The relative roles of various autoantibodies against IL-17-type cytokines in susceptibility to chronic mucocutaneous candidiasis (CMC) in patients with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) remain poorly defined. The purpose of this longitudinal study was to analyze the relationship between the occurrence of mucocutaneous candidiasis and levels of anti-IL-17A, anti-IL-17F and anti-IL-22 autoantibodies. We studied six APECED patients from four families with various disease manifestations. Clinical data were collected during regular follow-up. Anti-endocrine organ antibody levels and clinical chemistry and immunology parameters were determined in routine laboratory assays on freshly isolated serum. Levels of autoantibodies against IL-17A, IL-17F, IL-22, IFN-α, IFN-ω and TNF-α, and cytokine release by Candida-exposed blood cells were determined by ELISA. Mutations were analyzed by sequencing genomic DNA. Four patients carried the germline c.769C > T homozygous nonsense mutation, which results in R257X truncation of the AIRE protein, and two patients from the same family were compound heterozygous for the c.769C > T/c.1344delC mutation. We found persistently high levels of antibodies against IL-17A in the serum samples of one patient presenting CMC since infancy and low or undetectable anti-IL-17A antibody levels in the sera of five patients with no candidiasis or without severe candidiasis. By contrast, levels of autoantibodies against IL-17F and IL-22 were higher in all patients than in healthy controls. Release of IL-17-type cytokines by Candida-exposed blood mononuclear cells was low or negligible in all patients tested. We suggest that anti-IL-17A antibodies may play an important role in the predisposition to candidiasis of APECED patients. However, the lack of severe CMC in APECED patients with high levels of IL-17F and anti-IL-22 autoantibodies clearly calls into question the role of these antibodies as the principal cause of cutaneous and mucosal candidiasis in at least some APECED patients. These data also suggest that the impaired release of IL-17-type cytokines by blood cells may be an element of the immunopathology of CMC in APECED patients.

BACKGROUND:Chronic mucocutaneous candidiasis disease (CMCD) may result from various inborn errors of interleukin (IL)-17-mediated immunity. Twelve of the 13 causal mutations described to date affect the coiled-coil domain (CCD) of STAT1. Several mutations, including R274W in particular, are recurrent, but the underlying mechanism is unclear.OBJECTIVE:To investigate and describe nine patients with CMCD in Eastern and Central Europe, to assess the biochemical impact of STAT1 mutations, to determine cytokines in supernatants of Candida-exposed blood cells, to determine IL-17-producing T cell subsets and to determine STAT1 haplotypes in a family with the c.820C>T (R274W) mutation.RESULTS:The novel c.537C>A (N179K) STAT1 mutation was gain-of-function (GOF) for γ-activated factor (GAF)-dependent cellular responses. In a Russian patient, the cause of CMCD was the newly identified c.854 A>G (Q285R) STAT1 mutation, which was also GOF for GAF-dependent responses. The c.1154C>T (T385M) mutation affecting the DNA-binding domain (DBD) resulted in a gain of STAT1 phosphorylation in a Ukrainian patient. Impaired Candida-induced IL-17A and IL-22 secretion by leucocytes and lower levels of intracellular IL-17 and IL-22 production by T cells were found in several patients. Haplotype studies indicated that the c.820C>T (R274W) mutation was recurrent due to a hotspot rather than a founder effect. Severe clinical phenotypes, including intracranial aneurysm, are presented.CONCLUSIONS:The c.537C>A and c.854A>G mutations affecting the CCD and the c.1154C>T mutation affecting the DBD of STAT1 are GOF. The c.820C>T mutation of STAT1 in patients with CMCD is recurrent due to a hotspot. Patients carrying GOF mutations of STAT1 may develop multiple intracranial aneurysms by hitherto unknown mechanisms.

In July, 2011, a 47-year-old woman and her 16-year-old daughter presented to our services because of persistent and recurrent oral candidosis. The mother began experiencing genital candidosis at the age of 3 years and her nails became affected when she was 4 (see appendix). From the age of 5 years, she presented with herpetic vesicles two to four times per year, mainly affecting her face. She had chickenpox when she was 5 years old and 1 year later had shingles. At 21 years, she developed cutaneous herpes lesions over an 8×5 cm area on the right side of her face. At 30 years, she had another episode of shingles on her right trunk. At 45 years, retrosternal discomfort and pain on swallowing solid food developed. Gastroscopy showed 6 cm long narrowing of the oesophagus. Candida albicans was isolated. Except for one episode of pneumococcal pleuropneumonia with confluent lobar consolidation at the age of 38 years, no causal organisms of pneumonia were identified. The daughter had had recurrent oral and pharyngeal candidosis since infancy, and genital and nail candidosis from the ages of 12 and 16 years. She had chickenpox when she was 3 years old, but no primary episode of HSV infection had been reported. Recurrent infections with HSV and varicella began when she was 6. From that time, she had recurrent HSV infections mainly affecting the lips and mouth, mouth ulcers every 1–3 months since infancy, and two episodes of cutaneous varicella infection affecting the right thigh and knee. Department of ErrorTóth B, Méhes L, Taskó S, et al. Herpes in STAT1 deficiency. Lancet 2012; 379: 2500—In this Case Report (June 30), the title should have been: "Herpes in STAT1 gain-of-function mutation". This correction has been made to the online version as of Aug 31, 2012. Full-Text PDF

