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  • Title: Replicase-based DNA vaccines for allergy treatment.
    Author: Scheiblhofer S, Weiss R, Gabler M, Leitner WW, Thalhamer J.
    Journal: Methods Mol Med; 2006; 127():221-35. PubMed ID: 16988457.
    Abstract:
    Replicase-based vaccines were introduced to overcome some of the deficiencies of conventional DNA- and RNA-based vaccines, including poor efficiency and low stability. At ultra-low doses, these alphavirus-derived vectors elicit cellular as well as humoral immune responses. Additionally, replicase-based vectors induce "self-removal" of the vaccine via apoptosis of transfected cells. This chapter describes the construction of a replicon-based DNA vaccine vector from commercially available plasmids. We present protocols for monitoring cellular immune responses following replicase-based immunization including measurement of allergen-specific proliferation of splenocytes, ELISPOT, a FACS-based cytokine secretion assay providing information about T-helper subsets, and a cytokine fluorescent bead immunoassay.
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