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. 1963 Jun 1;17(3):465–486. doi: 10.1083/jcb.17.3.465

A HISTOCHEMICAL STUDY OF PHAGOCYTIC AND ENZYMATIC FUNCTIONS OF RABBIT MONONUCLEAR AND POLYMORPHONUCLEAR EXUDATE CELLS AND ALVEOLAR MACROPHAGES

I. Survey and Quantitation of Enzymes, and States of Cellular Activation

Arthur M Dannenberg Jr 1, Marvin S Burstone 1, Paul C Walter 1, June W Kinsley 1
PMCID: PMC2106214  PMID: 14024972

Abstract

The cytochrome oxidase (CO), aminopeptidase (AMP), succinic dehydrogenase (SD), acid phosphatase, esterase, and alkaline phosphatase of rabbit mononuclear (MN) and polymorphonuclear (PMN) peritoneal exudate cells and pulmonary alveolar macrophages (AM) - air dried on Mylar strips - were characterized by histochemical techniques with respect to stability, activators, inhibitors, and pH optima. A granule count method was established for the quantitation of these enzymes. For the acid phosphatase of MN, in which the most precise results were obtained, time, pH, substrate, and inhibitor curves resembled those commonly obtained biochemically. Five of these enzymes were usually more active in AM than MN, whereas the sixth, alkaline phosphatase, was not present in either cell type. AM also tended to consume more oxygen than MN and to divide more frequently. Since the most active cells in the population would be first involved in the host's defense against microbial agents, a comparison was made of the 10 per cent of the AM and MN with the highest enzymatic activities. No differences were found in the granule counts that were not reflected by the means. However, within a given AM population, cells containing ingested dust particles seemed to have higher enzymatic activities than those without particles. MN had greater acid phosphatase and SD activities than PMN and consumed more oxygen, but the CO, AMP, and esterase activites of both types of cells were of similar magnitude. PMN showed high alkaline phosphatase activity; MN showed none. A survey of the histochemical literature indicates that a positive correlation between the enzymatic and phagocytic activities of both MN and PMN exists in vivo.

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Selected References

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  1. ACKERMAN G. A. Histochemical demonstration of dehydrogenase activity in the cells of normal human blood and bone marrow. J Biophys Biochem Cytol. 1960 Sep;8:61–67. doi: 10.1083/jcb.8.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. ACKERMAN G. A. Histochemical demonstration of indoxyl acetate esterase activity in normal human blood and bone marrow. Lab Invest. 1960 Mar-Apr;9:298–304. [PubMed] [Google Scholar]
  3. ALLISON M. J., ZAPPASODI P., LURIE M. B. Metabolic studies on mononuclear cells from rabbits of varying genetic resistance to tuberculosis. II. Studies on cells from BCG-vaccinated animals. Am Rev Respir Dis. 1962 Mar;85:364–372. doi: 10.1164/arrd.1962.85.3.364. [DOI] [PubMed] [Google Scholar]
  4. BAKER B. L., KLAPPER Z. F. Oxidative enzymes in the foreign body giant cell. J Histochem Cytochem. 1961 Nov;9:713–714. doi: 10.1177/9.6.713. [DOI] [PubMed] [Google Scholar]
  5. BALOGH K., Jr, COHEN R. B. Histochemical demonstration of diaphorases and dehydrogenases in normal human leukocytes and platelets. Blood. 1961 Apr;17:491–496. [PubMed] [Google Scholar]
