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.
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.
Authors: Patricio Escalante; Tobias Peikert; Virginia P Van Keulen; Courtney L Erskine; Cathy L Bornhorst; Boleyn R Andrist; Kevin McCoy; Larry R Pease; Roshini S Abraham; Keith L Knutson; Hirohito Kita; Adam G Schrum; Andrew H Limper Journal: Am J Respir Crit Care Med Date: 2015-09-01 Impact factor: 21.405
Authors: Jennifer M Dan; Cecilia S Lindestam Arlehamn; Daniela Weiskopf; Ricardo da Silva Antunes; Colin Havenar-Daughton; Samantha M Reiss; Matthew Brigger; Marcella Bothwell; Alessandro Sette; Shane Crotty Journal: J Immunol Date: 2016-06-24 Impact factor: 5.422
Authors: Daniele E Mascarelli; Rhubia S M Rosa; Jessica M Toscaro; Isadora F Semionatto; Luciana P Ruas; Carolinne T Fogagnolo; Gabriel C Lima; Marcio C Bajgelman Journal: Front Cell Dev Biol Date: 2021-06-30