Replicase-based DNA vaccines for allergy treatment.
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.
Scheiblhofer
Sandra
S
Department of Molecular Biology, University of Salzburg, Salzburg, Austria.
Weiss
Richard
R
Gabler
Maximilian
M
Leitner
Wolfgang W
WW
Thalhamer
Josef
J
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology; D016207:Cytokines / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology / Q000628:therapy*; D007118:Immunoassay / Q000379:methods; D051379:Mice; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics / Q000276:immunology*; D012093:Replicon / Q000235:genetics / Q000276:immunology*; D012845:Sindbis Virus; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D014611:Vaccination; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology
D016207:Cytokines; D019444:Vaccines, DNA; D012324:RNA-Dependent RNA Polymerase