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A human skin window technique using micropore membranes.

A skin window technique using micropore membranes is described. Eight micrometre pore-size membranes are placed on abrasions made with a dental stone and covered with moist filter paper and impermeable film: adhesive tape is placed overall. The assembly is unobtrusive and stable and may be worn during normal activities. For clinical studies, a series of membranes used over 4-5 h gives information on distribution of leucocyte emigration sites, numbers and types of emigrant cells, their rate of locomotion in the membrane (solely under the influence of endogenous factors) and any difference in form, surface phenotype or function between cells at the dermal surface and those which have traversed the membrane. This system, using a 3-dimensional labyrinthine structured membrane, avoids errors inherent in previous techniques, e.g., differential adhesiveness, competition for space, no quantification of locomotion after emergence or indication of the productive area of the abrasion. Incidental findings include no major change in the nature of the emigrating population over 24 h, the presence of monocytoid cells from the earliest phase in normal subjects, the origin of emigrating cells in discrete foci, and an even distribution of cells in a fully infiltrated membrane. The skin window technique is capable of greater clinical precision and experimental elaboration than hitherto realised.

Cell Movement

The recruitment of inflammatory cells using the skin-window technique.

The chemotaxis of inflammatory cells induced by the skin-window technique using IgD as cytotaxigen or cytotaxinogen was studied in 16 patients with allergic contact dermatitis. Six patients with leg ulcer served as controls. By means of this method the recruitment of inflammatory cells with receptors for IgD could be shown.

Chemotaxis, Leukocyte

Cellular inflammatory response to fungal antigen studied with the skin window technique--correlation of the exudate with the evolution of the mycotic infection.

The skin window technique, using a coverslip, was applied to study the inflammatory exudate induced by killed pathogenic yeasts in patients with mycotic infections. The presence of eosinophils in the specific exudate after 3 hours seems to indicate specific IgE fixed in the tissues. This was demonstrated by the correlation of this specific eosinophilia with the eosinophilia induced by an anti-IgE serum and with the positive immediate intradermal reactions and by the correlation with serum-specific IgE detected with an immunoenzymatic technique. Increased eosinophilia is related to the active phase of the disease. The eosinophilia in the specific exudate seems to be more reliable than an in vitro technique for the detection of the specific IgE against fungal antigen. In this material, the eosinophilia demonstrated that yeast-specific IgE may be responsible for both immediate and delayed intradermal reactions.

Antigens, Fungal

Neutrophil migration in vivo: analysis of a skin window technique.

The applicability of a skin chamber technique for the quantitation of neutrophil migration in vivo was studied in healthy subjects. Skin lesions were generated on lower abdominal skin by separating epidermis from dermis by suction. In 23 subjects the mean chamber cell count at 24 h was 20.9 X 10(6)/cm2 of skin lesion area, and 97.5% of the cells were neutrophils. An analysis of the variance in up to 8 parallel chambers in 10 subjects supported the use of 4 simultaneous replicates. Reproducibility in consecutive experiments was studied by testing 9 persons twice; the results were found to agree closely. Kinetic studies were performed in 15 subjects. The method is useful for the in vivo study of diseases and medications thought to affect leukocyte function.

Adult

Experimental studies on phototoxic and photoallergic reaction by the skin window technique.

The differentiation of photoallergic contact dermatitis by 3,3', 4', 5-tetra-chlorosalicylanilide from phototoxic contact dermatitis by 8-MOP in guinea pigs was studied by skin window technique. Based upon the results, it may be concluded that the photoallergic reaction can be differentiated from phototoxic reaction by the appearing pattern of basophilic leukocytes.

Animals

Leucocyte function in Crohn's disease. Studies on mobilisation using a quantitative skin window technique and on the function of circulating polymorphonuclear leucocytes in vitro.

Leucocyte function was evaluated by mobilisation to skin windows with chambers and by the chemotactic, phagocytic, and nitro blue tetrazolium (NBT) reducing activity of circulating leucocytes in vitro in 20 patients with Crohn's disease, 21 healthy volunteers, and nine patients with sarcoidosis or tuberculosis. Leucocytes had been mobilised in significantly reduced numbers at 12, 24, 36, and 48 hours in Crohn's disease compared with healthy volunteers (P less than 0.01) and patients with sarcoidosis/tuberculosis (P less than 0.01). The leucocyte migration rate showed that mobilisation in Crohn's disease begins after a prolonged lag phase and is reduced compared with healthy volunteers (P less than 0.01) and patients with sarcoidosis/tuberculosis (P less than 0.02). The reduced mobilisation was not correlated with disease activity. In vitro random migration by leucocytes was slightly lower in Crohn's disease (P less than 0.05) than in healthy volunteers, but there was no difference after removal of the autologous plasma. Chemotactic response to casein did not differ between the groups studied. Serum independent and dependent phagocytosis did not differ from control groups. Serum independent phagocytosis was positively and significantly correlated to the disease activity (rho 0.4812, P less than 0.05). Resting leucocyte NBT reduction was increased in Crohn's disease and sarcoidosis/tuberculosis (P less than 0.01), but during phagocytosis a lower NBT reduction was found in Crohn's disease than in healthy volunteers (P less than 0.02). The inflammatory response in Crohn's disease, with reduced leucocyte accumulation, differs from patients with other granulomatous reactions and is independent of the disease activity. Our data suggest that the defect is not cellular. They support the hypothesis that a pathogenic factor in Crohn's disease may be foreign material that is normally eliminated remaining in the tissue and eliciting a chronic inflammatory response.

Adolescent