  6. BARKA T., SCHAFFNER F., POPPER H. Acid phosphatase and reticuloendothelial system. Lab Invest. 1961 May-Jun;10:590–607. [PubMed] [Google Scholar]
  7. BARRNETT R. J., SELIGMAN A. M. Histochemical demonstration of esterases by production of indigo. Science. 1951 Nov 30;114(2970):579–582. doi: 10.1126/science.114.2970.579. [DOI] [PubMed] [Google Scholar]
  8. BENDITT E. P., ARASE M. Enzyme kinetics in a histochemical system. J Histochem Cytochem. 1958 Nov;6(6):431–434. doi: 10.1177/6.6.431. [DOI] [PubMed] [Google Scholar]
  9. BRAUNSTEIN H., FREIMAN D. G., GALL E. A. A histochemical study of the enzymatic activity of lymph nodes. I. The normal and hyperplastic lymph node. Cancer. 1958 Jul-Aug;11(4):829–837. doi: 10.1002/1097-0142(195807/08)11:4<829::aid-cncr2820110422>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
  10. BRAUNSTEIN H., FREIMAN D. G., THOMAS W., Jr, GALL E. A. A histochemical study of the enzymatic activity of lymph nodes. II. Further investigation of normal and hyperplastic lymph nodes. Cancer. 1962 Jan-Feb;15:130–138. doi: 10.1002/1097-0142(196201/02)15:1<130::aid-cncr2820150118>3.0.co;2-y. [DOI] [PubMed] [Google Scholar]
  11. BURSTONE M. S. Esterase activity of developing bones and teeth. AMA Arch Pathol. 1957 Feb;63(2):164–167. [PubMed] [Google Scholar]
  12. BURSTONE M. S., FOLK J. E. Histochemical demonstration of aminopeptidase. J Histochem Cytochem. 1956 May;4(3):217–226. doi: 10.1177/4.3.217. [DOI] [PubMed] [Google Scholar]
  13. BURSTONE M. S. Histochemical comparison of naphthol AS-phosphates for the demonstration of phosphatases. J Natl Cancer Inst. 1958 Mar;20(3):601–615. [PubMed] [Google Scholar]
  14. BURSTONE M. S. Histochemical demonstration of acid phosphatases with naphthol AS-phosphates. J Natl Cancer Inst. 1958 Sep;21(3):523–539. [PubMed] [Google Scholar]
  15. BURSTONE M. S. The cytochemical localization of esterase. J Natl Cancer Inst. 1957 Feb;18(2):167–172. [PubMed] [Google Scholar]
  16. COHN Z. A., HIRSCH J. G. The isolation and properties of the specific cytoplasmic granules of rabbit polymorphonuclear leucocytes. J Exp Med. 1960 Dec 1;112:983–1004. doi: 10.1084/jem.112.6.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. DE SOUZA E. J., KOTHARE S. N. A method for the cytochemical demonstration of succinic dehydrogenase in human leucocytes. J Histochem Cytochem. 1959 Mar;7(2):77–79. doi: 10.1177/7.2.77. [DOI] [PubMed] [Google Scholar]
  18. DOYLE W. L. The distribution of phosphatases in the rabbit appendix after X-irradiation. Am J Anat. 1950 Jul;87(1):79–117. doi: 10.1002/aja.1000870104. [DOI] [PubMed] [Google Scholar]
  19. ELBERG S. S., MASCARENHAS P., FONG J. Response of peritoneal exudate cells to Brucella melitensis. Influence of nature ox inflammatory irritant. Proc Soc Exp Biol Med. 1960 Jun;104:295–297. doi: 10.3181/00379727-104-25814. [DOI] [PubMed] [Google Scholar]
  20. ESSNER E. An electron microscopic study of erythrophagocytosis. J Biophys Biochem Cytol. 1960 Apr;7:329–334. doi: 10.1083/jcb.7.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. FINDLAY G. H. The simple esterases of human skin. Br J Dermatol. 1955 Mar;67(3):83–91. doi: 10.1111/j.1365-2133.1955.tb12696.x. [DOI] [PubMed] [Google Scholar]
