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Flow Cytometry

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Separation of Immune Cell Subpopulations in Peripheral Blood Samples from Children with Infectious Mononucleosis

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Published on: September 7, 2022

Flow-based sorting of neonatal lymphocyte populations for transcriptomics analysis.

Ravi S Misra¹, Soumyaroop Bhattacharya², Heidie L Huyck¹

¹Department of Pediatrics, Division of Neonatology, University of Rochester Medical Center, Rochester, NY 14642, United States.

Journal of Immunological Methods

|July 21, 2016

Summary

This study developed a method to analyze T lymphocyte gene expression in preterm infants, identifying potential biomarkers for chronic lung disease. The validated procedure enables genome-wide expression data generation from newborn blood lymphocyte subsets.

Keywords:

BPD Flow sorting Lymphocytes PBMC Prematurity RNASeq T-Cell

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Published on: September 7, 2022

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Isolation of Precursor B-cell Subsets from Umbilical Cord Blood

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Published on: April 16, 2013

Area of Science:

Neonatal immunology
Respiratory medicine
Genomics

Background:

T lymphocytes play a role in chronic lung disease pathogenesis in preterm infants.
Assessing the lymphocyte transcriptome may reveal disease biomarkers and mechanisms.

Purpose of the Study:

To establish a validated procedure for generating genome-wide expression data from isolated lymphocyte subpopulations in newborn blood.
To identify potential biomarkers and understand mechanisms of chronic lung disease in preterm infants.

Main Methods:

Collected small peripheral blood volumes from preterm infants.
Isolated peripheral blood mononuclear cells (PBMCs) and sorted specific lymphocyte subsets using fluorescence-activated cell sorting.
Performed high-throughput sequencing (RNASeq) on isolated lymphocyte RNA.

Main Results:

Successfully obtained sufficient PBMCs for sorting and RNASeq from 67% of samples.
RNA of adequate quantity and quality was recovered from sorted cells for RNASeq analysis.
Demonstrated reproducible recovery of purified lymphocyte populations over time.

Conclusions:

A validated procedure exists for generating genome-wide expression data from isolated lymphocyte subpopulations from neonatal blood.
This method supports the identification of biomarkers and mechanisms in neonatal chronic lung disease.