Two Birds with One Stone: A Rabies Virus-based Vaccine against Filoviruses

so what I’m trying to do today is actually talk first a little bit about rabies virus because that’s you know the old laughs since 25 years and then go from rabies wise to Ebola virus which we work for about five six years now so rabies virus is another and neglected infectious disease it’s a well known disease actually to mankind but it’s a huge problem worldwide and what is probably not well known so this is actually also very simple virus it’s also negative stranded on a virus which are all very similar organized they have a host cell they have a host cell derived membrane so they all enveloped viruses they have a negative stranded on a as a genome which is only infectious and the infectious template when they ain’t kept sedated in the nucleoprotein they this is this protein therefore phosphoprotein all disruptor viruses which actually is a polymerase up unit also interfere achenes antagonist a matrix protein and a sing out like a protein here and and the beauty of these viruses is actually that you can actually replace functional disk like a protein here or you can add additional aren’t like a protein that’s basically the vaccine approach but before I actually come so all of you know this here classical rabies virus genotype 1 is responsible probably four ninety-nine percent of the human deaths but but you probably don’t know is there 16 other genotypes which go in to Philip groups and a lot of them are actually also emerging infectious disease and in reddit alter our viruses we don’t have a vaccine against and you attend today also in Africa and especially mccollough virus in South Africa is a problem and save us human death says no Sarah pions no vaccine so but all these viruses are very similar they basically you can distinct you by disease if it’s a classical rabies virus or Marco levels very similar disease just antigenically they different and most private eyes are highly nor dropping into infected house and cause a fatal and tefilin the lightest in mammals um and that’s without vaccine and actually rabies is after infections the bubble is the most lethal virus which kills a hundred percent of infected people and animals so hosts are all mammals it’s available mantra is at 40 to 70 thousand die from the disease each year probabilities and numbers are much higher this is space that in Africa we really don’t have a good reporting system and when they checked in Tanzania for so-called malaria and used encephalitis and children ten percent was actually rabies that tells you about how good that arm system is it surrounds the u.s. to post-exposure prophylaxis a good thing about rabies viruses that you normally know when you got exposed right you get bitten by animal so bad news is that’s a bad arrived rabies in the US a lot of people actually do not realize when they get exposed that’s why they tell you if you find the batteries in your bedroom catch it and you know let it analyzed or get rabies vaccination so lucky part about the US is again that all bad derived rabies why since the US is classical rabies virus of the vaccine actually will protect you oh that’s just to walk you a little bit Susan um boss those are different the u.s. is very special with rabies so we have different species we have raccoons skunks we have foxes and we have pets which transmit rabies virus and here’s the numbers I’m kind of emergency area for rabies I mean it’s uh it’s unbelievable how much rabies we have basically if you find a raccoon or skunk in your backyard it’s a good chance that they have rabies the numbers are really and and I have to say the USDA really fail to do anything vaccination wise because Europe got completely rid of rabies by vaccination where I have to say foxes are very easy to vaccinate with life Bates so with life virus it’s a little bit harder for skunks and raccoons this year shows analysis numbers here and more interesting is actually this is a little bit when you look you will find but that’s that’s bad

rabies virus and that’s the cases but you have to be careful because you really don’t know if you will be exposed or not and we want to study a little bit bad so bad in beds rabies is not as part of chanock sin in other mammals which it couldn’t because they live so close together if one has rabies every old thing would have rabies so that’s interesting but again worldwide distribution here you see rabies is actually everywhere except here it got actually eradicated by vaccination so the highest numbers are from India but that’s only because they have a relatively good reporting system Africa we really don’t know rabies virus infection is relatively simple you have normally transmission by bite you have no house response to the infection normally it goes we don’t know where it replicates first it’s a non lytic virus because it has to go up and motor neuron we know exactly how fast it’s about one millimeter each hour it revels up so you can calculate from that one it will reach your brain it moves to the cns and then it uses behavioral changes I mean it’s all pretty fascinating for 5 cheats you know it infects actually areas which normally are charge of your emotion and that makes a cursive behavior and then you have the two forms this is an older picture so if you have symptoms of rabies there’s really not a lot we can do for you they just retain people and so now we put them in a in a coma and you basically wait until they there’s some publication about therapeutics which I think about really working and they chose these the two phases so furious ray and it’s dumb we have a smaller topic that’s more like it is