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Effects of Milk Antibodies and other Proteins on the Commensal Flora
Vanessa Dunne-Castagna
Wednesday, Sept. 25, 2019 · 1:58 p.m. · 44m 28s
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bring back on a a first congratulations for making it this far you're still here but i do for you
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uh and it much appreciation for my opportunity to be here at two presents to this amazing group
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i've learned so much over the last two days and i'm glad to be able to um to do the best i can
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to try everything together so uh i am focused on that note
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secretary i ga and its impact on the command so microbe iota
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i would be talking to you about uh this complex here the commands all secretary i ga interaction
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as specifically in how command souls or these that you that i have uh been a yellow
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eyes thing in its association with secretary i ga the impact of this on the command so conversation
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uh both in vitro ending people animals model and uh this investigation as to you
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the ability of this complex secretary edgy increments alls to exclude pathogens
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i don't have to go into much detail one secretary actually we've heard
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quite a bit over the last couple days on this by active component
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um but just to remind you this is found in abundance in both ca last chime at about five make sperm no
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and also over lactation age which is thing in this chart still
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maintains high concentration at about one milligram per millilitre with a variation
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this molecule is heavily like costly that and that's one of the things that will focus on in this conversation
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this is an image from and let's get these review in got microbes that
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uh does diagram nicely these anything to like cans heavily decorating the secretary components well
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sorry the secretary component here uh the heavy chain on both i. g. a. one and i.
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g. two and also extra and like holding tank like ants on a secretary and you a one
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these like ants had been analysts quite extensively with a
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curly liberty is quite a biology lab at u. c. davis
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and this is a one sample of this a really amazing paper that looked at site specific like oscillation
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and uh in line is the relative abundance of these various and when to clay cans at each of these site
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the study was looking at three a collage and so help you help the women donating angle colossus from
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i did wanna just kind of bring your attention to these say like acid residues that we've heard discussed earlier today
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uh and and it looks like an collapsed um that i dislike of proteins
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are um heavily a sigh happily didn't so would you say like acid residues
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and over lactation aged does switch over to a more she costly
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did and so more uh a few cars residues on these molecules
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so their function in this is a from a review by nick
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mantis is group from a two thousand eleven in due course immunology
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that there are many many functions already know of secretary id a
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and just to highlight a few from this uh from this nice review
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we now know that not only are uh the secretary
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idea molecules important in f. a. b. specific reactions but
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these like cans also have a significant relevance in both in in surveillance
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uh instead directly interacting with epithelial cells to reduce inflammatory response
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including a reduction of the n. f. kappa b. pathway an increase in the probably
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merrick i. g. receptor which increases the amount of secretary i ga in the aluminium
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uh and even an increase in the um take junction binding
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proteins so promotion of them barrier protection in these epithelial cells
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we also know that with and without pathogens so this is even with the presence of simply commands all bacteria
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and that we have a significant amount of sampling of luna contents
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through l. and uh uh and cell receptors that are like and specific
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so requiring that like ants on the secretary idea module and this interaction allows us reverse
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translate to says of complexes subliminally antigens again
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both with comments or bacteria and also with pathogens
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uh to the is a invention nations where we have a danger excels
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expressing d. c. sign selecting receptor and and then menace
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residues on is quite go these quicker proteins that are
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involved in a binding to the standard excels and leads to an intolerance expression of in a look and and
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we also know that uh again this at at phoebe specific
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pathogen bindings this is a really classic immune exclusion or pathogen exclusion
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unknown for for decades about um the function of this molecule with that specificity
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but also many studies have not come out by recognising
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that like kansas also relevant in pathogen exclusion of this molecule
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including a study looking at the asked him created equal i
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inhibited by those that say like acid residues on music like cans
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costume toxin has been shown it to be inhibited simply by think like ends
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on the secretary component along so not even requiring that like the protein itself
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and then a number of other new cool so pathogens are also excluded due to look like and
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so and seemingly non specific interaction is a secretary i. g. molecule
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and finally numerous studies now have come out looking at the uh
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the impact of secretary i ga on the regulation of commands all conversation
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and again this could be due to bowl like an interactions as that's quite easy has demonstrated i'm back in two thousand eleven
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um but also nelson you studies have looked at this low affinity interaction of secretary
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i ga with comments of bacteria and this is really where my studies are focused
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so what the gaps in knowledge that uh our research groups both i did notice and but a lot about we've been looking at
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our first looking is simply the interaction between secretary i ga this is
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uh looking a milk circuitry i. j. so a very heterogeneous group of this
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like a protein interaction with select potential uh select
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commands alls and looking and it's increase in conversation potential
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so i've investigated the digestion of these mode of these onion complexes secretary
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and you come and so complexes adult in beach oh and in the val
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and the ability for this to improve the attachments both again in in future systems and also in a mouse model
