Robert D. Streilein

A topical antioxidant solution containing vitamins C and E stabilized by ferulic acid provides protection for human skin against damage caused by ultraviolet irradiation

BACKGROUND Skin cancer and photoaging changes result from ultraviolet (UV)-induced oxidative stress. Topical antioxidants may protect skin from these effects. OBJECTIVE We sought to determine whether a stable topical formulation of 15% L-ascorbic acid, 1% alpha-tocopherol, and 0.5% ferulic acid (CEFer) could protect human skin in vivo from substantial amounts of solar-simulated UV radiation. METHODS CEFer and its vehicle were applied to separate patches of normal-appearing human skin for 4 days. Each patch was irradiated with solar-simulated UV, 2 to 10 minimal erythema doses, at 2-minimal erythema dose intervals. One day later, skin was evaluated for erythema and sunburn cells, and immunohistochemically for thymine dimers and p53. UV-induced cytokine formation, including interleukin (IL)-1alpha, IL-6, IL-8, and IL-10, and tumor necrosis factor-alpha, were evaluated by real-time polymerase chain reaction. RESULTS CEFer provided significant and meaningful photoprotection for skin by all methods of evaluation. LIMITATIONS The number of patients evaluated was relatively small. CONCLUSION CEFer provided substantial UV photoprotection for skin. It is particularly effective for reducing thymine dimer mutations known to be associated with skin cancer. Its mechanism of action is different from sunscreens and would be expected to supplement the sun protection provided by sunscreens.

Neutrophil CD11b, L-selectin and Fc IgA receptors in patients with dermatitis herpetiformis

Summary Background  The skin lesions found in patients with dermatitis herpetiformis (DH) are characterized by the presence of neutrophils at the dermal papillary tips in areas where the diagnostic cutaneous IgA deposits are found. Although the presence of the skin lesions of DH is known to be associated with gluten‐sensitive enteropathy, the mechanisms that control the development of skin lesions are not known.

Increased E-selectin, IL-8 and IL-10 gene expression in human skin after minimal trauma

Abstract: While clinical observations suggest that trauma to the skin plays a critical role in the induction of skin lesions in some skin diseases, the mechanism by which these lesions are induced is not known. We have postulated that minor trauma to the skin may lead to the expression of critical adhesion molecules on epidermal endothelial cells (E‐selectin) and pro‐inflammatory cytokines, which would predispose these areas to the development of skin lesions. In order to test this hypothesis normal inner arm skin of 11 normal subjects was gently rubbed with a pencil eraser for 2 min. Four hours after rubbing, skin biopsies were obtained from the rubbed site and from adjacent normal, unrubbed inner arm skin. Expression of E‐selectin, intercellular adhesion molecules (ICAM‐1) and the mRNA of selected cytokines was studied utilizing real time polymerase chain reactions. Biopsies were also examined for the presence of an inflammatory infiltrate and for the presence of E‐selectin and ICAM‐1. No clinical or histologic changes were seen in the skin 4 h after this minor trauma. An increase in expression of mRNA of E‐selectin (median ratio rubbed skin expression/unrubbed skin expression = 9.0; range 0.9–161.0), ICAM‐1 (median rubbed skin expression/unrubbed skin expression = 3.2; range 0.9–19.8), IL‐8 (median rubbed skin expression/unrubbed skin expression = 6.6; range 2.6–57.3) and IL‐10 (median rubbed skin expression/unrubbed skin expression = 13.1; range 2.4–29.0) was noted. Immunohistochemistry revealed the presence of E‐selectin in the dermal blood vessels in three of four subjects 4 h after rubbing but not in the unrubbed skin. No changes in ICAM‐1 or HLA‐DR deposits were seen in the rubbed compared with the unrubbed skin. These findings demonstrate that minor trauma to skin may induce expression of E‐selectin, ICAM‐1 and IL‐8, which may make the skin a more permissive site for the development of inflammatory reactions. These findings may play an important role in the development of skin lesions in areas of minor trauma.

