Interleukin-10 production and DNA polymorphisms in patients with Hemolytic Uremic Syndrome (HUS).

G PINEDA; A SANTIAGO,; L ROSSETTI,; MV RAMOS; MA MONGELOS ; R EXENI; M ABELEYRO; M PALERMO,; A BRUBALLA; G FIORENTINO; C DE BRASI,

Mar del Plata

Congreso; LXVI Reunión Anual de la SAI/SAIC/SAFIS; 2018

SAI

Title: Interleukin-10production and DNA polymorphisms in patients withHemolytic Uremic Syndrome.Authors:Micaela Mongelos1,Gonzalo Pineda1, Andrea Bruballa1, Ramón Exeni2,Martín Abelleyro2, Liliana Rossetti2, Carlos DeBrasi2, Marina Palermo1, María Victoria Ramos1.1Patogénesis E Inmunología De Procesos Infecciosos,IMEX. 2Hospital del Niño, San Justo, La Matanza. 3Genética Molecularde la Hemofilia, IMEX.Introduction: Development of HemolyticUremic Syndrome (HUS) is associated with Shiga toxin and inflammatory response.However, anti-inflammatory mechanisms could also be triggered to limitinflammation in vivo, such asproduction of the regulatory cytokine IL-10. Individual differences in IL-10secretion associated with IL10-promotersingle-nucleotide polymorphisms (SNP), such as rs1800896 (Legacy notation: -1082A>G)in which A- and G-alleles show low and high IL10expression, respectively. Objective: The aim of this work was toevaluate the production of IL-10 and the specific SNP -1082A>G in patientswith HUS.Materials and Methods: Blood samples were collectedfrom HUS patients during the acute period and from Healthy Children (HC).Plasma and mononuclear cells (MNC) were isolated, and MNC were cultured withMedium or LPS (100ng/ml) for 20h. In parallel, absolute number of Monocytes(Mo) in MNC was calculated by flow cytometry. Protein expression of IL-10 wasevaluated in plasma and MNC-supernatants by ELISA. DNA was obtained from MNCand IL10 SNP -1082A>G analysis wasperformed by allele specific-PCR (as-PCR) using a PCR-control template on CTLA4.Results: Plasmatic IL-10 concentration was increased inHUS patients (Mean±SEM (pg/mL): HUS=270.9±117.8*, HC=82.1±7.1, n=15,*p<0.05). The absolute number of Mo from HUS and HC was calculated(Mean±SEM= (0.12±0.03/0.07±0.02).106 Mo/mL). However, the levels ofIL-10 secreted by MNC of both groups were similar after LPS stimulation(Mean±SEM (pg/mL: Basal/LPS (HUS=235±100/6470±1056*; HC=189±70/7630±1959*, n=5,*p<0.05 vs Basal). HUS(HC) DNA samples (n=8(8)) showed genotypes: [A/A]=2(4),[G/G]=1(0), [A/G]=5(4),  Conclusions: These preliminary resultsshow that plasmatic IL-10 is increased in HUS during acute period. Monocytes inMNC population suggests a tendency to be higher in HUS than HC according toprevious reports, however preliminary results show similar IL-10production.  Although carrying the at-risk-allele [G] of IL10 SNP rs1800896 prefigures abouttwice risk of HUS vs HC, largerpopulations are needed to confirm this association.Key words: HUS, SNP, IL-10