ABSTRACT
Lymphocytes carrying the receptor for helix pomatia lectin on their cytomembranes can be detected in lymphatic tissue and cutaneous T-cell lymphoma by means of peroxidase-labelled helix pomatia lectin. Cells positive for the HPL-receptor are identical with the population of T-lymphocytes, including their pathological variants, i.e. lymphoma cells and Sezary's cells.
Subject(s)
Lectins/analysis , Lymphoma/diagnosis , Receptors, Mitogen/analysis , Skin Neoplasms/diagnosis , T-Lymphocytes/analysis , Humans , Immunoenzyme Techniques , Mycosis Fungoides/diagnosisABSTRACT
Immunoperoxidase techniques, applied to formalin-fixed paraffin embedded tissues, provide a simultaneous histopathological and immunological diagnosis. Application of protein-A-peroxidase constitutes a new technical variant: The technique is relatively simple, decreases non-specific background staining, and binds mono- and polyclonal antibodies from different mammalian species.
Subject(s)
Immunoenzyme Techniques , Lymphoma/diagnosis , Skin Neoplasms/diagnosis , B-Lymphocytes , Humans , Isoenzymes , Lymphoma, Non-Hodgkin/diagnosis , Peroxidase , Peroxidases , Staphylococcal Protein ASubject(s)
Lymphocytes/ultrastructure , Lymphoma/pathology , Helix, Snails , Humans , Lectins , Lymph Nodes/pathology , Microscopy, ElectronABSTRACT
A receptor for Helix pomatia lectin (HPL) occurs on 71.5% of peripheral blood lymphocytes. With double incubation techniques we could show that the population of HPL-positive lymphocytes is by far (greater than 90%) identical with the population of T lymphocytes, as determined by "classical" markers (E rosetting) and newly developed T cell markers like OKT3 or anti-human Lyt3 monoclonal antibodies. In formaldehyde-fixed lymph node sections we could demonstrate the homing area of HPL-positive lymphocytes, the paracortical regions. Conventional histological counterstaining was readily possible. We have employed HPL-peroxidase for differentiation of lymphocytes in benign and malignant infiltrates in human skin. In reactive cutaneous infiltrates, as well as in cutaneous lymphomas, HPL-peroxidase staining allows a clear differentiation of the involved lymphatic cells, and may serve as a tumor marker in certain T lymphomas, where we could demonstrate the lymphoma cells to carry the HPL-receptor.
Subject(s)
Lectins , Lymph Nodes/analysis , Lymphocytes/analysis , Lymphoma/analysis , Receptors, Mitogen , Skin Neoplasms/analysis , Cell Membrane , Helix, Snails , Humans , Lymph Nodes/ultrastructure , Lymphocytes/ultrastructure , Lymphoma/ultrastructure , Skin Neoplasms/ultrastructureSubject(s)
Antigen-Antibody Complex/analysis , Immunoenzyme Techniques , Pemphigoid, Bullous/immunology , Skin Diseases, Vesiculobullous/immunology , Aged , Complement System Proteins/analysis , Female , Fluorescent Antibody Technique , Humans , Immunity, Cellular , Immunoglobulin A/analysis , Immunoglobulin G/analysis , Immunoglobulin M/analysis , Peroxidases , Staphylococcal Protein AABSTRACT
Pemphigus autoantibodies, bound in the epidermis during different stages of acantholysis, were demonstrated with a new technique for immuno-electron microscopy. Peroxidase-labeled Protein A was used as a new and specific tracer for tissue-bound antibodies of the IgG-type. Advantages were: (1) A small molecular weight of the tracer, (2) a rapid tissue penetration, and (3) shortened incubation times, thus better preserved tissue fine structures. Unspecific adsorption in tissues and on cells was found to be comparatively low.
Subject(s)
Autoantibodies/analysis , Epidermis/immunology , Pemphigus/immunology , Staphylococcal Protein A , Acantholysis , Antigen-Antibody Reactions , Epidermis/ultrastructure , Female , Humans , Microscopy, Electron , Middle Aged , Pemphigus/pathology , PeroxidasesABSTRACT
Peroxidase-labeled Protein A, stable immunoenzyme tracer of high reactivity and comparatively low molecular weight, has been applied in immuno-electron microscopy for detection of bound IgG-type pemphigus and bullous pemphigoid antibodies. Comparing Protein A-peroxidase with peroxidase-labeled immunoglobulins, we obtained similar morphological results in corresponding incubation techniques, but lower nonspecific adsorption of Protein A-peroxidase complexes in tissues. The Protein A-peroxidase molecules showed good tissue penetration abilities. Our rapid one-step incubation procedure led to enhanced preservation of tissue fine structures, without the need of prior tissue fixation. It seems that Protein A-peroxidase is able to replace peroxidase-labeled anti-IgG for immuno-electron microscopical purposes.
Subject(s)
Pemphigoid, Bullous/immunology , Pemphigus/immunology , Skin Diseases, Vesiculobullous/immunology , Staphylococcal Protein A/immunology , Humans , Immunoenzyme Techniques , Immunoglobulin G/analysis , Pemphigoid, Bullous/pathology , Pemphigus/pathology , Skin/immunology , Skin/ultrastructureABSTRACT
Protein A from Staphylococcus aureus, characterized by high affinity binding properties for IgG-type antibodies, was labeled with peroxidase to form a stable immuno-histological tracer molecule of comparatively low molecular weight. It has been used for demonstration of antibodies against tissue antigens, in direct and indirect techniques, and the findings were consistent with those in routine immunofluorescence and in staining with peroxidase-coupled anti-IgG. In comparison, the lowest unspecific tissue adsorption and staining was obtained with Protein A-peroxidase in buffer containing glucose and mannose. The immunohistological preparations were mounted and stored for documentation without apparent loss of staining.
Subject(s)
Immunoenzyme Techniques , Immunoglobulin G/analysis , Staphylococcal Protein A/immunology , Animals , Chromatography , Fluorescent Antibody Technique , Humans , Lupus Erythematosus, Systemic/immunology , Mice , Pemphigus/immunology , Protein Binding , Skin/immunology , Skin Diseases, Vesiculobullous/immunology , Staining and Labeling , Staphylococcus aureusABSTRACT
Protein A from Staphylococcus aureus, a cell wall protein with high affinity binding properties for IgG-type antibodies, has been labeled with peroxidase to form a stable immunohistological tracer molecule of relatively low molecular weight. It has been used for demonstration and titration of antinuclear antibodies in SLE sera on mouse liver sections in an indirect technique. The findings were consistent with those obtained by immunofluorescence and by staining with peroxidase-coupled anti-IgG. In contrast to immunofluorescence, the stained sections could be mounted and stored for documentation. In comparison, unspecific tissue adsorption and staining could be minimized by addition of glucose, galactose, and mannose as well as bovine serum albumine to the buffer containing Protein A-peroxidase.