Intercellular Pattern Research Articles

Keratinocytes in lesional skin of atopic eczema bear HLA-DR, CD1a and IgE molecules

Apparently normal, and lesional skin from patients with atopic eczema were investigated immunohistochemically with anti-HLA-DR, -CD1a and -IgE antisera. A CD1a+ intercellular pattern was observed in uninvolved skin in the majority of the patients whereas an HLA-DR+/CD1a+ network, mostly localized in basal and supra-basal areas, was shown in lesional skin of virtually all of them. Moreover, an HLA-DR+/CD1a+IgE+ intercellular pattern was observed in some of the patients only and was predominantly localized in those areas characterized by lymphocyte exocytosis, spongiosis or vesicle formation. Whether keratinocytes are able to synthesize CD1a antigen and Fc epsilon R or if these molecules are only produced and shed by CD1a+/IgE+ epidermal dendritic cells remains unclear.

Intra-epidermal neutrophilic IgA dermatosis with colitis

We report the association of an intra-epidermal blistering disease of the skin with colitis. Both the skin and large bowel epithelium were infiltrated by neutrophils and eosinophils, and showed intra-epithelial deposition of IgA in an intercellular pattern. The colitis and the blisters responded well to treatment with dapsone.

Epidermal Interleukin-1 is Increased in Cutaneous T-Cell Lymphoma

Interleukin-1 (IL-1) is a monocyte-derived polypeptide with immunoregulatory and proinflammatory functions. Although monocytes are the principle source of IL-1, other cells, such as keratinocytes, endothelial cells, renal mesangial cells, and neutrophils, produce a factor with IL-1 activity. The IL-1-like polypeptide produced by keratinocytes, epidermal-derived thymocyte-activating factor (ETAF), is similar on biological, biochemical, and molecular levels to monocyte-derived IL-1. Studies of IL-1 or ETAF have for the most part been undertaken using cell culture supernatants or cell lysates, and in situ localization of ETAF has not been demonstrated. Previous studies have suggested that ETAF is involved in the pathogenesis of cutaneous T-cell lymphoma (CTCL). To gain insights into the role of these cytokines in disease states, we investigated whether IL-1 could be localized in tissue sections using a direct immunofluorescence technique with a monoclonal antibody directed against IL-1. This monoclonal antibody partially inhibited ETAF activity and totally inhibited IL-1 activity in the co-stimulator assay and so could be used to detect IL-1 or ETAF. We studied skin biopsies from 10 healthy individuals, 10 patients with CTCL, and 11 patients with various inflammatory dermatoses. Intense epidermal fluorescence was demonstrated in all cases of CTCL, whereas minimal reactivity was visible in normal biopsies and the inflammatory dermatoses. Most patients with CTCL showed an intercellular pattern, while none of the normal controls or those with inflammatory dermatoses showed this pattern. An irrelevant IgM monoclonal antibody, used as a negative control, did not demonstrate epidermal staining. To further demonstrate specificity, we incubated the anti-IL-1 antibody with recombinant beta IL-1:Epidermal reactivity was completely blocked. In a separate experiment, COLO 16 cells, a squamous cell carcinoma cell line that constitutively produces ETAF, stained positively for the IL-1 antibody. We conclude that epidermal IL-1 is elevated in CTCL.

Immunohistologic Identification of Antigen-Presenting Cells in Cutaneous Sarcoidosis

Considerable evidence exists to show that activated T lymphocytes preferentially accumulate at sites of disease activity in sarcoidosis. Langerhans cells, which can be recognized by reactivity with an antibody to the T6 antigen are thought to play a primary role in T-lymphocyte activation by the skin, a tissue frequently involved in sarcoidosis. This immunohistologic study examined the distribution of OKT6-positive cells and surface expression of HLA-DR antigen in cutaneous sarcoid lesions. Skin specimens stained with an anti-HLA-DR antibody demonstrated diffuse staining of the granulomas. In addition, keratinocytes, which do not normally express HLA-DR antigens, were found to stain with monoclonal antibody to HLA-DR in an intercellular pattern. Examination of specimens for OKT6-reactive Langerhans cells revealed significantly greater concentrations in the epidermis overlying sarcoidal granulomas (33 +/- 7 cells/mm) than in the epidermis of age-, sex-, and race-matched controls (11 +/- 3 cells/mm, p less than 0.001). Of greater importance was the demonstration that significant numbers of OKT6-positive cells were present within the dermal sarcoid granulomas (19-208/mm2) in a distribution that paralleled that of Leu-3a-positive T lymphocytes. These data suggest that the epidermis may participate in activation of lymphocytes in cutaneous sarcoidosis, and implicate OKT6-positive cells in granuloma formation.

Effects of vitamin E on fibroblast fibronectin.

Fibronectin is a glycoprotein found mainly on fibroblast cell surfaces. Cultured human skin fibroblasts grow as monolayers in regular linear and whorl-shaped patterns. Exposure to a certain vitamin E analogue, D-alpha-tocopheryl-polyethylene-glycol-1000-succinate, induces concentration-dependent morphological and pattern changes. Using indirect immunofluorescence techniques, fibronectin, a cell protein associated with cell structure and intercellular patterns, was identified within the cells and the intercellular spaces. At a low concentration, the vitamin E analogue caused inhibition of the intercellular distribution of fibronectin without changes in intracellular fibronectin or significant disruption of the morphological patterns. At higher concentrations morphological changes ensued. These data suggest that the mechanism of vitamin E-induced morphological and intercellular pattern changes may not be manifested through inhibition of fibronectin deposition or secretion.