in humans you really don’t have the Furious disease in in humans there was a case of a lion with rabies and they get so furious disease that’s really not what you want the rabies infected line I guess so sorry so so basically pathogenicity of rabies very very shortly polygenic viruses replicate really slow spread fast they are less amano genic basically they don’t make a lot of protein they just sneaky they stay under the radar screen of your immune system they go into neuron and go up to your brain and we have two things which are also important new oral person para chanak rabies virus have a high specifically to infect neuronal cells and no other cells which make sense and then it’s neuro invasiveness this is a ability of virus to end us a CNS so strange thing is all these paired viruses are not really neurotropic but I highly neuroinvasive and we have no clue how that works so there’s still a lot of research to do now we come closer to the vaccine so rabies virus actually good vaccine so stories that I put it in your stomach 20 times it’s all not true in the meantime it’s I am like your flu vaccine you get three inoculation at a 07 and 21 to 28 send you normally immune for the rest of your life some people have to be boosted some don’t we get because we work with rabies we get letran every half year and tested for neutralizing antibodies which predict if you protect it or not good thing about rabies you even have stand out you have international units which predict if you protected and then about 30 to 50 million or so vexing get used which is argument for our combined vaccine post-exposure treatment you know over arm rabies is one of the diseases where you actually can do something after the fact what you do is active immunization and passive immunization at once so it’s a human arm i chi chi 0 so it’s really i’m from volunteer which they vaccinate and boost several times and it’s humans here as they are also some neutralizing antibody which may replace that it’s very expensive it’s two thousand five hundred dollars is one treatment with the serum and you have sir details and post-exposure treatment since the US alone so it’s good business I guess incubation times three to eight weeks in general there are description of cases after two years it’s true but in general mostly three to eight weeks the incubation time and after that mortality is a hundred percent again furious are

done which means you have this hyperx ability that’s aggression debts Hauser virus actually gets transferred by the aggression and biding in both cases you have debt by courier or respiratory arrest and what is really interesting on rabies too and I just briefly want to show that before I go actually 12x in approach you really do see not anything and save us I don’t know if you heard about it cases where I transmitted rabies two three or four people because they thought somebody died of a stroke and it ran splendid so organ and transplant and that person actually died of rabies and the reason I have to defend the doctors here a little bit you don’t see any pathogenic changes in the brain so if you look that looks like the normal here you bake a spade you have it full of rabies antigen here early on but you don’t be silly any apoptosis or damage of syneron so it’s really a hallmark of rabies virus and what a little bit research on that what we actually how does that happen how does the virus get cleared from the brain or how does it affect the neurons and save us want area neural cell that hypothesis or neural dysfunction and to analyze that we actually use that cream portimao system which has this gene so every cell is red and if you cut it out by expression of the three recombinase they turn to green from red to green so basically if in fact with the rabies virus expressing that protein you change to green this is a rabies virus with the query companies this is some reporter cells if you infect them with regular rabies virus they stay Brett and if you infect them it’s a tree they change to green the problem is really if a cell get cleared after the infection you won’t find it anymore right so you can’t look for the antigen so if if you use a not so pathogenic rabies virus and you look in a mouse two months later and it’s arrives you really don’t know what happened to sell so with this system we want to solve that problem and it actually worked better than I thought so that’s infected brain over time for my surviving so we use the vaccine strain of rabies virus a day 7 day 15 you see this infection so that’s all infected cells which change to green after 1.5 three months after six months you see all the cells are still there and we actually sorted for green and red cells and show that this neuron had our functional changes so what it means is actually that we want to look a little bit more into debt because that also tells you about something about therapy the damage may be already done in this neuron you probably can’t just restores a function but now I want to go to back to civ xine against the boiler virus I’m starting with this rabies was a worldwide risk but it’s all areas where you actually should get vaccinated if you go in this area I mean not in the cities but in remote areas and that’s where acquire rabies is a high risk and that was our starting point before we actually had the last Ebola outbreak because ever SPECT scenes out there but financially none of them were really attractive so we thought combining basically two vaccines in one would be a good way to address a problem Philip irises I really do not have to go in details through that because every guy really did a good job to explain all of that we have basically three main candidate Ebola virus sedan virus and malware virus they now separate list their separate virus before they wait together and as i said i’m also and when up viruses is that you know Seventeen’s about 15 kb the shows you such a baller virus of fascinating thing about Ebola virus that is a really long virus about four or five times longer and then rabies virus this is fiddle like structure and when we talk about vaccine icings a field in the meantime agree that really dis glycoprotein which is on the surface is the best effects in target I think that’s at least one was a field can agree on if we look a little bit into subliminal Chi I mean it’s not really

