Establishment of a healthy human range for the whole blood “OX40” assay for the detection of antigen‐specific CD4+ T cells by flow cytometry

R Sadler, EAL Bateman, V Heath… - Cytometry Part B …, 2014 - Wiley Online Library
R Sadler, EAL Bateman, V Heath, SY Patel, PP Schwingshackl, AC Cullinane, L Ayers
Cytometry Part B: Clinical Cytometry, 2014Wiley Online Library
Background Clinical investigation of antigen‐specific T cells in potentially immunodeficient
patients is an important and often challenging aspect of patient diagnostic work up. Methods
for detection of microbial exposure to the T‐cell compartment exist but are laborious and
time consuming. Recently, a whole blood technique involving flow cytometry and detection
of CD25 and OX40 (CD134) expression on the surface of activated CD4+ T cells was shown
to be accurate and concordant when compared with more traditional methods of antigen …
Background
Clinical investigation of antigen‐specific T cells in potentially immunodeficient patients is an important and often challenging aspect of patient diagnostic work up. Methods for detection of microbial exposure to the T‐cell compartment exist but are laborious and time consuming. Recently, a whole blood technique involving flow cytometry and detection of CD25 and OX40 (CD134) expression on the surface of activated CD4+ T cells was shown to be accurate and concordant when compared with more traditional methods of antigen‐specific T‐cell detection.
Methods
Whole heparinized blood was collected from healthy donors and set up using the “OX40” assay to detect antigen‐specific CD4+ T‐cell responses to Varicella Zoster Virus, Epstein‐Barr Virus (EBV), Cytomegalovirus, Candida albicans, and Streptococcus pneumoniae.
Results
The “OX40” assay technique was clinically validated for routine use in an NHS clinical immunology laboratory by analysis of incubation length (40–50 h), sample transport time (up to 24 h at room temperature), concordance with serology testing, proliferation and interferon‐gamma production. In addition, 63 healthy controls (age range 21–78) were tested for responses to generate a healthy control reference range.
Conclusions
The OX40 assay, as presented in this report, represents an economical, rapid, robust whole blood technique to detect antigen‐specific T cells, which is suitable for clinical immunology diagnostic laboratory use. © 2014 International Clinical Cytometry Society
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