1. Creating an Edible Vaccine for HIV
Alex Colello

2. What is HIV? Human Immunodeficiency Virus
Causes Acquired Immune Deficiency Syndrome
HIV is a retrovirus, which means its genome is made out of RNA, which can mutate much faster
Targets the immune system, especially helper T cells

3. Why Does This Matter?
Currently there are about 37 million people infected with HIV
In 2014 an estimated 2 million people became infected with HIV, and 1.2 million died from AIDS worldwide
70% of people living with HIV live in Africa

5. Advantages of Edible Vaccines
Significantly cheaper to produce than standard vaccines
Easier to distribute, especially in impoverished areas
Easy to administer

6. Why Bananas? It is very rare for people to be allergic to bananas
Bananas are easy to grow in tropical climates
It is a staple in the diet of many countries in need of a cheap HIV vaccine

7. Question
Can an edible preventative vaccine against HIV-1 be made by engineering banana plants to produce gp120?

8. Procedures
Obtain a Ti plasmid containing the sequence of gp120 in the T-dna region, ampicillin resistance as the bacterial selectable, and kanamycin resistance as the plant selectable marker

9. Procedures
Insert the plasmid into Agrobacterium tumefaciens using gel electroporation
Grow the bacteria on an agar plate containing ampicillin in order to select for only the transformed cells
Soak banana cotyledons in a bacteria solution, and grow the cotyledons on on an agar plate without antibiotics
Then transfer the banana cotyledons to a agar plate containing kanamycin to select for the transformed cells

10. Procedures
After collecting the banana fruit, use a Western blot to test if gp120 is present in the fruit
If gp120 is present, use 10 humanized mice that will be given the modified banana, and 10 mice that will be given unmodified banana
Every week, take plasma samples from the mice and use ELISA to determine if an antigen was produced by the mice

11. Expected Results The experiment will be successful if:
gp120 is found in the banana plant
Over the course of the HIV infection, mice that were given the banana produced more antibodies against HIV than those fed with normal bananas

12. Potential Problems The gp120 might be degraded in the stomach
It also might not be absorbed into the bloodstream and just pass through
The antigens may not be effective
HIV mutates very quickly
Antibodies produced by the mice might not do anything against HIV

13. Future Research
Perform the experiment with macaque, chimpanzees, and eventually humans
Increase the amount of gp120 produced by the bananas through regulatory sequences

14. Work cited
Kim, Tae-Geum, Andreas Gruber, and William H.r Langridge. "HIV-1 Gp120 V3 Cholera Toxin B Subunit Fusion Gene Expression in Transgenic Potato." Protein Expression and Purification 37.1 (2004): Web.
Joung, Y.h., J.w. Youm, J.h. Jeon, B.c. Lee, C.j. Ryu, H.j. Hong, H.c. Kim, H. Joung, and H.s. Kim. "Expression of the Hepatitis B Surface S and PreS2 Antigens in Tubers of Solanum Tuberosum." Plant Cell Rep Plant Cell Reports 22.12 (2004): n. pag. Web.
Zwick, Michael, and Dennis Burton. "HIV-1 Neutralization: Mechanisms and Relevance to Vaccine Design." CHR Current HIV Research 5.6 (2007): Web.
Vitti, Antonella, Maria Nuzzaci, Valentina Condelli, and Pasquale Piazzolla. "Simulated Digestion for Testing the Stability of Edible Vaccine Based on Cucumber Mosaic Virus (CMV) Chimeric Particle Display Hepatitis C Virus (HCV) Peptide." Methods in Molecular Biology Virus Hybrids as Nanomaterials (2013): Web.
Ahmad, Parvaiz, Muhammad Ashraf, Muhammad Younis, Xiangyang Hu, Ashwani Kumar, Nudrat Aisha Akram, and F. Al-Qurainy. "Role of Transgenic Plants in Agriculture and Biopharming." Biotechnology Advances 30.3 (2012): Web.
Forsman, A., E. Beirnaert, M. M. I. Aasa-Chapman, B. Hoorelbeke, K. Hijazi, W. Koh, V. Tack, A. Szynol, C. Kelly, A. Mcknight, T. Verrips, H. D. Haard, and R. A. Weiss. "Llama Antibody Fragments with Cross-Subtype Human Immunodeficiency Virus Type 1 (HIV-1)-Neutralizing Properties and High Affinity for HIV-1 Gp120." Journal of Virology82.24 (2008): Web.

15. Image Credits http://www.rkm.com.au/VIRUS/HIV/HIV-images/HIV-virus.jpg