IPPH 1: Immunology

Semester'1,''2013 ' ' ' ' ' ''''''''''''''''''''''''''''''VETS70005&

Lecture 13
Vaccines and vaccine development

A/Prof Jean-Pierre Scheerlinck

Phone: 8344 4441 Room Number: 103 Email: j.scheerlinck@unimelb.edu.au

Slide&1&

The'most'important'ba9les'did'not'occur'in'Verdun'or'the'rainforests'of'South?East' Asia'

1. 2. 3. 4. 5. 6. 7. 8. 9.

Overview&of&the&immune&system& Ab&eector&mechanisms& Recogni?on&of&An?gen&by&B&&&T&Cells& Thymus&development& B&cell&development& MHC&Restric?on& Cytokines& An?gen&Processing&& T/B&Collabora?on&

My'name'is'Jean?Pierre'Scheerlinck ' And'I'will'tell'you'about' these'ba9les ' 'but'occur'in'our'body'every'day'of'our'lives.'

10. Cell&mediated&immune&eectors& 11. Complement&system& 12. Tolerance&&&autoimmunity& 13. Vaccines&and&vaccine&development&

Slide&2&

The&power&of&vaccines:&Rinderpest&
Disease&
Signs:&fever,&oral&erosions,&diarrhea&and&lymphoid&necrosis& 95%&mortality&within&1Z2&weeks& Fragile&virus&quickly&deac?vated&by&heat&and&sunlight&
doesn't last for a long time unlike food and mouth disease which can spread via water.

Control&
Movement&control& Slaughter&of&aected&ca]le& Vaccina?on&1994Z2006& Last&case:&Kenya&2001& Eradica?on&declared&
8th&&August&2011&

South&Africa&1900&rinderpest&epidemic.&(New&York&Times)&

Slide&3&

WHOs&statement&on&ideal&vaccines&
how T and B cell epitopes relate to this

Aordable&worldwide& Heat&stable& Eec?ve&acer&a&single&dose& Applicable&to&a&number&of&diseases& Administered&by&mucosal&route& Suitable&for&administra?on&early&in&life&

Yen, Kenya 2010

Source: Goldsby et al, Immunology 5th edition! !

lonelyplane?mages.com&

Slide&4&

Future&and&underu?lized&vaccines&
Future& Vaccines&
HIV& Malaria& Meningococcus& TB& Dengue& HPV& Rotavirus& Pneumococcus&

Underu?lized& Vaccines&
Yellow&Fever&

Cholera& Inuenza& Hib&(conj)& Rubella& Jap&enc& Typhoid& Hep&B&

Tetanus&

Widely&used&Vaccines& Save&3&106&lives/year&
Diphtheria&

Measles& Pertussis& Polio&

1960&

1980&

2000&

Note:&the&halos&represent&deaths&currently&being&averted,&and&solid&circles&represent&deaths&es?mated&to&occur.&

immuniza?onbasics.jsi.com&

(WHOZIVR&August&2006)& Slide&5&

Most&vaccines&induce&immune&memory&

Boost&or&infec?on& Prime&

&Speed& &Magnitude& &Persistence& &Increased&avidity&

Second encounter -> immune memory, second immune response with these 4 characteristics.

0&&&&&&&&&&&2&&&&&&&&&&&&4&&&&&&&&&&&&6&&&&&&&&&&&&8& &&&&&&&&&&&&&Weeks&postZpriming&

Immunobiology&5,&Janeway&et&al& Slide&6&

Most&Vaccines&Rely&on&Immune&Memory&
Vaccination
Vaccination or primary infection

vaccination 'tricks' animal into thinking it already encountered the disease. 2nd time: big igG response

Infection
Infection after vaccination

Slide&7&

Comparing&Primary&and&Secondary&Immunity&

T cells direct what type of antibodies are made Higher afnity because of maturation of antibody Slide&8& response

