16 research outputs found
Primary alterations during the development of hidradenitis suppurativa
BACKGROUND: Hidradenitis suppurativa (HS) is a chronic, inflammatory disease of the apocrine glandârich (AGR) skin region. The initial steps of disease development are not fully understood, despite intense investigations into immune alterations in lesional HS skin. OBJECTIVES: We aimed to systematically investigate the inflammatory molecules involved in three stages of HS pathogenesis, including healthy AGR, nonâlesional HS and lesional HS skin, with the parallel application of multiple mRNA and proteinâbased methods. METHODS: Immune cell counts (T cells, dendritic cells, macrophages), Th1/Th17ârelated molecules (ILâ12B, TBX21, IFNG, TNFA, ILâ17, IL10, ILâ23A, TGFB1, RORC, CCL20), keratinocyteârelated sensors (TLR2,4), mediators (S100A7, S100A8, S100A9, DEFB4B, LCN2, CAMP, CCL2) and proâinflammatory molecules (IL1B, IL6, TNFA, ILâ23A) were investigated in the three groups by RNASeq, RTâqPCR, immunohistochemistry and immunofluorescence. RESULTS: Epidermal changes were already detectable in nonâlesional HS skin; the epidermal occurrence of antimicrobial peptides (AMPs), ILâ1ÎČ, TNFâα and ILâ23 was highly upregulated compared with healthy AGR skin. In lesional HS epidermis, TNFâα and ILâ1ÎČ expression remained at high levels while AMPs and ILâ23 increased even more compared with nonâlesional skin. In the dermis of nonâlesional HS skin, signs of inflammation were barely detectable (vs. AGR), while in the lesional dermis, the number of inflammatory cells and Th1/Th17ârelated mediators were significantly elevated. CONCLUSIONS: Our findings that nonâlesional HS epidermal keratinocytes produce not only AMPs and ILâ1ÎČ but also high levels of TNFâα and ILâ23 confirm the driver role of keratinocytes in HS pathogenesis and highlight the possible role of keratinocytes in the transformation of nonâinflammatory Th17 cells (of healthy AGR skin) into inflammatory cells (of HS) via the production of these mediators. The fact that epidermal TNFâα and ILâ23 appear also in nonâlesional HS seems to prove these cytokines as excellent therapeutic targets
Improvement of clinical and immunological parameters after allergen specific immunotherapy in atopic dermatitis
Background: Allergen immunotherapy (AIT) is considered a curative treatment in some atopic diseases, but in AD contradictory clinical results exist and the action of AIT has not been elucidated. In the literature there is no evidence for parallel investigations of permeability barrier, cutaneous, and blood immune responses after AIT in AD. Objectives: The objective was to investigate immune parameters in the blood and skin and to detect clinical, and barrier changes after AIT in AD. Methods: Mild-to-moderate AD patients (n=14) with concomitant allergic rhinitis to house dust mites were selected. All patients received topical treatment, while eight patients were randomly selected for adjuvant AIT also. At baseline and after 6 months clinical, barrier and immunological investigations (serum and skin tests) were performed. In selected patients, biopsies from atopy patch tests (APT) were analysed by immunohistochemistry for ADrelevant immune cells and mediators. Results: In the adjuvant AIT-group, clinical parameters and barrier functions improved significantly. Blood immune parameters displayed no significant changes. Post-AIT APT became negative in all patients in the AIT-group, but remained positive in the non-AIT group. Cutaneous dendritic cell and T cell recruitment decreased significantly after allergen challenge in the AIT-group, but no significant changes in skin or serum immunoglobulin E levels or prick test (SPT) reactivity were detected. Conclusions: AIT is a beneficial adjuvant treatment for sensitized AD patients. AIT improves not only clinical symptoms, but also permeability barrier functions. The effect of AIT on sensitization should be detected by APT, not by SPT
Newborn screening for Fabry disease by measuring GLA activity using tandem mass spectrometry
BACKGROUND: Fabry disease (FD) is an X-linked lysosomal storage disorder caused by the deficiency of alpha-galactosidase A (GLA). We evaluated a tandem mass spectrometry method to measure GLA activity. METHODS: One 3.2mm punch from a dried blood spot sample (DBS) was incubated with substrate and internal standard in the reaction buffer for 22 h. The resulting product was quantified against internal standard using MS/MS. RESULTS: The median GLA activity of male newborn DBS (N=5025) was 9.85 + or - 6.4 micromol/h/l (CI 95% is 9.67-10.02 micromol/h/l); The median GLA activity of female newborns (N=4677) was 10.2 + or - 6.3 micromol/h/l (CI 95% is 10.02-10.38 micromol/h/l). The difference between the two subgroups is within assay analytical variation. The GLA activities in the DBS samples from 9 juvenile and adult males with previously identified FD were below 1.64 micromol/h/l. The GLA activities from 32 juvenile and adult females with confirmed FD were below 4.73 micromol/h/l. In 5 (16%) females GLA activities were above the 0.5th percentile of lower limit of CI 95% at 3.18 micromol/h/l. CONCLUSIONS: The MS/MS method for Fabry disease newborn screening is robust and can be readily multiplexed with other lysosomal disorders such as Pompe, Gaucher, Niemann-Pick, and Krabbe diseases