Cellular kinetics of delayed hypersensitivity test reactions to topical glucocorticosteroids.

The phenotypes of the infiltrating cells in 13 patients with delayed hypersensitivity to topical glucocorticosteroids (GCS) were studied from sequential biopsies of positive epicutaneous test reactions by using the avidin-biotin-complex (ABC) technique. Monoclonal antibodies were used to identify the cells with the following phenotypes: T3, T4/T4a, T6, T8, T9, T11, M1, Ia1 (HLA-DR), interleukin-2 receptor/T26a, and dendritic reticular cell. The cellular kinetics of GCS hypersensitivity reactions were compared with delayed hypersensitivity reactions caused by allergens not related to GCS. In both GCS and non-GCS reactions the epidermal dendritic T6+ cells were more numerous than dendritic Ia1+ cells. There was a decrease in the number of both cell types during these reactions; in GCS reactions the decrease in the number of T6+ cells was seen later than in non-GCS reactions. Ia1+ keratinocytes were seen at sites near dermal infiltrates. Compared with the non-GCS delayed hypersensitivity reaction, there were fewer pan T (T11+/T3+) in the GCS reaction. The relative numbers of M1+ monocytes and the T4/T8 ratio were substantially lower in the latter; these findings can be explained as a GCS effect which modulates the delayed type hypersensitivity reaction.

Patch test reactions to inhalant allergens in atopic dermatitis.

To study whether inhalant allergens could induce eczematous reactions on normal skin of atopic patients we applied birch pollen and house dust mite antigens at 500 times the concentration used for prick testing as epicutaneous tests. Six out of 17 patients with atopic dermatitis in remission had positive delayed type reactions to birch pollen and three to house dust mite. Only one out of 13 atopic patients without history of atopic dermatitis but with seasonal allergic rhinitis had a positive patch test reaction to birch pollen and no patient had positive test reactions to house dust mite. No positive patch test reactions to birch pollen or house dust mite were seen in the ten healthy control subjects. In patients with positive test reactions biopsies from the test sites revealed epidermal spongiosis and vesiculation. Immunostaining of the epidermis revealed keratinocytes displaying both CD1 and HLA-DR. The present study suggests that inhalant allergens can exacerbate atopic dermatitis.

Effect of tissue processing on the immunostaining of inflammatory cell surface epitopes identified by monoclonal antibodies.

Optimal tissue processing conditions were defined for the immunohistochemical detection of inflammatory cell surface epitopes identified by OKT3, OKT4, OKT6, OKT8, OKIa1, OKM1, Leu7 and pan B cell antibodies. Snap-freezing in isopentan was superior to liquid nitrogen in preservation of morphology. Embedding of the tissues in Tissue-Tek II O.C.T. Compound diminished the intensity of immunostaining with all antibodies studied; however, the embedded tissues tolerated longer storage without drying. Optimal fixation with satisfactory preservation of morphology and immunogenicity was achieved with fixation of the frozen sections in acetone at 4 degrees C for 5 min. Blocking of the endogenous peroxidase with methanol-H2O2 treatment destroyed all epitopes studied except those identified with OKIa1, OKT6 and Leu7.

Erythromycin acistrate and erythromycin stearate in the treatment of non-gonococcal urethritis.

The antibacterial efficacy and tolerability of erythromycin acistrate (EA) and erythromycin stearate (ES) were compared in 100 male patients with non-gonococcal urethritis (NGU). The dosage of EA was 400 mg tid and that of ES 500 mg tid. Mean duration of treatment was ten days. When the final evaluation of the trial was made, the patient material was divided into two groups. One group consisted of patients with chlamydia-positive culture before treatment, the other of chlamydia-negative patients with signs of infection in the direct smear. There were 17 patients with chlamydial infection in the EA-group, and the microbiological cure rate was 100%. In the ES-group there were 21 patients with chlamydial infection and the microbiological cure rate was 95%. In the EA-group, the cure rate of chlamydia-negative NGU patients was 78%, and the corresponding figure in the ES-group was 86%. There was no difference in the cure rates between the two groups on either drug. In the EA-group, 25 patients (50%) reported side effects, in 22 these were gastrointestinal. In the ES-group, 26 patients (52%) reported side effects; in 22 these were gastrointestinal. Two patients in the EA-group discontinued the treatment because of gastrointestinal side effects. There were no differences between the groups in the frequency, severity and duration of side effects.

Local involvement of antigen-presenting cells and activated T cells in perilesional and clinically uninvolved skin in pemphigus vulgaris.

