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

tel-00827710, version 1 - 29 May 2013
Figure 17. Cross-priming efficiency is dependent on the route of immunization in the HY model.
C57BL/6 female mice were immunized i.d. or i.v. with 5x10 6 male splenocytes depleted for CD11c +
cells. On day 12 for i.d. immunization, and day 8 for i.v. immunization, the spleen and draining lymph
node were harvested and an IFNγ ELISPOT was performed. SFC, Spot forming cell.
In our model, we compared the influence of different routes of immunization on the
development of a CD8 + T cell response: specifically, the intravenous (i.v.) route, which
results in a systemic dissemination of antigen versus the intradermal (i.d.) route, leading to a
restricted local antigen dissemination.
We initiated our studies by developing and optimizing the tetramer-based enrichment strategy
for our cross-presentation model and testing it in combination with intracellular staining
techniques to incorporate the functional characterization of the stimulated T cell response.
Once these protocols were established, we compared the immune responses induced by the
administration of cell-associated antigen either i.d., or i.v. Uniquely, we chose a relatively low
dose of antigen for our immunization, as compared to what has been used in previous studies
with this technique (LPS + peptide) (Moon et al., 2007) in order to be relatively similar to
physiologic conditions relevant for interpretation of our results in the context of human
vaccination. Given this low dose of antigen, enrichment of tetramer-positive cells was
required to obtain a significant number of cells to study.
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