Assays-of-the-Immunocompetence.pdf - Assays of the...

Assays of the
Immunocompetence
Objectives:
1. Describe how T and B cells are
enumerated, isolated and how
function is assessed
2. Describe the approaches used to
measure phagocyte and natural
killer cell function.

Assays of Immune Competence
▪ Analysis of :
1. Phagocytic cells
>nitroblue tetrazolium test, microbicidal assay and chemotaxis
assay
2. B cells
> enumeration, B cell function,
lymphoproliferation assay, EAC rosette test
3. T cells
> enumeration, DTH skin test, E-rosette test,
lymphoproliferation assay

Analysis of Phagocytic cells
1. Nitroblue Tetrazolium Test
• Evaluate the ability to generate toxic molecules to
kill microorganisms
• NBT dye : yellow → dark blue : cytoplasm of
phagocyte

Analysis of Phagocytic cells
2. Microbiocidal Assay or Intracellular Killing
Assay
• Evaluate functional integrity
• Tube : bacteria + neutrophils : incubated for 30
minutes
• Normal : neutrophils will kill 85-90% of the
bacteria

Analysis of Phagocytic cells
3. Chemotaxis Assay
• Ability of neutrophils to move or migrate toward a
stimuli
• Migration occurs through nitrocellulose membrane
in an agar medium