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Primary immune deficiencies are usually attributed to genetic defects and, therefore, frequently referred to as inborn errors of immunity (IEI). We subjected the genomic DNA of 333 patients with clinical signs of IEI to next generation sequencing (NGS) analysis of 344 immunity‐related genes and, in some instances, additional genetic techniques. Genetic causes of the disease were identified in 69/333 (21%) of subjects, including 11/18 (61%) of children with syndrome‐associated IEIs, 45/202 (22%) of nonsyndromic patients with Jeffrey Modell Foundation (JMF) warning signs, 9/56 (16%) of subjects with periodic fever, 3/30 (10%) of cases of autoimmune cytopenia, 1/21 (5%) of patients with unusually severe infections and 0/6 (0%) of individuals with isolated elevation of IgE level. There were unusual clinical observations: twins with severe immunodeficiency carried a de novo CHARGE syndrome‐associated SEMA3E c.2108C>T (p.S703L) allele; however, they lacked clinical features of CHARGE syndrome. Additionally, there were genetically proven instances of Netherton syndrome, Х‐linked agammaglobulinemia, severe combined immune deficiency (SCID), IPEX and APECED syndromes, among others. Some patients carried recurrent pathogenic alleles, such as AIRE c.769C>T (p.R257*), NBN c.657del5, DCLRE1C c.103C>G (p.H35D), NLRP12 c.1054C>T (p.R352C) and c.910C>T (p.H304Y). NGS is a powerful tool for high‐throughput examination of patients with malfunction of immunity.

The genome structure of three ciprofloxacin-resistant Mycoplasma hominis clinical isolates was studied using next-generation sequencing on the Illumina platform. The protein sequences of the studied Mycoplasma strains were found to have a high degree of homology. Mycoplasma hominis (M45, M57, MH1866) was shown to have limited biosynthetic capabilities, associated with the predominance of the genes encoding the proteins involved in catabolic processes. Multiple single-nucleotide substitutions causing intraspecific polymorphism of Mycoplasma hominis were found. The genes encoding the efflux systems ABC transporters (the ATP-binding cassette superfamily) and proteins of the MATE (multidrug and toxic compound extrusion) family were identified. The molecular mechanism of ciprofloxacin resistance of the Mycoplasma hominis M45 and M57 isolates was found to be associated with the Ser83Leu substitution in DNA gyrase subunit A. In the Mycoplasma hominis MH1866 isolate it was related to the Lys144Arg substitution in topoisomerase IV subunit A.

BackgroundNLRP12-related autoinflammatory disease (NLRP12-AID) is an exceptionally rare autosomal-dominant disorder caused by germ-line mutations in NLRP12 gene. Very few patients with NLRP12-AD have been identified worldwide, therefore there is a scarcity of data on phenotypic presentation of this syndrome.ObjectivesHere we provide evidence that NLRP12-AID may have clinical manifestations characteristic for primary immune deficiencies (PID).Methods246 children with periodic fever (PF) of unknown origin were subjects to the next generation sequencing (NGS) analysis; 213 of these patients had signs of primary immunodeficiency (PID) manifested by recurrent infections, while 33 kids had isolated PF. The NGS panel was composed of 302 genes implicated in PID and/or AID.ResultsNLRP12 variants were detected in 20 cases (see the table 1). Median age of first AID-related fever episode was 12 months, ranging from 2 months to 13 years. Increased association with immune-mediated diseases (n=15, 75%) was observed. Nine patients demonstrated increased susceptibility to infection and two children suffered from Crohn’s disease. Administration of short courses of NSAID or corticosteroids resulted in resolution of the disease flare. In one severe case canakinumab (anti-interleukine-1β antibody) was successfully used.Abstract OP0098 – Table 1NLRP12 mutationID, sexNLRP12 related symptomsComorbid disease p.H304YPK, f RI, m AA, fPF, 1d/weekCyclic vomiting with hCRP since newborn Permanent aphtous stomatitis, PFAPA, APCVID, AIHA, pancytopenia, S, ILD Severe CD, food and drug allergyAsn715>SerZV, m KA, m VM, mPF (infections?) PF 5–7d/month with LA, redness, eye painCongenital red-cell aplasia, AIHA, S, LACVID, AIHA, S, LA, ILD, thrombocytopeniaChronic posterior scleritisp.F402LGA, f GE, mPFPFEpisodes of severe respiratory infection Severe CD, SThr861SerCA, m KS, mPF (infections)Episodic AP, diarrhoeaMeningitis, septic shock, otitis,Gln417LeuAR, m SK, m1d PFPersistent fever Severe recurrent MAS, ILDArg352CysDE, fPF, S, A, AP, urticaria, growth delay,p.Ala57SerAK, mRespiratory and intestinal infections with PFBacterial meningitisp.Ala5GluHT, m PFAPA-like syndrome headackes+eye painp.Arg723GlnBA, fPF, Ap.Val936LeuGA, fPF (infections?)Recurrent purulent sinusitis w\o feverp.Tyr246CysES, fFCAS, relapsed aseptic peritonitisp.Gly52Ser KV, fPF (infections?)p.R738fsSD, m1–4d of PF, cold-induced, 3–4/year,pneumonia, febrile seizures, brain oedemaAbbreviations: A-arthralgia, ABA-abatacept, AIHA-autoimmune hemolytic anaemia, AP-abdominal pain, CD-Crohn’s disease, CKB-canakinumab, CVID-common variable immunodeficiency, d-day, ILD-interstitial lung disease, LA-lymphadenopathy, PF-periodic fever, S-splenomegalyConclusionsSignificant number of patients with genetically assigned diagnosis of NLPR12-AID have clinical features which close resemble PID. This phenotypic overlap may result in underdiagnosis of NLPR12-AID among patients with PID.AcknowledgementsThis work has been supported by Russian Foundation for Basic Research (grant number RFBR 16–04–00 15...

