Lysosomal localisation of parallel tubular arrays in chronic lymphocytic leukaemia of T cell origin: an ultrastructural cytochemical study

An ultrastructural cytochemical study of lysosomal acid phosphatase was performed on leukemic cells in a case of chronic lymphocytic leukaemia of T cell origin (T-CLL). The cells showed inclusion bodies known as parallel tubular arrays, which often lay within acid phosphatase-positive, membrane-bound spaces. This suggests their lysosomal location.     

Lysosomal enzyme cytochemistry in acute leukaemia

A cytochemical study of the lysosomal enzyme beta-glucuronidase in 60 cases of acute leukaemia has shown a qualitative difference in the cytoplasmic distribution of the enzyme between blast cells of the lymphoid and myeloid cell series. This difference provides a useful additional method for cytochemical classification of cell type and is superior in this respect to the other lysosomal enzymes studied (aryl sulphatase and acid phosphatase). The beta-glucuronidase reaction is recommended in those cases of acute leukaemia in which the periodic acid-Schiff reaction is negative or equivocal.     

Stimulated intraphagosomal release of eosinophil peroxidase and acid phosphatase in the rats spontaneously infected with Mycoplasma pulmonis: a cytochemical study

In this study we performed a cytochemical comparison of peroxidase and acid phosphatase activities in peritoneal eosinophils from specific pathogen-free (SPF) and Mycoplasma pulmonis-infected rats. When eosinophils ingested polystyrene particles for 120 min, peroxidase- and acid phosphatase-positive specific granules, as well as small granules, fused to phagosomes. Unusual peroxidase activity was detected in some particle-containing phagosomes in 6.8% of eosinophils from control SPF rats and 31% of those from infected rats. Intense acid phosphatase activity was also demonstrated in some phagosomes in 4 and 20% of eosinophils from control and infected rats, respectively. The rate of peroxidase-positive and acid phosphatase-positive phagosomes to all ingested phagosomes was 8.1 and 7.3% in control rats and rose to 41.3 and 31.1% in those infected, respectively. The population of eosinophils (18%) in the control peritoneal cells did not change after infection (16.6%). These results suggest that the intraphagosomal release of lysosomal enzymes was significantly stimulated in peritoneal eosinophils of the rats spontaneously infected with M. pulmonis.     

Significance of lysosomes in the morphogenesis of coronaviruses

Summary Virus-containing electron-dense membrane-bound cytoplasmic bodies are described in tracheal epithelial cells of chickens infected with Infectious Bronchitis Virus and in intestinal epithelial cells of swine infected with Porcine Epidemic Diarrhea Virus.Using silver-methenamine staining, phosphotungstic acid staining and acid phosphatase enzyme cytochemical staining of ultra-thin sections, these bodies were shown to be virus-containing secondary lysosomes and residual bodies.The accumulation of viral particles in the lysosomes is suggested to possibly represent an intracellular defense mechanism. However, no morphological alterations were found indicating a destruction of the viruses by the lytic lysosomal enzymes.With 4 FiguresPart of this work was presented as a thesis before the faculty of Veterinary Medicine at the State University of Gent.     

Lysosomal response of a murine macrophage-like cell line persistently infected with Coxiella burnetii.

The lysosomal response of a murine macrophage-like tumor cell line (J774) during persistent infection with Coxiella burnettii was examined. By using acid phosphatase as a lysosomal marker, it was shown that phagosome-lysosome fusion occurred in J774 cells persistently infected with C. burnetii. This observation was verified using thorium dioxide, an electron-dense compound that is sequestered in secondary lysosomes. The phagolysosomes contained viable replicating rickettsiae. Spectrofluorometric analysis indicated that the phagolysosomal pH of persistently infected cells was acidic. In attempts to correlate rickettsial survival with lysosome function, the activities of several lysosomal enzymes were assayed in both infected and uninfected cells. Activities of acid phosphatase and beta-acetylglucosaminidase were not significantly altered during infection. However, infected cells appeared to display slightly higher intracellular lysozyme, beta-glucuronidase, and beta-galactosidase activities.     

