Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We herein describe an unusual case of acute myeloid leukaemia (AML) showing strong cytochemical reactivity for myeloperoxidase (MPO) but surprisingly no reactivity using flow cytometry for any of the lineage-specific cell surface markers, i.e. myelomonocytic antigens CD13, CD14 and CD33; or B-lymphoid antigens CD19, CD20 and immunoglobulins; or T-lymphoid antigens CD2, CD3 and CD5. The strong reactivity for MPO and the complete absence of reactivity for CD13 and CD14 was verified by an independent assay involving alkaline phosphatase-anti-alkaline phosphatase (APAAP). Our case is of interest for at least two reasons: First, a poorly differentiated variant of AML (negative for MPO but positive for one or more of the myeloid-lineage CD antigens) has been designated FAB M0. In terms of the expression of phenotypic markers, our case may be considered as an 'MPO (+), CD antigen (-) AML'. The CD antigens are known to be expressed very early during myeloid differentiation whereas MPO (in its functional form) is viewed as being expressed relatively late in the process. It is therefore intriguing from a biological standpoint why the supposedly early antigens (CD33 and CD13) remain unexpressed; this may represent an example of 'asynchronous differentiation' in leukaemia. Second, from a practical standpoint, the use of immunophenotyping as a first-line diagnosis would fail to detect such cases. This case strengthens the notion that immunophenotyping by flow cytometry does not eliminate the necessity of performing peroxidase cytochemical staining.
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PMID:Acute myeloid leukaemia with an unusual phenotype: myeloperoxidase (+), CD13 (-), CD14 (-) and CD33 (-). 138 46

A panel of monoclonal antibodies (mAbs) directed against B-cell and hairy cell leukaemia (HCL)-associated antigens was used to identify residual hairy cells in the peripheral blood and/or bone marrow samples from 20 patients with HCL, following treatment with interferon-alpha (IFN-alpha) or interferon-beta (IFN-beta). In all cases, hairy cells retained their characteristic phenotype, e.g. positivity for CD22, CD11c, CD25, CD32, and the HCL-associated trimeric protein (t-GP) recognized by the mAbs HML-1, B-ly7, LF61 and Ber-Act8. The most specific marker for identifying a small percentage of hairy cells in peripheral blood cytospins, was t-GP. In alkaline phosphatase/anti alkaline phosphatase (APAAP) stained preparations, t-GP+ hairy cells (provided with large cytoplasm and hairy surface) could be usually distinguished from t-GP+ normal lymphocytes (small-sized cells with smooth surface). In doubtful cases the percentage of residual hairy cells could exactly be estimated by double immunofluorescence staining for CD22 (B-cell marker) and t-GP. The rationale of the test is based on the finding that the small percentage (about 1%) of t-GP+ lymphocytes circulating in the peripheral blood of normal individuals are T-cells of the CD8 subset and not B-cells. The best markers for identifying residual hairy cells in routine bone marrow biopsies were CD45RA (mAb 4KB5) and CD20 (mAb L26). Immunohistological labelling was superior to morphological examination in picking up scattered hairy cells in bone marrow biopsies showing either severe hypoplasia or exuberant hyperplasia of normal haemopoietic series.
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PMID:Selection of a panel of monoclonal antibodies for monitoring residual disease in peripheral blood and bone marrow of interferon-treated hairy cell leukaemia patients. 170 9

