Protocol detail

Evaluating SUDV and EBOV Peptide-Specific T-cell Responses

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Multiscreen-IP plates (Millipore) were activated, coated and blocked according to the kit manufacturer’s instructions (Mabtech). 2 × 10⁵ freshly isolated splenocytes were added per well in triplicates with either 2 µg/ml the LYDRLASTV peptide present in both SUDV and EBOV GP^{38 (link)}, or the RPHTPQFLF peptide derived from SUDV GP^{38 (link)}, medium alone or Concanavalin A (Sigma-Aldrich). After 20 ± 2 h of incubation at 37 °C with 5% CO₂, plates were developed as recommended by the manufacturer (Mabtech). Plates were analyzed using the Immunospot analyzer and software (Immunospot).

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Öhlund P., García-Arriaza J., Zusinaite E., Szurgot I., Männik A., Kraus A., Ustav M., Merits A., Esteban M., Liljeström P, & Ljungberg K. (2018). DNA-launched RNA replicon vaccines induce potent anti-Ebolavirus immune responses that can be further improved by a recombinant MVA boost. Scientific Reports, 8, 12459.

Publication 2018

Multiscreen-IP plates Concanavalin A

Concanavalin a Peptide

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Variable analysis

independent variables

LYDRLASTV peptide present in both SUDV and EBOV GP
RPHTPQFLF peptide derived from SUDV GP
Medium alone
Concanavalin A (Sigma-Aldrich)

dependent variables

IFN-gamma production by freshly isolated splenocytes

control variables

2 × 10^5 freshly isolated splenocytes per well in triplicates
20 ± 2 h of incubation at 37 °C with 5% CO2
Multiscreen-IP plates (Millipore) activated, coated and blocked according to the kit manufacturer's instructions (Mabtech)

positive control

Concanavalin A (Sigma-Aldrich)

negative control

medium alone

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