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then investigating intolerance in the host and second looking at the complex itself
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so are there any specific functions of this commands all secretary i ga complex
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and the some looking at compensation resistance so potentially the
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ability for this um this complex to help reduce pathogen binding
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um perhaps in improved pathogen clearance and of course have improved barrier function on the host
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and this knowledge uh that we are gaining with the study is not only
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will help us investigate mechanisms as to what's happening in natural breast milk system
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but it surely in the development of a product that may improve
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the ability for probing attics um well it may improve administration probably addict
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this is looking at a possibility of creating some complex perhaps with remote carcass
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like the solace differed a bacterium some very common commands of bacteria of the infant
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and creating a a complex with secretary i ga and these bacteria
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that when administered to the infant may help protect the probing
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added from digestion in the ins in prince intestine and improve
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conversation once this complex reaches the uh
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infants colon and at the same time perhaps
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prevents pathogen infection with that f. a. b. u. specificity still with the secretary
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id molecules it's a big goal and i'll show you how far we've come
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so i just didn't use this list of commands alls i'm going too
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much detail on this but and these commercials deputies of if the bacterium
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as these these some of our steps p. c.s uh including being fancy as witches
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be long gone since pieces intent is be a long time since these these long gone
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these two guys appeared chosen due to eighty their ability to grow on specific
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you may not like a sack crimes it's you have chosen to use in my study are too because select also to a file
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and like two and the attach real sore l. and and see uh these
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guys up here these this these bogus uh i have also been extensively studied
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by the mills them so we have a extensive gentlemen characterisation about these organisms
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and uh we we have a lot of really fun side projects and i'm looking at their abilities to uh or is
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there gene expression alterations with the presence of their particular grows
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substrate and alterations and binding to this particular like approaching adventures
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i've also included three different like to the solace uh or like to say like here
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like the solace ram noses seven four six nine this is the uh essentially the same as the seas as an l. g. g.
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so i was looking at these as a currently marketed um probably attics
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i just kind of again looking at this concept of making us a chance little product
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so you will have seen the secretary i ga i just wanted to show the
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slide up here in case you're curious so this is the process of pure protein purification
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i used a donor milk for um women who were over three
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months of lactation age and uh collectively pull together numbers knock samples
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uh with this i uh creates cannot from here and um
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flow that's not through a capture selects i. g. affinity chromatography matrix
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which allowed a per really nice purification this is demonstration within s. yes page and was
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to much showing a really clear isolation of the spurting from milk with virtually no contamination
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so with this uh can share products i then was able to answer
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some of these questions initially looking at the interaction between secretary actually increments alls
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first is their concentration dependent binding of secretary i do to commence of bacteria
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and second how important are these and went quite cans so does the removal
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of these like ants lead to a reduction in the association of secretary i ga
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if you're familiar with i. g. a. and and secretary a. g. research you'll
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know that let's get busy has a ten to work has answered um the second
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question uh to an extent and so part of this was we could be
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cheerleading his studies and the other part of this was really as a novel investigation
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and so what i found uh and i use flow sitcom entry to analysts much of my secretary
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i. g. association and the spot you can see six i don't nine as made unique you later
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and uh i used a um a anybody actually that jeff gordon used also in study this is
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a lexus six forty five detection antibody for secretary i ga this is an example of a flow plot
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and this is that the bacterium infanticide be infanticide uh you're staying here where up
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at the top you can see an increase to my concentration of secretary i ga
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and uh hopefully you can see a very clear demonstration haven't increased association of my um bacteria
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with secretary i. g. a. with increased concentration so
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certainly evidence that this uh interaction is concentration dependent
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and i wanted to look at this a little bit more detail with some of the top binders
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so like to solace from this is that o. g. g. guy um beyond the mongol maybe longman finances all three had
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very robust binding under the highest physiological concentrations of secretary i ga
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and when i used a variations of this concentration you can see that
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uh these organisms performance a little bit differently well with their
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interactions with secretary i ga but interesting link all of them
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seem to reaches saturation and then type so interesting to me and perhaps those are you in the breast milk world because
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again the physiologically relevant concentration a secretary i ga in breast milk
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it's about one meg per mail for this uh perhaps demonstrates that in
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that press they'll knock environments we actually have low word saturation of our commence
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a bacteria that are found in a press not community and then then
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that they could be reaching 'em but more investigation is a record for this
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to answer the second part of this question how important are these and link quite cans at this interaction
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i utilise act like a cell high julie's that's derived from be infinite justices and o. b. i. wan
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and this clears that kind of bias score does leave behind a
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small residue but it but it efficiently does unclear these like onions
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from like uh proteins and allies my results of these uh digestion is with s.
