Abstract

Methods Peripheral blood samples were collected from premature infants at the time of hospital discharge at multiple sites and shipped to a central laboratory. Freshly purified PBMCs were isolated by Ficoll gradient centrifugation, sorted into individual lymphocyte cell types, and processed for total RNA. RNA isolated from CD8 lymphocytes (n=79) was used for RNA-Seq analysis using the Illumina HiSeq2500. Sequences were aligned using the SHRiMP algorithm and expression values were summarized using HTSeq. Normalized gene expression data were analyzed for significant changes in expression using various statistical approaches. Ingenuity Pathway Analysis software was used for gene set interpretation.

Highlights

  • We hypothesize that intrinsic and extrinsic factors associated with oxidative stress drive lymphocyte dysfunction contributing to lung disease in premature infants

  • Peripheral blood samples were collected from premature infants at the time of hospital discharge at multiple sites and shipped to a central laboratory

  • Purified PBMCs were isolated by Ficoll gradient centrifugation, sorted into individual lymphocyte cell types, and processed for total RNA

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Summary

Defining the effects of prematurity on the lymphocyte transcriptome

Soumyaroop Bhattacharya1*, Ravi Misra, Heidi Hyuck, Christopher Slaunwhite, Shannon Castiglione, Deanna Maffett, Anne Marie Reynolds, Gloria Pryhuber, Thomas Mariani. From International Conference on Human Genetics and 39th Annual Meeting of the Indian Society of Human Genetics (ISHG) Ahmadabad, India. From International Conference on Human Genetics and 39th Annual Meeting of the Indian Society of Human Genetics (ISHG) Ahmadabad, India. 23-25 January 2013

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