Lymph node cells from a patient with Hodgkin's disease (HD) were cultured without Epstein-Barr virus (EBV) or leukine adjuvant. A cell line (719-AB) emerged from the culture after four weeks. The cell line express CD20 (79%), CD 21 (30%), CD30 (63%), CD 35 (61%) antigens and weakly CD25 (19%). using Southern Blot technique, the existence of specific EBV DNA and polyclonal immunoglobulin genes rearrangement were observed in the cell line. In order to obtain a monoclonal antibodies (MoAb), mice Balb/C were immunized with this cell line. The splenic cells suspension of immunized animals were fused with the mouse myeloma NS1. Antibody IgM kappa from secreting clones 2B44 was studied using both indirect immunofluorescence with labeled anti-mouse immunoglobulin and immunohistochemistry based on alkaline phosphatase/antiphosphatase complex (APAAP) and ModAMeX technique on a panel of normal or pathological cells. Normal peripheral lymphocytes, monocytes, polymorphonuclear cells, and erythrocytes, did not react. The MoAb 2B44 recognized the dendritic reticulum cells and the smooth muscle cells of vessels on frozen section and paraffin section from HD or reactive lymph nodes. On specially processed paraffin sections (ModAMeX) Reed-Sternberg cells (RSC) were reactive with 2B44 MoAb (in 2 cases out of 5 tested). The molecular weight of the antigen recognized by 2B44 MoAb is of 37 kd. The description of a new epitope shared by different histological components might be of interest for defining a new cluster and better understanding the nature of RSC.
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PMID:Production of a monoclonal antibody (2B44) reactive on a shared epitope on dendritic reticulum cells, smooth muscle cells of vessels and Reed-Sternberg cells. 172 34

The aim of the present study was to compare the immunofluorescence technique (IF) with the immunoenzymatic (IE) alkaline phosphatase-antialkaline phosphatase method for the evaluation of the presence of lymphoid antigens (Ag) in 46 cases of acute myeloid leukemia (AML). The first technique allows detection of Ag expressed on the cytoplasmic membrane of living cells, whilst the second shows the presence of intracytoplasmic Ag on fixed cells. In general, the percentages of lymphoid Ag expression on AML cells are relatively low with both IE (15.2%) and IF (17.4%). We found a good correlation between the two methods for CD2 (4/4), CD7 (4/5), CD20 (1/1) and CD4 (2/2). The Ag CD19, CD21 and CD8 were negative in all cases, both with IE and with IF. CD3 (2 cases) and CD22 (1 case) were only evident with IE. CD10 was seen in 1 case with IF, whilst it was found more frequently with IE. For this reason, demonstration of CD10 with IF is more specific for the classification of acute leukemia.
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PMID:Incidence of lymphoid markers in acute myeloid leukemia. Alkaline phosphatase-antialkaline phosphatase versus immunofluorescence. 195 Mar 56

The B1 molecule (CD20) is a phosphoprotein found only on B lymphocytes. Multiple isoforms of the B1 molecule are expressed with Mr of 33,000, B1(33) and Mr of 34,500-36,000, B1(35). In this study it was found that nonproliferating B cells did not incorporate 32PO4 into B1 although phosphorylated class I histocompatibility molecules were easily detected. In contrast B1 isolated from proliferating or malignant B cells or B cell lines was heavily phosphorylated. Cross-linking B1 on the cell surface by antibody resulted in enhanced phosphorylation of B1 as did exposure to phorbol esters, and the membrane permeable diacylglycerol analog 1,2,-dioctanoylglyceron. B1(33) and B1(35) produced identical peptide maps following limited proteinase digestion. However, B1(35) contained both phosphoserine and phosphothreonine, while B1(33) only contained phosphoserine. In addition alkaline phosphatase was able to remove the phosphate residue(s) that resulted in generation of the B1(35) form of B1 but was unable to remove the phosphorylation of B1(33). These results suggest that phosphorylation of B1 molecules is associated with proliferation and that the different Mr forms of B1 result from the phosphorylation of B1 at different sites. Also, the finding that antibody binding to B1 generated a transmembrane signal may explain why antibody binding to B1 alters B cell function.
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PMID:Phosphorylation of the B1 (CD20) molecule by normal and malignant human B lymphocytes. 245 14