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t. s. page demonstrating a clear reduction in like like an ice of one's line here
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and found within you know for us is that the is the like oscillator proteins
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indeed have a reduced binding this is showing a picture of like to solicit with
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i'm a native secretary i. g. molecule and and here's like to solicit same concentrations with
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a a and a b. a. when she did michael we can see a significant loss of association
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of secretary i ga lease with him you know for us and send this detection in a body
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so certainly at some demonstration that the association of secretary i. g. a.
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i can be reduced with the depletion of these and link like ants
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with this information i i also wanted to look at this function so what what what purposes this interaction
00:13:01
and uh oh let's get these in one has really study showed that the secretary component alone
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was protective with the digestion of the idea molecule and so with this i hypothesise that perhaps
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commence a bacteria coded with the secretary coded i. g.
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a. and may also be protected from in future digestion
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and so i developed a model and kind of take a lot of trial and error to um
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to evaluate this question surpass bacteria two times a second time with the on the sugar substrate of interest
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i because for all bacteria lactose and then that preferred each most substrates for some buffer bacteria
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uh i checked the associations right i a call
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to these bacteria bounded with secretary i ga not right
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check the association with that fax came and then with this complex here
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i then subjected and this complex to i gastric digestion
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conditions using p. h. of four with percent perhaps and
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and then uh um intestinal indigestion with pink retain at each of seven
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with this complex i checked balls uh live counting with see if you um yes you're dilution and see if
00:14:06
you counting and then i also used to uh flows for said how much she with a set of toxicity essay
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and i evaluate this with with all organisms have interests in found variations in in
00:14:18
uh the protective capacity of secretary i j. but under all circumstances with every bacteria lines
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there did seem to be protective impact uh using secretary i ga so this is
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lawful or fold change over and these organisms digested but out that protected in a body
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in some cases every statistically significant uh and other cases very close to but um will be
00:14:41
repeating somebody's ass is to truly determine this that in all cases of i did a regression fine
00:14:45
uh using all different um concentrations the secretary a
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g. a. i certainly some uh i demonstrated correlation behind
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and the protective ability for secretary i ga to increase viability of bacteria postage action
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so this was an in beecher model and uh i wanted to see if this holds true in a mouse model
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and so on this for study this is a a um an investigation using biopsy mice
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this trial we were added a very short trial to investigate the survival uh and i choose to use the long gone
00:15:18
a long concerts pieces in finances and again this is one of these champion connoisseurs of the infant got so
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surely trying to um tease out what's happening uh in as best as i could with these in think human souls
00:15:30
for this trial by a collective baseline samples here to determine uh into that
00:15:34
there were no will be in fantasy since really no better bacterium in these mice
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i'll buy three days i've devices with the intent is either
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so she did or not associated with not secretary i ga
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i use that concentration i had found that was
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the um highest saturation rate for this particular bacteria
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and then collected people samples one they pose to garage to conceive on the second day
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and to determine the amount of bacteria that essentially were able to survive
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that passage and and persist for at least a short period of time
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and what i found was indeed that when the intent is was pretty associated with secretary i
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ga and this was knock i ga and that i had almost a complete lock fold increase
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of bacteria that we're able to passage through that and uh it has no content and um be sure to the feces
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now i will make one caviar that was that one of my colleagues brought to
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my attention this is fickle bacteria so i i cannot say with this plot alone that
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this demonstrates persistence in the intestine we do have buyer geography studies that certainly do show
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there is a still also same not quite about ten to that but see if you
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in almost every part of the intestine um post um post ingestion so at that day evan across the
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uh so i can i completely say this is due to just shedding of the bacteria
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uh or if it's evidence of survival of the bacteria but um but nonetheless again with by geography studies i can say that
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these little still did seem to be much higher in those um mice
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that were provided that the influences that was pretty associated with a. g. m.
