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

tel-00827710, version 1 - 29 May 2013
First, we determined the relationship between antigen dissemination and antigen presentation
by host accessory cells. We applied multiple experimental approaches to address this
question: both looking at the persistence of the MHC-peptide complexe on APCs, as well as
directly evaluating the proportion of injected cells that remain at different time points
following immunization.
A. Persistence of antigen cross-presentation
First, we assessed the persistence of antigen cross-presentation by evaluating the presence of
K b -SIINFEKL complexes in different lymphoid organs. In order to compare i.d. injection
performed on the flank and i.v. immunization, we looked in the spleen and in the inguinal
lymph node, as it is the draining lymph node for i.d. injection, and in the opposite inguinal
lymph node as a non draining lymph node control. Mice were immunized with K bm1 mOva
splenocytes and, at different time points, CFSE-labeled CD45.1 + OT-I splenocytes were
transferred as a means of assessing cross-presentation by host APCs (Figure 32).
Figure 32. Delayed but persistent antigen presentation in the local draining lymph node after
intradermal immunization. Mice were immunized i.d. or i.v. with 5x10 5 K bm1 mOva splenocytes. On
days 0, 3, or 21, 5x10 6 CD45.1 CFSE-labeled OT-I splenocytes were adoptively transferred into
immunized recipients. Three days later, the spleen, draining lymph node and a non-draining lymph
node were harvested and the dilution of CFSE staining of OT-I was determined. Results are
representative of three independent experiments.
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