new phila virus outbreak it was not predictable that it will such a big one but I mean 1976 we had the first out pregnant and we had outbreak every year a little bit it was predictable that at some point that will explode because most of this previous outbreak we’re really in remote areas which you could contain very well so the World Health Organization CDC went in and kind of make sure that everybody stays in this area so today with people more traveling and it’s actually well established that they actually check I probably started with one single two-year-old boy who got it probably from pets and then it spreads to funeral in a large area that choses area mostly Guinea Sierra Leone and Libya RIA well we’re certainly at the borders I mean every country really really wants to be I’m not having any Ebola cases but supporters are pretty often so I think there’s certainly some spread outside this as the newest number so it it seems to the number seems not to be rising as bad as initially was expected but I still pretty high that’s updated hear from you know chose you I can’t spell January so 20,000 cases about 80 s there and 13,000 confirmed case this is really important and I don’t know if people appreciated that the confirmation of these cases is so important because less Africa has a lot of hemorrhagic diseases and if you would live there and have Lassa fever which is not as deadly as Ebola or you have yellow fever or you have another hemorrhagic disease you have malaria of his high fever you really don’t want to get together with the people having Ebola so the diagnostic system is really really important that these people trust to go to the hospital for treatment okay that’s the case numbers briefly about the pathogenicity I think it’s velo stablished what happens with with Ebola virus that you have entry into macrophages and then it uses the lymphatic system to spread all over your body it replicates very well in lungs kidney liver at some point you just have this really really high viral load and you have this organ failure from the me and to liver and debt results and debt it’s not it also arms endothelial cells start to get leaky but it’s not always a hemorrhagic disease per se i think only fifty percent it doesn’t matter mostly organ failure which results to this it’s not airborne here’s the lungs of people’s it’s really another airborne disease you really have to get in contact with body fluids but at the end where the people there people die sir viral load are really extremely high you have ten to the seven viral particle I mean this is a lot so if you touch without protection do you have a good chance to get infected but it’s not airborne so this I really can skip because Erica already told you all about the receptor and macropinocytosis which which i think is interesting by itself because that’s not how virus is normally effect and then you need is cleavage shown here and this is actually the only it’s a npc one that’s the only identified receptor which is essential and this other one here initially this host factor binding that already tells you House factor binding is not very specific are all replaceable so that’s that’s really arms important binding but every cut didn’t mention it there’s also something else about Ebola enough for mouth but Ebola which tells you the importance of sandy buddy because survivors takes a lot of effort to make solar boat like a protein which may be Miss folded and which is also may be a target for anybody just to deplete antibody so basically eighty percent is that short soluble protein here which is just put used and pumped out and only twenty percent is a real protein and we know if you change that and you just have GP full-lengths which happened actually in some of the vaccine study you have a highly a tener

virus already so the soluble protein is important so in general debt to gather the data which i present it was making the field thinking that anybody against GP is probably how you can protect against ebola virus so i run you a little bit to say bola vaccines and i want to point out again antibodies you have virus neutralizing antibodies but you also have antibody-dependent cellular cytotoxicity which i think is actually a major a mechanism of protection you can have this antibody opsonization do you need antibodies do you need b cells are off alarm circulating antibodies enough or do you produce anybody and then certainly the biggest controversy on traversée is a function of cd8 t-cells do you need cd8 t-cells or did you meet just for clients so basically naive one which amplify and just clean up or did you do you need memory t-cells and it seems to be a little bit difficult I’m different for different vaccines and how well they work so frontrunner we have two frontrunner you all heard it so says a chimpanzee aetna virus vaccine cheez ke vaccine which which is basically mostly that work was initially all done by Gary Nabal and actually men Nancy force a driving force behind it