Inflammation

, Volume 7, Issue 4, pp 363–375 | Cite as

Demonstration of specific receptors for fluoresceinated casein on human neutrophils and monocytes using flow cytometry

  • Sharon L. Lewis
  • Dennis E. Van Epps
Original Articles

Abstract

-Casein is chemotactic for human neutrophils (PMNs) and monocytes. The binding of fluorescein (FITC) -conjugated casein (mixture ofα,β,and κ- casein) and purifiedα-casein to PMNs, monocytes, and lymphocytes was analyzed using flow cytometry. These studies demonstrate that 75–95% of PMNs and 46–85% of monocytes have membrane receptors for casein while lymphocytes lack these receptors. The binding of FITC-casein and FITC-α-casein was specific and was blocked only by unlabeled casein andα-casein, but not by ovalbumin, bovine or human serum albumin,β-casomorphin, C5a, or formyl-methionyl-leucylphenylalanine (fMLP). The binding of FITC-casein was reversible when PMNs were stained with this fluorescent agent and subsequently incubated with unlabeled casein. Double-labeling studies of mononuclear cells using FITC-casein and the OKM1 monoclonal antibody in conjunction with a rhodamine conjugated anti-Ig second antibody demonstrate that mononuclear cells binding FITC-casein also stain with the OKM1 monoclonal antibody, indicating a specificity for monocytes.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Dierich, M. P., andD. Wilhelmi. 1977. Essential role of surface-bound chemoattractant in leukocyte migration.Nature 270:351.Google Scholar
  2. 2.
    Glass, E. J., andA. V. Kay. 1980. Enhanced expression of human monocyte complement (C3b) receptors by chemoattractants.Clin. Exp. Immunol. 39:768.Google Scholar
  3. 3.
    Keller, H. U., andE. Sorkin. 1965. Studies in chemotaxis. II. The significance of normal sera for chemotaxis induced by various agents.Immunology 9:441.Google Scholar
  4. 4.
    Van Epps, D. E., A. D. Bankhukst, andR. C. Williams, Jr. 1977. Casein-mediated neutrophil chemotaxis: A parallel between surface binding and chemotaxis.Inflammation 2: 115.Google Scholar
  5. 5.
    Wilkinson, P. C. 1982. Chemotaxis and Inflammation, 2nd ed. Churchill Livingstone, Edinburgh. 22–23, 72–77, 96–98.Google Scholar
  6. 6.
    Golub, L. M., V. J. Iacono, G. Nicoll, N. Ramamurphy, andR. S. Kaslick. 1981. The response of human sulcular leukocytes to a chemotactic challenge.J. Peridont. Res. 16:171.Google Scholar
  7. 7.
    Wilkinson, P. C. 1972. Characterization of the chemotactic activity of casein for neutrophil leukocytes and macrophages.Experientia 28:1051.Google Scholar
  8. 8.
    Venge, P. 1979. Kinetic studies for cell migration in a modified Boyden chamber; Dependence on cell concentration and effects of the chymotrypsin-like cationic protein of human granulocytes.J. Immunol 122:1180.Google Scholar
  9. 9.
    Wilkinson, P. C., andG. R. Bradley. 1981. Chemotactic and enzyme-releasing activity of amphipathic proteins for neutrophils: A possible role for proteases in chemotaxis on substratum-bound protein gradients.Immunology 42:637.Google Scholar
  10. 10.
    Boyum, A. J. 1968. Isolation of mononuclear cells and granulocytes from human blood,Scand. J. Clin. Lab. Invest. 21(Suppl. 97):77.Google Scholar
  11. 11.
    Van Epps, D. E., andM. L. Garcia. 1980. Enhancement of neutrophil function as a result of prior exposure to chemotactic factor.J. Clin. Invest. 66:167.Google Scholar
  12. 12.
    Kabat, E. 1967. Estimation of protein with the Folin-ciocalteau phenol reagent.In Experimental Immunochemistry. E. Kabat and M. Mayer, editors. Charles C Thomas Co., Springfield, Illinois. 556.Google Scholar
  13. 13.
    Breard, J., E. L. Reinhesz, P. C. Kung, G. Goldstein, andS. F. Schlossman. 1980. A monoclonal antibody reactive with human peripheral blood monocytes.J. Immunol. 124: 1943.Google Scholar
  14. 14.
    Katagiri, T., I. Adachi, T. Terao, andT. Osawa. 1980. Alpha-casein binding proteins of guinea pig macrophage membranes and their possible roles in chemotaxis.J. Biochem,87:1421.Google Scholar
  15. 15.
    Wilkinson, P. C., andR. B. Allan. 1978. Binding of protein chemotactic factors to the surfaces of neutrophil leukocytes and its modifications with lipid-specific bacterial toxins.Mol. Cell. Biochem. 20:25.Google Scholar
  16. 16.
    El-Naggar, A. K., D. E. Van Epps, andR. C. Williams, Jr. 1980. Human B-and T-lymphocyte locomotion in response to casein, C5a, and f-Met-Leu-Phe.Cell. Immunol. 56:365.Google Scholar
  17. 17.
    O'Neill, J. G., andV. M. D. Parrott. 1977. I. Effect of culture and Con A on T and non-T lymphocytes.Cell. Immunol. 33:257.Google Scholar
  18. 18.
    Parrot, D. M. V., R. A. Good, G. J. O'Neill, andS. Gupta. 1973. Heterogeneity of locomotion in human T cell subsets.Proc. Natl. Acad. Sci. U.S.A. 75:2392.Google Scholar
  19. 19.
    Parrott, D. M. V., andP. C. Wilkinson. 1981. Lymphocyte locomotion and migration.Prog. Allergy 28:193.Google Scholar
  20. 20.
    Wilkinson, P. C., andR. B. Allan. 1978. Chemotaxis of neutrophil leukocytes towards substratum-bound protein attractants.Exp. Cell Res. 117:403.Google Scholar
  21. 21.
    Wilkinson, P. C., W. S. Haston, andJ. M. Shields. 1982. Some determinants of the locomotor behavior of phagocytes and lymphocytes in vitro.Clin. Exp. Immunol. 50:461 (review).Google Scholar
  22. 22.
    Brantl, V., H. Teschemacher, A. Henschen, andF. Lottspeich. 1979. Novel opioid peptides derived from casein (β-casomorphin). I. Isolation from bovine casein peptone.Hoppe- Seyler's Z. Physiol. Chem.360:1211.Google Scholar
  23. 23.
    Brantl, V., andH. Teschemacher. 1979. Material with opioid activity in bovine milk and milk products.Naunyn-Schmiedeberg's Arch. Pharmacol. 306:301.Google Scholar
  24. 24.
    Henschen, A., F. Lottspeich, V. Brantl, andH. Teschemacher. 1979. Novel opioid peptides derived from casein (β-casomorphin). II. Structure of active components from bovine casein peptone.Hoppe-Seyler's Z.Physiol. Chem. 360:1217.Google Scholar
  25. 25.
    Lottspeich, F., A. Henschen, V. Brantl, andH. Teschemacher. 1980. Novel opioid peptides derived from casein (β-casomorphin). III. Synthetic peptides corresponding to components from bovine casein peptone.Hoppe-Seyler's Z. Physiol. Chem. 361:1835.Google Scholar

Copyright information

© Plenum Publishing Corporation 1983

Authors and Affiliations

  • Sharon L. Lewis
    • 1
  • Dennis E. Van Epps
    • 1
  1. 1.Departments of Medicine and PathologyThe University of New MexicoAlbuquerque

Personalised recommendations