Alpha-naphtyl Acetate Esterase Research Articles

Effects of Aflatoxin on the Proportions of Peripheral Blood Leukocytes and Alpha-Naphtyl Acetate Esterase (ANAE) Positive Lymphocytes in the Mouse

Summary In this study detrimental effects of aflatoxin (AF) on the peripheral blood formula and the alpha naphthyl acetate esterase (ANAE) - positivity profile of peripheral blood lymphocytes (PBL) on the mice were determined. A total of 40 white mice were randomly divided in to 5 groups each having 8 animals. The first group served as a control and received standard diet. The remaining 4 groups were fed diets containing 200, 400, 800 and 1600 ppb AF, respectively. The results of this study have revealed that AF given by the food caused significant declines both in the proportion of PBL and in the percentages of ANAEpositive peripheral blood lymphocytes (T-lymphocytes) in a dose dependent manner. These results may be concerned as a haematological evidence for immuno-suppressive effect of AF on cell-mediated immunity.

Light and electron microscopic studies on alpha naphthyl acetate esterase activity of the peripheral blood T lymphocytes in van cats.

The purpose of this study was to determine the percentage of peripheral blood T lymphocytes and the localization of ANAE enzyme at the electron microscopic level in Van cats by using an alpha-naphtyl acetate esterase procedure. Peripheral blood samples taken from 20 Van cats were used. The percentage of ANAE positive lymphocytes was 83.0%. Neutrophilic granulocytes gave a negative reaction, whereas monocytes, eosinophilic granulocytes showed a diffuse granular positivity. In the electron microscopic examination, ANAE positive reactions were seen in lysosomal granules found in lymphocytes.

The Langerhans' cells.

The morphological, enzymatical, immunocytochemical and functional properties of Langerhans' cells are briefly reviewed. Langerhans' cells are located mainly in the squamous stratified epithelia, but are also present in the thymus and in superficial lymphnodes. At the ultrastructural level, they are characterized by unique cytoplasmic organelles, the Birbeck granules, whose function is still unknown. Langerhans' cells possess strong ATPase activity and are weakly positive for alpha-naphtyl acetate esterase and for acid phosphatase; they are immunoreactive for CD1a (T6), class II MHC antigens and S-100 protein. In some pathological conditions, like dermatopathic lymphadenopathy and Langerhans' cell histocytosis, Langerhans' cells also are characterized by the expression of monocyte-macrophage antigens. Langerhans' cells act as antigen-presenting cells to T lymphocytes; their functional capacity is strictly dependent on the levels of expression of class II MHC antigens. Langerhans' cells are of bone marrow origin and are derived from a circulating precursor which is probably related to the monocyte.

CHARACTERISTICS OF DENDRITIC CELLS IN RAT LIVER

The properties of rat liver dendritic cells (DC) were analyzed after collagenase digestion of the tissue and enrichment with density gradient centrifugation. The liver macrophages (Kupffer cells) were eliminated by adherence before the gradient centrifugation. The morphology of isolated DC in May-Grunwald-Giemsa (MGG) stained cytocentrifuge preparations resembled that of monocytes with certain dissimilarities. The expression of major histocompatibility complex (MHC) class II antigens (Ia) on DC was analyzed with the Staphylococcus aureus rosette method using a monoclonal antibody. The binding of anti-Ia antibody to rat liver DC was 3 times stronger than to passenger lymphocytes and 10 times stronger than to hepatocytes. All DC were Ia-positive tested with indirect immunofluorescence technique, and none of them were able to phagocytize antibody-coated human red cells. The DC did not express intracytoplasmic lysozyme, or surface Fc-receptors, and they all were negative in alpha-naphtylacetate esterase (ANAE) staining. Thus, although the dendritic cells of rat liver seem to belong to the monocytic series according to morphologic criteria, they were all negative when tested for typical monocyte/macrophage markers. The immunocenic potential of DC was analyzed by testing their ability to prime a naive recipient for graft rejection. The number DC needed for the priming was comparable with the number of spleen lymphocytes needed for an equivalent effect, indicating that the DC were highly immunogenic. The hepatocytes showed practically no immunogenic effect. Thus the immunogenic potential of the tested cells, i.e. their ability to induce accelerated transplant rejection, carries a good correlation with the expression of Ia-antigens on the cell surface.

Characterization of factor XIII containing-macrophages in lymph nodes with Hodgkin's disease.

A large number of cells containing subunit a of blood coagulation Factor XIII (FXIII) was detected by immunoperoxidase staining in lymph nodes with Hodgkin's disease. These relatively large, multipolar, mononuclear cells were often found in the immediate vicinity of malignant Hodgkin's cells. Intensive characterization of these cells carried out by immunofluorescent and enzymecytochemical techniques in double- and triple-labelling systems on the same sections clearly demonstrated that they represent tumour-associated macrophages (TAMs). FXIII containing-cells showed alpha-naphtyl acetate esterase (ANAE) positivity, and were labelled by monoclonal anti-Leu M3 antibody, a monocyte/macrophage marker, but not at all or only very weakly by anti-HLA-DR. Neither alkaline phosphatase (ALP) nor adenosine triphosphatase (ATPase) activity could be detected in these cells and surprisingly, they were consistently negative for acid phosphatase (AcP) as well. The presence of FXIII subunit a in tumour-associated macrophages suggests that this cell type might have an important role in the stabilization of fibrin deposits around tumour cells.

Lymphoid cells in the venous blood of 100 female patients with breast carcinoma. Follow-up studies

The following blood cells were examined in 100 patients with invasive carcinoma of the breast and 100 control patients without malignant disease: leukocytes, lymphocytes, monocytes and different sub-populations of lymphoid cells. Lymphoid cells were characterized by E-, EA- and EAC-rosetting technique, alphanaphtyl acetate esterase activity and by the ability of latex phagocytosis. Both groups showed no significant differences. In 81 patients undergoing postoperative radiation therapy lymphocytes and lymphoid cells were reduced considerably. These changes were still evident after 12 months (6 months after completion of radiotherapy). Mainly the T lymphocytes were effected by this decrease. A negative effect of the radiation induced reduction of immune competent cells must be discussed and examined further by clinical trials.

Complement receptor distinguishes between two subsets of large granular lymphocytes with different natural killer activity and cytochemical and ultrastructural features.

Human peripheral blood large granular lymphocytes (LGL)--that is, cells with intracytoplasmic azurophilic (electron-dense) granules, with a positivity for the cytochemical localization of certain acid hydrolases, and with avid surface receptors for the Fc portion of IgG--have been purified on Percoll density gradients. Approximately 30% of these cells expressed receptors for the third complement component (C3R). They were separated into C3R-positive and C3R-negative cells. C3R+ cells had a significantly greater natural killer (NK) activity against K562 target cells than C3R+ cells. This difference was unrelated to the presence in the C3R+ cells of a contaminant cell type incapable of NK activity, since cytochemical and ultrastructural analysis revealed that C3R+ and C3R- fractions contained comparable LGL numbers. Agarose cytotoxicity assays at the single-cell level demonstrated that C3R+ LGL contained a large number of cells that bound to but did not lyse the target. The remaining fully cytotoxic C3R+ LGL had, however, the same killing and recycling properties as the cells from the C3R fraction. Electron microscopy and cytochemical studies showed that C3R+ cells had fewer electron-dense granules than C3R cells and stained more faintly for the localization of alpha-naphtyl acetate esterase. In contrast to C3R cells, C3R+ LGL displayed morphological features suggesting that an active process of granule formation was taking place. Taken together, the data indicate that C3R+ cells represent a discrete subset or a maturational stage of LGL.

Demonstration of T Lymphocytes in Leprous Granuloma Using the Acid α Naphtyl Acetate Esterase Activity: An Attempt at Quantitative Analysis

In nine leprosy patients (1 TT, 1 BT, 4 BL and 3 LLp), esterase positive lymphocytes (T Lymphocytes) were studied in frozen sections of skin biopsies by alpha naphtyl acetate esterase pH 5.8 method (ANAE) Four patients had never been treated previously and five patients exhibited clinical and bacteriological evidence of relapse for inadequate therapy at first biopsy. There was an increase in ANAE (+) lymphocyte density in granulomas when second biopsies were done after efficient treatment, evaluated by bacillary index for the eight bacilliferous patients, and clinical improvement. The significance of T cells in granulomas is discussed.

