Abstract
Phagocytosis of erythrocytes was studied in vitro in an incubation system consisting of rat peritoneal macrophages and antibody-coated 59Fe-labeled erythrocytes. The system was characterized in terms of the rate and magnitude of erythrophagocytosis, determined by the interiorization of the 59Fe label. On incubation of 150 × 106 macrophages with 75 × 106 antibodycoated erythrocytes, erythrophagocytosis began within a few minutes and was essentially completed after 2 h when 50% of the offered red cells had been ingested by the macrophages. Heme oxygenase (HO) activity, which is very low in native macrophages, increased 4- to 10- fold in response to the ingested erythrocytes; this enzyme stimulation occurred with a delay of 3 h in relation to erythrophagocytosis. Actinomycin D or puromycin prevented the increase of HO activity without affecting erythrophagocytosis, which suggests that the enzyme stimulation was due to substrate-mediated enzyme induction.
Hydrocortisone (HC) (0.1 mg/ml medium) dissociated erythrophagocytosis from HO induction, leaving the former unimpaired but completely suppressing the latter. The suppressive effect of HC on the enzyme induction was completely prevented by 5 mg glucose and 0.02 U insulin/ml of the medium. In macrophages engaged in erythrophagocytosis. HC also lowered glucose removal from the medium and reduced formation of 14CO2 from [1-14C]glucose.
These results suggest that induction of HO in macrophages by the hemoglobin of ingested erythrocytes requires intact transport or metabolism of glucose. Glucose utilization appears to be impaired by HC, but is restored by additional glucose and insulin. The findings suggest that plasma steroid concentrations in the pharmacological range could reduce bilirubin formation in phagocytic cells in vivo without affecting the sequestration and degradation of erythrocytes. This provides a possible explanation for the observation that in patients with hepatogenous jaundice, steroids often lower the serum bilirubin level.
Full text
PDF










Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Axline S. G., Cohn Z. A. In vitro induction of lysosomal enzymes by phagocytosis. J Exp Med. 1970 Jun 1;131(6):1239–1260. doi: 10.1084/jem.131.6.1239. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baehner R. L., Gilman N., Karnovsky M. L. Respiration and glucose oxidation in human and guinea pig leukocytes: comparative studies. J Clin Invest. 1970 Apr;49(4):692–700. doi: 10.1172/JCI106281. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bissell D. M., Hammaker L., Schmid R. Liver sinusoidal cells. Identification of a subpopulation for erythrocyte catabolism. J Cell Biol. 1972 Jul;54(1):107–119. doi: 10.1083/jcb.54.1.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bunn H. F. Erythocyte destruction and hemoglobin catabolism. Semin Hematol. 1972 Jan;9(1):3–17. [PubMed] [Google Scholar]
- COHN Z. A., WIENER E. THE PARTICULATE HYDROLASES OF MACROPHAGES. I. COMPARATIVE ENZYMOLOGY, ISOLATION, AND PROPERTIES. J Exp Med. 1963 Dec 1;118:991–1008. doi: 10.1084/jem.118.6.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CROSBY W. H., FURTH F. W. A modification of the benzidine method for measurement of hemoglobin in plasma and urine. Blood. 1956 Apr;11(4):380–383. [PubMed] [Google Scholar]
- CROSBY W. H., HOUCHIN D. N. Preparing standard solutions of cyanmethemoglobin. Blood. 1957 Dec;12(12):1132–1136. [PubMed] [Google Scholar]
- Cline M. J. Leukocyte function in inflammation: the ingestion, killing, and digestion of microorganisms. Ser Haematol. 1970;3(2):3–16. [PubMed] [Google Scholar]
- Cohn Z. A., Ehrenreich B. A. The uptake, storage, and intracellular hydrolysis of carbohydrates by macrophages. J Exp Med. 1969 Jan 1;129(1):201–225. doi: 10.1084/jem.129.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Ehrenreich B. A., Cohn Z. A. Fate of hemoglobin pincytosed by macrophages in vitro. J Cell Biol. 1968 Jul;38(1):244–248. doi: 10.1083/jcb.38.1.244. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldstein G. W., Hammaker L., Schmid R. The catabolism of Heinz bodies: an experimental model demonstrating conversion to non-bilirubin catabolites. Blood. 1968 Mar;31(3):388–395. [PubMed] [Google Scholar]
- Greenwalt T. J., Flory L. L., Steane E. A. Quantitative haemagglutination. 3. Stuies of separated populations of human red blood cells of different densities. Br J Haematol. 1970 Dec;19(6):701–709. doi: 10.1111/j.1365-2141.1970.tb07015.x. [DOI] [PubMed] [Google Scholar]
- Huber H., Fudenberg H. H. Receptor sites of human monocytes for IgG. Int Arch Allergy Appl Immunol. 1968;34(1):18–31. doi: 10.1159/000230091. [DOI] [PubMed] [Google Scholar]
- JANDL J. H., FILES N. M., BARNETT S. B., MACDONALD R. A. PROLIFERATIVE RESPONSE OF THE SPLEEN AND LIVER TO HEMOLYSIS. J Exp Med. 1965 Aug 1;122:299–326. doi: 10.1084/jem.122.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- JANDL J. H., JONES A. R., CASTLE W. B. The destruction of red cells by antibodies in man. I. Observations of the sequestration and lysis of red cells altered by immune mechanisms. J Clin Invest. 1957 Oct;36(10):1428–1459. doi: 10.1172/JCI103542. [DOI] [PMC free article] [PubMed] [Google Scholar]
