elispot readers analyzers software  (Cellular Technology Ltd)

Journal: Vaccines

Article Title: A Single Dose of a Hybrid hAdV5-Based Anti-COVID-19 Vaccine Induces a Long-Lasting Immune Response and Broad Coverage against VOC

doi: 10.3390/vaccines9101106

Figure Lengend Snippet: Cellular immune response induced by Ad-vectored vaccines. BALB/c mice received 10 9 or 10 10 vp of an Ad-vectored vaccine (blue: CoroVaxG.5; red: CoroVaxG.3; grey: Ad.C; green: Naïve) and were sacrificed after 14 days (10 10 vp) or 140 days (10 9 vp). Cells secreting IFN-γ per million of splenocytes were determined by ELISPOT at ( a ) 14 days. ( b ) 140 days post-immunization. The samples were analyzed in duplicates. The results of each group are expressed as the mean of spot-forming units (SFU). For FACS analysis, splenocytes were stained with anti-CD8α, anti-CD62L and anti-CD44 fluorochrome-conjugated antibodies. Stained splenocytes were subjected to flow cytometry analysis to quantify memory T cells (TCM: CD44high CD62Lhigh and TEM: CD44high CD62LLow) ( c , d ). ( c ) Data are expressed as percentage of total CD8+ cells; unstimulated controls (black dots) were included. ( d ) Representative dot plots of each group. The gate shows TCM subpopulation. The box and whisker plots represent the median (mid-line), max and min (boxes) and range (whiskers). The percentage of TCM cells in each group is depicted in the upper part of the box. Vaccinated groups were always significantly different to the Naïve and Unstimulated group; * p < 0.05, ** p < 0.01; Kruskal–Wallis test with Dunn’s multiple comparisons a posteriori .

Article Snippet: The number of spots was determined using an automatic ELISPOT reader and image analysis software (CTL-ImmunoSpot ® S6 Micro Analyzer, Cellular Technology Limited (CTL), Cleveland, OH, USA).

Techniques: Vaccines, Enzyme-linked Immunospot, Staining, Flow Cytometry, Whisker Assay