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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)

The use of unlabeled antibody bridging technique with alkaline phosphatase monoclonal anti-alkaline phosphatase (APAAP) complexes makes it possible to solve the problem of short durability of immunofluorescent staining and the problem of nonspecific endogenous enzyme interference of blood cells with immunoperoxidase method. The technique of APAAP allows satisfactorily to demonstrate the cytoplasmic and surface membrane antigens of T-cells both in peripheral blood and bronchoalveolar lavage fluid (BALF). With the technique studied, the subsets of T-lymphocytes simultaneously in both peripheral blood and BALF of 26 patients with interstitial lung disease and of 16 apparently healthy subjects. The results showed: (1) In patients with interstitial pulmonary fibrosis (IPF) CD8 cells in BALF were higher in number than those in peripheral blood and BALF of normal subjects (P < 0.01). It is suggested that abnormalities of T-Lymphocytes might also play a role in the pathogenesis of IPF. (2) CD4 cells in BALF of patients with sarcoidosis were significantly higher in number than those in other groups (P < 0.01). However, CD8 cells in BALF of patients with sarcoidosis were lower in number than those in others (P < 0.01). The higher ratio of CD4/CD8 was found in sarcoidosis patients during active stage. The findings suggested that change of the ratio of CD4/CD8, as an immunoregulatory abnormalities in lung, could be regarded as one of parameters in assessing the activity in patients with sarcoidosis.
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PMID:[Immunoenzymatic labeling of monoclonal antibodies for surface antigens of T-cells using immune complexes of APAAP in patients with interstitial lung disease]. 130 80

Evaluation of blood cells to determine immunologic status is becoming an important clinical application of flow cytometric analysis. For a wider use of immunophenotyping technology in clinical laboratories, the authors developed a rapid method to detect monoclonal antibody-labeled cells using forward light scatter/absorption clinical flow cytometers such as the Technicon H*1 and Technicon H*2 differential complete blood count analyzers. Calf-intestinal alkaline phosphatase was conjugated to mouse monoclonal antibodies (anti-CD2, CD3, CD4, CD8, CD19) for direct immunoenzymatic labeling. The combination of 5-bromo-4-chloro-3-indolyl phosphate and nitroblue-tetrazolium salt in diethanolamine buffer at pH 9.6 was selected as buffer/substrate to yield stable, insoluble, and very intense purplish-blue precipitates on the surface of the cells labeled with monoclonal antibody-alkaline phosphatase conjugates. Endogenous alkaline phosphatase in granulocytes was inhibited with levamisole. Early mild fixation of the white cells permitted incubation at 38 +/- 1 degrees C, which accelerated each step of the reaction without disrupting the cells throughout the procedure. The method is competitive with the direct immunofluorescence whole-blood method used on fluorescence flow cytometers in speed, sensitivity, and accuracy, as demonstrated with alkaline phosphatase-conjugated anti-CD2, CD3, CD4, CD8, CD19 monoclonal antibodies.
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PMID:Subtyping lymphocytes in peripheral blood by direct immunoalkaline phosphatase labeling and light scatter/absorption flow cytometric analysis. 137

We have studied the phenotype and activation status of leukocytes in the bronchial mucosa in patients with isocyanate-induced asthma. Fiberoptic bronchial biopsy specimens were obtained from nine subjects with occupational (five toluene- and four methylene diisocyanate-sensitive) asthma, 10 subjects with extrinsic asthma, and 12 nonatopic healthy control subjects. Bronchial biopsy specimens were examined by immunohistology with a panel of monoclonal antibodies and the alkaline phosphatase-antialkaline phosphatase method. There was a significant increase in the number of CD25+ cells (interleukin-2 receptor-bearing cells, presumed "activated" T-lymphocytes; p less than 0.01) in isocyanate-induced asthma compared with that of control subjects. There were also significant increases in major basic protein (BMK-13)-positive (p less than 0.02) and EG2-positive (p less than 0.01) cells that represent total and "activated" eosinophil cationic protein-secreting eosinophils, respectively. In agreement with our previous findings, CD25+ (p less than 0.01), BMK-13 (p less than 0.03), and EG2+ (p less than 0.01) cells were also elevated in extrinsic asthma. No significant differences were observed in the numbers of T-lymphocyte phenotypic markers (CD3, CD4, and CD8) between subjects with asthma (isocyanate-induced and extrinsic) and control subjects. Similarly, no significant differences in immunostaining for neutrophil elastase (neutrophils) or CD68 (macrophages) were observed. The results suggest that isocyanate-induced occupational asthma and atopic (extrinsic) asthma have a similar pattern of inflammatory cell infiltrate. The results support the view that T-lymphocyte activation and eosinophil recruitment may be important in asthma of diverse etiology.
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PMID:Activated T-lymphocytes and eosinophils in the bronchial mucosa in isocyanate-induced asthma. 153 7

