Lysosomes and the "toxicity" of Rickettsiales. I. Cytochemical studies of macrophages inoculated in vitro with C. psittaci 6BC

1972 ◽  
Vol 18 (4) ◽  
pp. 457-464 ◽  
Author(s):  
Nonna Kordová ◽  
John C. Wilt
Keyword(s):  
Acid Phosphatase ◽  
Neutral Red ◽  
Immune Serum ◽  
Cytotoxic Action ◽  
Irreversible Damage ◽  
Lysosomal Acid ◽  
Control Serum ◽  
Mouse Macrophages ◽  
Rapid Destruction

Toxic doses of C. psittaci 6BC strain produced within 3–6 h after inoculation a rapidly progressive irreversible damage of mouse macrophages in vitro, an early leakage of a lysosomal enzyme, acid phosphatase, as well as a rapid disintegration of neutral red-stained cytoplasmic granules i.e. lysosomes. Pretreatment of the agent with immune serum neutralized the cytotoxic action of the agent and a reversible activation of lysosomes occurred. Pretreatment of the agent with undiluted control serum protected the macrophages from the early "toxic" injury; however, a cytopathic effect (CPE) occurred 48 h after inoculation and was accompanied by a release of lysosomal acid phosphatase. Heat-inactivated Chlamydiae produced an early agglutination of macrophages with leakage of lysosomal acid phosphatase, disintegration of neutral red-stained lysosomes, as well as a progressive irreversible damage of the macrophage. Concentrated, heat-inactivated agent-free, leuco-agglutinating material from 6BC infected L cells produced a similar injury of macrophages and their lysosomes. The findings are related to an earlier report of a rapid destruction of macrophages by C. psittaci in the intact animal (15). The possible mechanisms and factors of the "toxicity" of Rickettsiales are discussed.

Lysosomes and the "toxicity" of Rickettsiales. IV. Ultrastructural studies of macrophages infected with a cytopathic L cell-grown C. psittaci 6BC strain

1973 ◽  
Vol 19 (3) ◽  
pp. 315-320 ◽  
Author(s):  
Nonna Kordová ◽  
Jan Hoogstraten ◽  
John C. Wilt
Keyword(s):  
Acid Phosphatase ◽  
Neutral Red ◽  
Contributory Factor ◽  
Enzyme Release ◽  
High Concentration ◽  
Ultrastructural Studies ◽  
Cytoplasmic Organelles ◽  
L Cell ◽  
Mouse Macrophages

Mouse macrophages in vitro inoculated with a low dose of a cytopathic L cell-grown C. psittaci 6BC strain were harvested at times when a delayed cytopathic effect (CPE) was observed by light microscopy and when a cytochemical test for acid phosphatase revealed lysosomal enzyme release. An array of different lysosomal types was revealed by ultrastructural studies such as digestive vesicles with single or double outer membranes and whorls of membranes, dense residual bodies, aggregated and single scattered ferritin-like granules, and fragments of membranes. Enlargement of cysternae of endoplasmic reticulum, disintegration of cytoplasmic organelles, and thinning of ground cytoplasm were also observed. Cytoplasmic degeneration has been observed both within membrane-limited structures and in wider areas not limited by recognizable membranes.A high concentration of the agent resulted in an early toxic effect; this was accompanied by a rapid vacuolization of the cytoplasm of the macrophages which showed a poorly developed ultrastructure, and absence of mitochondria and cytoplasmic organelles. Cytochemical tests indicated that these alterations correlated with release of lysosomal acid phosphatase and disintegration of neutral red stained granules. Ultrastructural studies support the view that disintegration of lysosomes may be considered an important contributory factor in the pathology of chlamydial infection.

scholarly journals THE PARTICULATE HYDROLASES OF MACROPHAGES

1963 ◽  
Vol 118 (6) ◽  
pp. 1009-1020 ◽  
Author(s):  
Zanvil A. Cohn ◽  
Edith Wiener
Keyword(s):  
Acid Phosphatase ◽  
Yeast Cell ◽  
Cell Walls ◽  
Hydrolytic Enzymes ◽  
Neutral Red ◽  
Biochemical Properties ◽  
Extracellular Medium ◽  
Cell Content ◽  
Prolonged Incubation

