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. 1973 Mar 1;56(3):621–627. doi: 10.1083/jcb.56.3.621

LYSOSOMAL PHOSPHOLIPASES A1 AND A2 OF NORMAL AND BACILLUS CALMETTE GUERIN-INDUCED ALVEOLAR MACROPHAGES

Richard C Franson 1, Moseley Waite 1
PMCID: PMC2108922  PMID: 4347204

Abstract

A single intravenous injection of 0.1 mg of heat-killed Bacillus Calmette Guérin (BCG) in 0.1 ml of Bayol F produced an accumulation of activated alveolar macrophages (BCG induced). Cells were collected 3.5–4.0 wk after injection. Phospholipases A and three lysosomal marker enzymes (acid phosphatase, β-glucuronidase, and lysozyme) were measured in homogenates, and the distribution of the phospholipases A and lysosomal, mitochondrial, and microsomal marker enzymes were examined after sucrose gradient centrifugation of a postnuclear (1,000 g) supernatant. Homogenates of normal and BCG-induced macrophages contained phospholipases A1 and A2 which had optimal activity at pH 4.0 in the presence of 2.0 mM ethylenediaminetetraacetate (EDTA). These activities were inhibited 50–70% by 2.0 mM CaCl2. Homogenates of BCG-induced macrophages had specific activities of β-glucuronidase, acid phosphatase, and lysozyme, which were increased 1.5- to 3.0-fold over the controls, whether expressed as activity per mg protein or activity per 107 cells. The specific activities of the phospholipases A, on the other hand, were consistently lower than those of the control. Distribution of the phospholipases A and the lysosomal marker enzymes after sucrose gradient centrifugation suggested that the phospholipases A active at pH 4.0 in the presence of EDTA are of lysosomal origin since: (a) BCG treatment caused a selective increase in the density of particles which contained both the phospholipases A and three lysosomal marker enzymes; and (b) since the density of mitochondria and microsomes were not affected by BCG treatment. The increase in the density of lysosomes seen here may be related to previously described morphologic changes of BCG-induced alveolar macrophages.

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Selected References

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  1. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  2. Boak J. L., Christie G. H., Ford W. L., Howard J. G. Pathways in the development of liver macrophages: alternative precursors contained in populations of lymphocytes and bone-marrow cells. Proc R Soc Lond B Biol Sci. 1968 Feb 27;169(1016):307–327. doi: 10.1098/rspb.1968.0013. [DOI] [PubMed] [Google Scholar]
  3. COHN Z. A., WIENER E. THE PARTICULATE HYDROLASES OF MACROPHAGES. II. BIOCHEMICAL AND MORPHOLOGICAL RESPONSE TO PARTICLE INGESTION. J Exp Med. 1963 Dec 1;118:1009–1020. doi: 10.1084/jem.118.6.1009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Canonico P. G., Bird J. W. Lysosomes in skeletal muscle tissue. Zonal centrifugation evidence for multiple cellular sources. J Cell Biol. 1970 May;45(2):321–333. doi: 10.1083/jcb.45.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eibl H., Lands W. E. A new, sensitive determination of phosphate. Anal Biochem. 1969 Jul;30(1):51–57. doi: 10.1016/0003-2697(69)90372-8. [DOI] [PubMed] [Google Scholar]
  6. Elsbach P. Phospholipid metabolism by phagocytic cells. I. A comparison of conversion of [32P]lysolecithin to lecithin and glycerylphosphorylcholine by homogenates of rabbit polymorphonuclear leukocytes and alveolar macrophages. Biochim Biophys Acta. 1966 Dec 7;125(3):510–524. [PubMed] [Google Scholar]
  7. Franson R., Waite M., LaVia M. Identification of phospholipase A 1 and A 2 in the soluble fraction of rat liver lysosomes. Biochemistry. 1971 May 11;10(10):1942–1946. doi: 10.1021/bi00786a031. [DOI] [PubMed] [Google Scholar]
  8. Franson R., Waite M., Weglicki W. Phospholipase A activity of lysosomes of rat myocardial tissue. Biochemistry. 1972 Feb 1;11(3):472–476. doi: 10.1021/bi00753a028. [DOI] [PubMed] [Google Scholar]
  9. GIANETTO R., DE DUVE C. Tissue fractionation studies. 4. Comparative study of the binding of acid phosphatase, beta-glucuronidase and cathepsin by rat-liver particles. Biochem J. 1955 Mar;59(3):433–438. doi: 10.1042/bj0590433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Leake E. S., Myrvik Q. N. Changes in morphology and in lysozyme content of free alveolar cells after the intravenous injection of killed BCG in oil. J Reticuloendothel Soc. 1968 Feb;5(1):33–53. [PubMed] [Google Scholar]
  12. MYRVIK Q., LEAKE E. S., FARISS B. Studies on pulmonary alveolar macrophages from the normal rabbit: a technique to procure them in a high state of purity. J Immunol. 1961 Feb;86:128–132. [PubMed] [Google Scholar]
  13. Mizunoe K., Dannenberg A. M., Jr Hydrolases of rabbit macrophages. 3. Effect of BCG vaccination, tissue culture, and ingested tubercle bacilli. Proc Soc Exp Biol Med. 1965 Nov;120(2):284–290. doi: 10.3181/00379727-120-30513. [DOI] [PubMed] [Google Scholar]
  14. Sottocasa G. L., Kuylenstierna B., Ernster L., Bergstrand A. An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study. J Cell Biol. 1967 Feb;32(2):415–438. doi: 10.1083/jcb.32.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]

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