1. Classical Vaccines

2. Vaccines can eradicate pathogens and save lives

3. Vaccines have been made for 36 of >400 human pathogens Immunological Bioinformatics, The MIT press. +HPV & Rotavirus

4. Dodo Pathogenic Viruses Data derived from /www.cbs.dtu.dk/databases/Dodo. 1st column: log10 of the number of deaths caused by the pathogen per year 2nd column: DNA Advisory Committee (RAC) classification DNA Advisory Committee guidelines [RAC, 2002] which includes those biological agents known to infect humans, as well as selected animal agents that may pose theoretical risks if inoculated into humans. RAC divides pathogens into four classes. Risk group 1 (RG1). Agents that are not associated with disease in healthy adult humans Risk group 2 (RG2). Agents that are associated with human disease which is rarely serious and for which preventive or therapeutic interventions are often available Risk group 3 (RG3). Agents that are associated with serious or lethal human disease for which preventive or therapeutic interventions may be available (high individual risk but low community risk) Risk group 4 (RG4). Agents that are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available (high individual risk and high community risk) 3rd column: CDC/NIAID bioterror classification classification of the pathogens according to the Centers for Disease Control and Prevention (CDC) bioterror categories A–C, where category A pathogens are considered the worst bioterror threats 4th column: Vaccines available A letter indicating the type of vaccine if one is available (A: acellular/adsorbet; C: conjugate; I: inactivated; L: live; P: polysaccharide; R: recombinant; S staphage lysate; T: toxoid). Lower case indicates that the vaccine is released as an investigational new drug (IND)). 5th column: G: Complete genome is sequenced

5. Vaccines work by preparing the immune system so it can respond faster and stronger when challenged

6. The addaptive Immune system Figures by Eric A.J. Reits

7. Vaccination Administration of a substance to a person with the purpose of preventing a disease Traditionally composed of a killed or weakened micro organism Vaccination works by creating a type of immune response that enables the memory cells to later respond to a similar organism before it can cause disease

8. Early History of Vaccination Pioneered India and China in the 17th century The tradition of vaccination may have originated in India in AD 1000 Powdered scabs from people infected with smallpox was used to protect against the disease Smallpox was responsible for 8 to 20% of all deaths in several European countries in the 18th century In 1721 Lady Mary Wortley Montagu brought the knowledge of these techniques from Constantinople (now Istanbul) to England Two to three percent of the smallpox vaccinees, however, died from the vaccination itself Benjamin Jesty and, later, Edward Jenner could show that vaccination with the less dangerous cowpox could protect against infection with smallpox The word vaccination, which is derived from vacca, the Latin word for cow.

9. Early History of Vaccination II In 1879 Louis Pasteur showed that chicken cholera weakened by growing it in the laboratory could protect against infection with more virulent strains 1881 he showed in a public experiment at Pouilly-Le-Fort that his anthrax vaccine was efficient in protecting sheep, a goat, and cows. In 1885 Pasteur developed a vaccine against rabies based on a live attenuated virus A year later Edmund Salmon and Theobald Smith developed a (heat) killed cholera vaccine. Over the next 20 years killed typhoid and plague vaccines were developed In 1927 the bacille Calmette-Guérin (BCG vaccine) against tuberculosis vere developed

10. Vaccination since WW II Cell cultures Ability to grow cells from higher organisms such as vertebrates in the laboratory Easier to develop new vaccines The number of pathogens for which vaccines can be made have almost doubled. Many vaccines were grown in chicken embryo cells (from eggs), and even today many vaccines such as the influenza vaccine, are still produced in eggs Alternatives are being investigated

11. Vaccination Today Vaccines have been made for only 36 of the more than 400 known pathogens that are harmful to man. Immunization saves the lives of 3 million children each year, but that 2 million more lives could be saved if existing vaccines were applied on a full-scale worldwide

12. Categories of Vaccines Live vaccines Are able to replicate in the host Attenuated (weakened) so they do not cause disease Subunit vaccines Part of organism Genetic Vaccines Part of genes from organism

13. Vaccines have been made for 36 of >400 human pathogens Immunological Bioinformatics, The MIT press. +HPV & Rotavirus

14. Vaccines under IND in the US Venezuelan equine encephalomyelitisvirus Live Western equine encephalomyelitis virus Inactivated Estern equine encephalomyelitis virus Inactivated Coxiella burnetii Inactivated, Available in South Amerika; Lisenced vaccine in Australia Francisella tularensis Live Hantaan virus Live Rift Valley fever virus Inactivated, live Junin Virus Live, Available in South America

15. Live Vaccines Characteristics Able to replicate in the host Attenuated (weakened) so they do not cause disease Advantages Induce a broad immune response (cellular and humoral) Low doses of vaccine are normally sufficient Long-lasting protection are often induced Disadvantages May cause adverse reactions May be transmitted from person to person

16. Subunit Vaccines Relatively easy to produce (not live) Induce little CTL Viral and bacterial proteins are not produced within cells Classically produced by inactivating a whole virus or bacterium Heat Chemicals The vaccine may be purified Selecting one or a few proteins which confer protection Bordetella pertussis (whooping cough) Create a better-tolerated vaccine that is free from whole microorganism cells

17. Subunit Vaccines: Polysaccharides Polysaccharides Many bacteria have polysaccharides in their outer membrane Polysaccharide based vaccines Neisseria meningitidis Streptococcus pneumoniae Generate a T cell-independent response Inefficient in children younger than 2 years old Overcome by conjugating the polysaccharides to peptides Used in vaccines against Streptococcus pneumoniae and Haemophilus influenzae.