Immuno-alkaline phosphatase (AP) staining for T cell subsets (CD4 and CD8) of smears from fingerprick blood functioned well under tropical climatic conditions when smears were stored frozen with silica gel before being labelled. Unlabelled smears were stored for up to 12 months and could be transferred abroad without antigenic damage. Identical total CD4 and CD8 counts were obtained on venous and capillary blood, when compared using a FACS analyser. Although the AP method gave somewhat higher total CD4 and CD8 counts, the ratio remained the same. The major advantages of the method are: (i) no expensive equipment is required, (ii) only minute amounts of blood are needed, and (iii) slides can be stored for long periods before labelling and can be preserved for later reading. The method is suitable for community studies where there is a need for assessing the immune status of the population.
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PMID:Labelling of T cell subsets under field conditions in tropical countries. Adaptation of the immuno-alkaline phosphatase staining method for blood smears. 169 77

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

In a community study in Guinea-Bissau, West Africa, 47 HIV-2-seropositive cases and 87 matched controls were evaluated immunologically using immuno-alkaline phosphatase linked to avidin-biotin complex for the assessment of CD4 and CD8 status. HIV-2-seropositive individuals had significantly lower total numbers of CD4 cells and CD4/CD8 ratios, 38% having a total number of CD4 cells less than or equal to 0.5 x 10(9)/l and 36% having a CD4/CD8 ratio less than or equal to 0.8. Total numbers of CD4 cells less than or equal to 0.5 x 10(9)/l or CD4/CD8 ratio less than or equal to 0.8 were found in 53% of the HIV-2 seropositives compared with 11% among controls [odds ratio (OR) = 7.3; 95% confidence interval (CI): 3.1-17.1]. Lymphadenopathy was significantly more frequent among HIV-2 seropositives than among controls (OR = 3.4; 95% Cl: 1.5-7.6). HIV-2 seropositives with lymphadenopathy had significantly fewer lymphocytes (P = 0.008) and lower total CD4 (P = 0.029) and total CD8 number (P = 0.011) than HIV-2 seropositives without lymphadenopathy. This study indicates that HIV-2 has a significant immunosuppressive effect.
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PMID:Immunodeficiency in HIV-2 infection: a community study from Guinea-Bissau. 198 11

An enzyme-linked immunosorbent assay (ELISA) that can measure picogram quantities of human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein 120 (gp120) in cell culture medium or body fluids has been developed. Recombinant, soluble CD4 immobilized in microtiter trays was used to capture gp120, which was then detected with polyclonal sheep antibody to gp120 followed by biotinylated rabbit anti-sheep immunoglobulin G and an avidin-alkaline phosphatase indicator system. With a reference recombinant gp120, the assay showed a linear relationship between optical density and concentrations ranging from 60 to 6,000 pg/100-microliters well; precision of the assay varied with the concentrations and ranged from +/- 40% with amounts smaller than 200 pg to +/- 10% with amounts larger than 200 pg. In a group of coded samples containing 60 pg (approximately 10(7) molecules) of reference gp120, the assay correctly identified the samples as containing gp120 99% of the time, with no false-positive results recorded for blank samples. Recombinant gp120 prepared in another cell culture system demonstrated a binding coefficient 13-fold lower than that of reference gp120. Mixing standard amounts of reference gp120 with increasing concentrations of human sera reduced assay sensitivity, although the linear relationship between gp120 concentration and optical density remained. With this assay we were able to detect gp120 in HIV-1 suspensions prepared from cultured lymphoblastoid cells and in the sera of HIV-1-infected patients. This ELISA for gp120 should be useful for studying the biological role of gp120 in HIV infection.
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PMID:Enzyme-linked immunoassay for human immunodeficiency virus type 1 envelope glycoprotein 120. 199 48

