Voie d'immunisation et séquence d'administration de l ... - TEL

tel-00827710, version 1 - 29 May 2013
Figure 25. Intradermal immunization induces polyfunctional T cells. (A-C) Mice were immunized
i.d. or i.v. with 5x10 5 K bm1 mOva splenocytes. At the different time points, lymph nodes and spleen
were harvested and a K b -SIINFEKL tetramer-based enrichment was performed. The enriched fraction
was incubated for 4h with SIINFEKL-pulsed splenocytes, followed by surface and intracellular
staining. (A) Data from live CD3 + CD8 + T cells are shown. Cells producing IL-2 are highlighted in
red. The percentage of IFNγ + cells that produce either the three cytokines – IFNγ, IL-2 and TNFα (B)
or only one cytokine – IFNγ (C) were calculated. p-values were calculated using a Mann-Whitney test.
D. Memory potential of antigen-specific CD8 + T cells
Following from the T cell functionality results, we tested whether the route of immunization
impacts also secondary responses. As previously, mice were immunized i.d. or i.v., and 34
days later the same animals were re-stimulated by i.v. administration of a second dose of
5x10 5 K bm1 mOva splenocytes (Figure 26A). IFNγ-producing Ova-specific CD8 + T cells were
enumerated (Figure 26B). While the absolute number of tetramer-positive cells was similar in
both conditions, we detected a higher percentage of IFNγ-producing cells when the first
immunization was performed via the i.d. route (Figure 26C). We wanted to confirm these
Page 99 of 256