The influence of phagocytosis on the morphological and biochemical properties of macrophage hydrolase-containing granules has been studied in vitro. Following the uptake of large numbers of heat-killed bacteria, an intracellular rearrangement of hydrolytic enzymes occurred. This was associated with the solubilization of 50 to 60 per cent of the total cell content of acid phosphatase, cathepsin, lysozyme, beta glucuronidase, acid ribonuclease, and acid desoxyribonuclease and with a corresponding decrease in granule-bound enzyme. With more prolonged incubation the majority of the soluble intracellular pool of acid ribonuclease and lysozyme was lost to the extracellular medium. No change in the total content of any of the hydrolases was noted during 180 minutes of incubation in vitro. The morphological fate of the granules was studied by a histochemical method for acid phosphatase. After the phagocytosis of yeast cell walls there was a disappearance of acid phosphatase-positive granules and an accumulation of reaction product about the ingested particle. Experiments employing macrophages which were supravitally stained with neutral red also demonstrated the loss of neutral red-positive granules and the accumulation of the dye about the yeast cell walls. These results strongly suggest that lysis of macrophage granules occurs following phagocytosis and that a portion of the granule contents are then resegregated within the newly formed phagocytic vacuole.

scholarly journals Reduced Granule Cell Proliferation and Molecular Dysregulation in the Cerebellum of Lysosomal Acid Phosphatase 2 (ACP2) Mutant Mice

2021 ◽  
Vol 22 (6) ◽  
pp. 2994
Author(s):  
Xiaodan Jiao ◽  
Maryam Rahimi Balaei ◽  
Ejlal Abu-El-Rub ◽  
Filippo Casoni ◽  
Hassan Pezeshgi Modarres ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Acid Phosphatase ◽  
Granule Cell ◽  
Granule Cells ◽  
Mutant Mice ◽  
Lysosomal Acid ◽  
Protein Synthesis Machinery

Lysosomal acid phosphatase 2 (Acp2) mutant mice (naked-ataxia, nax) have a severe cerebellar cortex defect with a striking reduction in the number of granule cells. Using a combination of in vivo and in vitro immunohistochemistry, Western blotting, BrdU assays, and RT-qPCR, we show downregulation of MYCN and dysregulation of the SHH signaling pathway in the nax cerebellum. MYCN protein expression is significantly reduced at P10, but not at the peak of proliferation at around P6 when the number of granule cells is strikingly reduced in the nax cerebellum. Despite the significant role of the SHH–MycN pathway in granule cell proliferation, our study suggests that a broader molecular pathway and additional mechanisms regulating granule cell development during the clonal expansion period are impaired in the nax cerebellum. In particular, our results indicate that downregulation of the protein synthesis machinery may contribute to the reduced number of granule cells in the nax cerebellum.

scholarly journals THE INFLUENCE OF ERYTHROPHAGOCYTOSIS ON THE INTERACTION OF MACROPHAGES AND SALMONELLA IN VITRO

1966 ◽  
Vol 124 (2) ◽  
pp. 173-183 ◽  
Author(s):  
Fred A. Gill ◽  
Donald Kaye ◽  
Edward W. Hook
Keyword(s):  
Salmonella Typhimurium ◽  
Peritoneal Macrophages ◽  
Cytotoxic Action ◽  
No Inhibition ◽  
Mouse Macrophages ◽  
Mouse Erythrocyte ◽  
Mouse Peritoneal Macrophages

Phagocytosis and killing of Salmonella typhimurium by mouse peritoneal macrophages was inhibited when the bacteria and antibody-coated homologous erythrocytes or heterologous erythrocytes were simultaneously exposed to macrophages in vitro. No inhibition of phagocytosis or killing was observed in experiments employing uncoated or disrupted antibody-coated homologous erythrocytes. Degradation of S. typhimurium as measured by the loss of fluorescence from intracellular salmonella coated with fluorescein-labeled antibody was inhibited in macrophages which had previously ingested antibody-coated homologous erythrocytes. Anti-mouse-erythrocyte serum was found to have a cytotoxic action on mouse macrophages. However, the viability of macrophages was not altered by phagocytosis of antibody-coated homologous erythrocytes or uncoated heterologous erythrocytes.

Lysosomes and the "toxicity" of Rickettsials. II. Non-cytocidal interactions of egg-grown C. psittaci 6BC and in vitro macrophages

1972 ◽  
Vol 18 (6) ◽  
pp. 869-873 ◽  
Author(s):  
Nonna Kordová ◽  
Linda Poffenroth ◽  
John C. Wilt
Keyword(s):  
Acid Phosphatase ◽  
Cell Damage ◽  
Giant Cells ◽  
Host Cells ◽  
Marked Contrast ◽  
Lysosomal Acid ◽  
Cytoplasmic Vacuoles ◽  
L Cell

During the infection of cultured mouse peritoneal phagocytes with egg-grown C. psittaci 6BC strain, lysosomes retained their integrity and host cells were not damaged. Infected monocytes showed greater ability than uninfected monocytes to spread and transform into giant cells containing enlarged nuclei and masses of cytoplasm with clear cytoplasmic vacuoles. Chlamydial particles were released from the cytoplasm of infected phagocytes by pseudopodia-like extrusions. These events were in marked contrast to the effect of L cell grown C. psittaci 6BC strain which caused early leakage of lysosomal acid phosphatase into the cytoplasm of macrophages and induced a rapidly progressive irreversible cell damage (14).

