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Neutrophil Assessment Platform

Study Aim

This project aims to develop and use a platform that can evaluate neutrophil dysfunction in ME/CFS.

Investigators

  • Vanessa Velasco
  • Ronald Davis, PhD

Updates and Potential

  • The neutrophil assessment platform has been designed and tested. We achieved neutrophil isolation from unprocessed healthy control blood samples. With this device, we will compare how neutrophils from ME and HCs behave during an inflammation event and evaluate if ME neutrophils are dysfunctional.
  • Before an inflammation event (ME=3, HC=3): Neutrophils were monitored for 3 hr and assessed every 90 s. ME patients present higher averages of neutrophil dead and apoptotic populations from 0.25-2.5hr. Higher dead populations are observed until 3hr, but ME neutrophils undergoing apoptosis level with HCs at t=2.5-3hr. ME patient neutrophils have slightly lower populations of ROS for t=0-0.5 hr, then from t= 0.5-2hr they indicate higher ROS populations. After t=2hrs, ROS population decreases below the average of HCs. ME neutrophils migrate at lower average speeds, shorter distances, have smaller displacements and movement durations.
STUDY HYPOTHESIS AND DESCRIPTION

Neutrophil dysfunction (ND) is associated with chronic inflammation conditions. In ME/CFS, a small number of neutrophil studies indicated increased early apoptosis, reduced reactive oxygen species (ROS), up-regulation of a cytotoxic granule peptide emitted during neutrophil attacks, and decreased surface protein CD177, vital in neutrophil migration and bacteria killing activities. Yet, hallmarks of ND and possible sources of inflammation including abnormal migration and host attacks like neutrophil extracellular traps (NETosis) remain unexamined.

We propose to extend the current knowledge of ND in ME/CFS. Thus, we will perform a multi-parameter analysis evaluating apoptosis, migration, ROS, and NETosis in both pre-and post-inflammation neutrophil populations to potentially discover a unique disease signature and provide much needed insight into inflammation mechanisms in ME/CFS.

OBJECTIVES

Preparation for blood test. Beautiful young African woman doctor in white medical uniform ready to take blood sample from patient's vein

  1. Develop and implement a neutrophil assessment platform that facilitates neutrophil isolation from patient whole blood and artificially simulates an inflammation event.
  2. Before and after an inflammation event, compare ME/CFS and healthy patients’ neutrophil characteristics including: apoptosis, migration, Reactive Oxygen Species (ROS) and Neutrophil Extracellular traps (NETs).
Myalgic Encephalomyelitis / Chronic Fatigue Syndrome (ME / CFS) Post Treatment Lyme Disease Syndrome (PTLDS), Fibromyalgia Leading Research. Delivering Hope.Open Medicine Foundation®

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