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so certainly at least in the study here we do do see some evidence that secretary
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i. g. a. can improve that survival of commands a bacteria postage russian induced obscene mice
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and the last part of this initial investigation we just commences and secretary i ga
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was to look at this ability for um these commences to bind to build an inbuilt show 'em
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cockroaches this cool culture system up a human cullen aside and then also in the l. c. mice again
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so for the beecher trials i uh against humans trial and error i came
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upon this this concept of utilise thing h. t. twenty nine m. t. x. l.'s
00:17:49
which have a very high goblet cell productions i get a very significant because the layer
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um that i think better represents what's happening in the colon and i use k. could choose also says the cool culture
00:18:00
yeah for these ones i can use the in fantasy and i wanted to just do an evaluation of
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persisted be in fantasy after a short period of time sitting on
00:18:08
these coincides and look at any expression changes in the clan sites themselves
00:18:13
and again what i found was as i increase the association in this is
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pretty association before i put the bacteria on to the surface these cullen inside
00:18:21
so as i increase that percentage of population associated with the
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infinite jest i also so i've very clear um increase in
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the amount of bacteria there remain down to the surface of
00:18:31
these concise even after watching uh many times with p. b. s.
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and just a plot out to to that sloping give you clear number so for
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every one percent increase association for the spectre population you see about ten to the fourth
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see if you increase binding capacity so seem to be a very robust um binding
00:18:49
capacity with secretary i. g. m. and then again gene expression studies on these co cultures
00:18:56
when the intent as was provided in this is like fold or full changeover baseline with p. b. s. alone
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there's a significant alteration a number of different genes involved
00:19:04
in um information and also into a junction binding proteins
00:19:08
and when secretary i ga was associated with the
00:19:11
inventors prior to uh that interaction with the school cultures
00:19:15
essentially a brings things back to baseline uh if you did look over um
00:19:20
be in finances with out and be inferences with secretary i ga you could say
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that there was increased take junction binding increase the cost of production seem to be beneficial for
00:19:30
the introduction of this commence ultra first have the secretary edgy
00:19:34
association um prior to that interaction with these that cullen sites
00:19:39
and then again moving us into an in the persistence of looking at this inside amount
00:19:44
uh this was again in the l. c. might still see a very similar had time line here we wanted to collect
00:19:50
baseline samples to ensure that there was no b. b. influences in here initially device for three days with the intent is
00:19:56
in here we have the beginning of mike complex slide so we have four different groups to were provided water
00:20:01
um but you were provided with too few 'cause the lactose really increase the
00:20:05
ability for that be inferences to find its own little ecological niche and survive
00:20:10
we l. collected have people samples um they forty sixty eight and then across it on the nine to
00:20:16
look at persistence of the inferences and so what we found a break this down just a little bit
00:20:22
when the mice were provided with water and be in fantasy alone
00:20:27
there really was no detectable levels of influences in that
00:20:30
mouse on the sixty eight they were gone washed out
00:20:35
when the mouse was provided be in fantasy with that too because uh lactose allow
00:20:40
we now we're able to detect levels of persistence or all all these this trial
00:20:45
so in this case it looks like the uh that too few cars selectors alone will increase persistence of this the influences
00:20:51
now this is also true if you've been following any of the most work uh
00:20:54
we've also looked at this but you want them longer messy five nine six and found
00:20:58
uh actually the same thing so when when use h. t. mote consuming bacteria are provided essentially they're little suitcase
00:21:04
the fruit then uh they can seems to be that they can persist in even models where they normally would not
00:21:11
of course my question as the secretary idea do anything to this complex
00:21:16
so in this next group this is uh the intent is provided free associated with secretary i ga
00:21:22
and this is again back to that water model so whereas we initially thought no persistence
00:21:27
with the intent is provided alone now with just stop or vision of being intent is
00:21:32
a complex the secretary i ga you see them they pop back up again
00:21:38
if it seems that that's out a secretary i ga did indeed increase the persistence
00:21:42
a beam thing into this and this was all were or almost to
00:21:45
similar levels as just the provision of chief you can select those alone
00:21:49
so let's combine both of these together what happens when we give these mice
00:21:53
balls to picasso like those and the intent is associated with secretary i ga
00:21:58
it seems to be a combined effect so with these uh mine's
00:22:02
provided balls too because like those and previous or she'll be in finances
00:22:06
this like syllables up and i didn't put my significance on here but uh the script is that definitely
00:22:11
statistically um greater than or higher prevalence than any of the other groups uh together
00:22:17
so clear demonstration of the combined effect of both that specific h. m.
