Selective generation of CD8+ T-cell clones from the peripheral blood of patients with cutaneous reactions to beta-lactam antibiotics

The presence, phenotype, and functional characteristics of peripheral blood penicillin-specific T lymphocytes in individuals with cutaneous allergic reactions to penicillin were investigated using in vitro long-term culture techniques. Peripheral blood mononuclear cells from two penicillin-allergic patients were stimulated in vitro with penicillin, and T-cell blasts were clonally expanded by limiting dilution. Seven T-cell clones were derived, all of which were CD3⁺ CD4^? CD8⁺ HLA-DR⁺, and produced IL-2 and IFN-γ upon stimulation. T-cell proliferation required the presence of antigen and autologous, but not allogeneic, antigen-presenting cells. In addition to the parent compound, the T-cell clones also developed a proliferative response to penicilloyl, the major metabolite of penicillin. The cloned T-cell lines were found to exhibit marked suppressor activity for Con A mitogenesis. The observed suppressor activity required cell-to-cell contact, as supernatants from these T-cell clones had no comparable inhibitory effect. These findings indicate that there is a predominance of penicillin-specific CD8⁺ T cells in the peripheral blood of individuals sensitized to beta-lactam antibiotics.     

Effect of Subcutaneous Infiltration Anesthesia on Pain in Photodynamic Therapy: A Controlled Open Pilot Trial

BACKGROUND: Photodynamic therapy (PDT) has been established as a treatment option for nonmelanoma skin cancer, such as superficial basal cell carcinoma, actinic keratoses, or Bowen's disease. OBJECTIVE: A major drawback of PDT is pain during treatment that can cause extreme distress for some patients. METHODS: This study was a controlled, open trial comparing PDT in 16 patients on one side of the face with orally administered analgesics and PDT on the contralateral side of the face with subcutaneous infiltration anesthesia (SIA). The 5-aminolevulinic acid gel was applied 5 hours before treatment. Pain was assessed by the patient using a visual analog scale directly after treatment. RESULTS: Fifteen of 16 (94%) patients reported less pain during PDT after SIA compared to oral analgesics only. This effect was significant (Wilcoxon test for matched pairs, p= .001). No side effects due to SIA were observed. CONCLUSION: SIA is an effective and rather safe method for the control of PDT-associated pain.     

Successful Topical Treatment of Sorafenib-Induced Hand–Foot Skin Reaction in a Child with Hepatocellular Carcinoma

Abstract:  Orally active kinase inhibitors such as Sorafenib are known to elicit cutaneous side effects in the majority of adult patients, whereas specific cutaneous complications of this agent have not been described in children so far. We here present the first pediatric case of Sorafenib-induced hand-foot-skin reaction and its successful topical therapy facilitating continuation of kinase inhibitor treatment.     

C73R is a hotspot mutation in the uroporphyrinogen III synthase gene in congenital erythropoietic porphyria

Congenital erythropoietic porphyria (CEP) results from profoundly deficient activity of the fourth enzyme of the haeme biosynthetic pathway, uroporphyrinogen III synthase (UROIIIS). CEP is a rare, recessively inherited disorder, and mutations in the UROIIIS gene detected in CEP patients are heterogeneous. The notable exception to this rule is a single missense mutation, designated C73R, which represents over 40% of all mutant UROIIIS alleles. In this study, we investigated three separate families with CEP from different ethnic backgrounds. We performed haplotype analysis using two microsatellite markers that closely flank the UROIIIS gene on chromosome 10q24, spanning a region of 4 cM on the GB4 linkage panel. Haplotype analysis revealed the occurrence of C73R on different haplotypes in four out of four disease chromosomes studied. The results are consistent with the hypothesis that C73R is a hotspot mutation for CEP, and does not represent wide dispersion of a single ancestral mutant C73R allele.     

Inducibility of arylhydrocarbon-hydroxylase activity in human hair follicles by topical application of liquor carbonis detergens (coal tar)

Arylhydrocarbon-hydroxylase (AHH) is a cytochrome P-450-dependent polysubstrate mono-oxygenase which plays an important role in converting some compounds (e.g. benzo[a]pyrene) to highly reactive carcinogenic species. A simple AHH assay is described, using [3H]benzo[a]-pyrene as substrate. 7,8-Benzoflavone (10(-4)M) inhibits 92% of the measured enzyme activity. Liquor carbonis detergens (which contains coal tar) induces AHH activity in human hair follicles in vivo. We suggest that using this simple assay, hair follicles would be a very suitable tissue to test whether the AHH-controlling gene is of significance in producing cancer.     

CD8+ dermal T cells from a sulphamethoxazole-induced bullous exanthem proliferate in response to drug-modified liver microsomes

There is evidence that T lymphocytes play a critical role in the pathogenesis of drug-induced bullous exanthems. Sulphonamides are known to be among the most frequent aetiological agents in these severe drug-induced cutaneous hypersensitivity reactions. Several studies indicate that cytochrome P450-dependent metabolites of sulphonamides act as the nominal allergens. A 70-year-old woman with a severe blistering exanthem caused by cotrimoxazole (sulphamethoxazole and trimethoprim) was studied. We employed an in vitro approach to determine whether cytochrome P450-dependent enzymes activated drug-specific T lymphocytes from this patient. Immunohistochemical analysis of involved skin revealed a majority of epidermal CD8⁺ T lymphocytes, whereas the dermal infiltrate was composed of both CD4⁺ and CD8⁺ T cells. Dermal T lymphocytes isolated from lesional skin proliferated in response to sulphamethoxazole, but not to trimethoprim, in the presence of autologous mononuclear cells used as antigen-presenting cells. The antigen-specific response of sulphamethoxazole-specific T cells was significantly augmented in the presence of murine liver microsomes with P450-dependent catalytic activities. Our observations suggest that some cutaneous hypersensitivity reactions to sulphamethoxazole are due to drug-specific T lymphocytes. Cytochrome P450-dependent enzymes may play a critical role in the formation of the nominal antigen, which is recognized by antigen-specific T cells.