. S., Hammerschmidt D. E. Complement-induced granulocyte aggregation. Importance in myocardial infarction and shock lung. JAMA. 1981 May 22;245(20):2013–2017. doi: 10.1001/jama.245.20.2013. [DOI] [PubMed] [Google Scholar]
  17. Kawaoka E. J., Miller M. E., Cheung A. T. Chemotactic factor-induced effects upon deformability of human polymorphonuclear leukocytes. J Clin Immunol. 1981 Jan;1(1):41–44. doi: 10.1007/BF00915475. [DOI] [PubMed] [Google Scholar]
  18. Kraut E. H., Segal M., Sagone A. L., Jr Evaluation of the role of oxygen radicals in polymorphonuclear leukocyte aggregation. Inflammation. 1982 Jun;6(2):161–167. doi: 10.1007/BF00916240. [DOI] [PubMed] [Google Scholar]
  19. Lehmeyer J. E., Snyderman R., Johnston R. B., Jr Stimulation of neutrophil oxidative metabolism by chemotactic peptides: influence of calcium ion concentration and cytochalasin B and comparison with stimulation by phorbol myristate acetate. Blood. 1979 Jul;54(1):35–45. [PubMed] [Google Scholar]
  20. MACMILLAN D. C., OLIVER M. F. THE INITIAL CHANGES IN PLATELET MORPHOLOGY FOLLOWING THE ADDITION OF ADENOSINE DIPHOSPHATE. J Atheroscler Res. 1965 Jul-Aug;5(4):440–444. doi: 10.1016/s0368-1319(65)80081-3. [DOI] [PubMed] [Google Scholar]
  21. Mease A. D., Burgess D. P., Thomas P. J. Irreversible neutrophil aggregation. A mechanism of decreased newborn neutrophil chemotactic response. Am J Pathol. 1981 Jul;104(1):98–102. [PMC free article] [PubMed] [Google Scholar]
  22. Miller M. E. Cell elastimetry in the study of normal and abnormal movement of human neutrophils. Clin Immunol Immunopathol. 1979 Dec;14(4):502–510. doi: 10.1016/0090-1229(79)90102-8. [DOI] [PubMed] [Google Scholar]
  23. Niedel J., Wilkinson S., Cuatrecasas P. Receptor-mediated uptake and degradation of 125I-chemotactic peptide by human neutrophils. J Biol Chem. 1979 Nov 10;254(21):10700–10706. [PubMed] [Google Scholar]
  24. Oseas R. S., Allen J., Yang H. H., Baehner R. L., Boxer L. A. Mechanism of dexamethasone inhibition of chemotactic factor induced granulocyte aggregation. Blood. 1982 Feb;59(2):265–269. [PubMed] [Google Scholar]
  25. Oseas R. S., Boxer L. A., Butterick C., Baehner R. L. Differences in polymorphonuclear leukocyte aggregating responses among several species in response to chemotactic stimulation. J Lab Clin Med. 1980 Aug;96(2):213–221. [PubMed] [Google Scholar]
  26. Parness J., Horwitz S. B. Taxol binds to polymerized tubulin in vitro. J Cell Biol. 1981 Nov;91(2 Pt 1):479–487. doi: 10.1083/jcb.91.2.479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Redl H., Hammerschmidt D. E., Schlag G. Augmentation by platelets of granulocyte aggregation in response to chemotaxins: studies utilizing an improved cell preparation technique. Blood. 1983 Jan;61(1):125–131. [PubMed] [Google Scholar]
  28. Seligmann B. E., Fletcher M. P., Gallin J. I. Adaptation of human neutrophil responsiveness to the chemoattractant N-formylmethionylleucylphenylalanine. Heterogeneity and/or negative cooperative interaction of receptors. J Biol Chem. 1982 Jun 10;257(11):6280–6286. [PubMed] [Google Scholar]
  29. Seligmann B., Chused T. M., Gallin J. I. Human neutrophil heterogeneity identified using flow microfluorometry to monitor membrane potential. J Clin Invest. 1981 Nov;68(5):1125–1131. doi: 10.1172/JCI110356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Showell H. J., Freer R. J., Zigmond S. H., Schiffmann E., Aswanikumar S., Corcoran B., Becker E. L. The structure-activity relations of synthetic peptides as chemotactic factors and inducers of lysosomal secretion for neutrophils. J Exp Med. 1976 May 1;143(5):1154–1169. doi: 10.1084/jem.143.5.1154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sklar L. A., Oades Z. G., Jesaitis A. J., Painter R. G., Cochrane C. G. Fluoresceinated chemotactic peptide and high-affinity antifluorescein antibody as a probe of the temporal characteristics of neutrophil stimulation. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7540–7544. doi: 10.1073/pnas.78.12.7540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Smith C. W., Hollers J. C., Patrick R. A., Hassett C. Motility and adhesiveness in human neutrophils. Effects of chemotactic factors. J Clin Invest. 1979 Feb;63(2):221–229. doi: 10.1172/JCI109293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Smolen J. E., Korchak H. M., Weissmann G. Initial kinetics of lysosomal enzyme secretion and superoxide anion generation by human polymorphonuclear leukocytes. Inflammation. 1980 Jun;4(2):145–163. doi: 10.1007/BF00914161. [DOI] [PubMed] [Google Scholar]
  34. Snyderman R., Goetzl E. J. Molecular and cellular mechanisms of leukocyte chemotaxis. Science. 1981 Aug 21;213(4510):830–837. doi: 10.1126/science.6266014. [DOI] [PubMed] [Google Scholar]
  35. Spilberg I., Mehta J. Demonstration of a specific neutrophil receptor for a cell-derived chemotactic factor. J Clin Invest. 1979 Jan;63(1):85–88. doi: 10.1172/JCI109282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Williams L. T., Snyderman R., Pike M. C., Lefkowitz R. J. Specific receptor sites for chemotactic peptides on human polymorphonuclear leukocytes. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1204–1208. doi: 10.1073/pnas.74.3.1204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yasaka T., Boxer L. A., Baehner R. L. Monocyte aggregation and superoxide anion release in response to formyl-methionyl-leucyl-phenylalanine (FMLP) and platelet-activating factor (PAF). J Immunol. 1982 May;128(5):1939–1944. [PubMed] [Google Scholar]
  38. Yuli I., Tomonaga A., Synderman R. Chemoattractant receptor functions in human polymorphonuclear leukocytes are divergently altered by membrane fluidizers. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5906–5910. doi: 10.1073/pnas.79.19.5906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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]
  40. Zigmond S. H., Sullivan S. J. Sensory adaptation of leukocytes to chemotactic peptides. J Cell Biol. 1979 Aug;82(2):517–527. doi: 10.1083/jcb.82.2.517. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES