Alpha-naphthyl Acetate Esterase Reaction Research Articles

Atypical micromegakaryocytes, promegakaryoblasts and megakaryoblasts: a critical evaluation by immunohistochemistry, cytochemistry and morphometry of bone marrow trephines in chronic myeloid leukemia and myelodysplastic syndromes.

A morphometric analysis of bone marrow trephine biopsies has been performed to study the frequency and planimetric characteristics of so-called atypical micromegakaryocytes in chronic myeloid leukemia (CML) and myelodysplastic syndromes (MDS). In addition, an attempt was made to discriminate this particular cell population from small immature elements of megakaryocytopoiesis, such as promegakaryoblasts and megakaryoblasts. The staining reactions employed included periodic acid-Schiff (PAS), alpha-naphthyl acetate esterase (ANAE) and immunohistochemistry with a monoclonal antibody against platelet glycoprotein IIIa (Y2/51-CD61). Comparison of the various staining reactions applied to the different megakaryocytic elements together with morphometric measurements resulted in a clearcut identification of promegakaryoblasts. These were defined as the earliest immature and exclusively CD61-positive precursors. Atypical micromegakaryocytes were characterized by their dysplastic features and strong ANAE reactivity in addition to their positive CD61 staining. When stringent diagnostic criteria (diameter ranging between 10 to 15 microns, mean size about 12 microns) were applied, this abnormal cell population comprised less than 10% of total megakaryocytopoiesis in CML and MDS. It may be assumed that dysmegakaryocytic features in the latter disorders are partially generated by small to medium-sized megakaryocytes (diameter less than 30 microns). In conclusion, the relative frequency of promegakaryoblasts in the normal bone marrow (range 6-8%) is confirmed by evaluation of the immunohistochemical and cytochemical staining methods (CD61 and ANAE). Furthermore, the ANAE reaction facilitates the recognition of atypical micromegakaryocytes as well as small megakaryocytes. Thus cytochemistry provides a better insight into alterations of these cell lineages in various pathological conditions.

Cytochemical reactions in bovine and ovine lymphosarcoma.

Cytochemical reactions with a panel of special stains were explored in peripheral blood and lymph node cells of animals with the adult, calf, and thymic types of bovine lymphosarcoma as well as lymphosarcoma in sheep induced with the bovine leukosis virus. These results were compared with normal lymphoid cells. Cytochemical methods included Sudan black B, periodic acid-Schiff, acid phosphatase and three esterases. Cells from ovine lymphosarcomas and a thymic type of bovine lymphosarcoma were distinctively positive to the alpha naphthyl acetate esterase reaction. These cytochemical reactions which were obtained with standard cytochemical methods should aid in the characterization and classification of lymphoproliferative disorders in animals, especially when combined with immunologic surface markers.

Acid Phosphatase and Alpha-Naphthyl Acetate Esterase in Neoplastic and Non-Neoplastic Lymphocytes. A Statistical Analysis

Acid phosphatase and alpha-naphthyl acetate esterase reaction patterns were evaluated in lymphocytes from patients with a variety of neoplastic and non-neoplastic conditions: leukemia, 59; NHL, 53; and reactive follicular hyperplasia, 23. Fifteen individuals with normal peripheral blood were also studied. For both enzymes, statistical analysis showed a strong correlation between a globular reaction pattern and T lymphocytic origin in both non-neoplastic lymph nodes and normal peripheral blood specimens (P less than 0.0001). A similarly strong correlation was found between a granular acid phosphatase pattern and T lymphocytic origin in cell isolated from non-neoplastic lymph nodes (P less than 0.0001) but not in those obtained from normal peripheral blood where this pattern was observed with equal frequency in B, T, and null lymphocytes (P = 0.415). A granular alpha-naphthyl acetate esterase pattern was correlated with non-T lymphocytes from normal peripheral blood (P less than 0.0001), but was observed with equal frequency in B, T, and null lymphocytes fron non-neoplastic lymph nodes (P = 0.76). In the eight T cell neoplasias studied, a globular pattern was evident in the majority of cells for both enzymes. In the majority of the B cell neoplasias, however, a granular pattern was observed for both enzymes.

Human large granular lymphocytes contain an esterase activity usually considered as specific for the myeloid series.

AbstractA cytochemical marker such as alpha‐naphthyl acetate esterase (ANAE) has been found useful for the morphological identification of the subset of T lymphocytes having receptors for Fcμ (TM cells). ANAE reaction on TM cells gives a typical pattern of one to four positive spots, whereas this pattern is not found on T cells with receptors for Fcγ (TG cells). ANAE is abundant in monocytes but not detectable in granulocytes. Herein another type of esterase activity, naphthol‐AS‐D chloroacetate esterase (NCAE), is described; it is known to be abundant in granulocytes and was found to give a specific pattern of reactivity with the subpopulation of large granular lymphocytes (LGL). This pattern of fine granular staining was observed not only on LGL present in the TG cell subpopulation but also in LGL present in the non‐T, non‐B cells.Fractions of peripheral blood mononuclear cells which were ènriched up to 80% in LGL by Percoll discontinuous density gradient gave a similar percentage of specific NCAE pattern. In addition, among the different fractions from Percoll gradient, there was a good correlation (r = 0.94) between the number of NCAE‐positive cells and the natural killer activity against the natural killer susceptible K562 target cells. It will be important to determine whether or not this enzymatic activity plays a role in the cytotoxic activities of LGL.

Chromosome pattern in childhood acute nonlymphocytic leukemia (ANLL)

We studied the karyotype in 26 children with ANLL, which was diagnosed on the basis of the FAB classification. Clonal chromosome abnormalities were found in 21 of 26 patients. Four patients, including 3 with Down's syndrome, had AML(M1). Nine patients, including 3 with t(8;21), had AML(M2). All 3 patients with APL(M3) had t(15;17). Four patients had AMMOL(M4); 3 of these had a normal karyotype. Six patients had AMOL(M5); 5 and 11q rearrangements, and 3 of these had a break in 11q23. Only one patient had EL(M6), and he had a normal karyotype. One patient with t(11;19), classified as AML(M2) on Wright-Giemsa-stained cells, had a strong alpha-naphthyl acetate esterase reaction, indicating that the leukemic cells had a cytochemical feature characteristic of monocytes. Whereas t(8;21) and t(15;17) are uniquely associated with AML(M2) and APL(M3), respectively, the 11q rearrangements are also seen in AML(M1/M2), although they are more common in AMOL(M5) and AMMOL(M4). The case with t(11;19) suggests that cells with 11q rearrangements and with AML(M1/M2) may have both monocytic and granulocytic features. When we used our data and previous reports on 243 aneuploid patients (169 adults and 74 children) to correlate the chromosome abnormalities with patient age, we found differences in the chromosome pattern seen among various age groups. This suggests that different etiologic factors as well as changes in host susceptibility may influence the development of and the karyotypic pattern in the various types of leukemia. Moreover, the frequency of various chromosome abnormalities in childhood ANLL can provide a baseline for comparison of the frequency of the same abnormality in adults. The karyotypic analysis of childhood ANLL is important not only because of the information that can be obtained about childhood ANLL, but also because the data can provide substantial insight into the etiology of ANLL in adults.

Chromosome Pattern in Childhood Acute Nonlymphocytic Leukemia (ANLL)

Abstract We studied the karyotype in 26 children with ANLL, which was diagnosed on the basis of the FAB classification. Clonal chromosome abnormalities were found in 21 of 26 patients. Four patients, including 3 with Down's syndrome, had AML(M1). Nine patients, including 3 with t(8;21), had AML(M2). All 3 patients with APL(M3) had t(15;17). Four patients had AMMOL(M4); 3 of these had a normal karyotype. Six patients had AMOL(M5); 5 and 11q rearrangements, and 3 of these had a break in 11q23. Only one patient had EL(M6), and he had a normal karyotype. One patient with t(11;19), classified as AML(M2) on Wright-Giemsa-stained cells, had a strong alpha-naphthyl acetate esterase reaction, indicating that the leukemic cells had a cytochemical feature characteristic of monocytes. Whereas t(8;21) and t(15;17) are uniquely associated with AML(M2) and APL(M3), respectively, the 11q rearrangements are also seen in AML(M1/M2), although they are more common in AMOL(M5) and AMMOL(M4). The case with t(11;19) suggests that cells with 11q rearrangements and with AML(M1/M2) may have both monocytic and granulocytic features. When we used our data and previous reports on 243 aneuploid patients (169 adults and 74 children) to correlate the chromosome abnormalities with patient age, we found differences in the chromosome pattern seen among various age groups. This suggests that different etiologic factors as well as changes in host susceptibility may influence the development of and the karyotypic pattern in the various types of leukemia. Moreover, the frequency of various chromosome abnormalities in childhood ANLL can provide a baseline for comparison of the frequency of the same abnormality in adults. The karyotypic analysis of childhood ANLL is important not only because of the information that can be obtained about childhood ANLL, but also because the data can provide substantial insight into the etiology of ANLL in adults.

