Viral Vaccines-Vincent Ranciello Viruses are everywherewe are all currently infected with 10-12 different viruses.

We survive the barrage because we have great defense systems-our immune system. When we develop vaccines we are trying to emulate immune system. Vaccination takes advantage of memory and our immune system to protect us against infection. Vaccines wants to make memory so that we can be immune to pathogen when you encounter it.Vaccine breaks the transmission cycle of a host-host spread in a population. If you dont have enough susceptible individuals you break the transmission and that leads to breaking transmission cycle. Vaccines stimulate an immune response just like a natural infection does and then they provide memory. If you encounter virus later you have a very rapid or robust immunological response. Immune memory is real. In 1781 outbreak of measles on isolated island.no measles on the island for the next 65 yrsnot enough susceptibles on island to support reinfection and apparently no introduction from boats. In 1846 another outbreak and by then there were enough new susceptibles to sustain an outbreak, but during outbreak none of 1781 survivors of epidemic got infected.proving immunological memory.Immune memory lasts a long time and is maintained without re-exposure to virus. Memory lasts in the absence of infectionmemory usualy does not require stimulation to be maintained Immune MemoryT and b lymphocy Vaccine-induce memory without effects of pathogen. First Modern vaccine-JennerUsed technique called variolation.-take material from pox and inoculate another person with it.30% of people died but 70% lived and never got sick. Jenner realized that milkmaids who got cowpox on their hands never got smallpox.then Jenner decided to inoculate children with thatReplicates well in skin. In 1885 Pasteur made rabies vaccine Then in 1930s we had yellow fever vaccine. Anti-vaccine movement can be traced to Jenners days. We now use vaccines in massive immune campaign Polio =peak 1950sInactivated vaccine developed and coupled with oral vaccine introduced in early 1960s had eradicated Polio in US. Measles not completely eliminated due to pockets of susceptible individuals. Vaccines are now an integral part of our existencewe immunize people, pets even wild animals to prevent them from carrying rabies-release vaccine laced bait into environment.

Due to vaccines more productivity

Fundamental ideas -Maintenance of a critical level of immunity In order for vaccines to work you have to have a certain level of immunity in population. If you let immunity fall below a certain level you will get an epidemic. You have to keep immunization levels at a sufficient level to block transition -Herd Immunity You dont have to immunize everyone to protect population; you just have to immunize enough to prevent the virus spread. What is enough varies based on virus, population etc. Herd immunity-you can protect whole population by immunizing just a fraction of it.

What is the threshold? Depends on virus and populationpeople actually have to be immune, not just immunized.vaccines arent 100% effected. Virus spread will stop when probability of infection falls below a critical threshold. You could have the best vaccine in the world but if people dont use it, it is worthless.We have problems with large-scale vaccination programs due to societal attitudes. How do you make a vaccine? You need to know correleates of protection You need to balance Th1 and Th2 responses for r.g. Person you vaccinate must be protected against virusproblems ethical control group.Also have to make sure that anitbodies are protectivecant just show in the lab. Two kinds of vaccines Active and passive. Passive-put an immune product into the person e.g. antibodies or immune cells..e.g. rabies immune globulin from people who have been vaccinated against disease..Put rabies globulin at site of bite and then give person the vaccine. Works because rabies has long incubation period. When youre first born dont have a well developed immune system and you get antibodies from mother that protect you until youre immune system develops.

You are protected by mother up until around 6 months of age. By around a year, youre immune system has been fully developed. Active-Put modified form of pathogen into person so that they can develop immunity against pathogen Requirements Minimal side effects and safety, Protection must be long lasting Must induce protective immunity in the population Not everyone needs to be vaccinated due to herd immunity Low cost, genetically stablemutations causing pathogen to be safe cant revert, storage considerations, delivery (we would prefer oral because it doesnt require trained health care personnel and liked by more people.

Ways of making vaccines -Attenuate it-live natural virus given -Inactivated Inactvated virus vaccine Fractionation-Non-recombinant purified subunit vaccine Cloning Take viral genes and express them in a vector Tke virus DNA and inject it into muscle Make virus proteins

Express viral capsin proteins and make virus like particle proteins. For many viruses, the viral capsin will assemble when you just express one or a few proteins.

When you get chicken pox, 50 or so years later it can reactivate as zoster

Inactivated Vaccines Grow up virus, it is infectious but then inactivate it by chemicals etcyou thu have a non-infectious disease, but antigenicity is not compromised and so you will still get an immune response

Poliomyelitis Virion made up of four different viral proteins repeated around 60 times to form structure that protects virus RNA genes. The capsule is beautiful but just protects genime and allows it to go cell to cell. Polio develops as a result of ingesting it e.g. oral fecal. Virus can pass through stomach and intestinesa and gets in body through small intestiens.gets in blood and then into nerves through axonal transportreplicates in neurons in spinal cordand destroys them and then can get paralysis. As the virus gets replicated in small intestines, it is released in feaces also and thats how you can transmit to other people. First sizable outbreak in US-NY 1907barely on medical radar before this because incidences were so sporadic. Humans are only known reservoir of polio-important because we are trying to eradicate polio from globe and one of the requirements of eradication is you can only have people being the reservoir for the virus. Peak of disease warmer months Spread by fecal-oral transmission

Clinical features 90% of infections have no symptoms or some simple symptomsspinal paralytic and bulbar poliomyelitis which cause paralysis are rarer.