the chosen one that is it’s a replication deficient virus enter concern was as you know from the HIV at no.5 a lot of people have antibodies so the vaccine really doesn’t work well and if you put it in certain people at least for HIV it can even increase infections so not a good vaccine for Africa probably where HIV is prominent so people started an action that was initially done for the HIV vaccine to this chimpanzee at no virus and that chimpanzee aetna virus three and say as a sick c3 and shortly that was shown if you use that at nursery that’s the best one you give 10 to 11 that’s what I actually use into human trial to then you get a good response 10 to the 10 you don’t get such good response it’s a problem and that was recently actually published on this vaccine is that the antibodies go down over time so after six to eight months you lose basically your protection and you have to basically boost and you can actually boost with the same virus you need here the mva so that would be the approach I still have to say for emergency vaccine that probably would be okay 10 to say 11 but for long-term vaccine jsk has to work a little bit on that and introducing another viral vector won’t be that easy if you have to prime boost is two different one so the next one which you heard a lot about is probably the vesicular stomatitis virus vsv and rabies basically looks the same bsv is also a little bit nor dropping and you see here this extremely busy table that just is here to show it was extensively tested with stylus to 10 to seven and they always had protection they tested it against sudan they tested it and it is a very good vaccine but it is a replication-competent vector so as i told you before for rabies virus you can functional replace a flying a protein so that’s vesicular stomatitis virus you take own out is own clogger protein and put in arms a bowl out like a proteins and you have a replication vsv which uses a sable a cheapy to infect cells which works well but had some recently it was tested that incidents of joint inflammation was very high so it’s a second part of clinical trial now they go back from five times ten to the six to three hundred thousands of three times Centers for so certainly a hundred fold difference to that so we have to see how the antibody responses are but we don’t have any information on that yet and here are the other approaches because it’s a press you hear only about the two cells also I mean initially inactivated virus probably not a good idea and it really doesn’t work rallied up but they are certainly virus-like particles which you make from this protein which showed

armed protection DNA and recombinant adenovirus vaccinia virus actually didn’t work at all but say as a paramyxovirus human human cold viruses mostly which are used and worked well there are certain is a concern about pre-existing immunity vsv works well and say the rabies virus which I will spend some time now to talk about our vets in approach so in general for vaccine all chiefs the general slightest that’s our problem here so you have a replication-competent vectors they normally really really monogenic right replication deficient they less so because they can’t spread within the host and they keep on spreading until they cleared so they’re normally good vaccines mumps measles rubella is example for replication-competent viruses which work well replication deficient vectors I don’t think we have anything as current reticent but we certainly have deactivated viruses like for service virus or chemically killed virus where virus-like particle certainly vlp hepatitis B is a VLP produced in yeast what’s actually pretty good mostly you need to inoculation send VF DNA vaccines and we have protein itself which is mostly not really a monogenic without any active n because there’s no really good reason for your body to make a response against a single protein and then to get these things better all this here morimoto genex and you add at you and then you f ed event you go with it it’s a risk area again so it’s all the balance and try an arrow to make it with vaccine so our approach for rabies virus the sort was and as I said it was four or five years ago already to generate a recombinant rabies voice based vaccine vectors I’ve expressed glycoprotein of Ebola virus we thought that a desired vaccine Ebola or Marburg with a proof financially viable vaccine such as rabies vaccine would be a good idea justification at least twenty four thousand rabies test in Africa I mean if you think about that silly numbers that’s still very higher since I bola and the other idea of us live attenuated rabbits vector vaccine could be desperate Oh Africa arms are primates you know that primates are major victims of Ebola outbreaks the numbers are even higher one outbreak killed about 5,000 of them so oral vaccine for something like that is certainly something we would like to consider too and we saw that pre-existing vector immunity safety and high costs have which F in the previously every scene would be avoided by this approach so that’s our vaccine candidate rabies virus as I said five genes and p.m. and here’s a glycoprotein we feel so attenuated so this is already a vaccine vector used for all immunization in in Europe we put add additional are mutations that like a protein which prevents infection of neurons at least the preferential and then we just popped into a bullet like a protein into this vector we also made one without cheese similar to vsv ours didn’t spread for whatever reason I have not a good explanation for that and so this approach would be taking these variants inactivated by ppl which is a normal message to deactivates the current rabies vaccine and you spilled particle effects in similar to rabies virus the shows you some am studies of our particles we labeled with two different gold particles a larger one this would be the vector are directed against this actually is a small there’s a larger one against the vector and the smaller one here against GP so you get labeling of only glycoprotein and here you get labeling of rabies glycoprotein and Ebola GP so you see this protein is incorporated this is a little movie showing the same thing just enhanced by color two of this doesn’t want to play well sometimes it doesn’t work sorry anyhow you see here survival particles in green and you see here’s the rabies