Acid phosphatase and alpha-naphthyl acetate esterase in neoplastic and non-neoplastic lymphocytes. A statistical analysis.

Acid phosphatase and alpha-naphthyl acetate esterase reaction patterns were evaluated in lymphocytes from patients with a variety of neoplastic and non-neoplastic conditions: leukemia, 59; NHL, 53; and reactive follicular hyperplasia, 23. Fifteen individuals with normal peripheral blood were also studied. For both enzymes, statistical analysis showed a strong correlation between a globular reaction pattern and T lymphocytic origin in both non-neoplastic lymph nodes and normal peripheral blood specimens (P less than 0.0001). A similarly strong correlation was found between a granular acid phosphatase pattern and T lymphocytic origin in cell isolated from non-neoplastic lymph nodes (P less than 0.0001) but not in those obtained from normal peripheral blood where this pattern was observed with equal frequency in B, T, and "null" lymphocytes (P = 0.415). A granular alpha-naphthyl acetate esterase pattern was correlated with non-T lymphocytes from normal peripheral blood (P less than 0.0001), but was observed with equal frequency in B, T, and "null" lymphocytes fron non-neoplastic lymph nodes (P = 0.76). In the eight T cell neoplasias studied, a globular pattern was evident in the majority of cells for both enzymes. In the majority of the B cell neoplasias, however, a granular pattern was observed for both enzymes.

The origin of the dendritic reticulum cell. An experimental enzyme-histochemical and electron microscopic study on the rabbit spleen.

To obtain more information concerning the origin of dendritic reticulum cells, the development of germinal centers in the spleens of rabbits was investigated by conventional light microscopy, enzyme histochemistry, and electron microscopy. Washed sheep erythrocytes were used as antigen. Splenic tissue was examined on the 13th, 18th, 21st, 27th and 48th day after antigen administration. Electron microscopic investigations revealed transitional forms between typical fibroblastic reticulum cells, which formed the framework of the entire splenic white pulp, and typical dendritic reticulum cells. During this transformation, the enzyme histochemical pattern of alkaline phosphatase disappeared and a positive alpha-naphthylacetate esterase reaction appeared in the transformed cells. On the basis of these findings, it is highly likely that dendritic reticulum cells develop through transformation of fibroblastic reticulum cells during the development of germinal centers in rabbit spleens. The characteristic folding of the surface membrane of dendritic reticulum cells is probably caused by the conspicuous increase in size of the Golgi apparatus, the detachment of vesicles, and the uptake of such vesicles by the cell membrane observed electron microscopically during the cellular transformation. Receptors that are of significance in antigen trapping might reach the cell surface in this manner, i.e., with the Golgi vesicles.

A study of the nature of "hairy" cells, with emphasis on enzymatic markers.

Studies of peripheral blood, bone marrow, and spleen cells from three patients with hairy-cell leukemia were performed. Two of the three patients had well-organized cytoplasmic, ribosome-lamellar inclusions in their leukemic cells. Blast transformation and 3H-thymidine incorporation of lymphocytes seemed to fall within normal ranges when the findings were related to the absolute numbers of lymphocytes. The enzymatic markers demonstrated in hairy cells-strong acid phosphatase activity in endoplasmic reticulum and lysosomes, marked alpha-naphthyl acetate esterase reaction, and weak beta-glucuronidase activity-as well as their phagocytosis of latex particles, indicate a common origin with monocytes or histiocytes. No decisive results were obtained by immunofluorescence. Evaluation of the significance of the formation by hairy cells of mouse erythrocyte rosettes, as well as the presence of the typical hair-like projections, may require additional knowledge concerning the membrane of these cells.