Experimental infection and protection against ... - TI Pharma

Long-term protection against malaria after experimental sporozoite inoculation 201
Figure 2. Trial profile. Timelines of previous immunisation study in grey.
stained with IQTest CD45RO-phycoerythrin Texas Red (Beckman-Coulter), PerCP
anti-human CD3 (BioLegend, San Diego, CA, USA) and PE-Cy7 anti-human CD62L
(eBioscience) for 20 min at 4°C. After washing, cells were incubated for 15 min at
4°C with fixation Medium A (Caltag, S. San Francisco, CA, USA), washed and
stained with FITC anti-human IFNγ (eBioscience) and APC anti-human IL-2
(eBioscience) in permeabilization Medium B (Caltag) for 20 min at 4°C. 100,000
events in the lymphocyte gate, based on forward- and side-scatter, were
acquired on a CyAn ADP 9-color flow cytometer (Beckman-Coulter). Flow
cytometry analysis was performed using FlowJo software version 9.1. Analysis
was performed on CD3+ T cells and effector memory (EM) T cells. The IFNγ and
IL-2 gates were set based on negative control samples.
Statistical analysis
Data analysis was performed using SPSS software version 16.0. Differences in
cellular immune responses between subjects in the vaccine group and control
group were analyzed by the Mann–Whitney-U test. The correlation between
peak temperature and peak d-dimer values was assessed by Pearson correlation
analysis. A two-sided P value of less than 0.05 was considered to indicate
statistical significance.