Refine
Has Fulltext
- yes (98)
Is part of the Bibliography
- yes (98)
Year of publication
- 2015 (98) (remove)
Document Type
- Journal article (61)
- Doctoral Thesis (37)
Keywords
- cancer (4)
- membrane proteins (4)
- Aspergillus fumigatus (3)
- Vaccinia-Virus (3)
- biodiversity (3)
- gene (3)
- honeybee (3)
- in vitro (3)
- mushroom body (3)
- olfaction (3)
Institute
- Theodor-Boveri-Institut für Biowissenschaften (98) (remove)
Sonstige beteiligte Institutionen
ResearcherID
- D-1221-2009 (1)
- N-2030-2015 (1)
Each positive well in ELISPOT assays contains spots of variable sizes that can range from tens of micrometers up to a millimeter in diameter. Therefore, when it comes to counting these spots the decision on setting the lower and the upper spot size thresholds to discriminate between non-specific background noise, spots produced by individual T cells, and spots formed by T cell clusters is critical. If the spot sizes follow a known statistical distribution, precise predictions on minimal and maximal spot sizes, belonging to a given T cell population, can be made. We studied the size distributional properties of IFN-γ, IL-2, IL-4, IL-5 and IL-17 spots elicited in ELISPOT assays with PBMC from 172 healthy donors, upon stimulation with 32 individual viral peptides representing defined HLA Class I-restricted epitopes for CD8 cells, and with protein antigens of CMV and EBV activating CD4 cells. A total of 334 CD8 and 80 CD4 positive T cell responses were analyzed. In 99.7% of the test cases, spot size distributions followed Log Normal function. These data formally demonstrate that it is possible to establish objective, statistically validated parameters for counting T cell ELISPOTs.