How do dead and apoptotic cells affect ELISPOT results?
A recent CTL publication addresses the effects of dead and/or apoptotic cells on ELISPOT assay results.
2 separate questions are addressed herein:
Are measurements of frequencies of antigen-specific CD4+ or CD8+ T cells affected by the presence of dead and/or apoptotic bystander cells?
Where there is an observed increase of dead/apoptotic cells in stored PBMC – indicative of T cell damage - is this a reliable indicator of CD4+ or CD8+ T cell impairment?
Our findings revealed these answers:
As described in figures 3 & 4 in this paper, the presence of dead/apoptotic bystander cells does not affect CD4/CD8 frequency measurements at all.
Measurements of frequencies of Live/Dead/Apoptotic cells in PBMC samples before starting the assay as well as after the completion of the assay constitute a more reliable indicator of PBMC “health”.
This paper also introduces the high-throughput capable Live/Dead/Apoptotic cell counting feature of the ImmunoSpot® solution.
CTL continues striving to live up to its mission: Elevating ELISPOT to an Exact Science™.