ABSTRACT
T-cell receptor (TCR) gamma gene rearrangements which have been amplified in polymerase chain reactions (PCRs) and analysed by high resolution polyacrylamide gel electrophoresis have been used to investigate the clonal diversity of T-cells in joint effusions from 16 patients with rheumatoid arthritis, one with systemic lupus erythematosus and one with psoriatic arthropathy. Polyclonality was found in every case but an oligoclonal subset of dominant rearrangements was also demonstrated in all but the patient with psoriasis. Marked changes in the relative preponderance of the various clonotypes were observed in 29 of 48 paired tests from 12 cases before and after culture in media containing interleukin-2 (IL-2) showing that SF mononuclear cells cultured in vitro with IL-2 are not representative of those present in vivo.
Subject(s)
Arthritis, Psoriatic/immunology , Arthritis, Rheumatoid/immunology , Interleukin-2/pharmacology , Lupus Erythematosus, Systemic/immunology , Synovial Fluid/immunology , T-Lymphocytes/drug effects , Adult , Aged , Arthritis, Psoriatic/genetics , Arthritis, Rheumatoid/genetics , Base Sequence , Cells, Cultured , Clone Cells , Female , Humans , Lupus Erythematosus, Systemic/genetics , Male , Middle Aged , Molecular Sequence Data , Synovial Fluid/cytology , T-Lymphocytes/immunology , Time FactorsSubject(s)
Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor/genetics , Mouth Mucosa/immunology , Receptors, Lymphocyte Homing/genetics , Skin/immunology , T-Lymphocyte Subsets/immunology , Adult , Antibodies, Monoclonal , CD3 Complex/genetics , CD4 Antigens/genetics , CD8 Antigens/genetics , Cell Adhesion Molecules/genetics , Clone Cells , E-Selectin , Electrophoresis, Polyacrylamide Gel , Epitopes , Humans , Intercellular Adhesion Molecule-1 , Polymerase Chain ReactionABSTRACT
In a study of 49 biopsies from the margins of depigmented cutaneous lesions in 18 patients with vitiligo, highly significant overall increases were found in CD3+, CD4+, and CD8+ T cells, though cell numbers in individual cases were often within the normal range. Many of the T cells were activated (MHC class II+, interferon gamma+), of CD45RO (UCHL1+) memory subset, and many expressed the cutaneous lymphocyte-associated antigen (HECA-452+) typical of skin-homing T cells. Immunohistologically, the most intense epidermal T-cell infiltration was present within 0.6 mm of the edge of the lesion in 10 of 13 double-stained sections with a clearly defined zone of melanocyte depletion. In 40 lesions from 17 patients seen 11-64 weeks after biopsy, no apparent association was found between T-cell numbers and disease activity as assessed by Köbnerization of biopsy wounds or spread of depigmentation. These findings are consistent with the hypothesis that lesional T cells rather than circulating antimelanocytic antibody may be responsible for the supposedly autoimmune but characteristically patchy destruction of cutaneous melanocytes in vitiligo. Nevertheless, many of the infiltrating T cells are probably innocent bystanders attracted by upregulated cell adhesion molecules near sites of melanocyte damage.
Subject(s)
Skin/immunology , T-Lymphocytes/immunology , Vitiligo/immunology , Biopsy , CD3 Complex/immunology , CD4 Antigens/immunology , CD8 Antigens/immunology , Humans , Immunohistochemistry , Lymphocyte Activation , Melanocytes/pathology , Skin/injuries , Vitiligo/pathologyABSTRACT
The relationship between damage to cutaneous melanocytes and antimelanocyte autoimmunity in vitiligo is unclear. We have demonstrated abnormal expression of MHC class II molecules by perilesional melanocytes in 13/21 patients with vitiligo and a six-fold increase in the number expressing the intercellular adhesion molecule ICAM-1. These molecules have important roles in normal antigen presentation and activation of helper T lymphocytes, and their expression by melanocytes may contribute to the abnormal immune response in vitiligo. MHC class II is not expressed by melanocytes in psoriasis and is unlikely to be induced in vitiligo by cytokines released from activated non-melanocyte-specific T lymphocytes.
Subject(s)
Autoimmune Diseases/immunology , Cell Adhesion Molecules/immunology , Histocompatibility Antigens Class II/immunology , Melanocytes/immunology , Vitiligo/immunology , Humans , Immunohistochemistry , Intercellular Adhesion Molecule-1 , Psoriasis/immunologyABSTRACT
A sensitive method of screening for dominant T cell clones in small samples of DNA has been developed using the polymerase chain reaction to amplify and identify T cell gamma receptor gene rearrangements. It can detect such rearrangements in nanogram quantities of DNA from cultured T cell clones, even in the presence of 20-100 parts of polyclonal lymph node DNA, and works with DNA extracted from paraffin sections of cloned T cells which have been fixed in formalin. Presumptive clonal reactions have been obtained in preliminary tests on 8 of 10 unfixed T cell lymphomas but in 0 of 10 reactive lymph nodes.