Lysosomal activity in the aging rat liver: II. Morphometry of acid phosphatase positive dense bodies

Hepatocytes from 3-, 12- and 36-month-old rats were studied in situ with cytochemical and morphometrical techniques to obtain information about the intralobular and age-dependent variation in lysosomal volume. An increase with age was found for the lysosomal volume density. This increase is evident throughout the lobule, but occurs at different rates in the centrolobular and peripheral areas. It is most pronounced in the centrolobular area, resulting in a lysosomal volume density of 7.1% in old rats, compared to 3.6% in the peripheral area of the liver lobule. Both the size and the number of individual lysosomes contribute to the overall effect. Most dense bodies, including lipofuscin-containing aggregates in senescent animals, exhibit acid phosphatase activity. Our results suggest that the use of enzyme activity as a marker may yield higher values for lysosomal volume density than morphologic characteristics alone. The alleged homogeneity of the liver as a source of hepatocytes for studies on isolated cells must be questioned when lysosomal functions are concerned.     

Cytochemical properties of peripheral blood neutrophils in patients with hyperthyroidism]

In the cytoenzymatic investigations of peripheral blood neutrophils in patients with hyperthyroidism there was found the increase of acid phosphatase activity, beta-glucuronidase, leucine aminopeptidase, and catalase moreover there was found the decrease of the activity of alkaline phosphatase. After a two-week treatment with thiamazole (methimazole++) 50 mg in 24-hour dose there was observed the decrease of acid phosphatase activity in neutrophils. During incubation of plasma containing leucocytes, from healthy persons, with L-thyroxine there was observed the increase of the activity for acid phosphatase and beta-glucuronidase. In patients with hyperthyroidism there appear many changes of enzymic equipment of neutrophils which are concerned with lysosomal and connected with cell membrane enzymes. The results of cytochemical investigations after application of thiamazole and no difference, with exception of catalase, between patients with Graves-Basedow disease and with toxic goitre and the results of investigations in vitro with L-thyroxin point out, that there is the possibility of connection between the observed changes in the range of enzymic equipment of neutrophils and the hormonal state of the investigated group of patients.     

Cytochemical observations on lysosomal enzymes of rat lymphocytes at various ages

Light-microscopic cytochemical observations have been made in the presence of two lysosomal hydrolases (acid phosphatase and β-glucuronidase) in the lymphocytes of rats of various ages. The results indicate a different behaviour for the two enzymes which has been discussed by comparing the variable percentages of small and large lymphocytes.     

Acid phosphatase stimulation of the growth of Nocardia asteroides and its possible relationship to the modification of lysosomal enzymes in macrophages.

Lysosomal acid phosphatase levels are reduced in murine macrophages by virulent strains of Nocardia asteroides. At the same time, other lysosomal enzymes either remain unchanged or increase in activity, indicating that acid phosphatase is not lost because of degranulation or membrane leakage. This study shows that acid phosphatase was utilized as a sole carbon source by Nocardia asteroides and that acid phosphatase combined with glutamate as a carbon source enhanced nocardial growth. As a consequence, the inverse relationship that was observed between acid phosphatase activity and the bactericidal capacity of macrophages infected with N. asteroides appears to be due to the ability of N. asteroides to preferentially metabolize this lysosomal enzyme during growth within phagocytes.     

Lysosomes in Aortic Smooth Muscle Cells: Effects of Hypertension

Hypertension induces hypertrophy and increased turnover of aortic smooth muscle cells along with an accumulation of connective tissue in the aortic wall. We identified the lysosomes in normal and hypertensive aortic muscle cells by light and electron microscopy, utilizing cytochemical staining for acid phosphatase activity. Lysosomes were found to be more numerous in hypertensive vessels. Biochemical assays of two specific lysosomal enzymes revealed a doubling of acid phosphatase and a more than threefold increase in β-N-acetyl-glucosaminidase activities in hypertensive aortas.     

Evidence for phagosome-lysosome fusion in Mycobacterium leprae-infected murine Schwann cells.

Murine Schwann cells were infected with viable armadillo-derived Mycobacterium leprae in vitro, and the lysosomal marker enzyme, acid phosphatase, was stained by the Gomori reaction. Electron microscopic analysis revealed that Schwann cells infected with M. leprae possess acid phosphatase and that lysosomes fuse with infected phagosomes.     

