137 - Lymphocyte phenotype and activation in children at school-age following neonatal encephalopathy

Poster Number: 137
Publication Number: 137.362

Eman Isweisi, Trinity college Dublin, Saggart, Dublin, Ireland; Johana M. Isaza-Correa, Trinity College Dublin, Dublin, Dublin, Ireland; Arthur White, Trinity College Dublin, the University of Dublin, Dublin, Dublin, Ireland; Jason Wyse, Trinity College Dublin, Dublin, Dublin, Ireland; John Finbarr. Murphy, The National Maternity Hospital, Holles Street, Dublin2, Dublin 4, Dublin, Ireland; Afif EL-Khuffash, Royal College of Surgeons in Ireland, Dublin, Dublin, Ireland; Elaine Ní Bhraonáin, Irish Neonatal Health Alliance, Gorey, Wexford, Ireland; Mandy Daly, Irish Neonatal Health Alliance, BRAY, Wicklow, Ireland; John J. O'Leary, Trinity College Dublin Ireland, Dublin, Dublin, Ireland; Eleanor J. Molloy, CHI, dublin, Dublin, Ireland

Background: Neonatal Encephalopathy (NE) results in multi-organ dysfunction which may persist later in childhood

Objective: •To evaluate adaptive immune markers of inflammation in children at ages 2 to 5 years who experienced neonatal encephalopathy(NE)

• to determine if inflammation persist in childhood .

Design/Methods: •Lymphocytes phenotype and intracellular cytokine production was analysed by flow cytometry in children

     ages 2-5 years who experienced NE as new-born (n=18)           

     and control children(n=11,2-6years).    
•Changes in subpopulations of lymphocytes (CD3+, CD19+, CD56+,CD4+,D8+,TCRVdelta1+,TCRVdelta2+)were evaluated.

•Whole blood was stimulated without/with lipopolysaccharides (LPS) for 20 hours, followed by 4 hours of Monensin, Paramethoxyamphetamine (PMA) and Ionomycin to evaluate intracellular cytokines (IL-17a, TNF-α, IFN-γ) production and expression of lysosomal-associated membrane protein-1 (LAMP-1 or CD107a) in subpopulations of lymphocytes (Total CD3+, iNKT, γδ T cells, CD56+).

Results:

Evaluation of intracellular cytokines after stimulation, shows significant increase of degranulation marker CD107a in CD3+, iNKTs and γδ T cells in NE children (n=18) compared to controls (n=11). γδ T cells also expressed higher CD107a at basal level in the same group. TNF-α was seen increased in CD3+, iNKTs and γδ T cells in unstimulated samples of NE children compared to controls, and in stimulated iNKTs. IL-17a was significantly higher in iNKTs at basal level in NE compared to healthy children. No major differences in intracellular cytokines expression was found in NK cells. However, a trend of IFN-γ increase in NKs and γδ T cells

Conclusion(s):

Increased in degranulation markers and pro-inflammatory cytokine TNF-α might indicate persist inflammation mediated by lymphocytic populations in NE children.