A panel of 14 monoclonal antibodies (McAb) against hematopoietic cell surface antigens was applied on mononuclear blood or bone marrow cells from 40 cases of acute leukemia in order to compare immunoenzymatic staining (IE) (alkaline phosphatase) of fixed cells with immunofluorescence staining (IF) of unfixed suspended cells. According to the immunological results, 25 cases were phenotyped as ALL and 15 cases as AML. Cases with blast crisis secondary to chronic myelogenous leukemia (CML-BC) were not represented in this series. In all ALL cases the two methods gave an identical antigenic distribution. In 20 our of 21 cases of non-T-cell ALL, a B-cell progenitor origin was demonstrated by a positive staining reaction with the anti-CD19 McAb AB1 or HD37, and in 10 cases additionally with the anti-CD20 McAb B1 or 1F5. In contrast to the results obtained with IF, IE revealed a poor preservation of the AB1 epitope on CD19, whereas the HD37 epitope was equally well demonstrated by both methods. In 15 cases of AML the distribution of positive versus negative cells with IE or IF was identical for all McAb except J5 (anti-CALLA) (CD10) and B1 (CD20). Thus, 10/15 AML cases expressed CALLA with IE compared to 2/15 with IF. The corresponding figures for B1 were 5/15 and 0/15, respectively. Accordingly, normal myeloid precursor cells were CALLA-positive with IE but negative with IF. The discrepancy probably reflects the fact that, whereas both intracytoplasmatic and membrane-bound antigens are exposed in IE, only the latter are in IF. If the alteration of antigenic accessibility after fixation is considered, IE can safely be used for immunophenotyping of acute leukemia.
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PMID:Immunological typing of acute leukemias: immunoenzymatic staining of fixed cells compared with immunofluorescence staining of unfixed cells in suspension. 245 10

To clarify the histogenesis of B cell chronic lymphocytic leukemia (BCLL), clinicopathological and immunophenotypic studies were performed using a large panel of monoclonal antibodies on 12 cases with BCLL including three cases with prolymphocytic/chronic lymphocytic leukemia (CLL/PL). Immunophenotypically, CD19 and CD20 were positive for all cases of this series and CD5, CD21, CD22, CD23, CD25, CD38, Leu-8, KB-61, and bcl-2 protein were expressed in variable proportion from case to case. CD10, however, did not react. No alkaline phosphatase (ALP) positive cases were found. The phenotype of BCLL was similar to that of B cells of the mantle zone (MZ) of secondary follicle in the lymph node. It is therefore postulated that the neoplastic cells of BCLL in these cases might be derived from B cells of the MZ. Moreover, the cells possibly originated from the lymphocytes located in the inner layer of the MZ, since ALP+ B cells are usually observed in the outer layer of the MZ. The pseudofollicular (PF) pattern was observed in four biopsied lymph nodes among five cases tested, but no such a pattern in an aspiration clot of bone marrow. These four cases consisted of three cases with CLL and a case with CLL/PL. The immunohistochemical study showed that there were many proliferating cells showing Ki-67+ in the PF area of the lymph nodes. In these cases, leukemic cells might have developed from the PF area of the lymph node.
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PMID:Clinicopathological and immunophenotypic studies on 12 cases with B cell chronic lymphocytic leukemia. 783 79

Using the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique, plasma cells from multiple myeloma (MM, 23 cases), plasma cell leukemia (PCL, 2 cases) and reactive plasmacytosis (RP, 13 cases) were immunophenotyped with a panel of monoclonal antibodies (McAb). The results showed that McAbCD38 was strongly positive in high percentage of MM and RP cases and the CD9 was the next. 9/23 MM expressed CD10. Our results might indirectly support that CD10 is a malignant marker of MM with poor prognosis, a concept proposed by Durie. The results were (1) all RP but 1 acute monocytic leukemia related to RP were CD10 negative. (2) In our series 2 cases of plasma cell leukemia (PCL) expressed CD10; (3) 4 MM cases survived more than 2 years were CD10 negative. A few MM cases also expressed other surface markers of pre-B and B lymphocyte, such as CD19, CD20, CD22, HLA-DR, cytoplasmic mu chain. CD20 was positive in 4/21 MM and negative in all RP cases. 7/22 MM expressed HLA-DR, and 1/13 RP did so, among them there was a significant difference. HLA-DR seems to be another malignant marker of plasma cells. 1 MM expressed CD8, and 1 PCL highly expressed CD4 indicating PCL might be heterogeneous. Lymphoid stem cells may be involved in MM and PLC. We conclude that multiple myeloma cells have different immunophenotypes and CD10, CD20 and HLA-DR may help to differentiate MM from RP.
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PMID:[Preliminary study of immunophenotype of multiple myeloma cells]. 817 66