Expression of Antimicrobial Peptides in the Normal and Involved Skin of Patients with Infective Cellulitis

BACKGROUND Endogenous antimicrobial peptides participate in the innate defense of skin against a variety of pathogens. The systemic expression of these peptides in normal-appearing skin of patients with infective cellulitis is unknown. METHODS Study patients were adults with infective cellulitis and signs of systemic inflammation. Skin biopsy and serum specimens were obtained from patients and from control subjects with no active infection. Cathelicidin and human beta-defensin 2 mRNA expression were determined by real-time polymerase chain reaction. RESULTS Skin biopsy specimens from 11 patients and 4 uninfected control subjects were analyzed. The relative expression level for cathelicidin mRNA was elevated in both the involved and the distal normal-appearing skin of patients with cellulitis, compared with expression in the skin of control subjects (mean ratios, 39.46 vs. 1.32, P=.0059; and 21.41 vs. 1.32, P=.0059). Similarly, the relative expression level of human beta -defensin 2 mRNA was elevated in both the involved skin (mean ratios, 20,844 vs. 11.65; P=.0015) and in distal normal-appearing skin of patients with cellulitis (mean ratios, 201.1 vs. 11.65; P=.0103). DISCUSSION In response to cutaneous infection there is a local and distal increase in endogenous antimicrobial peptide mRNA in both involved and normal-appearing skin. These observations show, for the first time to our knowledge, that after infection the human body responds by increasing systemic innate immunity.

A multicentre randomized trial of the treatment of patients with pemphigus vulgaris with infliximab and prednisone compared with prednisone alone

Pemphigus vulgaris (PV) is a blistering disease and tumour necrosis factor‐α has a role in its pathogenesis.

Association of Serum B-Cell Activating Factor Level and Proportion of Memory and Transitional B Cells with Clinical Response after Rituximab Treatment of Bullous Pemphigoid Patients

TO THE EDITOR Bullous pemphigoid (BP) is an immunemediated disease with significant morbidity and mortality (Roujeau et al., 1998; Joly et al., 2012). Rituximab has been used to treat a number of immunemediated diseases including pemphigus vulgaris (Joly et al., 2007; Tony et al., 2011). We administered rituximab (1,000 mg; day 0 and 14) to seven BP subjects with recalcitrant disease to determine the safety, efficacy, and mechanisms related to rituximab treatment response and relapse. Seven BP subjects with persistent disease activity despite X17.5 mg per day prednisone were studied (five female subjects; two male subjects; mean disease duration 3.2 years) (Supplementary Table online), in accordance with the Declaration of Helsinki Principles and the Institutional Review Board of Duke University. Subjects were followed up for 12 months. Disease flares were defined as the development of any disease activity necessitating an increased dose of prednisone. All subjects tolerated the infusions well with no drug-related serious adverse events. All subjects experienced a cessation of new skin lesions and a prednisone dose of 25% of starting dose or 10mg at 6 months. Total disease activity significantly improved at 6, 9, and 12 months compared with baseline (P1⁄40.0156, Wilcoxon test). Two subjects experienced a flare of their disease activity at months 7 and 11.5. Subjects were evaluated by ELISA for IgG anti-BP180 and anti-BP230 autoantibodies (MBL, Woburn, MA) and IgE anti-BP180 using the NC16A domain of BP180 as described (Messingham et al., 2009). IgG anti-varicella zoster antibody levels (Trinity Biotech, Jamestown, NY) and serum B-cell activating factor (BAFF) levels (R&D Systems, Minneapolis, MN) were determined by ELISA. Analysis of peripheral blood B cells was conducted by flow cytometry as described (Levesque et al., 2009). Before rituxiamb treatment, six of seven subjects had serum IgG anti-BP180 antibodies (range: 40–3,556 U ml ), four of seven subjects had IgG anti-BP230 antibodies (range 14–73 U ml ), and five of seven subjects had IgE anti-BP180 antibodies (range: 2.6–164 U ml ) Supplementary Table online). There was a significantly lower level of IgG anti-BP180 at 6 and 9 months compared with baseline (Po0.05, Wilcoxon test). A decrease in the levels of IgG anti-BP230 and IgE anti-BP180 was noted, but it was not statistically significant at any time point compared with baseline (Figure 1). There were no significant differences in serum IgG anti-varicella zoster antibodies and total serum IgG and IgE levels after rituximab therapy compared with baseline (Figure 1). BAFF serum levels increased after rituximab treatment, peaking at month 9 at 3,454 pg ml 1 (mean, range1⁄4 387–5,287 pg ml ). In the two subjects who experienced disease flare, peak serum BAFF levels were lower than those in the five subjects who did not experience a disease flare (P1⁄40.0476, Mann–Whitney test, months 3, 4, and 12; Figure 2). Depletion of B cells was noted within 2 weeks after the first rituximab infusion and persisted until month 12. At baseline, most B cells (CD19þ ) expressed a naive (CD38þ /IgDþ /CD27 ; mean1⁄4 33%) or memory (CD38þ / /IgDþ / / CD27þ ; mean1⁄4 39%) phenotype. Analysis of B-cell phenotypes during B-cell recovery revealed that the proportion of B cells expressing naive or transitional (CD38þ þ /IgDþ /CD27 ) phenotypes was significantly increased (mean1⁄473%) at month 12 compared with week 0 (mean1⁄4 35%) (P1⁄40.0234, Wilcoxon test), with a decrease in memory B-cell populations at month 12 (mean1⁄4 17%) compared with baseline (mean1⁄439%; Supplementary Figure S1 online). Analysis of B-cell subsets during B-cell recovery in subjects who flared revealed that there was a greater increase in the proportion of memory B cells and plasmablasts compared with baseline (P1⁄4 0.0476, Mann–Whitney test) and a significantly lower proportion of transitional cells at months 9 and 12 when compared with those subjects who did not experience a clinical flare (P1⁄40.0476, Mann–Whitney test) (Supplementary Figure S2a, b online). We found that rituximab therapy for BP patients was associated with a decrease in autoantibody levels with a half-life of about 30 days, consistent with the half-life in the circulation of IgG. IgE anti-BP180 antibodies decline less with persistent autoantibodies during the follow-up period. The level of anti-varicella zoster IgG antibodies did not change, consistent with the presence of rituximab-resistant long-lived plasma cells. One interpretation of this Accepted article preview online 3 June 2013; published online 27 June 2013 Abbreviations: BAFF, B-cell activating factor; BP, bullous pemphigoid; VZV, varicella zoster virus RP Hall, III et al. Low Serum BAFF Predicts Flares after Rituximab