virus glycoprotein retsupurae GP so this this virus v be constructed I just show some part in ECG data before I show some monkey data so what you normally do if you have a virus and you want to eventually use it also as a live virus you do different inoculation for the potential of this virus to invade the cns and for spread and new Overland’s by direct inoculation into the brain and basically what can you see here is that after i pee inoculation so weight of all the mice goes in the right direction so there’s no patron icity same after i am and here it’s interesting this is now interserve repo I said told you before our vector off with our distress we stream mutation is still neurotropic even it’s a current Wildlife magazine and so you see if you put a direct in check the directness of brain so my svelte I lose weight and die but if you add this me 33 invitation you have a perfectly safe vector which doesn’t spread within the brain now we go to our arm first non-human primate experiment against the vectors and if you memorize the color it will be easier so bns p333 in black is always a vector the empty vector in red is a life replication-competent employs a replication deficient because i told you our GP virus doesn’t spread in fact by doesn’t spread and in green is basically surrett one which was inactivated by BPL this one we had four groups of monkeys three control monkeys which cuts a vector of five times ten to the seven group to five times center seventh our replication deficient 10 to seven that’s highest i love you on two hundred and fifty micrograms to inoculation this replication-competent you can do only one inoculation because after someone you have rabies immunogenicity and you can’t really do a second inoculation which is also an advantage of this because you can use that multiple times here you see prime boost and send V challenge because we used rhesus macaques we challenged to controls a little bit earlier just to make sure that our challenge virus really kilts this monkeys and here is the other control and select cities and then we follow basically by standard on the animals with blood draw and analyze the immune responses that comes here and this actually controls here sometimes monkeys like humans it’s rare but they survived Ebola infection so that was from heinz Feldman a controlled CRF our Eliza essays which I show you here at day 07 14 28 35 42 and that’s groups of the monkey so this you would sing if you reach that level you should be fine because it was a surviving animal at least if antibodies are some major effector cells here and you see a day zeros as nothing’s and we immunize at a seven you see a little bit goes up at a 14 day 28 and this is Miss antigen immune responses just keep on going up over time because the antigens are pretty stable this viral particle here day 28 we boosted this screen group is was a replication deficient you see it reaches that what made me happy at least for some days I tell you the end of the story soon so but that we would think that’s about the similar antibody responses we need for protection as stay 42 that’s when they were transferred for challenged so now the progression expectancies you immunize and send your challenge and serene out is survival right and that’s what we did until you see post challenge day so they were all played at the day of the challenge and here I giveaways arms results survivor is a redbox and what you can tell here alter our vector immunized animals didn’t survive as expected all the life viral vector animals arrived here you see this two survived of the replication deficient and of the killed only tues arrived and two died so if you follow them longer you see day 16 after

challenge to d 28 you really get nice immune responses you challenge and say moon response goes up and up and up and protected antibodies are animals end up with a lot of antibodies so that seems to be really some measurement of protections a problem boss oh yeah and invite before i show that other I just recently found that paper which I think it’s interesting because it lines really nicely up survivor load so if you do a nonhuman primate experiment you know if your monkeys reaches tied around 10 to the fifth ten to the seven you probably won’t save them it’s maha man actually to sacrifice at this level and if you look here it’s really survival load this is clearly an indicator of pathogenicity and survival so this is actually these viral loads are from human and you see survival rate from about thirty percent if you never detect to tie the highest intent to the fifth and really hide that rate if the virus goes higher and that’s pretty much what you see non-human primates to this year’s completely unrelated to my talk but I saw that’s interesting too because you get all these messages how easy some people safe human and see other war not and it may it’s really 8h related you know as younger you are as more likely you survive so I have to