00:22:21
o. and secretary i ga and again just to kind of bring this back
00:22:25
to what we're talking about in the fall group made this is this
00:22:27
is the infant that we're looking at um that something similar to an environment
00:22:32
or the interns are exposed to commence the bacteria
00:22:35
that perhaps are associated with secretary i ga if you're
00:22:38
looking at this in the breast milk environments and of course have the presence of many different h. m. o.s
00:22:43
in that same environment so perhaps this combined effect does really improve
00:22:47
the ability for commences to get into the infants and to columnist inside
00:22:55
oh so in summary for this first section secretary i. g. a. i can say certainly increases the causation of
00:23:00
select commercials this is not sure with every commence evaluated
00:23:03
but certainly would be infanticide few the others that we evaluated
00:23:07
angry including increase protection from a tight digestion
00:23:10
improved conic retention and also the improved host response
00:23:16
so with this information i wanted to then look at the function of this complex in pathogen exclusion
00:23:22
which shows to we we actually um collaborated with the white person
00:23:25
or at hannah in new york with their model of seven l. invasion
00:23:29
and so we use seminal attack miriam anyone to investigate
00:23:32
the act essentially the functional cross reactivity of a monoclonal recombinant
00:23:38
secretary i. g. a. called sound for this is specific to a non pay for it all
00:23:42
salmonella and the complex in this would be infanticide looking at the first the prevention
00:23:47
of the centre half join button and we just system and also in these bouncy mice
00:23:53
so i'm just gonna uh take a minute explain this um this pretty cool model that we adopted from the white person or
00:23:59
uh they generously provided these two strains as seminal attack miriam
00:24:04
one is the news jane and one of the wild type
00:24:07
well how can my son resistance which allows us to um go these
00:24:10
on selective actor especially in a mouse systems we can isolate these guys out
00:24:15
um but that that type has this all five engine that is specific to the self or secretary i ga
00:24:21
in a body is is men extensively studied in them and to slap and also with liz could they see
00:24:26
um and shown to reduce the ability for this
00:24:29
seminar tech miriam to invade a and persist inside
00:24:33
i'm an animal model and then that's what type has a and do our deletion in a gene that's encoding the
00:24:40
assimilation of the o. five antigen so in this case we have no self or interaction at least not as specific
00:24:47
sell for interaction i do have evidence that this still does buy into this guy
00:24:50
but does not seem to be um to to that specificity of f. a. b.
00:24:54
these guys also have that lacks the gina reporter gene that allows them to turn blue under blue white screening actors and
00:25:01
good way for us to differentiate one to the other we chose therefore to use that
00:25:07
competitive index this is paul for our in future models and also in our mouse models
00:25:12
uh to evaluate the impact of the specificity of the self or in a body and
00:25:16
for these competitive in that indices we just did a while type divided by mutant bacteria
00:25:21
and this is uh looking at those that were able to invade cullen slides or
00:25:25
to migrate to the priors patches in a mouse model and the idea here are
00:25:30
competitive index of one essentially means an equal number of all the while type and onions
00:25:35
um and a combat index anything less than one indicate self
00:25:38
or specificity where we had a fewer number of the wild type
00:25:42
and then no change in the mutant so we're looking at this as
00:25:45
our um as a baseline markers to evaluate effectiveness of self or specificity
00:25:51
and also to get away from the inherent variability in mouse models and to look at seminal an invasion
00:25:58
parent teacher models we use again we actually for both we use the in fantasy
00:26:03
and after treatment groups we have four groups here first
00:26:06
is the same analysis or positive control just seminal alone
00:26:11
second is the seminal and make sure so we had you come over concentrations essentially the well type and you
00:26:16
and this um they could control we have ourself for specific in a body
00:26:20
that should be able to inhibit the well take the not the mutant bacteria
00:26:25
and then our question marks of the intent is provided as
00:26:28
a precondition to these cullen asides prior to seminal invasion or infection
00:26:33
and then finally this uh actually a question mark here does this complex itself or with the intent is
00:26:39
have any specificity at preventing invasion seminar and what we've counts
00:26:46
first just looking at total while tight bacteria able to invade
00:26:50
excellence they'd make sure this is circle culture system this is
00:26:53
our positive and negative control so we do indeed see a
00:26:56
reduction it with that sell for specificity of our while type bacteria
00:27:01
wendy in fantasy was provided with no self or are we still sound
00:27:05
levels that were the same as in our positive control which was expected
00:27:10