We report a novel strategy, called end-product (EP) amplification, capable of enhancing the sensitivity of immunohistochemical procedures by about an order of magnitude or more. The strategy employs an antibody (anti-EP) to the product generated by the action of horseradish peroxidase on 3,3'-diaminobenzidine (DAB), and can be extended to the products of other enzymes as well, e.g., alkaline phosphatase. Amplification is the consequence of the ability of anti-EP to detect the multiplicity of product moelcules resulting from the turnover of substrate by a single enzyme molecule. The subsequent detection of anti-EP was by biotinylated goat anti-rabbit antibody, followed by avidin-peroxidase and DAB or by avidin-alkaline phosphatase and Vector Red. Further amplification can be accomplished by repeated cycles of the protocol. Anti-EP was produced by immunization with a bovine serum albumin (BSA) conjugate of a soluble polymer of DAB, prepared by a carefully controlled reaction of DAB with horseradish peroxidase and hydrogen peroxide. Coupling to BSA (and to RSA) was accomplished with glutaraldehyde. The titer of anti-EP was established by ELISA. Formalin-fixed, paraffin-embedded sections of five cases of Hodgkin's disease and five tonsils with follicular hyperplasia were immunolabeled for the following lymphoid markers: CD3, CD20, CD30, CD45RA, and CD68. EP amplification with anti-EP was also applied to cases of CMV pneumonia and cerebral toxoplasmosis to determine whether this procedure could improve detection of the infectious agents. Immunolabeling of the primary antibody was performed by the avidin-biotin-peroxidase technique with DAB as the reaction substrate. The specificity of EP amplification was tested by demonstrating binding of anti-EP with Vector Red with the generation of a fluorescence end-point. There was complete congruence in the distribution of the DAB signal and the red immunofluorescence representing EP amplification. The intensity of the DAB signal was increased as much as 16-fold by EP amplification, making possible a reduction in the amount of the primary antibody by as much as 85-90%. Sensitivity also increased with respect to weakly expressed antigens and low concentrations of infectious agents.
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PMID:A strategy for immunohistochemical signal enhancement by end-product amplification. 875 54

Endometrium was collected from 20 high-dose progestogen-treated patients and examined for leukocyte populations by immunohistochemistry and phloxine-tartrazine staining. A labelled streptavidin-biotin-alkaline phosphatase technique was used with antibodies against leukocyte common antigen (LCA), T cells (CD3), neutrophils (NP57), macrophages] (CD68, KP1) and B cells (CD20). The numbers of LCA (1070 +/- 117/mm2), CD3 (459 +/- 60/mm2), CD68 (129 +/- 21/mm2) positive cells and endometrial granulated lymphocytes (EGL) (236 +/- 41/mm2) were significantly higher than those in the control group (P < 0.001). Of these, EGL increased most (6.7 times). NP57 positive (NP57+) neutrophils were present in five out of 20 progestogen-treated samples and NP57 negative (NP57-) neutrophils in another six out of 20; while a neutrophil was only identified in one control tissue (P = 0.002). Three progestogen-exposed endometrial samples had either focal or extensive necrosis, and many NP57+ and NP57- neutrophils were present in the necrotic areas. EGL, neutrophils and macrophages are known to release a number of cytolytic and cell toxic molecules which may play a role in the initiation or acceleration of progestational endometrial necrosis.
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PMID:Effects of high dose progestogens on white cells and necrosis in human endometrium. 892 Nov 21


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