In between the rotavirus and multiple sclerosis projects.

1. Immunity vs. Tolerance: Production of a vaccine antigen and autoantigen in tobacco and lettuce chloroplasts. Wesley Schonborn PI: Dr. Henry Daniell

3. Bombardment

8. Mucosal Immunology

12. MS Project Vector Construction IL-4/10, MBP PCR Add SmaI and XbaI IL-4/10, MBP SmaI XbaI IL-4/10, MBP pCR Blunt II Topo pBR322 Cut with XbaI and SmaI Ligate into vector with CTB IL-4/10, MBP SmaI XbaI NdeI NotI SmaI XbaI CTB psbA(T)

13. MS Project Vector Construction IL-4/10, MBP SmaI XbaI NdeI NotI Cut with NdeI and NotI IL-4/10, MBP XbaI NotI SmaI NdeI SalI Ligate into LsLf vector CTB psbA(T) CTB psbA(P) psbA(T)

16. Vector Map NSP4

17. 4.3 kb 2 kb 5 kb

18. Western/Quantification There is some degradation over time (in older leaves). Young and bottom full-sized leaves were separated since they were several weeks apart in age. 50kDa 25kDa 15kDa Young Mature Old Also want to compare tobacco species.

19. Age of Leaf Quantification Young Mature Old

24. References Holmgren J & C Czerkinsky. Mucosal Immunity and Vaccines. Nature Medicine 11:S45-S53 (2005). Holmgren J, N Lycke & C Czerkinsky. Cholera toxin and cholera B subunit as oral-mucosal adjuvant and antigen vector systems. Vaccine 11:1179-1206 (1993). Inobe J-i, AJ Slavin, Y Komagata, Y Chen, et al. IL-4 is a differentiation factor for transforming growth factor- secreting Th3 cells and oral administration of IL-4 enhances oral tolerance in experimental allergic encephalomyelitis. European Journal of Immunology 28:2780-2790 (1998). Slavin AJ, R Maron & HL Weiner. Mucosal administration of IL-10 enhances oral tolerance in autoimmune encephalomyelitis and diabetes. International Immunology 13:825-833 (2001). Sun J-B, J Holmgren & C Czerkinsky. Cholera toxin B subunit: an efficient transmucosal carrier-delivery system for induction of peripheral immunological tolerance. PNAS 91:10795-10799 (1994).