- KATZ R., DUCCI H., ALESSANDRI H. Influence of cortisone and prednisolone on hyperbilirubinemia. J Clin Invest. 1957 Sep;36(9):1370–1374. doi: 10.1172/JCI103536. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Landaw S. A., Callahan E. W., Jr, Schmid R. Catabolism of heme in vivo: comparison of the simultaneous production of bilirubin and carbon monoxide. J Clin Invest. 1970 May;49(5):914–925. doi: 10.1172/JCI106311. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Makman M. H., Dvorkin B., White A. Influence of cortisol on the utilization of precursors of nucleic acids and protein by lymphoid cells in vitro. J Biol Chem. 1968 Apr 10;243(7):1485–1497. [PubMed] [Google Scholar]
- Marikovsky Y., Danon D. Agglutination of young and old human red cells by blood group antibodies. Vox Sang. 1971 Feb;20(2):174–177. doi: 10.1111/j.1423-0410.1971.tb00548.x. [DOI] [PubMed] [Google Scholar]
- Munck A. Metabolic site and time course of cortisol action on glucose uptake, lactic acid output, and glucose 6-phosphate levels of rat thymus cells in vitro. J Biol Chem. 1968 Mar 10;243(5):1039–1042. [PubMed] [Google Scholar]
- North R. J. The localization by electron microscopy of acid phosphatase activity in guinea pig macrophages. J Ultrastruct Res. 1966 Sep;16(1):96–108. doi: 10.1016/s0022-5320(66)80025-4. [DOI] [PubMed] [Google Scholar]
- OSTROW J. D., JANDL J. H., SCHMID R. The formation of bilirubin from hemoglobin in vivo. J Clin Invest. 1962 Aug;41:1628–1637. doi: 10.1172/JCI104620. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petrik P., Collet A. J. Electron microscopic study of the in vivo erythrophagocytosis by alveolar macrophages of the cat. II. Late period: digestion. Z Zellforsch Mikrosk Anat. 1971;116(4):477–486. doi: 10.1007/BF00335053. [DOI] [PubMed] [Google Scholar]
- Pimstone N. R., Engel P., Tenhunen R., Seitz P. T., Marver H. S., Schmid R. Inducible heme oxygenase in the kidney: a model for the homeostatic control of hemoglobin catabolism. J Clin Invest. 1971 Oct;50(10):2042–2050. doi: 10.1172/JCI106697. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pimstone N. R., Tenhunen R., Seitz P. T., Marver H. S., Schmid R. The enzymatic degradation of hemoglobin to bile pigments by macrophages. J Exp Med. 1971 Jun 1;133(6):1264–1281. doi: 10.1084/jem.133.6.1264. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RAABO E., TERKILDSEN T. C. On the enzymatic determination of blood glucose. Scand J Clin Lab Invest. 1960;12(4):402–407. doi: 10.3109/00365516009065404. [DOI] [PubMed] [Google Scholar]
- Rabinovitch M. Phagocytosis of modified erythrocytes by macrophages and L2 cells. Exp Cell Res. 1969 Aug;56(2):326–332. doi: 10.1016/0014-4827(69)90021-4. [DOI] [PubMed] [Google Scholar]
- Rabinovitch M. The dissociation of the attachment and ingestion phases of phagocytosis by macrophages. Exp Cell Res. 1967 Apr;46(1):19–28. doi: 10.1016/0014-4827(67)90405-3. [DOI] [PubMed] [Google Scholar]
- Rifkind R. A. Destruction of injured red cells in vivo. Am J Med. 1966 Nov;41(5):711–723. doi: 10.1016/0002-9343(66)90032-5. [DOI] [PubMed] [Google Scholar]
- Schiff L. The use of steroids in lier disease. Medicine (Baltimore) 1966 Nov;45(6):565–573. doi: 10.1097/00005792-196645060-00023. [DOI] [PubMed] [Google Scholar]
- Tenhunen R., Marver H. S., Schmid R. Microsomal heme oxygenase. Characterization of the enzyme. J Biol Chem. 1969 Dec 10;244(23):6388–6394. [PubMed] [Google Scholar]
- Tenhunen R., Marver H. S., Schmid R. The enzymatic catabolism of hemoglobin: stimulation of microsomal heme oxygenase by hemin. J Lab Clin Med. 1970 Mar;75(3):410–421. [PubMed] [Google Scholar]
- Tenhunen R., Marver H. S., Schmid R. The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. Proc Natl Acad Sci U S A. 1968 Oct;61(2):748–755. doi: 10.1073/pnas.61.2.748. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WILLIAMS R., BILLING B. H. Action of steroid therapy in jaundice. Lancet. 1961 Aug 19;2(7199):392–396. doi: 10.1016/s0140-6736(61)92481-3. [DOI] [PubMed] [Google Scholar]
- Weed R. I., Reed C. F. Membrane alterations leading to red cell destruction. Am J Med. 1966 Nov;41(5):681–698. doi: 10.1016/0002-9343(66)90030-1. [DOI] [PubMed] [Google Scholar]
- Weiss L., Tavassoli M. Anatomical hazards to the passage of erythrocytes through the spleen. Semin Hematol. 1970 Oct;7(4):372–380. [PubMed] [Google Scholar]
- Werthamer S., Hicks C., Amaral L. Protein synthesis in human leukocytes and lymphocytes: 1. Effect of steroids and sterols. Blood. 1969 Sep;34(3):348–356. [PubMed] [Google Scholar]
- Wiener E., Marmary Y., Curelaru Z. The in vitro effect of hydrocortisone on the uptake and intracellular digestion of particulate matter by macrophages in culture. Lab Invest. 1972 Feb;26(2):220–226. [PubMed] [Google Scholar]
- Young D. A. Glucocorticoid action on rat thymus cells. Interrelationships between carbohydrate, protein, and adenine nucleotide metabolism and cortisol effects on these functions in vitro. J Biol Chem. 1969 Apr 25;244(8):2210–2217. [PubMed] [Google Scholar]