Enumeration and Characterization of Human Memory T Cells by Enzyme-Linked Immunospot Assays
Figure 2
Epstein-Barr virus (EBV)-specific T-cell responses determined by standard and cultured ELISPOT assays. Peripheral blood mononuclear cells (PBMCs) from (a) an EBV seropositive immunocompromised patient (an hematopoietic stem cell transplant recipient suffering from a EBV-associated posttransplant lymphoproliferative disorder) and (b) an EBV seropositive healthy subject were analyzed by standard and cultured ELISPOT assays in response to peptide pools (15 amino acids in length with an 11 amino acid overlap) representing the full-length lytic (BZLF1 and BMRF1) and latent (EBNA1, EBNA3a, EBNA3b, EBNA3c, LMP1, and LMP2) EBV proteins. The mean number of spots from duplicate wells was adjusted to million PBMCs. Results are shown as net spots/million PBMCs calculated by subtracting the mean number of spots in wells with cultured medium only from the mean number of spots in wells from each EBV peptide pool. Results from the cultured ELISPOT were adjusted per proliferation index (number of antigen-stimulated cells after 10 days of culture divided by the number of medium-stimulated cells after 10 days of culture). Error bars represent the standard error of the mean. T-cell subsets indicated are at the time of ELISPOT analyses.