stay way I would be dead very eighty-five percent so H is a major factor and they say in certain areas they have more younger patients so we have to be a little bit careful if we say says really now a therapy for Ebola okay and now we come back to survival load and what I really liked about our that’s a complete viral load over time but if I go here you see supports out these three bold lines they never had any viral load one of that had no viral load and this one never had virus so they were completely protected if you look here this which has a bit little bit lower antibody this is this here and this is this one really doesn’t line up too much but DC lines up so the second one has previous short and if you don’t see good antibodies that what happens and you get a lot of viral load and that was animals which had to be sacrified however now comes if you look to when we went into the challenge in this animal here and here and here they are very similar this antibody levels right I mean this year what is the difference and that was what puzzles are why do to survive and why it could be CDL responses but I really believe monster antibody and we looked a little bit more and Sandy buddy and what we really saw here if you look to the igg want to I 222 in humans and monkeys igg1 is th one responses and tth one responses are necessary to actually have a DCC so if we look to that over time after challenge then you actually see that all the animals arriving here they go more to th one versus the other one go more to th2 so we really believe that the antibody response is a major and it’s not significant if you take the replication fusion but if you take this to this arriving replication-competent and killed it’s it’s highly significant that th one bias is important and I don’t want to say if you don’t certainly the level of antibody is very important to but I think the quality of the antibody is very important to when you need more th1 response so what else are we doing I’m almost done I’m still on time I guess how do we increase th one buys for Sabo la GP so for humans we really want to kill tracks and everything else is I would you know it’s just rabies now get hard to get ever proved so what we think so we have two hypotheses one is Ebola virus is controlled enlarge and end by anybody with anybody dependent cell-mediated psychosis playing a major role that’s that’s our overall hypothesis servicing we don’t say neutralizing antibodies are not important but I think you need both neutralizing and non neutralizing antibody we know that rabies virus she has a complete th1 buyers response not completely but a very very strong and

once area it’s kind of a desperate one we think that’s a high density on sarabia values chi answer surveys is important there’s really no no good basis in immunology I still think that structure recognition of structure is very important any moon responses at least it makes sense and in any case having more of your antigen of choice in there would be a good thing so we thought can we improve cooperation of GP and send getting a more th1 bias or and a higher immune response and what we did for that was a codon optimization something I always thought won’t have a dramatic effect but it has so we codon amazed arms sigh at GP if all our GP this is our vaccine which are just showed in this experiment and this is a code and optimized expressed and we get about four fold more increase in fact now in survival particle which I really happy about this is rabies chi here and this is Ebola GP and they equal it’s about ten percent now within the party go and we did one quick experiment which is also a little bit confusing because we had to use at event basically I got four monkeys from Z NIH but I wanted to test new at events and they said without at events we don’t test the vaccine and said okay what can you do for non-human primate we reduce the amount of antigen so that’s worse a new particle one used the tlr 12 agonist 10 tlr3 it’s from a company so i can tell you what it exactly is they were immunized twice this 100 micro gram and we had a hundred percent protection after challenge so situation it certainly can be gathered without at events which would be the preferred way to go because i learned that at your end this really makes your life very difficult with the FDA so in conclusion I think we identified the best candidate at least for for rabies virus with a codon optimized we think that’s a PP ppl inactivated vaccine is the best we can use we actually dis started to be in society of a grant and IDT biological is a contract on Germany we already produce arms a year Sudan and malbork in parallel and they were all recovered on a proofed cell line so we can go in production soon but in the meantime due to the outbreak and it’s NIH a certainly interested in multiple different vaccines we got some funding to produce her arm cesario strain house as a real Ebola virus of the current outbreak and that’s done end of this month most likely master seed we have we started just a big non-human primate study was 24 animals we look for those dependency and the need of a key event and we also prepare the first cling of drug also NIH sponsored hopefully we now by mid of this this year yeah you never know things take time and you still have to be careful i think some approaches may be pushed a little bit too hard before it was too slow so i hope i could actually convince couldn’t in Vince this guy but could convince you that you actually this one bullet can kill two birds and I want to thank all the people involved it’s a huge amount of people who actually involved in such studies most of my work is now done at integrate research facility at Fort Detrick which is a facility just for you for researcher to perform experiment with PL for pathogen we work with Heinz Feldman beach that last animal study at the Rocky Mountain lab we have the production team for the vaccine it was disrespecting was now licensed to XL bio to drive the commercial part of it and this group is in my lab who works on the more basic research thank you wonderful thank you hearing backside be very promising we have any questions from the floor there’s tributes and Mike’s going on which I can’t see do you foresee the possibility of using this

