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Clearance of the stimulating antigen from this organ by the later time points. In contrast, animals immunized i.d. had the majority of initial response localized to the cervical lymph nodes nearest to the site of immunization, while cells at only later divisions were detected in spleen. By day 7, little proliferation was observed with a very few CFSElow F5 cells detected using this MVA.HIVANP dose and route. Using i.m. immunization with this lower dose of MVA.HIVA-NP, very little if any antigen-specific cell division was induced at NDLN and spleen at any time post-delivery. This is consistent with our and others previous observations [38, 39] indicating that at least for the detection of antigen-specific T cells in the spleen, the i.m. route of immunization is the least effective delivery of the three routes investigated here. Thus, comparison of the routes of immunization suggested that the priming of the response was highly localized with very little cell division occurring on day 3 in distant lymphoid organs and that the initiation of proliferation was transient, lasting a maximum of 7 days. The MVA.HIVA-NP dose affected frequencies of cells entering division, but not the division rate One of the important factors affecting the strength of an immune response is the amount of immunogen, i.e. the vaccine dose. Thus, using the i.d. route of vaccination, we investigated the effect of MVA.HIVA-NP dose on the F5 cell division in the DLN, spleen and NDLN over the first 7 days after vaccine delivery Fig. 3 ; . The lowest dose of 5 103 PFU of MVA.HIVA-NP did not induce any appreciable responses in any of the analysed lymphoid organs, suggesting that a minimum threshold of antigenic stimulus has to be reached to trigger the immune response. For the higher doses of 5 105 and 5 107 PFU, a response was initiated in the LDN and moved over time and vivelle.
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