The&Persistence&of&Memory&
Memory&B&cells&live&much&longer& than&Ab&secre?ng&cells&
Immune&memory&last&a&long& ?me&(some?mes&as&long&as& animal&lives)& Ab&response&diminishes&over& ?me&in&a&ma]er&of&months&
Persistence&of&memory&(S&Dali&Z&1931)&

Persistence&of&memory&may& require&the&con?nuous&presence& of&an?gen&(uncertain)&&

retain high level of antibody in the serum but declines after several years t and b cells remain at high level of memory
Slide&9&

Induc?on&of&immune&responses&&
second infection
Cytotoxic T cell Cytotoxic T cell CD8 MHC I Antigen T helper cell CD4 TCR MHC II Dendritic cell T helper cell B cell CD4
Slide&10&

Infection IFN-" Antigen T helper cell CD4 TCR MHC II

CD8

t cell in periphery

Antibody

Ab binds to Ig, taken up and presented to T cell

Vaccines&Against&Hidden&An?gens&
Vaccine&against&gut&an?gen&of&ca]le&?ck& (BM86)&
Tick&feeds&on&ca]le&blood& An?bodies&bind&gut&an?gen&in&?ck& Tick&gut&damaged&

Animal&not&exposed&to&an?gen&during& infec?on&
No&boost&of&immune&response&during& infec?on& Relies&on&presence&of&preZexis?ng&Ab&

Requirement&for&regular&booster&
Ab&levels&in&serum&decline& Memory&is&present&but&is&of&no&use&since& infec?on&does&not&result&in&exposure&to& an?gen&

dependent on the presence of antibodies in the serum. ticks - bite through skin. require regular booster to Slide&11& h]p://www.moa.gov.zw/& keep serum levels high.

Types&of&vaccines:&Life&a]enuated&&
Passage&of&disease&in#vitro#or&in& nonZnatural&species&&
Mutants&occur&that&are&no&longer&as& pathogenic&(i.e.&a]enuated)& Ocen&unknown&what&has&changed& Random&process& For&example&poliomyeli?s&vaccine&&

Ra?onal&muta?on&of&pathogenicity& genes&
Need&to&be&iden?ed& Each&is&single&muta?on&so&poten?al&for& reversion/complementa?on& most used. loses pathogenicity in culture if single point mutation -> could potentially revert back to pathogenicity
Minor&P,&Nat&Rev&Microbiol.&2004&Jun;2(6):473Z82.& Slide&12&

A]enuated&vaccines&

Have&many&of&the&characteris?cs&of&the&pathogen& since live attenuated can still get sick Ocen&provides&good&immunity&& with the same symptoms. lasts for a Some&induce&diseaseZlike&symptoms& long time Can&cause&disease&in&immunoZcompromised&animals& Slide&13&

Issues&with&life&a]enuated&vaccines:&Recombina?on&
Two&vaccine&strains&can&recombine&to&form&a&pathological&strain&
Recent&research&from&Melb&Uni&Veterinary&School& Infec?ous&Laryngotrachei?s&Virus&(ILTV)& Two&poultry&vaccine&strains&in&Australia&
SA2/A20&produced&by&Pzer& Serva&produced&by&Intervet&

Recombina?on&event&leads&to&virulent&strain&

do not use both vaccines at the same time

Vaccine&strain&A20/SA2& Vaccine&strain&Serva&

Virulent&strain& SangZWon&et&al,&Science,&2012,&337:188&Slide&14&

Types&of&vaccines:&Inac?vated&
Killed&whole&organisms&(viruses,&bacteria)&
For&example&inuenza&vaccine&produced& &in&eggs& Toxoid&vaccines&(i.e.&inac?vated&toxin,&for&& &&&&&&example&tetanus)&& Very&commonly&in&veterinary&medicine& Majorly&induce&an?body&responses&

& Advantages:&
Safe& Standardized&successful&technology&

Disadvantages:&
reduced&immunogenicity&>&mul?ple&vaccina?ons,&adjuvants&

Slide&15&

Types&of&vaccines:&Recombinant&protein&
Cloned&protein&from&pathogen&
For&example&Hepa??s&B&vaccine&
Surface&protein&of&Hepa??s&B&produced&in&yeast& Spontaneous&assembly&into&a&par?cle&