Demonstration of extracellular acid phosphatase activity in the involuting,antimesometrial decidua in fed and acutely fasted mice by combined cytochemistry and electron microscopy

An ultrastructural cytochemical study of acid phosphatase activity in the antimesometrial decidua on days 9–11 of pregnancy was performed in fed and acutely fasted mice. Specimens were fixed in a buffered mixture of paraformaldehyde and glutaraldehyde and were incubated in a buffered medium containing sodium β-glycerophosphate and cerium chloride for ultrastructural localization of acid phosphatase activity. Fed and fasted animals showed extracellular acid phosphatase reaction product in the decidual-trophoblast interface, in the region of loosely and tightly packed, mature decidual cells, and in the region of predecidual cells. Reaction product was absent in the region of nondecidualized stromal cells. Extracellular acid phosphatase activity was more conspicuous in the region of mature decidual cells in fasted mice than in fed mice, and it was apparently similar in the region of predecidual cells in both fed and fasted mice. Acid phosphatase reaction product was also observed in lysosomes in all cells studied. Because acid phosphatase activity reflects the presence of lysosomal hydrolases in general, our results suggest that there is matrix degradation by lysosomal enzymes in both fed and fasted mice. These events may be part of the process of tissue remodeling in regions of predecidual cells and mature decidual cells. However, it is also possible that, in the region of mature decidual cells, breakdown of matrix constituents is a mechanism to provide nutrients for the growing fetus. This mechanism is probably enhanced in fasted mice. Anat. Rec. 252:1–7, 1998. © 1998 Wiley-Liss, Inc.     

Purple acid phosphatase of human brain macrophages in AIDS encephalopathy

Brains from AIDS patients with an HIV-induced encephalopathy but without opportunistic infections or indications for an inflammation were studied by immuno- and enzyme-histochemical methods. It was found that the macrophages of these brains expressed a lysosomal tartrate-resistant acid phosphatase which gave a good immunological cross-reaction with an antibody to the well-characterized iron-containing bovine spleen purple acid phosphatase, belonging to the group of purple phosphatases, which are regarded as a marker for a special phenotype of activated macrophages. It was discussed that the numerous brain macrophages found in AIDS encephalopathy derive from latently infected monocytes which are believed to be drawn to the brain from the bloodstream.     

Osteoclast Origin of Giant Cells in Giant Cell Tumors of Bone: Ultrastructural and Cytochemical Study of Six Cases

To clarify the histogenesis of giant cells appearing in giant cell tumors of bone (GCTB), an ultrastructural and cytochemical study of six cases was performed with both acid phosphatase (ACPase) and tartrate-resistant acid phosphatase (TRACPase) as marker enzymes. TRACPase is considered a specific marker for osteoclasts. ACPase was demonstrated in the macrophagelike stromal cells, the multinucleated giant cells, and the infiltrating macrophages. The enzyme reaction was localized in lysosomal dense bodies and Golgi areas. Intense TRACPase activity was demonstrated in the multinucleated giant cells, whereas a weak reaction was found in the macrophagelike stromal cells. The multinucleated giant cells and macrophagelike stromal cells resembled osteoclasts with regard to the subcellular localization of TRACPase. The present results suggest that the giant cells in GCTB are indeed derived from osteoclasts.     

Osteoclast Origin of Giant Cells in Giant Cell Tumors of Bone: Ultrastructural and Cytochemical Study of Six Cases

To clarify the histogenesis of giant cells appearing in giant cell tumors of bone (GCTB), an ultrastructural and cytochemical study of six cases was performed with both acid phosphatase (ACPase) and tartrate-resistant acid phosphatase (TRACPase) as marker enzymes. TRACPase is considered a specific marker for osteoclasts. ACPase was demonstrated in the macrophagelike stromal cells, the multinucleated giant cells, and the infiltrating macrophages. The enzyme reaction was localized in lysosomal dense bodies and Golgi areas. Intense TRACPase activity was demonstrated in the multinucleated giant cells, whereas a weak reaction was found in the macrophagelike stromal cells. The multinucleated giant cells and macrophagelike stromal cells resembled osteoclasts with regard to the subcellular localization of TRACPase. The present results suggest that the giant cells in GCTB are indeed derived from osteoclasts.     

Alkaline and Acid Phosphatase in Murine Leukemia

Alterations for acid and alkaline phosphatase levels and their pattern of splenic and lymph node activity in normal and virus-induced lymphoblastic leukemia were studied. Enzyme levels were examined by using both cytochemical and biochemical procedures. The GC leukemia virus, a ribonucleic acid murine virus antigenically related to the Rauscher-Moloney viruses, was used to stimulate acid and alkaline phosphatase by producing lymphomaceous disease in Ha/ICR mice. With the Burstone and Gomori cytochemical procedures, both enzymes were found in higher than normal levels in lymphomaceous spleen and lymph nodes. Confirmation of the cytochemical studies was obtained by enzyme assay of cell-free homogenates in each case with the exception of spleen acid phosphatase. The discrepancy between the cytochemical tests which showed significant elevation of spleen acid phosphatase and the enzyme assays which failed to reveal such elevation could be due to a labile acid phosphatase isozyme which is lost on cellular disruption during homogenate preparation. A significant spleen alkaline phosphatase specific activity elevation above normal was found with a 50% incidence only when leukemic spleen wet weight increased nearly threefold its normal value. This result suggests that alkaline phosphatase elevation is a secondary event occuring after the onset of disease and is not a fundamental metabolic alteration concerned with the onset of murine lymphoblastic leukemia.     