Subject(s)
Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor , Polymerase Chain Reaction/methods , T-Lymphocytes/cytology , Base Sequence , Cell Line , Clone Cells/cytology , DNA/analysis , Electrophoresis, Polyacrylamide Gel , Humans , Lymphoma/genetics , Molecular Sequence DataABSTRACT
Patients with malignant lymphoma may have cutaneous and subcutaneous involvement that exhibits a striking symmetry about the coronal axis. The symmetry of these lesions may be caused by site-specific migration from the circulation, preferential proliferation by lymphocytes of the neoplastic clones at defined anatomical sites, or both mechanisms. Similar behaviour by benign lymphocytes may explain the symmetry and selective anatomical distribution of lesions in other skin diseases.
Subject(s)
Ear Neoplasms/pathology , Ear, External , Lymphoma/pathology , Skin Neoplasms/pathology , Adult , B-Lymphocytes/cytology , Biopsy , Clone Cells/physiology , Evaluation Studies as Topic , Female , Forearm , Hodgkin Disease/complications , Hodgkin Disease/pathology , Humans , Leg , Male , Middle Aged , T-Lymphocytes/cytologyABSTRACT
A strategy is described for detecting large T-cell clones among small numbers of lymphoid cells in fresh or formalin-fixed tissue samples using the polymerase chain reaction (PCR) to amplify and identify rearrangements of the V and J genes of the T-cell gamma receptor. Following hybridization of primers with gene-specific sequences at judiciously selected locations on each of the eight potentially active V and five potentially active J genes, the PCR can theoretically amplify DNA segments which span the join between rearranged V and J genes and are of approximately 384 different sizes, each segment size reflecting different gamma gene clonal rearrangements. Large monoclonal populations of T lymphocytes, indicated by excessive amounts of particular PCR segment sizes, can be further characterized by direct nucleotide sequencing of the hypervariable N regions of these segments, and the presence of such clones can be confirmed directly in tissue sections by in situ hybridization with N region-specific oligonucleotide probes.
Subject(s)
Clone Cells , Gene Amplification , T-Lymphocytes/pathology , Base Sequence , DNA-Directed DNA Polymerase/metabolism , Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor , Humans , Molecular Sequence DataSubject(s)
DNA-Directed DNA Polymerase/metabolism , DNA , Gene Amplification , Animals , Base Sequence , DNA/biosynthesisSubject(s)
Immune System Diseases/etiology , Lymphocyte Activation , T-Lymphocytes , Clone Cells , HumansABSTRACT
Immunohistochemical staining of formalin fixed paraffin sections with the mouse monoclonal antibodies E29 and CAM 5.2 effectively distinguish epithelium (antigen positive) from stroma (antigen negative) in normal endometrium. These antibodies were used as epithelial markers in the study of eight malignant mixed Müllerian tumours (MMT) of endometrium and gave normal reactions with well differentiated neoplastic glands; in contrast, negative or very weak staining was observed in poorly differentiated epithelial cells, present in large numbers in three cases. Abnormal antigen-containing cells were observed in the stroma of seven MMT. In some cases these are probably due to stromal invasion by malignant epithelium but in others they may represent an early stage of epithelial differentiation in mesenchymal cells of the malignant stroma.
Subject(s)
Antigens, Neoplasm/analysis , Neoplasms, Germ Cell and Embryonal/immunology , Uterine Neoplasms/immunology , Cell Transformation, Neoplastic/pathology , Epithelium/immunology , Female , Humans , Immunoenzyme Techniques , Neoplasms, Germ Cell and Embryonal/pathology , Uterine Neoplasms/pathologySubject(s)
Pigmentation Disorders/genetics , Albinism/genetics , Albinism/pathology , Animals , Chimera , Female , Humans , Lentigo/genetics , Lentigo/pathology , Male , Melanins/genetics , Melanins/physiology , Mice , Peutz-Jeghers Syndrome/genetics , Peutz-Jeghers Syndrome/pathology , Pigmentation , Pigmentation Disorders/embryology , Pigmentation Disorders/pathology , Vitiligo/genetics , Vitiligo/pathologyABSTRACT
Patterns of depigmentation in vitiligo are illustrated which appear to form maps of obscure local anatomical subdivisions of the human skin. These are distinctive but as yet poorly defined and do not generally correspond to known anatomical structures or boundaries.