Journal: Scientific Reports
Article Title: Protective immunity differs between routes of administration of attenuated malaria parasites independent of parasite liver load
Figure Lengend Snippet: Unprotected mice after ID immunization show reduced liver T cell effector responses compared to protected mice. ID immunized mice were challenged followed by artesunate treatment, and distinguished as protected (p, i.e. luciferase negative) or unprotected (unp, i.e. luciferase positive). Hepatic CD8 and CD4 T cells were analyzed at day 7 after challenge. (A) IFN-γ concentration in supernatant of total leukocytes after culture for 36 h with CSP and sporozoites as measured by ELISA. (B) Representative FACS plots of CD8 + gated T cells for intracellular expression of IFN-γ or TNF after culture for 4 h with CSP and brefeldin A, and for surface expression of CD107a after 4 h culture with CSP, brefeldin A and monensin. Numbers indicate the frequency of the gated cell population. Graphs show a summary of 2 experiments with 8–14 mice per group for frequency of IFN-γ ( C ), TNF ( D ), or CD107a ( E ) -expressing CD8 T cells and CD4 T cells. The dotted line indicates the mean cytokine or expression level for protected IV immunized mice (N = 9–16). Significant difference by Mann-Whitney test is indicated by *p
Article Snippet: For the detection of intracellular IL-10 it was required to culture cells with CSP peptide (10 µg/ml) plus P. yoelii sporozoites (Py wt-GFP-Luccon , 25,000/ml) for 36 h including PMA, ionomycin and brefeldin A in the last 4 h. For flow cytometric detection of surface-exposed CD107a, cells were cultured for 4 hours with 0.25% GolgiStop (BD Biosciences), 10 µg/ml brefeldin A and anti-mouse CD107a-AlexaFluor488 (eBioscience, Vienna, Austria).
Techniques: Mouse Assay, Luciferase, Concentration Assay, Enzyme-linked Immunosorbent Assay, FACS, Expressing, MANN-WHITNEY