Biopsies obtained from both the perilesional areas and clinically uninvolved skin of patients with pemphigus vulgaris (PV) were studied for antigen-presenting cell and lymphocyte phenotype and/or activation phenotype using monoclonal antibodies in avidin-biotin-peroxidase complex staining. Perilesional PV skin contained CD4+ and CD8+ T lymphocytes as the predominant cell type, but cells with a potential antigen-presenting function displaying CD11b phenotype of monocyte/macrophages and, in particular, CD1 phenotype of Langerhans cells were also present. The number of mononuclear inflammatory cells was greater in perilesional than in clinically uninvolved PV skin, and so were the proportions of CD4+, CD8+, CD25+, Ia+ cells (p less than 0.01), and CD1+ Langerhans cells and transferrin receptor positive cells (p less than 0.05). These findings confirm and extend earlier observations on local involvement of immunocompetent cells in PV.

Immunocompetent cells of fixed drug eruption.

The positive provocation test reactions of the skin of six patients with fixed drug eruption (FDE) were studied from timed skin biopsies taken between 2 hours and 9 days after the appearance of FDE. Monoclonal antibodies to the following immunocompetent cell surface epitopes were used: T3, T4, T6, T8, T9, M1, Ia1, Drc, Leu7 and B cell. The dermal infiltrate comprised 60-80% of T lymphocytes at all the times studied. Cells with T4 and T8 epitopes were displayed in similar numbers. A transient decrease in the number of T6+ cells of the epidermis could be detected with a simultaneous and also transient increase of the T6+ cells in the dermis, which suggests a possible traffic of Langerhans' cells from the epidermis to the dermis. The epidermal Ia1+ cells showed changes similar to but less marked than the T6+ cells. The number of the dermal Ia1+ cells increased continuously. In the late biopsies these Ia1+ cells comprised up to 90% of the infiltrating cells. Except for the finding of a reduction of T6+ and Ia1+ epidermal cells, the cellular kinetics of FDE are similar to those seen in both cutaneous immunological and irritant reactions.

Eczematous reactions in atopic patients caused by epicutaneous testing with inhalant allergens.

To determine whether inhalant allergens could induce eczematous lesions we studied 17 patients with atopic eczema (with or without allergic rhinitis), 13 patients with allergic rhinitis without atopic eczema and 10 healthy control subjects. The allergens, birch pollen (Betula verrucosa) and house dust mite (Dermatophagoides pteronyssinus), were applied in aluminium chambers for 48 h on clinically normal skin. In 17 patients with atopic eczema, six epicutaneous test reactions of the delayed type to birch pollen and three to house dust mite were seen at 48 or 72 h. In 13 patients with allergic rhinitis without eczema there was one delayed reaction to birch pollen and none to house dust mite. No delayed type test reactions to either allergen were seen in the controls. Biopsies of the positive test sites revealed an eczematous reaction with epidermal spongiosis and microvesiculation. Immunostaining of cryostat sections showed dermal cell infiltrates consisting of mainly T lymphocytes (ratio of T4:T8, 2-6:I) and to a lesser degree Langerhans and indeterminate T6+ cells. 50-90% of the cells were Ia+. The numbers of basophils and mast cells did not exceed 10-15%.

Delayed hypersensitivity to topical corticosteroids.

Topical hypersensitivity to corticosteroids was studied by epicutaneous testing using the Finn Chamber technic. The steroids were tested in both ethanol and white petrolatum and, in certain cases, in dimethyl sulfoxide. Additionally, commercial preparations were tested. Three groups of patients were studied: (1) patients with a history of hypersensitivity to at least two topical preparations (five of ten patients studied showed a positive patch test reaction for corticosteroids), (2) patients in whom topical corticosteroid hypersensitivity was suspected because of treatment-resistant eczema (seven of twenty-five patients showed a positive patch test reaction), and (3) dermatologic inpatients and outpatients undergoing epicutaneous testing for suspected topical hypersensitivity. Hydrocortisone-17-butyrate (H-17-B) was included in the standard patch test series; of 450 patients tested, two showed a positive patch test reaction. All the patients with corticosteroid hypersensitivity had a positive reaction to H-17-B. In six patients, additional hypersensitivities to one or several other steroid preparations were seen. Use testing was performed as an open test, with 0.1% or 1% H-17-B in ethanol on normal skin of the flexor side of the upper extremities. A positive test reaction was seen in only one of nine patients. Results of use testing with the commercial 0.1% H-17-B (Locoid) ointment were always negative. Our study suggests that the sensitivity of patch tests for corticosteroid hypersensitivity can be increased by using ethanol as vehicle.