Immunofluorescence in psoriasis: studies of immunoglobulins, complement deposits, and three membrane markers.

Sixteen psoriatic patients and 11 control subjects were investigated by immunofluorescence for skin immunoglobulins (IG) and complement (c) deposits and for keratinocyte membrane markers with anti beta 2 microglobulin (beta 2 m) antibodies (Ab), Con A, and pemphigus Ab. IG and/or c deposits were almost constant in involved epidermis. Three patterns were associated: (1) parakeratotic nuclear (dots-dashes), (2) stratum corneum (SC) intercellular (lamellar pattern), (3) vascular (granular or linear deposits in papillary dermal vessels). In uninvolved epidermis nuclear or vascular deposits could occasionally be present but intercellular pattern was never found in SC. Control specimens were always negative. The three surface markers investigated (beta 2 m, Con A receptor, Pemphigus antigen) could be demonstrated on psoriatic keratinocytes with only slight differences in distribution when compared with controls. Thus, by this methodology no important abnormality could be found in psoriatic keratinocyte membrane antigens.

Ia-like antigens in lichen planus.

The occurrence of Ia-like antigens in the skin of patients with lichen planus was studied by an indirect immunofluorescence (IIF) technique with use of immunosorbent-purified anti-Ia antibodies. Normal skin from the patients and skin from healthy persons showed dermal histiocytes and epidermal suprabasal dendritic cells expressing these antigens. In lichen planus lesions Ia-like antigens were found on virtually all mononuclear cells in the dermal infiltrate and showed an intercellular pattern in the epidermis. The finding of cells expressing Ia-like antigens in the dermal infiltrate, most probably activated T lymphocytes, renders likely a cell-mediated type of reaction in the pathogenesis of lichen planus.

Stage specific larval, pupal, and adult cuticles in the tracheal system of Sarcophaga bullata.

Successive tracheal cuticles of the dorsal longitudinal trunks are studied with the electron microscope. Minor differences seen at the light microscope level are seen as major qualitative and quantitative ones at the ultrastructural level. The larval and pupal cuticles are secreted by similar epithelial cells; these possess large polytene chromosomes. Cell division and possibly cell replacement occur prior to adult cuticle secretion. The findings are discussed in terms of cell specificity, intra- and inter-cellular pattern formation. This simple epithelium, the individual cells of which are capable of producing different cuticles, is interesting since the system is also shown to be responsive to hormone application.

Epidermal Intercellular Staining with Fluorescein-Conjugated Phytohemagglutinins

Fluorescein-conjugated phytohemagglutinins, Concanavalin A and Phytohemagglutinin P were used to stain frozen-cut sections of human skin to determine the presence and distribution of carbohydrate-containing substances in the epidermis. An intercellular staining pattern was observed which appeared identical to that found in various forms of pemphigus. The intercellular staining could be inhibited, or abolished, by α-methyl-D-mannopyranoside or α-methyI-D-glucopyranoside in the case of Concanavalin A, and by N-acetyl-D-galactosamine in the case of Phytohemagglutinin P, but not by a variety of other monosaccharides tested. A similar pattern of staining could be produced in vivo in mouse and guinea-pig skin by intradermal injection of fluorescein-conjugated Concanavalin A. Staining with fluorescein isothiocyanate alone produced a staining pattern which was a negative image of that seen with the fluorescein-conjugated phytohemagglutinins, namely cytoplasmic staining, but absent intercellular staining.

The argyrophilic mosaic of the internal limiting membrane of the retina

It has been known for many years that when the surface of the retina is stained with silver a characteristic mosaic pattern appears. Several authors have offered explanation for its origin but no unanimity of opinion exists. We have re-investigated this problem using the additional techniques of electron microscopy and staining with ruthenium red. We conclude that the argyrophilic mosaic pattern is due to the selective uptake of silver at the cell borders of the Müller and accessory glial cells. Viewed from the surface these silver lines are seen in depth and stand out in contrast to the lighter silver deposition on the internal limiting membrane. This intercellular pattern is closely comparable to that seen in silver staining of many cellular membranes. Positive staining with ruthenium red shows a similar distribution in the retina to that of silver; possible explanations for these findings are briefly considered.

Lymphocyte-mediated transport of aggregated human gamma-globulin into germinal centre areas of normal mouse spleen.

GERMINAL centres develop early in the immune response, and although knowledge of these structures is far from complete, they are known to be intimately associated with the capacity to produce specific antibodies after antigenic stimulation1. The localization of antigen in these centres coincides with the appearance of specific serum antibody2, which is found in a close relationship to reticular cells in these areas3. It produces a characteristic dendritic intercellular pattern of immunofluorescent staining or autoradiography when isotopically labelled. This localization of antigen in germinal centres did not occur in rabbits rendered tolerant to haemocyanin or human serum albumin4, but injected immune complexes produced an identical pattern of localization of antigen in germinal centres in normal rat lymph nodes5.

Pemphigus and Other Diseases

The intercellular pattern of fluorescence was demonstrated in 24 of 75 patients with diseases other than pemphigus. A relationship was found between the presence of anti-B isohemagglutinins and intercellular fluorescence as demonstrated by the indirect immunofluorescent (IIF) technique, using fetal rhesus monkey ( Macaca mulatta ) esophagus as the substrate. When the anti-B isohemagglutinin titer was reduced or abolished, after absorption with B substance, the intercellular pattern was reduced considerably or abolished. The importance of the blood type of the substrate employed in the IIF technique and the blood type of the patient is stressed so that these factors which may give rise to error in the interpretation of intercellular fluorescence are excluded.