Epicutaneous immunization with ovalbumin and CpG induces TH1/TH17 cytokines, which regulate IgE and IgG2a production

Background: Subcutaneous allergen-specific immunotherapy is a standard route for the immunotherapy of allergic diseases. It modulates the course of allergy and can generate long-term remission. However, subcutaneous allergen-specific immunotherapy can also induce anaphylaxis in some patients, and therefore additional routes of administration should be investigated to improve the safety and tolerability of immunotherapy. Objective: We sought to determine whether epicutaneous treatment with antigen in the presence of a Toll-like receptor 9 agonist can suppress TH2-mediated responses in an antigenspecific manner. Methods: Epicutaneous immunization was performed by applying a skin patch soaked with ovalbumin (OVA) plus CpG, and its suppressor activity was determined by using the mouse model of atopic dermatitis. Finally, adoptive cell transfers were implemented to characterize the regulatory cells that are induced by epicutaneous immunization. Results: Epicutaneous immunization with OVA and CpG reduces the production of OVA-specific IgE and increases the synthesis of OVA-specific IgG2a antibodies in an antigen-specific manner. Moreover, eosinophil peroxidase activity in the skin and production of IL-4, IL-5, IL-10, and IL-13 are suppressed. The observed reduction of IgE synthesis is transferable with T-cell receptor (TCR) ab1CD41CD252 cells, whereas IgG2a production is dependent on both TCRab1 and TCRgd1 T cells. Further experiments show that the described phenomenon is myeloid differentiation primary response 88, IFN-g, and IL-17A dependent. Finally, the results suggest that epicutaneous immunization with OVA and CpG decreases the synthesis of OVA-specific IgE and skin eosinophil peroxidase activity in mice with ongoing skin allergy. Conclusion: Epicutaneous application of protein antigen in the presence of adjuvant could be an attractive needle-free and self-administered immunotherapy for allergic diseases

Epicutaneous Immunization with TNP-Ig and Zymosan Induces TCRαβ+ CD4+ Contrasuppressor Cells That Reverse Skin-Induced Suppression via IL-17A

BACKGROUND: Our previous work showed that epicutaneous (EC) immunization with protein antigen e.g. TNP-conjugated mouse immunoglobulin (TNP-Ig) in the form of a patch prior to hapten sensitization inhibits Th1-mediated contact hypersensitivity (CHS) in mice. We also found that suppression of CHS was mediated by TCRαβ(+) CD4(+) CD8(+) T suppressor (Ts) cells producing TGF-β. The aim of the current study was to investigate the role of innate immunity in suppression of CHS. METHODS: Mice were immunized by applying gauze patches containing protein antigen alone or in the presence of zymosan and then tested for CHS response. Adoptive cell transfer experiments were used to study mechanisms involved in reversal of skin-induced suppression. Influence of EC immunization on cytokine production by lymph node cells was measured by ELISA. RESULTS: We found that EC immunization with TNP-Ig and zymosan before TNP-Cl sensitization reverses skin-induced suppression as demonstrated in vivo and in vitro. The reversal of skin-induced suppression was transferable by antigen-specific TCRαβ(+) CD4(+) T contrasuppressor (Tcs) cells. Furthermore, we showed that the contrasuppression was IL-17A dependent and TLR2 and MyD88 independent. CONCLUSIONS: Our work strongly suggests that EC immunization with protein antigen and zymosan reverses skin-induced suppression and that this approach may be a potential tool to increase immunogenicity of weekly immunogenic antigens

Dystrofia twarzowo-łopatkowo-ramieniowa – korelacje elektrofizjologiczno-kliniczne

Background and purpose In recently published reports, electrophysiological findings were analysed, in some facioscapulohumeral muscular dystrophy (FSHD) cases without genetic disease confirmation. In several reports, some electrophysiological findings were described, not specific for myopathy. The aim of study was to analyse electrophysiological findings in a genetically homogeneous FSHD group to find possible relationships between electromyography (EMG) abnormalities and clinical symptoms. Material and methods 37 patients with genetically proven FSHD (23 men and 14 women) aged 7–58 years (mean 28.8 years) were studied. Electromyographic examinations were done according to a uniform scheme for FSHD. Quantitative EMG examination was performed in vastus lateralis, tibialis anterior, deltoid and biceps brachii muscles. Results There was no correlation between clinical features and electrophysiological findings. EMG confirmed myopathic changes in all patients with most advanced changes in tibialis anterior and deltoid muscles. Some of these changes were unspecific for myopathy and the degree of their intensity differed in particular muscles. The most advanced changes were observed in the tibialis anterior and deltoid muscles. The usefulness of the size index for myopathic processes assessment was confirmed. Analysis of so-called outliers for motor unit activity potential parameters did not show any new data for evaluation of the myopathic process. Myopathic changes in our material were not as advanced as those described in classical dystrophies. Histopathological examinations of skeletal muscle were normal in about 1/3 of patients. Conclusions We established that myopathic changes are clearly present in FSHD, with different degrees of intensity, most pronounced in tibialis anterior and deltoid muscles. There was no correlation between electrophysiological findings and clinical features. The size index provided the highest motor unit potential diagnostic sensitivity in FSHD.Wstęp i cel pracy W dotychczas publikowanych doniesieniach analizowano zjawiska elektrofizjologiczne u chorych z dystrofią twarzowo-łopatkowo-ramieniową (facioscapulohumeral muscular dystrophy – FSHD), niekiedy bez genetycznego potwierdzenia rozpoznania. W niektórych badaniach wykazano obecność nieswoistych dla miopatii zmian elektro-fizjologicznych. W pracy podjęto próbę oceny zmian elek-tromiograficznych (EMG) w genetycznie homogennej grupie chorych na FSHD. Przeanalizowano zmiany elektrofizjologiczne i wyłoniono ewentualne korelacje elektrofizjologiczno–kliniczne w FSHD. Materiał i metody Materiał stanowiła grupa 37 pacjentów z genetycznie potwierdzonym rozpoznaniem FSHD (23 mężczyzn i 14 kobiet) w wieku 7–58 lat (średnia wieku: 28,8 roku). Badania EMG przeprowadzono wg jednolitego schematu. Wykonywano ilościowe badanie EMG mięśni: obszernego bocznego uda i piszczelowego przedniego, naramiennego, dwugłowego ramienia. Wyniki Nie wykazano korelacji pomiędzy stopniem nasilenia objawów klinicznych a parametrami EMG. Badania pozwoliły na potwierdzenie miopatycznego charakteru zmian. Zmiany były nieswoiste, a stopień ich nasilenia różny w poszczególnych mięśniach. Największe zmiany obserwowano w mięśniu piszczelowym przednim i naramiennym. Potwierdzono przydatność wskaźnika wielkości w ocenie procesu miogennego, analiza tzw. outliers dla parametrów potencjałów jednostki ruchowej nie wniosła nowych danych. Zmiany miopatyczne w analizowanym materiale nie były tak zaawansowane jak te w klasycznych dystrofiach. Badanie histopatologiczne mięśnia szkieletowego w ok. 1/3 przypadków było prawidłowe. Wnioski W FSHD badania elektrofizjologiczne potwierdzają miopatyczne uszkodzenie mięśni, o różnym stopniu nasilenia w poszczególnych mięśniach. W największym stopniu zmiany obecne są w mięśniu piszczelowym przednim i naramiennym. Parametry elektromiograficzne nie wykazują korelacji ze stopniem zaawansowania objawów klinicznych. W opracowaniu autorów największą diagnostyczną czułość dla zmian miopatycznych wykazywał wskaźnik wielkości potencjałów czynnościowych jednostek ruchowych