Inflammation Markers and Major Depressive Disorder in Patients With Chronic Heart Failure: Results From the Sertraline Against Depression and Heart Disease in Chronic Heart Failure Study.

Background Major depressive disorder (MDD) and chronic heart failure (CHF) have in common heightening states of inflammation, manifested by elevated inflammation markers such as C-reactive protein. This study compared inflammatory biomarker profiles in patients with CHF and MDD to those without MDD. Methods The study recruited patients admitted to inpatient care for acute heart failure exacerbations, after psychiatric diagnostic interview. Patients with Beck Depression Inventory (BDI) scores lower than 10 and with no history of depression served as the nondepressed reference group (n = 25). MDD severity was defined as follows: mild (BDI 10-15; n = 48), moderate (BDI 16–23; n = 51), and severe (BDI ≥ 24; n = 33). A Bio-Plex assay measured 18 inflammation markers. Ordinal logistic models were used to examine the association of MDD severity and biomarker levels. Results Adjusting for age, sex, statin use, body mass index, left ventricular ejection fraction, tobacco use, and New York Heart Association class, the MDD overall group variable was significantly associated with elevated interleukin (IL)-2 (p = .019), IL-4 (p = .020), IL-6 (p = .026), interferon-&ggr; (p = .010), monocyte chemoattractant protein 1 (p = .002), macrophage inflammatory protein 1&bgr; (p = .003), and tumor necrosis factor &agr; (p = .004). MDD severity subgroups had a greater probability of elevated IL-6, IL-8, interferon-&ggr;, monocyte chemoattractant protein 1, macrophage inflammatory protein 1&bgr;, and tumor necrosis factor &agr; compared with nondepressed group. The nondepressed group had greater probability of elevated IL-17 (p < .001) and IL-1&bgr; (p < .01). Conclusions MDD in patients with CHF was associated with altered inflammation marker levels compared with patients with CHF who had no depression. Whether effective depression treatment will normalize the altered inflammation marker levels requires further study. Trial Registration: ClinicalTrials.gov NCT00078286.

Expression of interleukin-4 and interferon-gamma in the small bowel of patients with dermatitis herpetiformis and isolated gluten-sensitive enteropathy.