  22. FLECK L. La leukergie et son role dans l'immunité. Ann Inst Pasteur (Paris) 1957 Mar;92(3):380–395. [PubMed] [Google Scholar]
  23. FLECK L., MURCZYNSKA Z. The phenomenon of leukergy. Arch Pathol (Chic) 1949 Mar;47(3):261–272. [PubMed] [Google Scholar]
  24. FLECK L. Recent investigations on leukergy. Tex Rep Biol Med. 1956;14(4):424–431. [PubMed] [Google Scholar]
  25. FLECK L. Uber Leukergie. Acta Haematol. 1952 Nov;8(5):282–293. doi: 10.1159/000204171. [DOI] [PubMed] [Google Scholar]
  26. FOLLETTE J. H., VALENTINE W. N., REYNOLDS J. A comparison of human leukocyte phosphatase activity toward sodium beta-glycerophosphate, adenosine 5'-phosphate and glucose 1-phosphate. Blood. 1959 Apr;14(4):415–422. [PubMed] [Google Scholar]
  27. FREI J., BOREL C., HORVATH G., CULLITY B., VANNOTTI A. Enzymatic studies in the different types of normal and leukemic human white cells. Blood. 1961 Sep;18:317–327. [PubMed] [Google Scholar]
  28. GEDIGK P., BONTKE E. Uber die Enzymaktivität im Fremdkörpergranulationsgewebe. Virchows Arch Pathol Anat Physiol Klin Med. 1957;330(5):538–568. doi: 10.1007/BF00956748. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. GOMORI G. Chromogenic substrates for aminopeptidase. Proc Soc Exp Biol Med. 1954 Dec;87(3):559–561. doi: 10.3181/00379727-87-21443. [DOI] [PubMed] [Google Scholar]
  31. GROGG E., PEARSE A. G. E. The enzymic and lipid histochemistry of experimental tuberculosis. Br J Exp Pathol. 1952 Dec;33(6):567–576. [PMC free article] [PubMed] [Google Scholar]
  32. HANKS J. H., WALLACE J. H. Determination of cell viability. Proc Soc Exp Biol Med. 1958 May;98(1):188–192. doi: 10.3181/00379727-98-23985. [DOI] [PubMed] [Google Scholar]
  33. HANKS J. H., WALLACE R. E. Relation of oxygen and temperature in the preservation of tissues by refrigeration. Proc Soc Exp Biol Med. 1949 Jun;71(2):196–200. doi: 10.3181/00379727-71-17131. [DOI] [PubMed] [Google Scholar]
  34. HANNIBAL M. J., NACHLAS M. M., SELIGMAN A. M. Histochemical localization of cytochrome oxidase in human tissues and neoplasms. Cancer. 1960 Sep-Oct;13:1008–1016. doi: 10.1002/1097-0142(196009/10)13:5<1008::aid-cncr2820130522>3.0.co;2-2. [DOI] [PubMed] [Google Scholar]
  35. HOFFMANN G. T., ROTTINO A., STERN K. G. Demonstration by the Nadi reaction of cytochrome oxidase activity in cells of the lymphoid and myeloid series obtained from normal individuals and patients suffering from Hodgkin's and other diseases. Blood. 1951 Nov;6(11):1051–1058. [PubMed] [Google Scholar]
  36. HOSODA S., NAKAMURA K. A new method for histochemical observation of the subcutaneous connective tissue. J Histochem Cytochem. 1960 Jan;8:72–73. doi: 10.1177/8.1.72. [DOI] [PubMed] [Google Scholar]
  37. HOWARD J. G. Activation of the reticulo-endothelial cells of the mouse liver by bacterial lipopolysaccharide. J Pathol Bacteriol. 1959 Oct;78:465–470. doi: 10.1002/path.1700780212. [DOI] [PubMed] [Google Scholar]
  38. HOWARD J. G., ROWLEY D., WARDLAW A. C. Investigations on the mechanism of stimulation of non-specific immunity by bacterial lipopolysaccharides. Immunology. 1958 Jul;1(3):181–203. [PMC free article] [PubMed] [Google Scholar]
  39. KAPLOW L. S. A histochemical procedure for localizing and evaluating leukocyte alkaline phosphatase activity in smears of blood and marrow. Blood. 1955 Oct;10(10):1023–1029. [PubMed] [Google Scholar]