Inactivated Polio vaccine (IPV) Treat polio virus whith formaldehyde or formalin inactivates it, then can inject it in the muscleJonas Salkused until early 60s 10 days after vaccine was releasedpeople came down with polio..traced to certain lots of vaccines Cutter labs didnt execute Jonas protocol properly and virus wasnt completely inactivated and some kids were injected with infectious virus. IPV injected, make antibodies in your blood.antibodes in blood attach infectiou agent in blood.

INFLUENZA Very different from polio vaccine. Has ENVELOPE-LIPID MEMBRANE AROUND GENETIC INFOembedded in it are HA and NA Three types of influenzaA, B and COnly A causes pandemics, though B also causes similar illnessC very mild illness Pandemic-global epidemic of infection

Flu virus varies from year to year, hence we have to keep updating vaccine Get infected by inhaling virus, enter respiratory tract through mouth or nose etcvirus replicates in epithelial layer of the tract and can replicate anywhere from upper to lower tract. Sometimes it can infect upper tract or lower tract-fatal cases of influenza due to pneumonia. Short incubation period 1-5 days Main vaccine we use in the US is grown in embryonated chicken eggs-formalin inactivated or detergent or chemically disrupted virions. Then inoculated intramuscularly . Protection correlates to antibodies to HA and NA proteins in virion itself. Virus different from polio in that as it multiplies in hostit changes genetically and antigenically. Every two or three years we have to reformulate vaccine, because it no longer matches predominant circulating strains. Strains must be selected in the first few months for manufacture Polio and influenza-Inactivated vaccines

Subunit vaccine Break virus into components and use purified components to immunize, or clone an appropriate viral gene and express in cells whether bacteria, insect cells etc, then purify protein . Antigen used is usually a capsid or a membrane protein. Advantages-Because made by recombinaent DNAwont be any infectious agent Disadvantages- Can be expensive, also antigenicity can be poor becuause you are using a singe viral protein that doesnt replicate at all, usually just induces production of antibody, so if your virus requires production of t cells for protectionusually not a good approach, we dont have a good way of delivering them have to use injection . Problem that they dont replicate, and thus they dont induce innate immune system so that it can prime for adaptive.these dont cause inflammation, so for many of these we basically have to add adjuvant, a chemical which will induce inflammationproteins themselves dont produce inflammation.

Many years ago an inactivated vaccine to respiratory syncytial virus was made but it didnt work at all in fact those that got it, got worse diseaewhen rsv infects cell, engages TLR 4 and cytoplasmic RNA censorsthat gives a very good adaptive response, affinity maturationfor vaccine it doesnt replicate and does not engage TLR 4 makes no RNA and you dont get good adaptive response,

very crappy antibodies..low affinity.and get a lot of inflammatory reaction in lung which makes worse diseaseVirus needs to be pro-inflammatory. Adjuvant helps cause inflammation, make a very good acquired immune response, present antigen as particles, help localize antigen and stimulate inflammation. These all help improve antibody response

Hep B virus is a subunit vaccine capsid protein produced by yeast -Assembles into empty particles HPV vaccine L 1 PROTEIN can be cloned and expressed then purified these will be reassembled spontanreoudly into empty virus particles. Together with adjuvant gives a very good response.

Infectious Attenuated (live) vaccines Manipulate virus in some way so that it no longer causes disease..in inactivated you have to get multiple doses to get good response eventually. With infectious you can give one dose and it will amplify itself because it is infectious. Typically made by passaging viruses grows in human cells transfer to monkey cells hopefully acquire mutations that allow it to grow well in monkey cells but not in human cells..thus not caussign disease. Another kind of polio vaccine is infectious attenuated (OPV). He took virulent polio virus and passd it in a variety of monkey cells until eventually coming up with mutant that would no longer cause mutations in people IPV gives you immunity only in blood, OPV gives you immunity in gut and blood.

The OPV can sometimes can cause polio-vaccine associated paralytic polioyou excrete more virulent virus than IPV.switched back to IPVthus no vaccine associated polio and no wild type polio We get vaccine associated poliowhen you ingest vaccine.ends up in gutmutations revert within 2 or 3 days..and they become virulent and you shed virulent virussome kids react to virulent virus and develop poliomyelitis.

Flu Mist-Live attenuated These are cold adapted and temperature sensitive mutantsso prefer to replicate in upper tract

GENETIC SHIFT AND DRIFT SHIFT diversity that arises after reassortment of genome with a different strain relatively rare Drift diversity arising from copy error snad immune selection can happen each time genome replicates.

Genetic drift occurs by point mutations in HA..thus eventually virus will not be recognized by antibody and you have to change the vaccineInfluenza virus RNA in 8 pieces in virion when these virus infect cell.you can get mixtures of progeny if for example cell had been infected by two viruses 2009-brand new strain emerged from swinereassortment of swine and avian and human strainsrna segments from various virusesreasortmen tis thus a big problem. They are trying to express HA like particles in plantsthey have been seen to be immunogenic and this is a very cheap alternative to chick egg, Also it takes longer to formulate and produce a vaccine in egg.for plants you can get nucleotide sequence on day 1 and then plant based vaccine would be ready for first wave of pandemic. Temperature Stability Mix with silk stabilizes vaccine Microneedle Carbohydrate glass-stabilizes just like silk fit on syringeas buffer passes through, melts carbohydrate glass Can we eradicate viral disease? Humans must be only host Vaccination must induce lifelong immunity

Problem with using OPV.can cause outbreaks.if you stop immunization you will still have circulating vaccine derived poliohave to vaccinate against vaccine