Broad-spectrum sunscreens containing the TriAsorB™ filter: In vitro photoprotection and clinical evaluation of blue light-induced skin pigmentation.

BACKGROUND: Blue light (BL), particularly high-energy visible (HEV) light (400-450 nm), can cause skin damage and pigmentation. Therefore, effective sunscreens should offer photoprotection beyond ultraviolet (UV) radiation to also prevent or limit BL-induced cutaneous effects. OBJECTIVES: To evaluate the in vitro BL photostability and photoprotection properties of nine sunscreens containing the broad-spectrum UV/BL phenylene bis-diphenyltriazine (PBDT or TriAsorB™) filter, together with three other organic UV filters, and to assess the in vivo photoprotection level provided by two of these products against BL-induced skin pigmentation. METHODS: In vitro BL photostability and photoprotection factors, comprising the percentage of BL radiation stopped by the product (%BL) and the critical wavelength extended to BL (BL-CW), were determined by spectrophotometry. The in vivo photoprotection provided by two representative sunscreens (i.e. similar formulations, one non-tinted and one tinted) was assessed in two open randomized studies (20 and 16 women, respectively) after exposure of two test areas (with and without sunscreen) on the back of each subject to a 412-nm irradiation dose at 50 J/cm2 , using instrumental and clinical measurements of skin pigmentation. The percentage sunscreen photoprotective effectiveness (%PPE) was calculated by comparing intrasubject post-exposure pigmentation changes between the with and without sunscreen test areas. RESULTS: In vitro, the nine PBDT-containing products were highly photostable and had a BL-CW ≥385 nm and a %BL ≥30% (range: 30%-50%), thus allowing effective BL photoprotection. In vivo, both representative sunscreens prevented BL-induced immediate skin pigmentation (1 and 24 h post-exposure) with %PPE values ranging from 50.7% to 75.5% for colorimetric assessments (p < 0.001) and from 31.2% to 72.7% for visual scores (p ≤ 0.001). CONCLUSIONS: All PBDT-containing sunscreens were considered effective at absorbing BL radiation in vitro. The two representative broad-spectrum sunscreens tested in subjects significantly reduced BL-induced immediate skin pigmentation following single exposure to monochromatic BL radiation.

Discontinuous epitopes on the C2 domain of coagulation Factor VIII mapped by computer-designed synthetic peptides.

The occurrence of alloantibodies against Factor VIII (FVIII) is the main iatrogenic complication in haemophilia A (HA). Anti-FVIII autoantibodies may also spontaneously appear in non-HA patients, leading to acquired haemophilia A. In both contexts, the antibody response against FVIII is complex and difficult to analyse due to the lack of suitable tools. Our purpose was to comprehensively map, at the amino acid level, discontinuous epitopes of the C2 domain of FVIII targeted by patients' antibodies. We synthesized 33 synthetic peptides, which were predicted by the bioinformatic algorithm PEPOP to mimic C2 domain discontinuous epitopes. Using an inhibition assay based on the x-MAP technology, we evaluated their ability to block the binding to the C2 domain of anti-C2 domain antibodies from pooled plasma samples. Nine peptides were thus selected and tested again in individual plasma samples. Our results support the view that C2 domain epitopes are organized as an epitopic mosaic distributed around the molecule, showed that each patient displayed a specific anti-C2 epitopic profile, and confirmed the complexity and variability of the immune response against the C2 domain of FVIII. This ability to finely map epitopes could be further used to follow the antibody specificity modifications over time.

Simultaneous detection and epitope mapping of anti-factor VIII antibodies.

The development of antibodies (Abs) against infused factor VIII (FVIII) is currently one of the most serious complications in the treatment of patients suffering from haemophilia A. Improved prevention and eradication of these anti-FVIII Abs remain a challenge for both clinicians and scientists. Here we describe an immunoassay to simultaneously detect and map the epitope specificity of haemophilia A patients' inhibitors by screening plasma against both heavy and light chains (HC and LC) of human plasma-derived FVIII (pFVIII). The format used was a two-site sandwich assay, where one monoclonal antibody (mAb) specific for the HC or LC was first immobilized on beads, and then incubated with the different forms of pFVIII. After incubation with patients' plasma samples, binding was revealed by a phycoerythrin-labeled secondary Ab. Samples from haemophilia patients with autoantibodies (autoAb) or alloantibodies (alloAb) were screened in this format. The former preferentially recognized the LC, whereas the latter were directed against both LC and HC. This technology appears attractive as it is fast and requires only 100 microl of patient's plasma. Furthermore, not only are anti-FVIII Abs detected, but information on their epitopic specificity is also obtained.

Characteristics, mechanisms of action, and epitope mapping of anti-factor VIII antibodies.

The development of anti-factor VIII (FVIII) antibodies (Abs), also called inhibitors, is currently one of the most serious complications arising during the treatment of hemophilia A patients. Improved prevention and eradication of these Abs remain a challenge both for clinicians and scientists. Numerous studies in the literature have reported on their epitope specificity, on their mechanism of FVIII inactivation, as well as on the methods used for their detection. In this review, we summarize the current knowledge on the nature (isotypes, kinetic properties), epitope properties, and mechanisms of action of anti-FVIII Abs. Furthermore, we present methods for detection and epitope characterization of anti-FVIII Abs with emphasis on the Luminex technique susceptible to facilitate the monitoring of changes in the epitope specificity of these Abs.

Production and characterization of mouse monoclonal antibodies reactive to Chikungunya envelope E2 glycoprotein.

Chikungunya fever is an arbovirosis of major impact in public health in Asia and Africa. Chikungunya (CHIK) virus is member of the genus Alphavirus and belongs to the Semliki Forest (SF) antigenic complex. We describe for the first time a panel of monoclonal antibodies (MAbs) reactive to CHIK envelope E2 glycoprotein. For the screening of E2-specific MAbs, we expressed a recombinant soluble CHIK E2 protein in Drosophila S2 cells. Analyzed by immunological methods, MAbs 3C3, 3E4, and 8A4 were selected on the basis of their reactivity. Their epitopes are located to the outer surface of CHIK virion. These MAbs have no cross reactivity with related members of SF antigenic complex with the notable exception of Igbo-Ora virus. Anti-CHIK E2 MAbs 3C3, 3E4, and 8A4 should be helpful for studying the biology of CHIK virus and pathogenesis of disease. The combination of 8A4 and 3E4 is suitable for developing a specific antigen-capture ELISA.