Risk factors for invasive infections by heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) may involve resistance to opsonophagocytosis and bacterial killing. hVISA strains typically have a thickened cell wall with altered peptidoglycan cross-linking. To determine whether hVISA may be endowed with an increased resistance to phagocytosis, this study assessed the characteristics of uptake and killing by granulocytes of three hVISA strains. All isolates were analysed by multilocus sequence typing and staphylococcal chromosome cassette mec typing. One of the strains belonged to the Hungarian meticillin-resistant S. aureus (MRSA) clone ST239-MRSA-III and the other two to the New York/Japan MRSA clone ST5-MRSA-II. In the presence of 10 % normal serum, the extent of phagocytosis and killing by blood granulocytes was equivalent for hVISA, MRSA and meticillin-sensitive S. aureus (MSSA) strains. Using granulocytes and serum from one patient who survived hVISA infection, the rate of phagocytosis and killing was also found to be comparable to that by control cells in the presence of 10 % serum. However, phagocytosis and killing of hVISA and MRSA (ATCC 25923) strains by normal granulocytes was markedly decreased in the presence of low concentrations (1 and 2.5 %) of serum from the patient who survived hVISA infection compared with that found with normal human serum. These data suggest that hVISA and MRSA isolates may be more resistant to opsonophagocytosis and bacterial killing than MSSA isolates, at least in some cases.

