Abstract
We studied the in vivo killing and degradation of Mycobacterium aurum, a nonpathogenic, acid-fast bacillus, within macrophages after inoculation into the peritoneal cavity of CD-1 mice. The degradative process could be divided in five successive steps that were characterized on ultrastructural and cytochemical grounds and the relative contributions of which were determined by quantitative electron microscopy of samples taken at different times. The main ultrastructural alterations observed during the degradative process were ribosome disaggregation, coagulation of the cytoplasmic matrix, and change in the membrane profile from asymmetric to symmetric, with loss of the polysaccharide components from the outer layer, followed by membrane solubilization and intracellular clearing, followed by digestion of the innermost (peptidoglycan) layer of the cell wall, and at the end of the process, disorganization and collapse of the remaining layers of the cell wall. The correlation between viability and morphology indicated that the first ultrastructural signs of viability loss are cytoplasmic coagulation, change in the membrane geometry, and disappearance of ribosomes. The labeling of lysosomes of peritoneal macrophages with ferritin or by the cytochemical demonstration of inorganic trimetaphosphatase showed that fusion of lysosomes with phagosomes containing mycobacteria occurs in the phagocytes in the mouse peritoneal cavity and is already extensive as soon as 1 h after the inoculation of the bacilli.
MeSH Terms
Animals
Immunity, Cellular
Lysosomes/ultrastructure
Macrophages/immunology,ultrastructure
Membrane Fusion
Mice
Microscopy, Electron
Mycobacterium/immunology,ultrastructure
Mycobacterium Infections/immunology
Peritoneal Cavity/immunology,microbiology
Phagocytosis
Phagosomes/ultrastructure
Time Factors
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Silva M T
Appelberg R
Silva M N
Macedo P M
References (24)
24 references, click to expand
-
Bacteriolysis.
Annu Rev Microbiol. 1965;19:79-104
PMID: 5318449
-
Electron microscopic aspects of membrane alterations during bacterial cell lysis.
Exp Cell Res. 1967 May;46(2):245-51
PMID: 4961137
-
A method for counting acid-fast bacteria.
Int J Lepr Other Mycobact Dis. 1968 Jan-Mar;36(1):78-82
PMID: 4869698
-
Electron-transparent zone of mycobacteria may be a defence mechanism.
Nature. 1970 Nov 28;228(5274):860-1
PMID: 4097724
-
Ultrastructural and chemical alterations induced by dicumarol in Streptococcus faecalis.
Biochim Biophys Acta. 1971 Oct 12;249(1):114-21
PMID: 4110917
-
Ultrastructural alterations induced by moist heat in Bacillus cereus.
Appl Microbiol. 1972 Sep;24(3):463-76
PMID: 4116891
-
Characterization of autolysins from Mycobacterium smegmatis.
J Bacteriol. 1977 Feb;129(2):750-5
PMID: 14109
-
Oxygen-dependent microbial killing by phagocytes (first of two parts).
N Engl J Med. 1978 Mar 23;298(12):659-68
PMID: 24176
-
Changes in pH within the phagocytic vacuoles of human neutrophils and monocytes.
Lab Invest. 1978 Sep;39(3):179-85
PMID: 30859
-
Effects of local anesthetics on bacterial cells.
J Bacteriol. 1979 Jan;137(1):461-8
PMID: 104970
-
The subcellular distribution and biochemical properties of peroxidase in monocytes and macrophages.
Front Biol. 1979;48:463-514
PMID: 387465
-
Oxygen metabolism and the toxic properties of phagocytes.
Ann Intern Med. 1980 Sep;93(3):480-9
PMID: 6254418
-
Differences in response among inbred mouse strains to infection with small doses of Mycobacterium bovis BCG.
Infect Immun. 1981 Apr;32(1):42-7
PMID: 7012036
-
Temporal changes of lysosome and phagosome pH during phagolysosome formation in macrophages: studies by fluorescence spectroscopy.
J Cell Biol. 1981 Jun;89(3):645-52
PMID: 6166620
-
Ultrastructure of Mycobacterium leprae and other acid-fast bacteria as influenced by fixation conditions.
Ann Microbiol (Paris). 1982 Jul-Aug;133(1):59-73
PMID: 6810742
-
Ultrastructural alterations of mycobacterium leprae in skin biopsies of untreated and treated lepromatous patients.
Ann Microbiol (Paris). 1982 Jul-Aug;133(1):75-92
PMID: 7051933
-
Electron microscopic study of Mycobacterium leprae membrane.
Int J Lepr Other Mycobact Dis. 1983 Jun;51(2):219-24
PMID: 6352528
-
The interpretation of the ultrastructure of mycobacterial cells in transmission electron microscopy of ultrathin sections.
Int J Lepr Other Mycobact Dis. 1983 Jun;51(2):225-34
PMID: 6352529
-
Intracellular location of Mycobacterium leprae in macrophages of normal and immune-deficient mice and effect of rifampin.
Infect Immun. 1983 Nov;42(2):802-11
PMID: 6358034
-
Ultrastructural characterization of normal and damaged membranes of Mycobacterium leprae and of cultivable mycobacteria.
J Gen Microbiol. 1984 Feb;130(2):369-80
PMID: 6427399
-
Cytochemical study of macrophage lysosomal inorganic trimetaphosphatase and acid phosphatase.
J Ultrastruct Res. 1985 Jan;90(1):80-8
PMID: 2999416
-
Oxygen-dependent and oxygen-independent mechanisms of microbicidal activity of neutrophils.
Immunol Lett. 1985;11(3-4):159-63
PMID: 3910565
-
Evidence for inhibition of fusion of lysosomal and prelysosomal compartments with phagosomes in macrophages infected with pathogenic Mycobacterium avium.
Infect Immun. 1986 Apr;52(1):252-62
PMID: 2870027
-
A SIMPLIFIED LEAD CITRATE STAIN FOR USE IN ELECTRON MICROSCOPY.
J Cell Biol. 1965 May;25:407-8
PMID: 14287192