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. 1981 Jan 1;153(1):182–195. doi: 10.1084/jem.153.1.182

Identification of alpha-naphthyl butyrate esterase as a plasma membrane ectoenzyme of monocytes and as a discrete intracellular membrane- bounded organelle in lymphocytes

PMCID: PMC2186051  PMID: 7452152

Abstract

A reaction for an esterase, with a nonhalogenated, short-chain naphthyl ester (alpha-naphthyl butyrate or alpha-naphthyl acetate) as the substrate, has been used to identify mononuclear phagocytes by light microscopy. By analyzing techniques used in the collection, separation, fixation, processing, and embedding of human blood leukocytes for electron microscopy, we adapted the light microscopic method for use in determining the fine structural localization of this reaction. In monocytes, the reaction product covered the external surface of the plasma membrane. This distribution indicated that monocytic esterase is an ectoenzyme. The addition of NaF completely inhibited the monocytic reaction. In lymphocytes, the reaction product was localized in membrane-bounded intracellular organelles, similar to those previously shown to contain phospholipid and called Gall bodies. These organelles correspond with the punctate densities or focal reaction product observed by light microscopy. Other investigators believe that this distribution of enzyme in lymphocytes marks a subset of T cells, the Tmicro. The lymphocytic reaction was not inhibited by NaF.

