High expression of CD14 CD16 , CD14 CD16 , CD14 CCR2 , and CD14 CD62L cells in childhood TB, and monocyte-derived cytokines reflected both pro- and anti-inflammatory profiles. Higher sCD163 and CD14 CD163 monocytes might help physicians in the differential diagnosis of pulmonary TB in children. Pediatr Pulmonol. 2017;52:675-683. © 2016 Wiley Periodicals, Inc.

Background: Chronic colonization with Pseudomonas (P.) aeruginosa worsens the prognosis of cystic fibrosis (CF) patients. This study aims to analyze the functional properties of neutrophils in CF patients with P. aeruginosa colonization.Methods: Patients with CF (n = 16) were grouped by positivity of P. aeruginosa in sputum culture, as positive (P.+) or negative (P.−), then compared with age and sex matched healthy controls (n = 8). Adhesion molecules, apoptotic index, intracellular CAP-18, interleukin 8 (IL-8), and tumor necrosis factor α (TNF-α) levels of neutrophils, following P. aeruginosa and lipopolysaccharides (LPS) stimulation, were analyzed by flow cytometry. IL-1β, IL-6, TNF-α, and IL-17 plasma levels were determined by Luminex.Results: Patients with CF had increased phagocytosis of Escherichia coli and P. aeruginosa, upregulated oxidative burst and chemotaxis. Increased neutrophil apoptosis was noted in CF patients. In unstimulated conditions, higher levels of CD16 + TNF-α + and CD16 + IL-8 + neutrophils were determined, whereas bacteria and LPS stimulation significantly decreased secretion of CAP-18 from CD16 + neutrophils of CF patients. Plasma levels of IL-1β, TNF-α and IL-17 in P.+ patients were higher than in P.− group. Conclusion:Our findings confirm inadequate neutrophil defense towards pathogens in CF. A significant difference in migration, phagocytosis, oxidative burst, percentage of IL-8 producing neutrophils, IL-1β, TNF-α, and IL-17 secretions were noted among CF patients according to their colonization status, which might induce a further destructive effect on airways, resulting in an unfavorable prognosis for children with CF who also have colonization.

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Background: Obesity-associated asthma (OA) is a difficult to treat asthma phenotype due to its severity and poor response to inhaled steroids. Early-onset allergic (EoOA) and late-onset non-allergic (LoOA) OA are suggested subtypes of this phenotype.Natural Killer (NK) cells are key elements of innate immunity involved in cytotoxicity and immune regulation, with uncertain role in OA pathogenesis. Methods: Early-onset allergic and LoOA patients together with obese non-asthmatic (ONA) controls have been enrolled in the study. Peripheral blood samples have been collected for analysis. Percentages of total NK cells, CD3 − CD56 dim and CD3 − CD56 bright NK cell subsets, cytotoxic activity, intracellular interferonγ, interleukin (IL)-10, IL-13, IL-17 secretion and activatory receptors (NKG2D, NKp46i and NKp44) have been investigated by flow cytometry. The effect of IL-12 and IL-23 stimulation on NK cells and intracellular cytokines in different groups have also been analysed and compared with unstimulated conditions.Results: Results of ONA (n = 5, age 42 ± 8), EoOA (n = 5, age 42 ± 10) and LoOA (n = 8, age 46 ± 8) patients have analysed. Body Mass Index has been found to be negatively correlated with CD69 (p = .022, r = −0.534). NKG2D receptor has been significantly low in CD56 dim cells of asthma population (p = .046). NKp44 receptor expression has increased after IL-12 stimulation in EoOA and control group (p = .02). Intracellular IL-10 content has increased in LoOA and control subjects (p = .018, p = .03) but not in the EoOA group. Intracellular IL-17 level has found be higher in allergic OA group.LoOA patients showed a decreased NK cytotoxicity compared with the early-onset asthma group (p = .05). Conclusion:Our study suggests an impaired NK receptor expression, activation and reduced cytotoxicity in OA patients together with variances between different subtypes of this phenotype. This data would be beneficial for tailoring a more personalized treatment strategy combatting steroid resistance and frequent exacerbations in this group of patients.

5056 Objective: Most patients with myeloproliferative neoplasms (MPN) have mutations that lead to constitutive activation of the tyrosine kinase signal transduction pathways. JAK2V617F mutation is present in about 97% of polycythemia vera (PV), and in 60% of essential thrombocythemia (ET) and primary myelofibrosis (PMF) patients. JAK2-exon-12 mutations, MPL exon-10-mutations are also seen in MPNs. In addition to this mutations, some patients with MPN may acquire mutations of other genes, such as in genes involved in negative regulations of signaling pathways (LNK, CBL, SOCS1, SOCS2, SOCS3) and epigenetic regulations (TET2, ASXL1, DNMT3A, or EZH2). IKZF1 deletions and TP53 mutations are mainly found post-MPN acute myeloid leukemia (AML). Loss of function mutations in TET2, ASXL1 and EZH2 genes, may be early events preceding JAK2V617F but may also secondary genetic event. These mutations are more frequently observed in PMF than in PV and ET. They are also present in other myeloid malignancies such as myelodysplasia and AML. Moreover several cases have been described that are positive for BCR-ABL and JAK2V617F mutation. It is thought that the mutagenic phenotype existing inherently in patients with MPN can generate new mutations. With the contribution of these mutations, PV, ET and PMF might evolve into secondary myelofibrosis or leukemia at different ratios. Activation induced cytidine deaminase (AID) is required for a B cell terminal differentiation mechanism shared by somatic hypermutation and class-switch recombination. Aberrant expression of AID can induce imperfect mutations in non-immunoglobulin genes and was detected in human lymphomas, lung cancer and gastric or intestinal metaplasia. High AID expression is associated with poor prognosis in follicular lymphomas, chronic lymphocytic leukemia and promotes B lymphoid blast crisis in chronic myeloid leukemia. Current study aims to investigate AID expression levels in myeloproliferative neoplasms and their relationship to genetic instability in MPN patients. Material and methods: The patient group consisted of 120 patients with bcr-abl negative myeloproliferative neoplasms (PV: 32 patients, ET: 62 patients and PMF: 18 patients, post-ET or post-PV myelofibrosis: 5 patients, unclassified MPN: 1 patient, MPN-AML: 2 patients). The patients were admitted to Istanbul University, Istanbul Medical Faculty, Division of Hematology Outpatient Clinics. The control group consisted of 69 healthy persons. We evaluated AID expression levels of the patients and of the control group. RNA was extracted from peripheral blood samples of patients and controls, RNA was converted to complementary DNA (cDNA). Using the primers and probes of the target gene (AID) and reference gene HPRT (hypoxanthine phosphoribosyltransferase), cDNA was amplified by realtime polymerase chain reaction and expression levels of the genes were evaluated. Results: We found that AID expression levels in the patient group were significantly higher than in the control group (Target-AID/Reference-HPRT, mean value: 0. 1747 vs 0. 0417, respectively, p<0. 001). We found that the patients age, blood hemoglobin levels, history of thrombosis, bone marrow reticulin levels, serum LDH levels, organomegaly and having JAK2V617F mutation were not correlated with AID expression levels. Conclusion: We showed that AID expression levels in patient with myeloproliferative neoplasms were significantly higher than in healthy control group. The role and the biological significance of high AID expression levels in genetic instability in MPNs need to be elucidated. Disclosures: No relevant conflicts of interest to declare.