  40. KARRER H. E. The ultrastructure of mouse lung: the alveolar macrophage. J Biophys Biochem Cytol. 1958 Nov 25;4(6):693–700. doi: 10.1083/jcb.4.6.693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. KOLER R. D., SEAMAN A. J., OSGOOD E. E., VANBELLINGHEN P. Myeloproliferative diseases; diagnostic value of the leukocyte alkaline phosphatase test. Am J Clin Pathol. 1958 Oct;30(4):295–301. doi: 10.1093/ajcp/30.4.295. [DOI] [PubMed] [Google Scholar]
  42. LEONARD B. J., ISRAELS M. C., WILKINSON J. F. Alkaline phosphatase in the white cells in leukaemia and leukaemoid reactions. Lancet. 1958 Feb 8;1(7015):289–292. doi: 10.1016/s0140-6736(58)91031-6. [DOI] [PubMed] [Google Scholar]
  43. MALATY H. A., BOURNE G. H. The histochemistry of simple esterase. Acta Anat (Basel) 1955;23(4):337–350. doi: 10.1159/000141005. [DOI] [PubMed] [Google Scholar]
  44. 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]
  45. MYRVIK Q. N., LEAKE E. S., FARISS B. Lysozyme content of alveolar and peritoneal macrophages from the rabbit. J Immunol. 1961 Feb;86:133–136. [PubMed] [Google Scholar]
  46. NACHLAS M. M., SELIGMAN A. M. The comparative distribution of esterase in the tissues of five mammals by a histochemical technique. Anat Rec. 1949 Dec;105(4):677–695. doi: 10.1002/ar.1091050405. [DOI] [PubMed] [Google Scholar]
  47. NACHLAS M. M., TSOU K. C., DE SOUZA E., CHENG C. S., SELIGMAN A. M. Cytochemical demonstration of succinic dehydrogenase by the use of a new p-nitrophenyl substituted ditetrazole. J Histochem Cytochem. 1957 Jul;5(4):420–436. doi: 10.1177/5.4.420. [DOI] [PubMed] [Google Scholar]
  48. NOVIKOFF A. B., ESSNER E. The liver cell. Some new approaches to its study. Am J Med. 1960 Jul;29:102–131. doi: 10.1016/0002-9343(60)90011-5. [DOI] [PubMed] [Google Scholar]
  49. PEARSON B., DEFENDI V. A comparison between the histochemical demonstration of non-specific esterase activity by 5-bromoindoxyl acetate, alpha-naphthyl acetate and naphthol AS acetate. J Histochem Cytochem. 1957 Jan;5(1):72–83. doi: 10.1177/5.1.72. [DOI] [PubMed] [Google Scholar]
  50. QUIGLEY H. J., DAWSON E. A., HYUN B. H., CUSTER R. P. The activity of alkaline phosphatase in granular leukocytes during pregnancy and the puerperium: a preliminary report. Am J Clin Pathol. 1960 Feb;33:109–114. doi: 10.1093/ajcp/33.2.109. [DOI] [PubMed] [Google Scholar]
  51. SHNITKA T. K., SELIGMAN A. M. Role of esteratic inhibition on localization of esterase and the simultaneous cytochemical demonstration of inhibitor sensitive and resistant enzyme species. J Histochem Cytochem. 1961 Sep;9:504–527. doi: 10.1177/9.5.504. [DOI] [PubMed] [Google Scholar]
  52. STAHELIN H., SUTER E., KARNOVSKY M. L. Studies on the interaction between phagocytes and tubercle bacilli. I. Observations on the metabolism of guinea pig leucocytes and the influence of phagocytosis. J Exp Med. 1956 Jul 1;104(1):121–136. doi: 10.1084/jem.104.1.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. STEIGLEDER G. K., SCHULTIS K. Die Bedeutung des Nachweises unspezifischer Esterasen in Bindegewebszellen der Haut. Arch Klin Exp Dermatol. 1957;204(5):448–456. [PubMed] [Google Scholar]