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Selected References

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  1. BRAUNSTEIN H. Esterase in leucocytes. J Histochem Cytochem. 1959 May;7(3):202–202. doi: 10.1177/7.3.202. [DOI] [PubMed] [Google Scholar]
  2. Bainton D. R., Golde D. W. Differentiation of macrophages from normal human bone marrow in liquid culture. Electron microscopy and cytochemistry. J Clin Invest. 1978 Jun;61(6):1555–1569. doi: 10.1172/JCI109076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bodel P. T., Nichols B. A., Bainton D. F. Appearance of peroxidase reactivity within the rough endoplasmic reticulum of blood monocytes after surface adherence. J Exp Med. 1977 Feb 1;145(2):264–274. doi: 10.1084/jem.145.2.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bodel P. T., Nichols B. A., Bainton D. F. Differences in peroxidase localization of rabbit peritoneal macrophages after surface adherence. Am J Pathol. 1978 Apr;91(1):107–117. [PMC free article] [PubMed] [Google Scholar]
  5. Buchmüller Y., Mauel J. Inhibitors of serine-esterases enhance lymphokine-induced microbicidal activity in macrophages. J Reticuloendothel Soc. 1980 Jan;27(1):89–96. [PubMed] [Google Scholar]
  6. Carson F. L., Martin J. H., Lynn J. A. Formalin fixation for electron microscopy: a re-evaluation. Am J Clin Pathol. 1973 Mar;59(3):365–373. doi: 10.1093/ajcp/59.3.365. [DOI] [PubMed] [Google Scholar]
  7. DePierre J. W., Karnovsky M. L. Ecto-enzyme of granulocytes: 5'-nucleotidase. Science. 1974 Mar 15;183(4129):1096–1098. doi: 10.1126/science.183.4129.1096. [DOI] [PubMed] [Google Scholar]
  8. FISCHER R., SCHMALZL F. UBER DIE HEMMBARKEIT DER ESTERASEAKTIVITAET IN BLUTMONOCYTEN DURCH NATRIUMFLUORID. Klin Wochenschr. 1964 Aug 1;42:751–751. doi: 10.1007/BF01487816. [DOI] [PubMed] [Google Scholar]
  9. GOMORI G. Chloroacyl esters as histochemical substrates. J Histochem Cytochem. 1953 Nov;1(6):469–470. doi: 10.1177/1.6.469. [DOI] [PubMed] [Google Scholar]
  10. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  11. Grossi C. E., Webb S. R., Zicca A., Lydyard P. M., Moretta L., Mingari M. C., Cooper M. D. Morphological and histochemical analyses of two human T-cell subpopulations bearing receptors for IgM or IgG. J Exp Med. 1978 May 1;147(5):1405–1417. doi: 10.1084/jem.147.5.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. HEMPELMANN L. H., KNOWLTON N. P., Jr The nature of neutral red staining refractive particles in the lymphocytes of the blood of normal humans. Blood. 1953 Jun;8(6):524–535. [PubMed] [Google Scholar]
  13. Jaubert F., Monnet J. P., Danel C., Chretien J., Nezelof C. The location of non-specific esterase in human lung macrophages. An ultrastructural study. Histochemistry. 1978 Dec 29;59(2):141–147. doi: 10.1007/BF00518509. [DOI] [PubMed] [Google Scholar]
  14. Kaplow L. S. Cytochemical heterogeneity of human circulating monocytes. Acta Cytol. 1975 Jul-Aug;19(4):358–365. [PubMed] [Google Scholar]
  15. Kitagawa S., Takaku F., Sakamoto S. Evidence that proteases are involved in superoxide production by human polymorphonuclear leukocytes and monocytes. J Clin Invest. 1980 Jan;65(1):74–81. doi: 10.1172/JCI109662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Knowles D. M., 2nd, Holck S. Tissue localization of T-lymphocytes by the histochemical demonstration of acid alpha-naphthyl acetate esterase. Lab Invest. 1978 Jul;39(1):70–76. [PubMed] [Google Scholar]
  17. Kraut J. Serine proteases: structure and mechanism of catalysis. Annu Rev Biochem. 1977;46:331–358. doi: 10.1146/annurev.bi.46.070177.001555. [DOI] [PubMed] [Google Scholar]
  18. Kulenkampff J., Janossy G., Greaves M. F. Acid esterase in human lymphoid cells and leukaemic blasts: a marker for T lymphocytes. Br J Haematol. 1977 Jun;36(2):231–240. doi: 10.1111/j.1365-2141.1977.tb00644.x. [DOI] [PubMed] [Google Scholar]
  19. Lee M. J., Waters H. C., 3rd Inhibition of monocyte esterase activity by organophosphate insecticides. Blood. 1977 Nov;50(5):947–951. [PubMed] [Google Scholar]
  20. Li C. Y., Lam K. W., Yam L. T. Esterases in human leukocytes. J Histochem Cytochem. 1973 Jan;21(1):1–12. doi: 10.1177/21.1.1. [DOI] [PubMed] [Google Scholar]
  21. MOLONEY W. C., MCPHERSON K., FLIEGELMAN L. Esterase activity in leukocytes demonstrated by the use of naphthol AS-D chloroacetate substrate. J Histochem Cytochem. 1960 May;8:200–207. doi: 10.1177/8.3.200. [DOI] [PubMed] [Google Scholar]
  22. Manconi P. E., Marrosu M. G., Paghi L., Correale G., Zaccheo D. Alpha-naphthyl acetate esterase activity in human lymphocytes: distribution in lymphocyte subpopulations and in mitogen-activated cells. Scand J Immunol. 1979;9(2):99–104. doi: 10.1111/j.1365-3083.1979.tb02711.x. [DOI] [PubMed] [Google Scholar]
  23. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  24. Mueller J., Brun del Re G., Buerki H., Keller H. U., Hess M. W., Cottier H. Nonspecific acid esterase activity: a criterion for differentiation of T and B lymphocytes in mouse lymph nodes. Eur J Immunol. 1975 Apr;5(4):270–274. doi: 10.1002/eji.1830050411. [DOI] [PubMed] [Google Scholar]
  25. Pangalis G. A., Waldman S. R., Rappaport H. Cytochemical findings in human nonneoplastic blood and tonsillar B and T lymphocytes. Am J Clin Pathol. 1978 Mar;69(3):314–318. doi: 10.1093/ajcp/69.1.314. [DOI] [PubMed] [Google Scholar]
  26. Radzun H. J., Parwaresch M. R., Kulenkampff C., Staudinger M., Stein H. Lysosomal acid esterase: activity and isoenzymes in separated normal human blood cells. Blood. 1980 Jun;55(6):891–897. [PubMed] [Google Scholar]
  27. Remold H. G. The enhancement of MIF activity by inhibition of macrophage associated esterases. J Immunol. 1974 Apr;112(4):1571–1577. [PubMed] [Google Scholar]
  28. Schmalzl F., Braunsteiner H. The cytochemistry of monocytes and macrophages. Ser Haematol. 1970;3(2):93–131. [PubMed] [Google Scholar]
  29. Schmalzl F., Braunsteiner H. [On the origin of monocytes]. Acta Haematol. 1968;39(3):177–182. doi: 10.1159/000208959. [DOI] [PubMed] [Google Scholar]
  30. Uusitalo R. J., Karnovsky M. J. Surface localization of 5'-nucleotidase on the mouse lymphocyte. J Histochem Cytochem. 1977 Feb;25(2):87–96. doi: 10.1177/25.2.14210. [DOI] [PubMed] [Google Scholar]
  31. WACHSTEIN M., WOLF G. The histochemical demonstration of esterase activity in human blood and bone marrow smears. J Histochem Cytochem. 1958 Nov;6(6):457–457. doi: 10.1177/6.6.457. [DOI] [PubMed] [Google Scholar]
  32. Ward P. A. Leukotaxis and leukotactic disorders. A review. Am J Pathol. 1974 Dec;77(3):520–538. [PMC free article] [PubMed] [Google Scholar]
  33. van Furth R., Raeburn J. A., van Zwet T. L. Characteristics of human mononuclear phagocytes. Blood. 1979 Aug;54(2):485–500. [PubMed] [Google Scholar]

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