s of the 16th International Symposium on Infections in the Immunocompromised Host s1 predicting a p.K219fsX234 change in the nibrin protein. The patient presented with progressively growing, painless, erythematous and indurated plaques and ulcers on the right leg at 6 years of age. Methods: Mutational analysis of genomic NBS1 was performed using the BigDye Terminator Cycle sequencing kit. Mycobacterium DNA in skin specimens was detected by PCR using Mycobacterium genus and M. tuberculosis species-specific primers. Biopsy specimens were used for culture. Sections of the formalin-fixed paraffin-embedded skin biopsy samples were stained with Ziehl-Neelsen and H&E stainings. Results: Both culture and Ziehl-Neelsen staining gave negative results but PCR testing with Mycobacterium genusand M. tuberculosis species-specific primers confirmed the diagnosis of CTB. A unique feature of this patient was the presentation of CTB with no pulmonary or systemic manifestation of TB. Conclusions: This is the first report describing the association of NBS with CTB clearly showing the importance of genetic testing to define mycobacterial etiology of CTB. Acknowledgements: Supported by a TÁMOP 4.2.2-08/1-2008-0015 Grant P3 Dectin-1-Mediated Immunity is Redundant for Host Defense against Mucocutaneous Candidiasis Beáta Tóth, Péter Gogolák, Szilvia Taskó, László Maródi. University of Debrecen, Medical and Health Science Center, Debrecen, HU. Background: Two groups have recently reported that genetic deficiency of Dectin-1 due to mutation of CLEC7A (Tyr238X/Tyr238X or Tyr238X/WT) or premature termination of the caspase recruitment domain-containing protein 9 resulting from CARD9 mutation (Gln295X/Gln295X) may predispose patients to chronic mucocutaneous candidiasis CMC (1-3). Materials: We studied the frequency of CLEC7A mutation in 51 Hungarian healthy individuals. Genomic DNA was isolated from blood leukocytes. Blood monocytes and monocye-derived dentritic cells (MDDCs) were used to study phenotypic expression of Dectin-1. Methods: CLEC7A was sequenced with the Big Dye Terminator cycles sequencing kit and analyzed using an ABI 3130 capillary sequencer. Mixed mononuclear cells (MCs) were isolated from heparinized venous blood. MDCCs were obtained by culture of monocytes isolated by using immunomagnetic cell separation assay, in AIMV medium supplemented with 75 ng/mL GM-CSF and 100 ng/mL IL-4 for 5 days. Measurement of receptor expression was performed by using a FACSCalibur flow cytometer. Secretion of IL-17A by MCs Immunodeficiency Syndromes P1 Bypassing Cytokine and Cytokine Receptor Defects: Using Interferon Alpha to Replace Interferon Gamma Elizabeth Sampaio, Hannelore Bax, Ervand Kristosturyan, Li Ding, Li Madison, Sarah Browne, Steven Holland. NIH, Laboratory of Clinical and Infectious Diseases, Bethesda, US. Background: Patients with mutations in the genes encoding the interferon gamma receptor (IFN-gR) and signal transducer and activator of transcription 1 (STAT1) often present with extensive and therapy resistant mycobacterial infections. However, not much is known about STAT1 activation by IFNα in the treatment of mycobacterial infections and in situations in which IFN-γ signalling is impaired. We evaluated the effect of IFN-γ and/or IFN-α treatment on cellular responses in vitro in cells from patients with mutations in the IFN-γR, STAT1, or with high titer autoantibodies to IFN-γ. Materials: Purified elutriated monocytes and EBV-transformed B cells were used in the experiments. Antibodies were purchased from Cell Signaling and Becton&Dickinson. Methods: Immunoblots, co-immunoprecipitation, flow cytometry, and real time PCR were performed on cells from normal donors and from patients with IFN-g signal defects. Cells were treated in vitro with IFN-γ(200U/ml), IFN-α(1,000U/ml) and IFN-γ/α co-treatment for 15-30min (STAT phosphorylation) and 3h (gene expression). Results: IFN-α activated STAT1 phosphorylation in cells lacking a functional IFN-γR. Both IFN-γ and IFN-α activated STAT1 in patients with STAT1 mutations, but usually to a lesser extent than in normal cells. Clinically, we used IFN-α in patients with refractory mycobacterial infections who had mutations in IFN-γR1, IFN-γR2, or had high level autoantibodies to IFN-γ. IFN-α used subcutaneously three times weekly or pegolated interferon once weekly were well tolerated. IFN-α was more effective against M. avium complex infections than M. abscessus, but whether this reflects a dose-dependence of treatment of M. abscessus or differential activity is still unclear. Four patients with mutations affecting the IFN-γR had objective improvements in infection burden and clinical condition after initiating IFN-α. The combined use of IFNγ and IFNαα was associated with increased STAT1-mediated response and activation of typical IFN-γγ genes (IP10, CXCL9, IRF1). Increased protein levels and enhanced STAT1/STAT3 association were also observed. Conclusions: An underlying mechanism involving the crosstalk between STAT1 and STAT3 is proposed implying the balance between these opposing regulators to be central to the effects induced by IFNs. Our results suggest that the overlap of cytokine signal transduction pathways can be exploited clinically to treat severe infections. P2 Association of Nijmegen Breakage Syndrome with Primary Cutaneous Tuberculosis Melinda Erdős, Beáta Tóth, Mohamed Mahdi, Imre Veres, Mária Kiss, Éva Remenyik, László Maródi. University of Debrecen, Medical and Health Science Center, Debrecen, HU; Szent-Györgyi University, Szeged, HU. Background: Cutaneous tuberculosis (CTB), a very rare manifestation of mycobacterial infections tipically occurs with pulmonary or systemic manifestations of tuberculosis. Materials: We report here on a 7-year-old Caucasian girl with clinical and immunological manifestations of Nijmegen Breakage syndrome (NBS) caused by a c.657del5bp deletion mutation of the NBS1 gene Figure 1. Facial characteristics of NBS (up) and lupus vulgaris on the right

The aim of this study was to identify mutations in the gene encoding for lysosomal beta-glucocerebrosidase (GBA; gene symbol, GBA) in Hungarian patients with Gaucher disease (GD), and to study genotype-phenotype relationships. Genotypes and allele variations in 27 patients with type I GD of 25 unrelated families were studied. Of the 54 mutant alleles, we detected 38 frequent (N370S, 22/54; RecNciI, 8/54; L444P, 8/54) and 9 rare (N188S, R257Q, R285C, G377S, R120W, T323I, 84GG, 1263-1317del and 1263-1317del/RecTL) mutations. In addition, we identified two novel mutations. The N370S/RecNciI genotype found in 8 patients and the N370S/L444P genotype found in 5 patients were the most frequent genotypes in this cohort. In 22 patients the mutations occurred in heterozygosity with the N370S sequence variant, and one patient was homozygous for the L444P mutation. These data suggest that N370S, RecNciI, and L444P are the most prevalent mutations in Hungarian patients with GD. This mutation profile is characteristic for a Caucasian (non-Jewish) population. The c.260G>A and c.999G>A missense mutations are described here for the first time in GD patients contributing to the panel of reported GBA mutations.