scholarly journals LYSOSOMAL ACID HYDROLASES IN MICE INFECTED WITH BCG

1965 ◽  
Vol 121 (5) ◽  
pp. 727-738 ◽  
Author(s):  
Kazuhisa Saito ◽  
Emanuel Suter
Keyword(s):  
Acid Phosphatase ◽  
Liver Homogenate ◽  
Subcellular Fractions ◽  
The Other ◽  
Acid Phosphatases ◽  
Acid Hydrolases ◽  
Lysosomal Acid

Experiments are reported dealing with the increase of lysosomal acid hydrolases induced by BCG infection. Acid hydrolases were determined quantitatively in peritoneal MP, liver homogenate, and plasma of normal and BCG-infected mice. A significant increase of acid phosphatase, ß-glucuronidase, and cathepsin was found in MP and liver homogenate of BCG-infected mice. In plasma also a significant increase of acid phosphatase and ß-glucuronidase was noticed. The results of the determination of the enzymes in centrifugally separated subcellular fractions of liver homogenate indicated clearly that the acid hydrolases associated mainly with the "large granular" fraction, which consists of mitochondria, lysosomes, and microsomes and that infection with BCG caused significant increase of the enzymes specifically in this fraction. Differences in the pattern of location among centrifugally separated fraction of liver homogenate were observed between acid phosphatase and the other two acid hydrolases. MP cultured in vitro doubled their acid phosphatases content within 24 hours, whereas ß-glucuronidase rather decreased in the same cells.

scholarly journals THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES

1965 ◽  
Vol 121 (1) ◽  
pp. 153-170 ◽  
Author(s):  
Zanvil A. Cohn ◽  
Belinda Benson
Keyword(s):  
Acid Phosphatase ◽  
Specific Activity ◽  
Total Content ◽  
Progressive Increase ◽  
Neutral Red ◽  
Mononuclear Phagocytes ◽  
In Vitro Differentiation ◽  
Dense Granules

The in vitro differentiation of homogeneous populations of monocyte-like cells from the unstimulated mouse peritoneal cavity is described. Under the conditions employed, a progressive increase in cell size occurs without significant cell division. This process is characterized morphologically by the accumulation of phase-dense and neutral red-positive granules, mitochondria, and lipid droplets. The phase-dense granules react strongly for acid phosphatase. Biochemical determinations indicate marked increases in the total content and specific activity of acid phosphatase, cathepsin, and ß-glucuronidase. The production of acid phosphatase is more rapid and extensive than that of the other two hydrolases. From these data it appears that the conversion of a monocyte-like cell to a mature macrophage is accompanied by the formation of increased numbers of lysosome-like cytoplasmic organelles. Mouse peritoneal phagocytes stimulated in vivo with a bacterial lipopolysaccharide undergo a similar series of morphological and biochemical events.

The Demonstration of Human Prostatic Acid Phosphatase (Pap) With Phosphorylcholine

1976 ◽  
Vol 34 ◽  
pp. 88-89
Author(s):  
José A. Serrano ◽  
Hannah L. Wasserkrug ◽  
Anna A. Serrano ◽  
Arnold M. Seligman
Keyword(s):  
Acid Phosphatase ◽  
Prostate Gland ◽  
Prostatic Acid Phosphatase ◽  
Human Prostate ◽  
Rat Tissues ◽  
Specific Substrate ◽  
Acid Phosphatases ◽  
Lysosomal Acid ◽  
Cacodylate Buffer

As previously reported (1, 2) phosphorylcholine (PC) is a specific substrate for prostatatic acid phosphatase (PAP) as opposed to other acid phosphatases, e.g., lysosomal acid phosphatase. The specificity of PC for PAP is due to the pentavalent nitrogen in PC, a feature that renders PC resistant to hydrolysis by all other acid phosphatases. Detailed comparative cytochemical results in rat tissues are in press. This report deals with ultracytochemical results applying the method to normal and pathological human prostate gland.Fresh human prostate was obtained from 7 patients having transurethral resections or radical prostatectomies. The tissue was fixed in 3% glutaraldehyde- 0.1 M cacodylate buffer (pH 7.4) for 15 min, sectioned at 50 μm on a Sorvall TC-2 tissue sectioner, refixed for a total of 2 hr, and rinsed overnight in 0.1 M cacodylate buffer (pH 7.4)-7.5% sucrose.

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