  54. THORBECKE G. J., OLD L. J., BENACERRAF B., CLARKE D. A. A histochemical study of acid and alkaline phosphatase in mouse livers during various conditions modifying activity of the reticuloendothelial system. J Histochem Cytochem. 1961 Jul;9:392–399. doi: 10.1177/9.4.392. [DOI] [PubMed] [Google Scholar]
  55. TRUBOWITZ S., FELDMAN D., BENANTE C., HUNT V. M. The alkaline phosphatase content of human leukocytes following marrow suppression by nitrogen mustard. J Lab Clin Med. 1958 Aug;52(2):259–265. [PubMed] [Google Scholar]
  56. TRUBOWITZ S., FELDMAN D., MORGENSTERN S. W., HUNT V. M. The isolation, purification and some properties of the alkaline phosphatase of human leucocytes. Biochem J. 1961 Aug;80:369–374. doi: 10.1042/bj0800369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. TRUBOWITZ S., MOSCHIDES E., FELDMAN D. Alkaline phosphatase activity of the polymorphonuclear leukocyte in rapidly induced leukopenia and leukocytosis. J Lab Clin Med. 1961 May;57:747–754. [PubMed] [Google Scholar]
  58. VACCARI F., SABOTTO B., MANZINI E. Alkaline phosphatase activity of leukocytes in shock. Blood. 1955 Jul;10(7):730–734. [PubMed] [Google Scholar]
  59. VALENTINE W. N., BECK W. S. Biochemical studies on leucocytes. I. Phosphatase activity in health, leucocytosis, and myelocytic leucemia. J Lab Clin Med. 1951 Jul;38(1):39–55. [PubMed] [Google Scholar]
  60. VALENTINE W. N., FOLLETTE J. H., HARDIN E. B., BECK W. S., LAWRENCE J. S. Studies on leukocyte alkaline phosphatase activity: relation to stress and pituitary-adrenal activity. J Lab Clin Med. 1954 Aug;44(2):219–228. [PubMed] [Google Scholar]
  61. VALENTINE W. N., FOLLETTE J. H., SOLOMON D. H., REYNOLDS J. The relationship of leukocyte alkaline phosphatase to stress, to ACTH, and to adrenal 17-OH-corticosteroids. J Lab Clin Med. 1957 May;49(5):723–737. [PubMed] [Google Scholar]
  62. VOGEL F. S. A lipolytic enzyme in reactive histiocytes of guinea pigs with experimental encephalomyelitis. J Exp Med. 1951 Apr 1;93(4):305–312. doi: 10.1084/jem.93.4.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. WACHSTEIN M., MEISEL E. A comparative study of enzymatic staining reactions in the rat kidney with necrobiosis induced by ischemia and nephrotoxic agents (mercuhydrin and DL-serine). J Histochem Cytochem. 1957 May;5(3):204–220. doi: 10.1177/5.3.204. [DOI] [PubMed] [Google Scholar]
  64. WACHSTEIN M., MEISEL E., FALCON C. Histochemistry of thiolacetic acid esterase: a comparison with nonspecific esterase with special regard to the effect of fixatives and inhibitors on intracellular localization. J Histochem Cytochem. 1961 May;9:325–339. doi: 10.1177/9.3.325. [DOI] [PubMed] [Google Scholar]
  65. WACHSTEIN M., MEISEL E. Influence of experimental renal damage on histochemically demonstrable succinic dehydrogenase activity in the rat. Am J Pathol. 1954 Jan-Feb;30(1):147–165. [PMC free article] [PubMed] [Google Scholar]
  66. 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]
  67. WEISS L. P., FAWCETT D. W. Cytochemical observations on chicken monocytes macrophages and giant cells in tissue culture. J Histochem Cytochem. 1953 Jan;1(1):47–65. doi: 10.1177/1.1.47. [DOI] [PubMed] [Google Scholar]
  68. WILTSHAW E., MOLONEY W. C. Histochemical and biochemical studies on leukocyte alkaline phosphatase activity. Blood. 1955 Nov;10(11):1120–1131. [PubMed] [Google Scholar]

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