The Neutrophil Monitoring Laboratory assesses neutrophil dysfunction in patients with different primary immunodeficiencies. Our CLIA-certified laboratory develops and performs high-complexity assays to analyze and evaluate biological specimens for neutrophil activity and various inflammatory mediators or cytokines. We provide clinical, translational, and basic research to support the Laboratory of Clinical Immunology and Microbiology at the National Institute of Allergy and Infectious Diseases. 

High-complexity assays for diagnosing patients

The laboratory develops and performs high-complexity, CLIA-certified assays used to diagnose patients with:   

  • Chronic granulomatous disease, a genetic immunodeficiency characterized by recurrent bacterial and fungal infections, caused by a defect in reactive oxygen species production.

  • Leukocyte adhesion deficiency, a defect that prevents neutrophils from migrating to infection sites.

We employ multiplex analysis platforms to monitor levels of inflammatory cytokines and other biomarkers in biological specimens or culture supernatants.

 

Providing resources through assay development, biorepository access

We develop state-of-the-art assays to evaluate diverse neutrophil functions, such as reactive oxygen species production, chemotaxis, cell-shape change and spreading, exocytosis, and alterations in surface antigen expression. The Neutrophil Monitoring Laboratory maintains an extensive repository of patient plasma, cryopreserved peripheral blood mononuclear cells, Epstein-Barr virus–transformed B-cell lines, dermal fibroblasts, DNA, and RNA, all of which represent a valuable resource for investigators.  

Diagnosis of chronic granulomatous disease and its subtypes 

Our laboratory develops and performs high-complexity, CLIA-certified assays used to diagnose patients with chronic granulomatous disease, a genetic immunodeficiency characterized by recurrent bacterial and fungal infections, caused by a defect in reactive oxygen species production. 

  • Evaluate reactive oxygen species production

  • Assess phagocyte oxidase protein expression by Western blotting or fluorescence-activated cell sorting analysis of proteins in permeabilized neutrophils 

  • Identify genetic defects using Sanger sequencing or droplet digital polymerase chain reaction 

  • Distinguish p47phox CGD subtypes and determine the number of GTGT versus delta GT at the start of exon 2 by using ddPCR

Neutrophil dysfunction associated with actinopathies 

We study increased micropinocytosis, neutrophil vacuolization, and changes in neutrophil surface-antigen expression and activation states. 

  • Assess chemotaxis using a modified Boyden chamber (Neuro Probe) or CHEMOTAXIScan instrumentation 

  • Evaluate neutrophil morphology alterations using light and electron microscopy 

Analysis of inflammatory cytokines and other biomarkers in biological specimens 

We analyze serum and plasma from COVID-19 patients, patients with rare genetic immunodeficiencies, and patients undergoing hematopoietic stem-cell transplantation. 

  • Analyze biomarkers using enzyme-linked immunosorbent assay-based or bead-based multiplex platforms 

  • Detect neutrophil trap formation via image analysis and fluorescence microscopy or fluorescence spectrophotometry monitoring of DNA release into the extracellular space 

  • Characterize neutrophil chemotaxis using image analysis