To determine the potential role of the placenta in transmission of human immunodeficiency virus (HIV) from mother to fetus, the ability of human placental tissue to support HIV type 1 (HIV-1) infection was examined. HIV-1-seronegative first-trimester placentas were maintained in culture and infected with HIV-1. Virus production, measured by HIV-1 antigen release into the supernatant, and HIV-1 DNA, identified by polymerase chain reaction, were detected for at least 12 days postinfection. Western immunoblot analysis showed Gag proteins, precursor p55, and cleavage products p24 and p17 in HIV-1-infected tissues. Double labeling of placental villi with antibodies to CD4 and placental trophoblast-specific alkaline phosphatase indicated that trophoblasts express CD4 antigen. Additionally, immunostaining of HIV-1-infected tissues with anti-p24 antibodies demonstrated HIV-1 protein expression in placental trophoblasts. Evaluation of human chorionic gonadotropin and progesterone production by the placental cultures indicated that there was a 90% decrease in human chorionic gonadotropin and a 70% decrease in progesterone production in HIV-1-infected cultures in comparison with controls. These data demonstrate that trophoblastic cells of human placenta tissue express CD4 and are susceptible to HIV-1 infection; also, placental endocrine function is decreased by HIV-1 infection. Thus, the placenta may serve as a reservoir of HIV-1 infection during pregnancy contributing to infection of the fetus, and decreased placental hormone production may result in impaired fetal development.
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PMID:Human immunodeficiency virus type 1 infection of human placenta: potential route for fetal infection. 201 57

The phenotypes of lymphocytes infiltrating the epithelium of the jejunal and ileal Peyer's patches in foetal sheep at about 130 days gestation and 2-month-old lambs were examined using indirect immunoperoxidase histochemistry, a panel of monoclonal antibodies and enzyme histochemistry. Computer-assisted morphometric analysis enabled the relative size of reactive areas within epithelia to be estimated. The comparison of the intraepithelial lymphocyte populations associated with structurally developed Peyer's patches of foetal sheep and those of lambs allowed assessment of the impact of extrinsic factors from which the sheep foetus is shielded. The study confirmed the postnatal expansion in the villous intraepithelial lymphocyte population and showed that this expansion involved the CD8 and gamma delta phenotypes. CD4 lymphocytes did not appear in the follicle-associated epithelium until after birth. Unlike the villous epithelium, the follicle-associated epithelium had a high frequency of IgM+ and MHC II+ cells, which was dramatically reduced after birth. This postnatal reduction was particularly prominant in the follicle-associated epithelium of the jejunal Peyer's patch, where the frequency of IgM+ cells fell from 12.4% in foetal sheep to 0.7% in lambs. Double staining for alkaline phosphatase in the jejunal Peyer's patch suggested that clusters of IgM+ cells were associated with M cells.
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PMID:Computer-assisted morphometric analysis of absorptive and follicle-associated epithelia of Peyer's patches in sheep foetuses and lambs indicates the presence of distinct T- and B-cell components. 202 45

Lymph node aspirates from 18 peripheral T-cell lymphomas (PTLs) were analyzed. Cytologic and immunocytologic studies were performed on Cytospin preparations using the alkaline phosphatase-antialkaline phosphatase method with a panel of monoclonal antibodies (CD3, CD4, CD8, CD19 and CD30). The cytologic diagnosis was confirmed by histologic investigation. Nine lymph node aspirates from patients with Lennert's lymphoma, angioimmunoblastic (AILD)-type PTL and pleomorphic small-cell-type PTL were composed predominantly of small-to-intermediate-sized lymphocytes. An admixture of plasma cells, eosinophils, neutrophils, lymphocytes with an irregular nucleus, granula in the cytoplasm or abundant cytoplasm was also seen. Nine lymph node aspirates from patients with T-immunoblastic lymphoma, pleomorphic large-cell-type PTL and large-cell anaplastic (Ki-1+) lymphoma showed marked cytologic heterogeneity. Immunocytologic investigation of the aspirates using the antibodies CD3, CD4, CD8, CD19 and CD30 was helpful for the differentiation of PTLs from reactive lymphadenopathy and other malignant lymphomas. A strong predominance of CD3+ cells was found in only seven cases. The aspirates expressed a helper/inducer phenotype in 11 cases and a suppressor/cytotoxic phenotype in 4 cases. A T-cell phenotype not corresponding to the normal T-cell phenotype was found in nine cases. In 15 of the 18 cases, the number of CD19+ cells was found to be less than 15%. The large cells of the large-cell anaplastic (Ki-1+) lymphoma expressed the antigens CD30 and CD45 and were negative for CD15. These findings indicate that immunocytologic studies can be used in improving the cytologic diagnosis of PTLs.
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PMID:Cytologic and immunocytologic studies of peripheral T-cell lymphomas. 204 31


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