Majorly&induce&an?body&responses&

Advantages:&
Safe& Standardized&successful&technology&

Disadvantages:&
Reduced&immunogenicity&>&mul?ple&vaccina?ons,& adjuvants& Expensive&>&few&veterinary&applica?ons&

Slide&16&

Types&of&vaccines:&DNA&
Injec?on&of&DNA&>&animal&produces&the&vaccine&
Fish&vaccine&is&only&commercial&DNA&vaccine& No&purica?on&of&protein& No&classical&adjuvants& Can&use&immunoZmodulators&incorporated&into&DNA&vaccine&

Advantages:&
Can&induce&cytotoxic&T&cell&responses&

Disadvantages:&
Very&low&level&of&DNA&expression& Generally&not&very&ecacious& Expensive&>&few&veterinary&applica?ons&
genengnews.com& animal muscle cell makes its own protein goes into mhc1 pathway good for inducing cytotoxic t cell response. mhc2 presentation as well..?

Slide&17&

Vaccine&formula?ons:&summary&
Live& a]enuated& Immune&response& Humoral& Cellular& & Memory& & Manufacturing& & & & Safety& & B&cells& CD4+& CD8+& Humoral& Cellular& & Ease&of&produc?on& Cost& Transport/storage& & +&+&+& +&/&&Th1& +&+&+& +&+&+& +&+&+& & +& +& +& +&+& Killed/protein& subunit& & +&+&+& +&/&&Th1& & +&+&+& +&/&& & +&+& +& +&+&+& +&+&+&+& DNA&vaccine& & +&+&+& +&+&+&Th1& +&+& +&+&+& +&+& & +&+&+&+& +&+&+& +&+&+& +&+&+&

Source: Gurunathan et al, Annu. Rev. Immunol., 2000

Slide&18&

Vaccine&components&
determines which specic antigen is recognised.

An?gen&=&Adap?ve&immune&response&
Specic&molecule&from&pathogen&against&which& immune&response&is&induced& Need&to&be&neutralising& Ocen&on&surface&of&pathogen&(an?body&accessible)&

Adjuvant&=&Innate&immune&response&
Increases&immune&response&to&an?gen& Increase&in&terms&of&be]er&memory&response& not&necessarily&primary&response& Longer&term&immune&response/memory& Denes&TYPE&of&immune&response&induced& No&an?gen&specic&response&to&adjuvant& Ocen&works&through&TollZlike&receptor& Vaccine&vector&can&be&seen&as&adjuvant& An?gen&dose&sparing& ^ e.g if used as live attenuated vaccine
kecat.site88.net&

ISCOMATRIX:& &&&&&&&&&cholesterol,&lipids,&saponin&

Slide&19&

Choice&of&an?gen&
Trial&and&error&with&some&ra?onal&&
If&an?bodyZmediated&needs&to&be&on&cell&surface& If&T&cell&mediated&no&requirement&to&be&on&surface& Preferably&boostZable&during&infec?on&

An?gen&varia?on&and&selec?on&
Trypanosome&(sleeping&sickness),&successive&waves&of& parasites&with&dis?nct&surface&an?gens& HIV&>&rapid&muta?on&and&selec?on&&

pressure on pathogen -> mutation to try and evade immune system

Male&and&female&Schistosoma&mansoni&in&copula,& transverse&sec?on&

www2c.cdc.gov&

microbiologybytes.com&

Slide&20& ncbi.nlm.nih.gov&

Choice&of&adjuvant&
Dene&immune&response&required&
CTL& Th&(which&type,&Th1/Th2/Th17)& Ab&(which&isotype,&IgG,&IgE,&IgA)&

Choices&are&limited&
Alum&is&most&widely&used&in&humans&and&& animals&
Good&Th2&but&poor&Th1&adjuvant&
dpi.qld.gov.au&