and then i would be in finances that was a preacher greater precision you would sell for
00:27:15
we saw in the even there was a reduction in the wild
00:27:18
type not exactly to the same levels but still statistically significantly reduced
00:27:23
uh for the well type bacterium and again to measure the specificity of the self for anybody to see if this really was
00:27:29
that anybody that was doing this reduction down to to the surface
00:27:32
of the site comments of bacteria to look at the competitive index
00:27:37
a positive control had no difference this is just seminar
00:27:41
provided alone no difference in wild type versus new bacteria
00:27:45
when provided that sell for free conditioning the wild type was inhibited and the um that and you didn't
00:27:50
really was not so we had a significant drop about fifty percent less while time bacteria then you wouldn't
00:27:57
when the intent is was provided as a preachy men here we start back to baseline
00:28:01
no difference in whatever says onions and now the question of selectivity that sell for anybody
00:28:06
indeed we star that would be intent is was pretty associated with that
00:28:10
specific sell for the specificity of that anybody was still there was able
00:28:15
to selectively inhibit the wound tight bacteria over the noon indeed to even
00:28:21
greater levels then when self or was specifically to provided to that complex
00:28:26
this was great indication that are hypothesis help through the intent is complex was
00:28:31
sector e. i. g. a. is effective in reducing seminal invasion and indeed was effective
00:28:36
with that specificity so the f. a. b. specific binding was still present even when
00:28:41
that be in theatres was associated with that come with the us actually actually i'm
00:28:47
this ad uh looking at gene expression um this is just and
00:28:50
a million cullen sites so we saw that the in thing into solo
00:28:54
uh was able to reduce in looking eight productions the pro inflammatory site kind
00:28:59
this is salmon now and this is the inferences it did not seem to be secretary i. g. had a significant impact your
00:29:05
but when secretary i j. was free associate would be infanticide leave some tight junction binding
00:29:10
proteins were increased with the presence of secretary i ga uh versus when it was known
00:29:16
and then finding i'll start finding looking at this moving this into a mouse model
00:29:21
uh again the same kind of model here we had three days at being in fantasy bondage
00:29:26
and in this case we uh when it is to look
00:29:28
at the ability for that specific style for response to salmon now
00:29:32
uh this was our first control group or we had um
00:29:35
salmonella provided along with no be intent is as a negative control
00:29:40
a seminal are provided with that sell for as our class are positive control kind of something that's around
00:29:45
and then these mice were provided that the inventors with or without sell for and
00:29:50
uh infected with stamina well one day or three days post uh be inferences coverage
00:29:56
and again we had different groups here provided either um water for these control groups here or in the
00:30:02
intent is was provided we uh give these guys too because like tools to improve persistence of being in fantasy
00:30:08
and what we found first just looking at total invaded salmonella with balls about type and the mutants
00:30:13
no difference no difference in any of these groups with total seminal invaded
00:30:17
into the parts patch but again looking at competitive index was more our question
00:30:23
the competitive and index when there's no be intent is
00:30:25
just seminal alone or competitive in texas was close to one
00:30:30
uh and then when that a seminal and make sure was pretty associated with self or we saw that competitive
00:30:35
index drop as expected showing you the specificity of that
00:30:38
self or in a body against the while type bacteria
00:30:42
so this was what we're comparing all over treatment groups to
00:30:45
see if anything showed the same level of specificity reduction innovation
00:30:50
our first group when we just infected with salmonella one day
00:30:53
post crash there really was no difference from that initial competitive index
00:30:57
but here when we had the um the advances that was precision without
00:31:02
four and we go vice with salmonella today's post um being in fantasy
00:31:06
we did actually see a reduction in the competitive index indicating that this self
00:31:10
for that was presented with the intent is in like that of a mouse
00:31:15
three days after the presence of this with the introduction of this was able to selectively reduce
00:31:20
the white type salmonella bacteria from uh invading that the priors patch
00:31:25
so indeed we did see it the same uh or similar results as
00:31:29
we signed the job that this complex does indeed prevent that seminal invasion
00:31:33
into the priors patches similar to the provision of that self or alone
00:31:39
so again there's i there's complex here this idea of the function of
00:31:42
this complex we certainly can see that it was a functional in preventing interrogation
00:31:47
uh and then overall just looking at this are the concept of this function increase
00:31:52
the persistence of the comments of bacteria and perhaps prevents other into her pathogenic infections