Cellular changes associated with Triton WR-1339 accumulation in rat hepatocytes. I. Autophagy.

One of the major changes in hepatocyte morphology resulting from Triton WR-1339 treatment is an apparent increase in autophagy. There is a rapid response with autophagic vacuole formation involving the Golgi apparatus and smooth endoplasmic reticulum occurring within 1 hr post-injection. Positive cytochemical localization of several lysosomal hydrolases confirmed the lysosomal nature of these autophagic structures. Between 6 and 12 hr post-injection there was a proliferation of acid phosphatase and aryl sulfatase positive myelin figures in the hepatocyte cytoplasm. By 12 hr autophagic vacuole formation had diminished and by 1 day post-injection there was an obvious reduction in myelin figures. Morphological observations suggested that this reduction might occur by clearance at the plasmalemma or by fusion with Triton filled lysosomes. Myelin figures were observed with decreasing frequency through 5 days post-injection at which time the Triton WR-1339 filled lysosomes predominated.     

Fluorescent cytochemical detection of estrogen and progesterone receptors in breast fine-needle aspirates

Estrogen and progesterone receptors were studied in fine-needle aspiration biopsy specimens of 56 patients with primary, recurrent, or metastatic breast carcinoma. The ligands, 17 B-estradiol-6-carboxymethyloxine-bovine serum albumin fluorescein isothiocyanate (FITC-BSA estradiol) and hydroxyprogesterone hemisuccinate bovine serum albumin tetramethyl rhodamine isothiocyanate (TMRITC-BSA progesterone), were used in the fluorescent cytochemical method. The findings obtained from the aspirated cells with the use of the fluorescent cytochemical technique were compared with results obtained from the cell population of the same tumor after removal with the use of both the fluorescent cytochemical technique and the biochemical dextran-coated charcoal (DCC) assay. For the needle aspirates, there was 89% concordance for estrogen receptor and 86% concordance for progesterone receptor between biochemical and cytochemical results. A high degree of correlation was also demonstrated between fine-needle aspirates and imprint preparations with the use of the cytochemical technique. This study suggests that the fluorescent cytochemical technique is an effective tool in assessment of estrogen and progesterone receptor content in fine-needle aspirates of primary and metastatic breast cancer. The fluorescent cytochemical technique can be performed easily at community hospitals and is well suited for specimens of insufficient size for biochemical assay.     

Tartrate-resistant, purple acid phosphatase in Gaucher cells of the spleen. Immuno- and cytochemical analysis

Bioptic material from the spleen of a three-year-old boy with a type 1 Gaucher disease was studied by immuno- and cytochemical methods with special regard to the macrophage-derived Gaucher cells. These cells were positive with PAS and Prussian blue staining, and were immuno-positive with the monoclonal 25 F9 antibody, specific to mature, non-inflammatory macrophages. Large Gaucher cells and their postulated small precursor cells revealed strong tartrate-resistant acid phosphatase (TRAP) and unspecific carboxylate esterase activities. Using a polyclonal antibody to bovine spleen purple phosphatase, a lysosomal TRAP from splenic macrophages, the TRAP of the Gaucher cells could be identified to belong to this group of iron-containing, purple acid phosphatases immunocytochemically. The origin of splenic Gaucher cells from blood monocytes and their further development are discussed.     

Cytochemical study of granulocyte enzymes in germ-free animals with adenovirus infections

The activity of some dehydrogenases and hydrolases was studied by cytochemical methods in the peripheral blood neutrophils of germ-free guinea pigs infected with adenoviruses. The gnotobiotic animals were obtained by hysterotomy in an operation isolation room after which they were transferred into manipulation isolation room and infected with human adenovirus type 1. A depression of enzymes of alpha-glycerophosphate shunt and NADP-H2-diaphorase in neutrophils two days after infection and activation of lactate dehydrogenase and acid phosphatase at 4 days were demonstrated. The pattern of changes in the enzymatic status of intact and infected gnotobiotic animals allowed a diagnosis of adenovirus infection in most cases.