vaccine to deplete the bat reservoir not it will be very hard i mean if you to immunize beds i mean that was assault with rabies i mean you you could you could try but that would be challenging i mean noop you you could paint despaired swiss be certain fruits code of this virus but i think it’s only could be made but realistically it would be very expensive probably and in this area you probably want to put your resources more and so humans i would sink because yeah would be interesting i would love to drive but i think you have a lot of challenging to find some money for that but it’s not impossible to all immunize bad miss rabies we can do that in your preclinical studies so far have you been able to look at route of immunization like intradermal to optimize dendritic cell presentation now we doubt we didn’t that’s that’s a good idea to because in India they do inter-terminal immunization with the rabies vaccine just for cost factors it’s a little bit challenging in India it works well because they have a lot of nurses which can do is a TB test but yes only that something for later right now we want to try probably just two doses of the vaccine arm for safety interface one an aside question from your vaccine research I was curious on an early slide you showed various wild animal species in the US I was curious does Canada have rabies surveillance program and what are the factors up there it’s update in Canada I’m very successful immunized with recombinant adenovirus this yeah you can do that too and they had some success with that system service in the USA for some drivers with recombinant vaccinia virus explorer expressing rabies rabies Chi but there are some incidents that somebody got unfortunately infected this rabies I mean I have to defend the USDA a little bit this may be the problem is ter most abilities that we work on that too in Europe it’s not as hot you know like intimate vest during the summers are a business not to stables virus because I think so all rabies virus is still pretty good see other thing with all immunization is the problem for example if human eyes stock state they don’t you a lot foxes are too good sure so if they find a bite they they chew it up as if you have a doc you know how they eat they just swallow you know so that doesn’t help so there’s are certainly challenging saying skunks are incredible difficult to immunize nobody knows why you can put in 10 to the 10 into oral cavity and data on sero convert so yes there’s a lot of work to be done question the differential disease penetrates of the Ebola with age which we see in so many infections with West Nile for example one what do you think is the underlying basis for that presumably immune response capacity and the other the response to these vaccines are there going to be age related as well older people respond less well and yeah that’s that’s a good question that certainly sinks you have to look into if why it is we really don’t know no i mean so normal thing is a very young it’s a very old have but it’s that H is not very old so so i really like i think nobody knows why that is perhaps it’s we will learn from the clinical study if that is Africa specific to co-infection to multiple infection you know do these people have already other infection I mean they have quite a lot of viruses their selves but that that certainly has to be analyzed and and certainly we suspect since there needs to be some risk analyzes too because we have people with AIDS and and things like that so there’s certainly all concerns so I was reading on the most of the antibodies produced for a for a against a glycoprotein are against the museum region of the glycoprotein have you tried building a vaccine without the museum ricans see if you get more neutralizing antibodies and what’s the difference between up now we didn’t I talked to Heinz Feldman they made the vsv visa news in delay I’m deleted GP and it was horrible didn’t work

whatsoever so I didn’t awesome what we did is actually for our Eliza plate we have two different cheapies we have one visa Muse in domain and we have one without some using domain as a coding antigen and we really didn’t see significant difference in say Liza tied us so we probably have a different antibody so in debt we didn’t see but it would be really good to to look a little bit more to cell population what you get what we try to do is going now and humanized mice to actually look what kind of antibodies we may get there so do you have any kind of toxicology studies have you look into the monkey studies with your ever love X in candidate aadmi toxicology studies in monkeys do you have any chance to look into that with your candidate vaccines we we have a new study now but that just started two or three weeks ago so i will probably now in two months we will have two three months we will have to challenge data then I know more but right now we’re not even analyzed Sierra a lot of the meats and deletions in the field are badly designed and unfold the protein so we just don’t have a good answer okay thank you