Use of commercially available monoclonal antibodies for immunoenzyme double staining

Loos, C. M. Vand Der; Das, Pranab K.; Houthoff, H. J.

doi:10.1007/bf01002426

Cited by 24 publications

(12 citation statements)

References 14 publications

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“…The basic concept for the immunoenzyme double staining techniques has been described previously [25,26]. Its detailed performance was dependent on the combination of the primary antibodies.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Costimulatory molecules in human atherosclerotic plaques: an indication of antigen specific T lymphocyte activation

et al. 1997

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Atherosclerotic plaques contain inflammation, composed largely of macrophages and lymphocytes. A proportion of lymphocytes shows signs of activation, but the question arises whether they are activated in an antigen specific way. The expression of costimulatory molecules-receptors that provide accessory signals during antigen-specific activation is a prerequisite for such a condition. This aspect of inflammation in atherosclerotic lesions has not been investigated. Human arterial segments with diffuse intimal thickening, fatty streaks and atherosclerotic plaques were studied with immuno-single and double staining methods. Macrophages and T lymphocytes were stained with CD68 and CD3, respectively, and pan-B cell markers CD19 and CD22 were also used. Costimulatory molecules B7-1 and B7-2, together with their common ligand CD28, and CD27 with its ligand CD70, were stained with specific monoclonal antibodies. The results show that most T lymphocytes were CD27 positive and that only a subpopulation of these (5-15%) was positive also for B7-1, CD28 and CD70. Macrophages expressed B7-1, B7-2, CD28 and CD70, while macrophages positive for CD28 and CD70 have not been reported yet. The expression of costimulatory molecules was most pronounced in the superficial layers at the fibrous cap, but decreased towards the lipid core. This study shows, therefore, that atherosclerotic plaques provide costimulatory signals generally accepted as a prerequisite for adequate T cell stimulation. In addition, this study reveals that only approximately 5 -15% of the lymphocytes appears actively involved in the inflammatory reaction. © 1997 Elsevier Science Ireland Ltd.

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Section: Immunohistochemistrymentioning

confidence: 99%

Costimulatory molecules in human atherosclerotic plaques: an indication of antigen specific T lymphocyte activation

et al. 1997

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“…Alkalinephosphalase activity was detected with Napthol AS-MX, fast red violet sail and levamisole added sequentially to veronal acetate buffer (Boorsma, 1984). Horseradish peroxidase activity was detected according to van der Loos et al, (1988) using disodium sulphur succinatc. trimethyl benzidine, citrate-phosphate buffer and pcrhydrol.…”

Section: Double Stainingmentioning

confidence: 99%

Macrophage heterogeneity in human fetal tissue. Fetal macrophages

Oliver

1990

Clinical and Experimental Immunology

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SUMMARYThe immunophenotype of the macrophage population in human fetal tissue was studied, using a panel of monoclonal antibodies against cells of the macrophage/monocyte lineage. Using a doublelabelling technique two main populations were observed in tissue from 14 weeks of estimated gestational age (EGA); EBMl 1 ' DR* and EBMl 1 ' DR cells of which a small proportion were also RFD7'. Most macrophages were negative with 3.9, an antibody specilic for the adhesion molecule P150.95 and LP9 which is specific for a lysosomal enzyme. The exception to this was a small population of positive cells in the thymus. Small numbers of 3.9* cells were also infrequently observed In tissue at and above 17 weeks of EGA. while occasional RFD9' cells were only observed in most tissues, before this time. The higher percentage of macrophages were DR' DQ DP , with a few DQ* eells appearing at 15 weeks of EGA. In the thymus. DQ' cells outnumbered DP* cells especially in the medulla. These results indicate the heterogeneous and Immature nature ofthe fetal macrophage population and point to the importance of age. tissue-specific factors and probable immune mediators in macrophage differentiation.

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“…naphthol-AS-MX phosphate in combination with Fast Blue or Fast Red) showed a lower sensitivity than the NBT/ BCIP detection method (De Jong et al, 1985). (2) Detecting HRP activity as a red precipitate with aminoethylcarbazole or a green precipitate with tetramethylbenzidine shows a sensitivity comparable with the use of DAB, but the red precipitate dissolves in ethanol and the green reaction product is not heat resistant ( Van der Loos et al, 1988), both conditions essential to the in situ DNA-hybridization protocol.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous application ofin situ DNA hybridization and immunohistochemistry on one tissue section

et al. 1989

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Combined application of a non-radioactive in situ DNA hybridization procedure and the immunoperoxidase technique on one tissue section is described. Of six potential protocols, only one proved to be successful. First, the immunohistochemical procedure including visualization of enzyme activity is performed; the in situ DNA hybridization protocol is then applied. Using this protocol, several antigens, detected with monoclonal antibodies, and target DNAs, detected by using biotinylated human cytomegalovirus or human papilloma virus type 16 DNA probes, could be distinguished by their peroxidase activity (brown precipitate) and alkaline phosphatase activity (purple-blue precipitate) respectively. The method allows immunophenotyping of virus-infected cells as well as simultaneous visualization of two viral parameters. This technique has important implications for research and diagnostic purposes.

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Use of commercially available monoclonal antibodies for immunoenzyme double staining

Cited by 24 publications

References 14 publications

Costimulatory molecules in human atherosclerotic plaques: an indication of antigen specific T lymphocyte activation

Costimulatory molecules in human atherosclerotic plaques: an indication of antigen specific T lymphocyte activation

Macrophage heterogeneity in human fetal tissue. Fetal macrophages

Simultaneous application ofin situ DNA hybridization and immunohistochemistry on one tissue section

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