00:31:59
but that i just wanted to say a great thank you to my enormous a secretary i. g. team that
00:32:03
i've collected over time and then both the nose lab and i wanted a lab where i do all my work
00:32:09
but that's not that's not any questions are much for most some questions from looked over
00:32:26
a lot of problems more so formidable more point for what to call commercial
00:32:33
oh that's a good question um so we've chosen get and i and i use the term comments
00:32:38
on most of the time this conversation so commences are those that are derived from uh the infant
00:32:43
essentially this one thing in ten million um call wanting tracks and able to essentially persists inside that
00:32:50
um that uh organism so these commands those the ones that
00:32:54
i've chosen here we're all isolated out of a healthy infant
00:32:58
um and use because we see them in the human population
00:33:04
good good work or the bars don't know that's what hope
00:33:11
whole shallow level and holes in real question
00:33:20
what you call rather common
00:33:25
uh oh about two bottles don't ah so we've i've looked at this a little bit and we
00:33:30
looked at association let's knock secretary i ga with some uh now uh with the hack with kevlar batter
00:33:36
and do you see that it can bind it to these organisms and uh it seems to be
00:33:41
more preventative then it is a increased increase persistence
00:33:46
so these in in future system that we're looking at
00:33:48
when i free associate seminal our preset jihad for those of course it's a little bit different model with someone
00:33:53
i can see that actually knock secretary i j. alone
00:33:57
is inhibitory to invasion of similar to these con side
00:34:01
and uh with the heck i see a reduction in adhesion
00:34:04
ivy hackman pretty associated with as secretary i ga derive from no
00:34:08
so does not seem to be doing the same thing it does bind non non specifically but but but what
00:34:13
a little more blue mood followers is not for the
00:34:19
golden for shoes for to to loop holes filled interruption points
00:34:26
yeah but bowls full solid pretty long remembered two
00:34:29
more books uh_huh popular should yellow receiving something different
00:34:36
caucus we just call it an interest of your commercials i don't know i i
00:34:44
think that we should investigate that yeah and we we do see a similar effect
00:34:48
i mean i i did look i i said three different uh inter pathogens in all seem to be doing
00:34:53
uh seem to be holding true that no secretary i. g. still seems
00:34:57
to be somewhat protective even with them and other you're you're absolutely right that
00:35:01
the of the little cells are not it's angle it it must still pass through those at the little cells to get into
00:35:07
out there in the tissue and the and since that that is actually what we're looking at is just the prevention of that younger
00:35:13
usable especially for your for the problems you yeah that doesn't really
00:35:19
don't it's probably good reason how does it know is the quality program
00:35:25
that's a good question i mean of course we're looking at in general overlooking a gram
00:35:29
positive versus gram negative years well all the pathogens that we're looking at here a gram negative
00:35:33
and the commences all the ones we've chosen or gram positive and
00:35:37
even let's get easier originally when he began this investigation side difference and
00:35:41
used a equal by the nasa versions so this commence a legal lie and
00:35:46
saw that it did not perform the same as the gram positive comments of bacteria
00:35:50
so perhaps this this this he is more due to just take the graham nature of the bacteria not necessarily to its pathogenic nature
00:35:56
but in a different goes to draw the last one proved most of the state
00:36:03
i saw a lot of solo and it's yeah i'll go on
00:36:10
record well but who loves commercials yeah function differently yeah they mine
00:36:19
i'm gonna point
00:36:21
of course or box and blurted it from white boy or geology
00:36:27
um hum also yeah some other mechanism by which you can infer
00:36:31
uh oh gee look prevention of of of the destruction boy yeah oh by
00:36:38
the host yeah i i'm one thing that i have not had the op
00:36:42
to do you get is you any mechanism based investigation i mean the cell
00:36:45
for anybody has then thoroughly investigated by the make make notices group and others
00:36:50
and looking at its ability to actually calls or the bacterial cells all changing it's fashion mentality et cetera
00:36:56
and uh i i actually found a reduction and even though while time with
00:37:00
self without for that specific anybody still the i'm sorry the mutant bacteria was still able to you
00:37:06
fine to that specific in a bunny and still reduce its
00:37:09
invasions of something else is that work besides that's specific interaction
00:37:13
oh yeah with using embodies the be a beer a very interesting study to do more mechanism based research
00:37:20
the water
00:37:23
h. o. o. one goals for those who are important yeah
00:37:32
shell shell will probable shaw sold quite sure helps to
00:37:36
go into the organisation will rush will show you how close
00:37:42
one call them our somewhat shorten people we have talked about that but we've