Although possessing a morphologically similarsmall bowel abnormality to patients with isolatedgluten-sensitive enteropathy (GSE), patients withdermatitis herpetiformis (DH) have few gastrointestinal symptoms and exhibit blistering skin lesionsand cutaneous IgA deposits. To determine whetherclinical discrepancies between these gluten-sensitiveconditions might be the result of different patterns of small bowel cytokine expression, duodenalbiopsies were obtained from eight DH patients and nineisolated GSE patients. Biopsies were evaluated forinterleukin-4 (IL-4) and interferon-γ(IFN-γ) expression by reverse-transcriptase polymerasechain reaction (message) and immunohistochemistry(protein). In DH patients, most of whom had no gutsymptoms, IFN-γ mRNA expression was significantly less than in isolated GSE patients withsymptomatic gut disease. Conversely, IL-4 mRNAexpression in DH patients was greater than that foundamong isolated GSE patients. These findings suggest thatthe different clinical phenotypes of glutensensitivity may be caused by variation in cytokineexpression in the small bowel response togluten.

RNA-Seq and ChIP-Seq Reveal SQSTM1/p62 as a Key Mediator of JunB Suppression of NF-κB-Dependent Inflammation

Mice with epidermal deletion of JunB transcription factor displayed a psoriasis-like inflammation. The relevance of these findings to humans and the mechanisms mediating JunB function are not fully understood. Here, we demonstrate that impaired JunB function via gene silencing or overexpression of a dominant negative mutant increased human keratinocyte cell proliferation but decreased cell barrier function. RNA-seq revealed over 500 genes affected by JunB loss-of-function which included an upregulation of an array of proinflammatory molecules relevant to psoriasis. Among these were TNFα, CCL2, CXCL10, IL6R and SQSTM1, an adaptor protein involved in NF-κB activation. ChIP-Seq and gene reporter analyses showed that JunB directly suppressed SQSTM1 through binding to a consensus AP-1 cis-element located around 2 Kb upstream of SQSTM1-trasncription start site. Similar to JunB loss-of-function, SQSTM1-overexpression induced TNFα, CCL2 and CXCL10. Conversely, NF-κB-inhibition genetically with a mutant IκBα or pharmacologically with PDTC prevented cytokine, but not IL6R, induction by JunB-deficiency. Taken together, our findings indicate that JunB controls epidermal growth, barrier formation and proinflammatory responses through direct and indirect mechanisms, pinpointing SQSTM1 as a key mediator of JunB-suppression of NF-κB-dependent inflammation.

TCR Vbeta expression in the small bowel of patients with dermatitis herpetiformis and gluten sensitive enteropathy. Limited expression in dermatitis herpetiformis and treated asymptomatic gluten sensitive enteropathy.

Abstract: Dermatitis herpetiformis (DH) is a blistering skin disease characterized by cutaneous deposits of IgA and an associated, most often asymptomatic, gluten sensitive enteropathy (GSE). Gluten sensitive enteropathy is also seen in patients that do not have skin disease or cutaneous IgA deposits, but do have significant gastrointestinal (GI) complaints. Patients with DH and with GSE without skin disease have similar small bowel morphologic changes and HLA associations and both the skin disease and the GI symptoms can be controlled by a gluten free diet. It is not known what factors allow almost all patients with DH to continue to eat gluten and not develop symptomatic gastrointestinal disease. We have examined the expression of the Vβ T‐cell receptor (TCR) in the small bowel of patients with DH (n=11) and of patients with both symptomatic (n=10) and asymptomatic (n=7) GSE without skin disease to determine if differences in the pattern of TCR Vβ expression are associated with differences in the clinical manifestations of these diseases. TCR Vβ expression was analyzed using RT‐PCR from small bowel biopsies. Patients with DH and those with GSE without skin disease that were on a gluten free diet and asymptomatic were found to express 6.6 and 5.6 out of 20 Vβ families respectively, with no single family preference. Examination of peripheral blood lymphocytes from these patients did not reveal any restriction of TCR Vβ family expression. In contrast, patients with symptomatic GSE expressed 12.6 Vβ families (P< 0.05), with no consistent preferential expression of any single Vβ family between patients. Patients with DH, who are continuing to ingest wheat, show a more restricted pattern of TCR Vβ utilization, similar to that of treated patients with GSE without skin disease, and significantly different from GSE without skin disease patients eating gluten. These findings suggest that the restricted nature of the TCR Vβ expression may play a role in the different clinical manifestations of dermatitis herpetiformis and isolated gluten sensitive enteropathy.