Dendritic reticulum cell sarcoma? Four cases of a lymphoma probably derived from dendritic reticulum cells of the follicular compartment

This paper describes the histological picture of four tumours of the follicular compartment of the lymph node, in which the proliferating cell appeared to be the dendritic reticulum cell (DRC). This assumption was based on the results of light microscopical, ultrastructural, immunological, and enzymehistochemical investigations. The tumour cells resembled DRC's closely in (1) the striking pattern of interdigitations and occasional tight junction-like contacts between the neoplastic cells on electron microscopical analysis; (2) presence of receptors for the activated third component of complement on the membrane of the cells; (3) absence of monoclonal immunoglobulins and T-cell antigen on the surface and of lysozyme, alpha 1-antitrypsin or alpha 1-antichymotrypsin in the cytoplasm of the neoplastic cells. Moreover, (4) the tumour cells showed moderate alpha-naphtyl acetate esterase, weak to absent acid phosphatase and (with one exception) strong 5-nucleotidase activity. Furthermore, (5) the neoplastic cells expressed Ia-like antigens on the surface in all four cases. The relation with follicle centre cell lymphomas, the differential diagnosis and clinical data are discussed.

Colostral T lymphocytes detected by intracytoplasmic and membrane markers.

Colostral lymphocytes were studied using two established T-cell markers: intracytoplasmic alpha-naphtyl-acetate esterase (ANAE) staining and membrane receptors for sheep erythrocytes (E rosettes). ANAE staining allowed counting and identification of T-cell subsets independently of the status of membrane structures and receptors frequently altered in colostral cells. The fact that a sizeable number of colostral lymphocytes had the same phenotype as the majority of mature circulating peripheral blood lymphocytes supports the hypothesis that colostral lymphocytes may play a role in protecting neonates against infections, in transferring immune information to the newborn, or in modulating the immune response via release of soluble factors. A considerable percentage of colostral T lymphocytes are ANAE-negative. This phenotype is similar to that observed among thymocytes.

Lymphocyte morphology in middle ear effusions.

Cytological studies were performed on 157 middle ear effusion specimens, 54 of which belonged to the serous and 103 to the mucoid category. The cytocentrifuged samples were stained with May-Gruenwald-Giemsa stain and divided into the three groups on the basis of cellular pattern: granulocytic, lymphocytic-monocytic and mixed. The T-lymphocytes were counted after incubation for six hours using alpha-naphtyl-acetate esterase (ANAE) marker and counterstaining with toluidine blue. In 57 samples, reliable assessment of lymphocytes could not be made because the absolute number of white blood cells was small or the lymphocytes were less than 100. In the remaining 40 serous and 60 mucoid samples, lymphocytes were present in sufficient numbers. The mean percentages of ANAE-positive lymphocytes were 62% serous and 57% mucoid. The results indicate that there is no numerical defect of the T-cells in chronic middle ear effusions.

Cytochemical profile of human haematopoietic biopsy cells and derived cell lines.

Twenty-three human haematopoietic cell lines, normal and mitogen stimulated peripheral blood lymphocytes and tumour material from fresh leukaemias, myelomas and lymphomas were investigated with a panel of cytochemical reactions. Normal and mitogen stimulated lymphocytes, non-neoplastic lymphoblastoid cell lines (LCL), lymphoma lines with B-lymphocyte characteristics, chronic lymphocytic leukaemia and fresh lymphocytic lymphomas reacted weakly or negatively with all stains. T-lymphocyte acute leukaemia lines were PAS and alpha-naphtyl acetate esterase positive. Myeloma lines and fresh myelomas were strongly beta-glucoronidase positive. A histiocytic lymphoma cell line was strongly esterase positive with naphtol AS-D acetate esterase inhibited by NaF. The three fresh histiocytic lymphomas, however, reacted as the lymphocytic lymphomas suggesting a lymphoid origin. A myeloid leukaemia line was strongly positive for acid phsophatase. No major disagreement was noted between the reactivity of established neoplastic lines and the corresponding fresh biopsy cells indicating an unaltered qualitative expression of enzyme production after prolonged in vitro culture.