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It has been demonstrated recently in mycosis fungoides and lichen planus that T lymphocyte subsets may be identified in cutaneous lymphocytic infiltrates using the immunoperoxidase technic in conjunction with monoclonal antibodies produced by the technic of Kohler and Milstein. This communication describes the application of this technic to cutaneous lymphoid infiltrates of parapsoriasis in which T cell predominance has been demonstrated previously. The lymphoid infiltrates of six patients with atrophic parapsoriasis were examined by the indirect immunoperoxidase technic using monoclonal antibodies (from two commercial sources) directed against "helper" and "suppressor" T cell subsets. Both "helper" and "suppressor" cells (as defined by a positive reaction with monoclonal antibodies) could be identified in cutaneous infiltrates. "Helper" cells predominated, but in varying degrees among patients. The relevance of these findings in relation to the possible development of clinical mycosis fungoides from atrophic parapsoriasis is discussed. In addition, factors causing difficulty in the consistent identification of cell subtypes are discussed. These factors suggest that in the present state of imperfection, difficulty will be experienced in using this technic for the accurate quantification of percentages of lymphocyte subsets in tissue sections.U

A study was performed using monoclonal antibodies and the immunoperoxidase technic to identify T-cells and T-subsets in human tonsil sections. The in situ topographic identification of T-cells, helper cells, and suppressor cells was achieved. This model may therefore be applied to other normal lymphoid tissues. Its potential pathologic importance lies in the fact that certain disorders may be associated with depletion of lymphocyte subsets from their normal areas of "homing," and that lymphoma cells may mirror their putative normal counterparts in their selective metastatic migration pattern. The improved knowledge of normal lymphocyte subset microenvironment afforded by this technic may therefore be of value in applied pathology.

A monoclonal immunocytochemical method with 25 monoclonal antibodies was used to study the distribution in human tonsil of determinants expressed on T cells (mature and immature), Langerhans cells, B cells, killer/natural killer cells, macrophages, immature myeloid and lymphoblastic leukemia cells. Many of the respective determinants were found to have a discrete topographic distribution in normal reactive tonsil. The common acute lymphoblastic leukemia antigen and a determinant found on myeloblastic leukemia cells (My10) were not found in the specimens of the tonsil examined.

Tissues from atrophic large plaque parapsoriasis were examined using the indirect immunoperoxidase technique with a monoclonal T cell antibody as primary antibody, and affinity isolated peroxidase conjugated goat anti-mouse IgG as second stage antibody. The "T" cell nature of the dermal infiltrate and of single exocytic epidermal lymphocytes was demonstrated. The results obtained suggest that the method utilized will be useful for the demonstration of lymphoid cell subpopulations in a variety of cutaneous disorders in which a lymphocytic infiltrate forms a significant component of the histopathology observed. This technique also has the potential of providing information on the topographic localization and differentiation of lymphocytes within the various levels of the skin. Certain technical manipulations which may result in an improvement in the quality of results obtained are also discussed.

Experimental work on animal models suggests that various lymphoid subpopulations (K/NK, lymphocytes, macrophages, T-cells) may play a role in immunity against neoplasia, including lymphomas. The incidence and possible role of these cells in spontaneously occurring human tumors is less clear. The prevalence of these lymphoid subpopulations was studied in 60 cutaneous biopsy specimens using in situ immunocytochemistry. Monoclonal HNK-1 (Leu 7), which detects human granular lymphocytes with K/NK function, was used to detect K/NK lymphocytes; macrophages were identified by diffuse cytoplasmic esterase activity; T-cells were identified using monoclonal antibodies OKT3 and Leu 1. HNK-1+ lymphocytes, esterase-positive macrophages, and infiltrating T-cells were identified in varying numbers in the cases studied. The demonstration of HNK-1+ lymphocytes, macrophages, and T-lymphocytes in cutaneous lymphoma lesions is of potential therapeutic application. Whatever the naturally occurring role of these immune cells, it is possible that they may be manipulated pharmacologically for the benefit of the patient.

This report describes the in situ demonstration of "T" lymphocytes and "T" lymphocyte subsets in tissue sections using a case of lichen planus as a model. The technique utilized is the indirect immunoperoxidase method with monoclonal antibodies to "T" cells and "T" cell subsets. In the case examined a predominance of helper "T" cells was found. The potential application of this method in the immunopathology of skin and internal organs is discussed.

A monoclonal antibody, OKT 6, has been developed against an antigen which is expressed by immature T cells as part of their normal intrathymic differentiation, but not by mature peripheral T cells. It was though that a search for the expression of such an antigen might be worthwhile in prelymphomatous conditions. This communication describes the investigation of lymphocytic infiltrates of atropic parapsoriasis, lymphomatoid papulosis, and a small group of miscellaneous skin conditions with OKT 6 and the immunoperoxidase technic. OKT 6-positive cells were identified in the dermis in varying numbers in four cases of atrophic parapsoriasis and in one case of lymphomatoid papulosis, but not in any of the other disorders. Positive epidermal staining was noted in all tissues examined. The pattern obtained suggested that epidermal dendritic cells may react with OKT 6. The findings indicate that OKT 6-positive cells may be found outside the thymus in certain conditions. The observations in epidermis cast doubt on the exact nature of the positively reacting cells observed in dermis, suggesting that may either be immature thymocytes or possibly Langerhans cells.

An immunoperoxidase study was carried out on human tonsil (15 specimens) and human lymph node (5 specimens) using OKT6, a monoclonal antibody which was raised against a determinant on immature thymocytes. OKT6-positive cells were identified in the crypt epithelium of all tonsils examined and in occasional clusters in the interfollicular areas of two lymph nodes. OKT6 has recently been shown to react with epidermal dendritic cells (Langerhans' cells). This study confirms that OKT6-reactive cells may be found outside the thymus. The pattern of staining obtained suggests that OKT6 reactivity belongs to a dendritic subpopulation. The significance of the finding in relation to physiology and pathology is discussed. These physiological findings may also be relevant to the immunotherapy of T-cell lymphomas.

"Parapsoriasis" is a term used to include a heterogeneous group of conditions, one variant of which, at least, eventuates in mycosis fungoides in a substantial percentage of cases. The T-cell origin of mycosis fungoides is well established. The lack of similar information on lymphoid cell types in parapsoriasis prompted an immunoperoxidase study using a specific antihuman T-cell serum in a group of seven patients with parapsoriasis. Our findings demonstrated a preponderantly T-cell infiltrate in the categories of parapsoriasis examined.