MF59&(Squalene)&approved&for&human&use&in&Europe&but&not&US&
Novar?s,&Inuenza&vaccine&

AS04&(monophosphoryl&lipid&A&(MPL))&a&TLR4&ligand,&licensed&in&Europe&
Combined&with&alum,&in&GlaxoSmithKlines&Cervarix&and&Hepa??s&B&vaccines&

Quil&A&approved&for&animals&but&not&humans&
Good&mixed&response&Th1/2&but&side&eects&make&it&unsuitable&for&human&use&

cytotoxic response, hypersensitivity mucosal - want iga large parasite - want mast cells, ige compliment xation - igg best one

Slide&21&

Current&adjuvants&and&how&they&work&
Depot&for&Ag&&
alum,&incomplete&Freunds&adjuvant&

Induce&inamma?on&

Inammasome&(alum)& A]ract&cells&to&the&site&of&injec?on&(Alum,&complete&Freunds&adjuvant)& Target&Ag&to&dendri?c&cells&(CTLAZ4&targe?ng)& Ac?vate&dendri?c&cells&(see&TLR&below)&

Target&Ag&to&specic&Ag&processing&pathways&
ISCOMs&

Increase&access&of&immune&system&to&Ag&
Nanobeads&

Ac?vates&Toll&like&receptor&(TLR)&pathways&
&complete&Freunds&adjuvant& CpG&mo?ves&in&DNA&vaccines&

Slide&22&

Vaccine&delivery&
Depends&on&species&
10,000&chicken&in&shed&
In&water,&aerosol& Single&delivery!& Injec?on&OK& No&needles&(thank&you!)&
organidad.com&

Ca]le&on&outback&property& Humans&

Dierent&vaccine&vectors&
Delivery&depends&on&vector&used& Need&for&cold&chain?&

Route&of&immunisa?on&aects& immune&response&induced&
TH1/Th2&balance&
Slide&23& visualphotos.com&

Mucosal&vaccines&
Mucosal&surfaces&are&important&pathogen&entry& points&
Lung,&gut,&vagina,&mouth,&nose& Very&large&area&exposed&to&many&poten?al& pathogens& Early&destruc?on&of&pathogen&prevents&infec?on& of&other&organs&& mucosal surfaces weak point

Mucosal&immunity&requires&mucosal&vaccina?on&
Lymphocyte&recirculate&to&mucosal&surface& Some&degree&of&protec?on&between&mucosal& surface&
Vaccina?on&at&one&mucosal&site&may&protect&other& mucosal&sites&
myoo.com&

Lung&surface&area&=&area&of&a&tennis&court&

Specic&adjuvants&are&ocen&very&important&
Oral&vaccina?on&requires&specic&adjuvants& otherwise&we&would&mount&immune&responses&to& food&an?gens&

mucosal administration -> more likely to give mucosal response

Slide&24&

Vaccine&ecacy&
Protec?on&from&disease&symptoms&
Animal&is&a&carrier&and&can&s?ll&transmit&disease& but&is&not&sick&

Reduc?on&in&transmission&
Animal&has&a&reduced&risk&of&passing&on&infec?on& Eect&can&be&sucient&to&bring&R0<1&&&

Sterilising&immunity&
Pathogen&is&eliminated&from&animal& Animal&can&no&longer&be&infected&
mo?fake.com&

more than 1, disease will spread, R0 >1 disease will die out.

Slide&25&

Eect&of&vaccina?on&on&herd&immunity&
When&sucient&animals&within&a&popula?on& are&immune,&so&that&disease&can&not&become& established&
Gene?cally&resistant&popula?on& Vaccina?on&of&large&propor?on&of&animals&

Even&if&some&animals&are&not&protected&they& are&unlikely&to&be&exposed&to&disease&because& the&disease&is&absent&from&popula?on&

Some&parents&do&not&vaccinate&their&children&against& whooping&cough&because&herd&immunity&makes&this& disease&so&rare& If&too&many&people&do&this&whooping&cough&will& become&prevalent&again&

www.niaid.nih.gov&

Slide&26&