00:37:48
not done that yeah in part um no i i've not done that yeah
00:37:53
um but i think i well it has been done just milk and commence of bacterial just one
00:38:01
yeah
00:38:05
yeah uh_huh job you're more water or certainly but there are so many other factors and no click and also the binding of
00:38:13
you know this to secretary i. g. a. and they're different
00:38:16
physiological functions or or a genetic alterations that happen with that bacteria
00:38:21
in that in lieu of not so it would be difficult even interesting just look to
00:38:25
see what happens that to figure out why it's happening would be a a big question
00:38:30
and and part i mean not only as a study looking at um what's happening in all but
00:38:34
also what could happen if we take this out of that not system and we're trying to introduce
00:38:39
probably addicts uh maybe preterm infants right and and making sure that we are not stimulating of robust
00:38:44
inflammatory response with these guys 'cause obviously the inference don't know what's good and
00:38:48
what's bad so we can protect these on the way and it might be beneficial
00:38:55
just questions from the lip service
00:38:58
you know the very most presentation my question was a little bit of carried total funding to the human
00:39:05
um a lot of partial but the majority of the trilogy is produced under small well
00:39:13
it's sort of it is that we should trilogy is alive and
00:39:17
working in the human column and no degraded boy to i probably will
00:39:25
you know so people will probably be the most boring but
00:39:35
so probably build more very crucial hello cumbersome for a big hole now
00:39:45
oh wow what a remote what probably possible with him you know what if you look at the logo
00:39:54
the most popular will probably remember what
00:39:58
rule one thousand rule avoid all home you'll
00:40:03
go out local book well off of what the real who who who who who
00:40:12
also doubles of our um drop out of all uh what uh what i remember
00:40:19
cool culture or culture or what most horrible syllables
00:40:23
for all was walking this oh bought something well but
00:40:32
i know oh show because all the lines but for all
00:40:38
the other shows probably probably the whole or so while reliever
00:40:47
the the most boring but bush movable oh sorry functional
00:40:58
oh
00:40:59
cool illegals will will work well probably because he was brought work yeah there's
00:41:04
a startling you portion if there's absolutely evidence of i have active secretary idea
00:41:11
are numerous areas we're looking at the animal models with deficiency and secretary i share secretary molecules and you see
00:41:17
that what normally used in a colon with that which is the conversation of that out i'm you cause
00:41:21
a layer uh that those bacteria able to really penetrate that our mucus a layer an enter either into that
00:41:27
the the alkaline cells with out secretary i ga or secretary i. g. m. present
00:41:32
so certainly does it now only found but um most important in the not the colon
00:41:39
one of the things i wanted to just look at all oh
00:41:43
yeah also yes absolutely do they might never but yeah well
00:41:54
to move
00:41:58
oh oh oh i'm probably of types or because i'm sorry your volume shoreham
00:42:09
read your consortium hopper true from those for um
00:42:18
hum yeah don't work well oh well not much choice
00:42:25
yeah but you could avoid grumbles fish a lot for the school
00:42:32
because yeah absolutely so we we've not followed i would not in our laboratory done
00:42:39
i'm a levels of investigation as jeff gordon doesn't
00:42:41
identifying groups of bacteria associated with the particular phenotype but
00:42:46
the idea of creating um second concoction essentially of different organisms
00:42:50
has then uh something that we would really like to look at and i'm in
00:42:54
particular i mean we looked at like to sew sent to put a bacterium in part
00:42:57
because it's the radical that like to the solace are more smart has
00:43:01
no and the for the bacteria warmer climates so if we can create
00:43:05
something that might essentially be beneficial all the way down
00:43:07
the intestinal tract especially for looking at prevention of interior pathogens
00:43:11
so many are small intestinal and gifts might not have an impact on the small intestine pathogens
00:43:16
but like to design my so might be really interesting investigation
00:43:20
to take this a concoction of of balls monticello in chronic
00:43:24
bacteria associate them always secretary i jane sort of put them into a model
00:43:28
with both a silent assigned colony pathogens to see if it does the same thing
00:43:34
its name a case of i. g. i. g. u. h. m. c. i. g.
00:43:42
a. diffusion see if id and he's covering all told him of purple of of
00:43:49
yeah but i thought i was the identity the set of i think that was a paper looking
00:43:53
at secretary i. g. m. association right i i
00:43:56
forgot who did that analysis probably just because remember what
00:44:02
the boy who who was the people ah okay yeah
00:44:07
but i i do know yeah they do associates or mobiles with numerous members of the microphone and stuff
00:44:15
okay
00:44:17
no further questions thank you very much for most without the
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