18. Subunit Vaccines: Toxoids Toxins Responsible for the pathogenesis of many bacteria Toxoids Inactivated toxins Toxoid based vaccines Bordetella pertussis Clostridium tetani Corynebacterium diphtheriae Inactivation Traditionally done by chemical means Altering the DNA sequences important to toxicity

19. Subunit Vaccines: Recombinant The hepatitis B virus (HBV) vaccine Originally based on the surface antigen purified from the blood of chronically infected individuals. Due to safety concerns, the HBV vaccine became the first to be produced using recombinant DNA technology (1986) Produced in bakers’ yeast (Saccharomyces cerevisiae) Virus-like particles (VLPs) Viral proteins that self-assemble to particles with the same size as the native virus. VLP is the basis of a new vaccine against human papilloma virus (HPV) For more information se: http://www.nci.nih.gov/ncicancerbulletin/NCI_Cancer_Bulletin_041205/page5http://www.nci.nih.gov/ncicancerbulletin/NCI_Cancer_Bulletin_041205/page5

20. Genetic Vaccines Introduce DNA or RNA into the host Injected (Naked) Coated on gold particles Advantages Easy to produce Induce cellular response Disadvantages Low response in 1st generation Genes can also be carried by genetically modifies viruses or bacteria

21. Epitope based vaccines Advantages (Ishioka et al. [1999]): Epitope based vaccines Can be more potent Can be controlled better Can induce subdominant epitopes (e.g. against tumor antigens where there is tolerance against dominant epitopes) Can be designed to break tolerance Can target multiple conserved epitopes in rapidly mutating pathogens like HIV and Hepatitis C virus (HCV) Can overcome safety concerns associated with entire organisms or proteins Epitope-based vaccines have been shown to confer protection in animal models ([Snyder et al., 2004], Rodriguez et al. [1998] and Sette and Sidney [1999])

22. Passive Immunization Antibodies harvested from infected patients or animals are then used to protect against disease [Marshall et al., 2003] Used in special cases against many pathogens: Cytomegalovirus Hepatitis A and B viruses Measles Varicella Rubella Respiratory syncytial virus Rabies Clostridium tetani Varicella-zoster virus Vaccinia Clostridium botulinum Corynebacterium diphtheriae

23. Therapeutic vaccines Vaccines to treat the patients that already have a disease Targets Tumors AIDS Allergies Autoimmune diseases Hepatitis B Tuberculosis Malaria Helicobacter pylori Concept suppress/boost existing immunity or induce immune responses.

24. Cancer vaccines Strategies Induce immune response (break tolerence) against overexpressed self antigens Target mutated proteins Therapeutic cancer vaccines can induce antitumor immune responses in humans with cancer Antigenic variation is a major problem that therapeutic vaccines against cancer face Tools from genomics and bioinformatics may circumvent these problems Se also: http://cis.nci.nih.gov/fact/7_2.htmhttp://cis.nci.nih.gov/fact/7_2.htm

25. Allergy vaccines Increasing occurrence of allergies in industrialized countries The traditional approach is to vaccinate with small doses of purified allergen Second-generation vaccines are under development based on recombinant technology Genetically engineered Bet v 1 vaccine can reduce pollen-specific IgE memory response significantly Example of switching a “wrong” immune response to a less harmful one. Figure by Thomas Blicher.

26. Therapeutic Vaccines against Persistent Infections For example for preventing HIV-related disease progression Most of the first candidate HIV-1 vaccines were based entirely or partially on envelope proteins to boost neutralizing antibodies Envelope proteins are the most variable parts of the HIV genome Vaccines composed of monomeric gp120 molecules induce antibodies that do not bind to trimeric gp120 on the surface of virions A number of recent vaccines are also designed to induce strong cell- mediated responses. Escapes from CTL responses are associated with disease progression and high viral loads Some CTL epitopes escape recognition quickly because they are not functionally constrained, others might need several compensatory mutations because they are in functionally or structurally constrained regions of HIV-1

27. Vaccines Against Autoimmune Diseases Multiple sclerosis T cells specific for mylein basic protein (MBP) can cause inflammation of the central nervous system. The vaccine uses copolymer 1 (cop 1), a protein that highly resembles MBP. Cop 1 competes with MBP in binding to MHC class II molecules, but it is not effective in inducing a T cell response On the contrary, cop 1 can induce a suppressor T cell response specific for MBP, and this response helps diminish the symptoms of multiple sclerosis A vaccine based on the same mechanisms is developed for myasthenia gravis More information: http://www.ninds.nih.gov/disorders/multiple_sclerosis/detail_multiple_sclerosis.htm, http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12667659&query_hl=4 http://www.ninds.nih.gov/disorders/multiple_sclerosis/detail_multiple_sclerosis.htm http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12667659&query_hl=4

28. Vaccines Market The vaccine market has increased fivefold from 1990 to 2000 Annual sales of 6 billion euros Less than 2% of the total pharma market Major producers (85% of the market) GlaxoSmithKline (GSK), Merck, Aventis Pasteur, Wyeth, Chiron Main products (>50% of the market) Hepatitis B, flu, MMR (measles, mumps, and rubella) and DTP (diphtheria, tetanus, pertussis) 40% are produced in the United States and the rest is evenly split between Europe and the rest of the world [Gréco, 2002] It currently costs between 200 and 500 million US dollars to bring a new vaccine from the concept stage to market [André, 2002] More information: Gréco, 2002 André, 2002 Gréco, 2002 André, 2002

29. Trends From Whole live and killed organisms Problems Adverse effects Production To Subunit vaccines Genetic vaccines Challenges Enhance immunogenecity