Abstract
Tissue sections of frozen biopsy specimens obtained from normal and hyperplastic human lymphoid tissues, 33 cases of non-Hodgkin lymphomas as well as various forms of immunoregulatory disorders (angioimmunoblastic and dermatopathic lymphadenopathy) were analysed in immunofluorescence tests (using red TRITC and green FITC double-labelling). A panel of antisera including well-characterized conventional reagents to immunoglobulin classes, T lymphoid and Ia-like antigens, and monoclonal antibodies was used. In selected cases the results were compared with the observations of membrane-marker staining on viable cells in suspension. the findings show that the immunological methods can give a very accurate analysis of the normal and malignant lymphoid cells, and can provide complementary information to conventional histology. The investigator can choose the reagent combinations which give answers to various specific questions: e.g. antisera to light chains establish the monoclonality of lymphomas, whilst staining combinations for human T and Ia-like antigens are particularly useful in various immunoregulatory disorders. Monoclonal antibodies will be particularly useful in various immunoregulatory disorders. Monoclonal antibodies will be particularly useful reagents for analysing the tissue distribution of lymphoid subpopulations and ancillary cells in tissue biopsy specimens.
Highlights
Summary.-Tissue sections of frozen biopsy specimens obtained from normal and hyperplastic human lymphoid tissues, 33 cases of non-Hodgkin lymphomas as well as various forms of immunoregulatory disorders were analysed in immunofluorescence tests
Interchangeable filter sets used on the modern microscopes facilitate the simultaneous application of antibodies labelled with fluorescein-isothiocyanate (FITC-green) and tetraethylrhodamine-isothiocyanate (TRITC-red)
Previous investigators have analysed the "monoclonality" of Jg (K or A light chain) expression of non-Hodgkin lymphomas (NHL) in tissue sections with antisera to K or A labelled with different fluorochromes (Levy et al, 1977; Warnke & Levy, 1978)
Summary
Surgical biopsy specimens were divided; one part was fixed in 10% formol saline and processed for paraffin embedding, the second part was embedded in Ames OCT Compound (Miles Lab), frozen in liquid N2 within 3 h of excision and stored at - 70°C until sectioning. Goat anti-Ia-like serum reacts with cells of B antiserum to human IgM (,u-specific) coupled lymphoproliferative diseases, non-T non-B to TRITC or to FITC The reagent reacts with nuclear TdT in antisera to K and A light chain were coupled common ALL and Thy-ALL, in normal cor- to TRITC and FITC respectively At this high concentration the antibody saturates the binding sites but gives no non-specific staining Both markers were detected by goat anti- E rosettes (a T-lymphocyte and thymocyte mouse IgG coupled to FITC or TRITC marker).-These were formed by mixing 106. Cytoplasmic Ig staining.-This was performed on cytocentrifuge preparations fixed in ethanol These smears were stained with appropriate dilution of FITC-coupled antisera against K, A, ,, y, cx and 8 chains, or with the reagent combinations described above as 4, 5 and 6. Both indirect IF and PAP labelling were successful (see below)
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