1. Vaccines H.Sidra Yasin (BIOT 412)

2. Learning objectives What are the Methods to produce the vaccines How we can modify the Vaccines What are the Routs of administration of vaccines What are the Types of vaccines What is Reverse vaccinology and its purpose Summary of all topics with conclusion

3. General Method to produce the vaccine

4. Immunisation Department, Centre for Infections Vaccine composition Following are: ComponentPurposeExample Adjuvantsenhance the immune response to a vaccine aluminium salts Preservativesprevent bacterial or fungal contamination of vaccine thiomersal Additivesstabilise vaccines from adverse conditions such as freeze-drying or heat, thereby maintaining a vaccine’s potency gelatine Residuals from manufacturing process Inactivating agents Antibiotics - prevent bacterial contamination during manufacturing process Egg proteins- some vaccine viruses are grown in chick embryo cells Yeast proteins formaldehyde neomycin, streptomycin, polymyxin B influenza, yellow fever HepB vaccine

5. Modifiers of vaccines

6. Adjuvants Boosters

7. Adjuvants Chemical substance that can be added to a vaccine in order to enhance the immune response to the vaccine.

8. Types 1. Freund’s Adjuvant 2. Aluminum Hydroxide 3. Aluminum Phosphate (Alum)

9. Boosters Periodic “booster” administration must be given in order to strengthen and lengthen the duration of immunity

10. Routs of administration

11. Routes Intradermal administration. ◦Three types are; intravenous intramuscular subcutaneous. Oral administration. ◦Vaccine is usually given in liquid form. ◦Foods Intranasal administration.

12. Types of vaccines

13. TraditionalRecombinant vaccines 1. Types A. Inactivated B. Live C. Attenuated 2. Pathogens A. Bacteria B. Virus C. Parasites 1. Subunit Vaccines 2. peptide vaccines 3. Attenuated Vaccines 4. Vector Vaccines 5. Bacterial Antigen Delivery Systems

14. Traditional vaccines

15. Live, Attenuated Vccines Act like natural infection Live, but weakened, viruses or bacteria Altered organisms, either genetically or chemically but non pathogenic Example: Attenuated virus vaccine for yellow fever, which utilizes the YF17D strain, a weakened form of the wild virus.

16. Live, Attenuated vaccines AdvantagesDisadvantages Single dose sufficient to induce long-lasting immunity Strong immune response Local and systemic immunity Others… Polio and Adeno Potential to revert to virulence Contraindicated in immunosuppressed patients Interference by viruses or vaccines and passive antibody Poor stability Potential for contamination

17. Inactivated Vaccines  Either: Suspensions of whole intact killed organisms ◦e.g. whole cell Pertussis, Influenza, Rabies, HepA  Or: Acellular and sub-unit vaccines ◦e.g. Acellular Pertussis vaccine contains between 2-5 components of the whole cell Pertussis bacteria

18. Inactivated vaccines

19. 1.whole actual pathogen killed, either by a heat treatment or chemically Salk vaccine for polio, which utilizes whole polioviruses that have been inactivated by formaldehyde.

20. 2.Fractional

21. Protein based; Toxoids  Stimulates the antibody mediated response Exotoxins  Toxoids are vaccines which consist of exotoxins  Immunity against the toxins, but not necessarily the bacteria that produce the toxins.  Examples: Botulinum antitoxen Diphtheria antitoxen Pertusis Tetanus toxoids

22. Protein based; Subunit Pathogenic agent Use components of pathogenic organism instead of whole organism Advantage: no extraneous pathogenic particles i.e DNA Disadvantage: Costly Examples o HSV Method of production o Tuberculosis o Foot -and-Mouth Disease virus (FMDV)

23. Polysaccharide based; pure pure cell wall polysaccharide from bacteria

24. Polysaccharide based; conjugate Polysaccharide linked to a carrier protein More potent lacks long term immunological memory Protect against: o Hib o Pneumococcal diseases o Tetanus o Diphtheria

25. Inactivated vaccines AdvantagesDisadvantages Stable Constituents clearly defined Gives sufficient humoral immunity if boosters given No mutation or reversion Can be used with immuno- deficient patients Many vaccinees do not raise immunity Shorter lasting immunity Boosters needed Need several doses Adjuvant needed Failure in inactivation and immunization with virulent viruses Others…

26. Possible Limitations of Traditional Vaccine Production Not all infectious agents can be grown in culture Animal/human cell culture expensive if needed Yield of viruses from cultures can be low Safety precautions for culture of live agents Insufficient killing/attenuation of agents Reversion of attenuated agents Traditional vaccines are less diverse

27. New Strategies Delete virulence genes Use live nonpathogenic carriers for immunization (unrelated pathogenic agent) Clone antigenic determinants into alternative host Address autoimmune system response/problems

28. So!!!

29. Recombinant vaccines

30. Types 1. Subunit Vaccines 2. Peptide vaccines 3. DNA Vaccines 4. Vector Vaccines

31. Peptide vaccines Use discrete portion (domain) of a surface protein as Vaccine. These domains are ‘epitopes’  antigenic determinants  are recognized by antibodies Use highly immunogenic carrier molecule

32. With carrier proteins

33. DNA Vaccines DNA vaccines consist of plasmids that contains genes for certain types of antigens. Once administered, the plasmid is taken up by the target cell and the genes are expressed. The cell then either excretes the antigen or displays it on an MHC-I molecule.

34. Genetic Immunization Delivery of a gene for the antigen to a host organism Use vector containing cDNA from viral protein/ eukaryotic promoter Inject into muscle POTENTIAL Eliminates purification of antigen Protein is modified post-translationally

35. Chimeric Vaccines Consist of attenuated viruses have been engineered to carry antigens from multiple types of pathogens. The yellow fever vaccine has been engineered to carry antigens from HIV, different types of bacteria, malaria, even cancer.  immunity against several different diseases with one administration.

36. Human Diseases for Which Recombinant Vaccines Are Currently Being Developed

37. Vaccine Production methods

38. Vaccine Production Methods Manufacturing strategies: ◦In-vivo ◦In-vitro ◦Chemical Synthesis  Some vaccines can be produced using any one of the three methods while for other vaccines, only one method will work.

39. In-Vitro Here, using recombinant DNA technology, vaccines can be produced in yeast cultures, bacterial cultures, or cell cultures. Recombinant vaccines, such as chimeric and Attenuated virus/bacteria vaccines, are produced in this manor.

40. In-Vivo Embryonated Chicken eggs are commonly used as in producing flu vaccines. Vaccines, like anti-idiotype, can also be produced in lab animals, such as mice.  There are even some species of plant, such as bananas, that have been genetically engineered to produce a vaccine.

41. Chemical Synthesis Produced in a lab. Vaccines that utilize synthetic peptides as well as conjugated lipids and polysaccharides Used in combination with either in-vivo or in-vitro production.

42. Summary!!!

43. Thank you!