Background:Chronic non-bacterial osteomyelitis (CNO) is an immune-mediated disease associated with cytokine dysbalance.Objectives:The aim of our study was to evaluate the cytokines levels in CNO and compare to juvenile idiopathic arthritis (JIA) – disease with immune-mediated mechanism.Methods:The diagnosis of CNO made with criteria, proposed by Jansson (2007, 2009), after the exclusion of other causes of bone disease [1]. We included 42 patients with NBO, 28 patients with non-systemic juvenile idiopathic arthritis (JIA). We evaluated plasma levels of 14-3-3 protein, S100A8/S100A9-protein, interleukine-6 (IL-6), interleukine-18 (IL-18), interleukine-4 (IL-4), interleukine-17 (IL-17), interleukine-1β (IL-1 β) and tumor necrosis factor-α (TNFα) in 2 groups by the ELISA. Statistical analysis was carried out with Statistica 10.0 software. We utilized descriptive statistics (Me; IQR), Mann-Whitney tests.Results:We have found differences in the proinflammatory biomarkers between CNO, JIA. Patients with NBO had lower levels of studied cytokines, exclude14-3-3-protein, S100A8/S100A9 and interleukin-6 compare to JIA patients (table 1).Table 1.Comparison the cytokine levels between CNO, JIA NParameterNBO (n=42)JIA (n=28)pHemoglobin, g/l112 (104; 124)120 (114.5; 126.0)0.02WBC x 109/l7.9 (7.0; 10.5)8.0 (6.7; 10.0)0.86PLT x 109/l347 (259; 408)336.5 (274.0; 390.5)0.98ESR. mm/h25.0 (9.0; 46.0)8.5 (2.5; 13.0)0.013CRP, mg/l6.1 (0.6; 2.4)1.8 (0.4; 11.9)0.02714-3-3, ng/ml21.4 (18.5; 27.1)19.9 (18.0; 27.8)0.77S100A8/S100A9, ng/ml5.9 (5.2; 6.5)5.9 (5.0; 6.2)0.76IL-6, ng/ml126,2 (112.8; 137.5)132.4 (117.4; 142.9)0.16IL-18, ng/ml270.1 (200.1; 316.1)388.3 (373.9; 405.1)0.0000001IL-4, ng/ml15.3 (11.5; 18.2)18.7 (16.2; 20.2)0.003IL-17, ng/ml83.1 (71.1; 97.3)99.2 (87.3; 115.8)0.003IL-1b, ng/ml47.4 (42.0; 51.3)70.8 (65.3; 73.6)0.0000001TNFa, ng/ml19.4 (17.8; 21.3)23.1 (20.2; 25.9)0.0006Conclusion:Patients with CNO had less proinflammatory activity then JIA patients, besides IL-6 and S100A8/S100A9. Further investigations required for finding new more precise biomarkers and finding possible molecular targets for treatment.This work supported by the Russian Foundation for Basic Research (grant № 18-515-57001)References:[1]Jansson AF, et al. Clinical score for nonbacterial osteitis in children and adults. Arthritis Rheum. 2009;60(4):1152-9.Disclosure of Interests:None declared