Antigen Immunization schedule Adujvant Immunizing a mouse Antigen

Antigen & Immunization & schedule Adujvant Immunizing a mouse Antigen sources DNA vaccine Carrier effect Epitope or antigenic determinant Conformational epitope “Hot” epitope Amino acid sequence on epitope

小白鼠免疫流程 BALB/c 0 2 4 6 8 Antigen (50 mg/mouse) 加佐劑製成乳劑 Emulsified in 0. 5 m. L Freund's Complete Adjuvant At least three booster shots, same dose in 0. 5 m. L Freund's Incomplete Adjuvant Booster shots might be reduced if Titer. Max is use as adjuvant Trial Bleeding Titer Determination 10 Final booster (soluble antigen) 12 Cell fusion 14 wk 試採血

Ref: Weiner & Kennedy (1999) Scientific American July: P 34, Genetic Vaccines DNA Vaccine 抗原基因 X Vector 抗體免疫 Cytokins B Cytokins Antigen Presenting Cell X X X 肌肉或肝臟細胞 Tc HMC 1 X 細胞免疫 Proteolysis Kill

Carrier 蛋白質對免疫反應的影響 – Carrier Effect B cell TH cell DNP BSA hapten 牛血清 白蛋白 卵白 蛋白 OA DNP 免疫小鼠 取出脾臟細胞 移植到新個體 (以 X-ray 照射過) DNP 抗原促進注射 DNP 抗體反應 BSA ++++ + DNP OA OA ++++ Adapted from Roitt et al (2001) Immunology. p. 134

Male et al (1987) Advanced Immunology. 8. 2 蛋白質分子 表面有許多 抗原決定基

Conformational Epitopes Roitt et al (1985) Immunology. 6. 4

“Hot” Epitopes Male et al (1987) Advanced Immunology. 8. 5

蛋白質某些胺基酸對抗原性有重大影響 Myoglobin peptide 133 136 140 145 N K A L E L F R K D I A A K Y E L F R K D I A A K Y N K A L E L F R K D I A A K Reactivity with T cells ++ ++ + + ++ - Adapted from Male et al (1987) Advanced Immunology. 8. 6