Research on an AIDS vaccine has been ongoing for years. Very few positive results have been achieved. More recently the focus has been on broadly neutralizing antibodies (bNAbs). Broadly neutralizing antibodies show up naturally in a person infected with HIV only later in the infection. These antibodies have been shown to be effective in neutralizing the HIV virus but only if they appear early in HIV infection. There is some discussion about developing bNAbs to fight the flu or ebola. Also, there is discussion about isolating these antibodies from large numbers of people and pooling them to also make a passive immunization vaccine.
For HIV there is some investigation in trying to develop a vaccine which elicits bNAbs.
In your investigation of this problem, determine why the vaccines developed in the past have largely been ineffective in preventing AIDS.
Also, answer the following questions.
Why are bNAbs effective?
How do bNAbs work at the molecular level at the antibody binding site?
How do bNAbs work at the molecular level at the antibody binding site?
Why are they more effective than other antibodies in early infection?
Why are they produced later in HIV infection?
Could bNAbs be isolated and purified and used in a passive vaccine? If so how could they be harvested?
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The vaccines developed to treat AIDS have been ineffective in preventing AIDS because the AIDS virus is constantly changing and mutating so a typical antibody can only bind to a single strain of it. According to William Schief, professor of immunology at the Scripps Research Institute, “everyone’s effectively infected with a unique strain and there’s really 30 million different viruses.” Due to the uniqueness of the AIDS virus, antibodies that bind to a single specific antigen can’t attack it. However, in patients infected with HIV, that were late in their infection, three stains of broadly neutralizing antibodies were discovered. These bNAbs are effective because they bind to multiple strains of antigens and can effectively neutralize up to 90% of AIDS virus strains. These bNAbs are an incredible find but there isn’t any know reason to why the immune system creates them.
ReplyDeleteBNAbs work at the molecular level by neutralizing any effects that the antigen has on the cells and can then block the virus from entering the host CD4 cells. At the antibody binding site we can see that the binding site can bind to several epitopes and have longer loops allowing them to bind to a larger variety of antigens. Overall bNAbs are like extra “sticky” antibodies so they can stick to more strains of HIV than a regular antibody that binds to a single antigen.
BNAbs are more effective than other antibodies early in the infection because bind to more than one strain of HIV. BNAbs also mutate to bind to more antigens and they not only block the virus from entering T-cells but also tags them to be destroyed. So, they are more effective because they will prevent the destruction of the immune system thus fighting the infection more effectively.
It isn’t fully clear why 20% of people infected with HIV develop bNAbs later in their infection, but studies have shown that the bNAbs can take years to mature and become effective neutralizers. In some patients bNAbs develop early on but were non neutralizing. BNAbs also spend time mutating and changing their binding sites to bind effectively.
There isn’t any known effective way to harvest bNAbs but research is still being done to find a way to isolate and purify bNAbs and used in a passive vaccine. As of now, most bNAbs are being harvested from blood of donors infected with HIV. The antibodies are then purified through centrifugation and filtering. I would think that these broadly neutralizing antibodies could be made in an inoculated animal or organism that can host HIV that could induce the same conditions as a human and then harvested and purified from that serum. This would still take years because there isn’t a clear reason for the emergence of bNAbs and it isn’t clear how long it can take for the bNAbs to mature.
Sources used:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4249098/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3218537/
http://betablog.org/using-naturally-occurring-antibodies-to-treat-hiv/
http://tagbasicscienceproject.typepad.com/tags_basic_science_vaccin/2015/04/broadly-neutralizing-antibody-suppresses-hiv-in-clinical-trial.html
http://science.sciencemag.org/content/329/5993/856.full
http://www.cell.com/cell/abstract/S0092-8674(15)00686-8
https://www.buzzfeed.com/azeenghorayshi/where-is-the-hiv-vaccine?utm_term=.ejjyLv4wy#.djOYDNnWY
Sarah, your comment on how bNAbs that were naturally developed in patents early in the infection were non-neutralizing is very interesting. I wonder how this might affect how the body would react to a bNAbs vaccine early in on the infection. This would need to be considered when developing a bNabs HIV-1 vaccine.
DeleteI find it so interesting that there is so much variability within this virus (30 million?!). Obviously that poses a problem, just as you mentioned, in creating an effective vaccine though. You mentioned that bNAbs can take years to fully mature and become effective neutralizers; I wonder what exactly triggers their creation and stimulates their maturation. This would definitely give more insight into why only some HIV patients have them.
DeleteI really hope that one day we could be able to harvest bNabs, hopefully sometime in our lifetime. Its crazy how this virus has become to malignant and has adapted into such a rapid growing disease
DeletebNABs neutralize a broad array of HIV stains and are effective as therapeutics. Antigen structures that co-crystallize with bNAB are used as a template for HIV vaccines. (http://bnaber.org/).
ReplyDeleteMolecularly, bNABs bind onto the protein Env on the virus surface. Env is responsible for allowing HIV to enter CD4 cells. Therefore, when bNAB binds to that protein it blocks HIV from targeting CD4 cells and it also tags the virus for the immune system to destroy. (http://betablog.org/using-naturally-occurring-antibodies-to-treat-hiv/).
In addition to Sarah's comment that bNABs are more effective because they tag the virus, they also can prevent infection in mucosal tissue. (http://jvi.asm.org/content/early/2016/10/14/JVI.01762-16.abstract).
There is no current way to create a vaccine through purification of bNABs. I think it could be done through creating a serum by injecting them into an animal.
DeleteThere was an idea to inject people with bNABS, but because they are not producing them on their own they would need to be injected every month for a lifetime. This would be an impractical solution to such a wide spread and rapid disease.
What has been shown to be effective is the injection of bNAB genes into muscle cells of monkeys, however we do not know if this would last long enough or is safe for humans.
(https://www.buzzfeed.com/azeenghorayshi/where-is-the-hiv-vaccine?utm_term=.ejjyLv4wy#.djOYDNnWY).
All of the vaccines developed in the past to prevent AIDS have used one antibody, which poses a problem since no single antibody can cover all of the HIV-1 variants. Now with the discovery of broadly neutralizing antibodies, there is another, more effective way to prevent AIDS. bNAbs can neutralize up to 90% of HIV-1 variants and are effective than other antibodies because they can target epitopes on multiple HIV-1 variants.
ReplyDeletebNAbs target three different epitopes on HIV-1 antigens, which are the CD4-binding site, the membrane proximal external region (MPER), and the N-linked glycerin containing epitopes. The bNAbs’ ability to target the N-linked glycerin containing epitopes is especially important because it enables bNAbs to avert the glycerin shield of the HIV-1 envelope spike and reach specific protein epitopes. The HIV-1 envelope spike is covered with glycans which effectuate host cell recognition and entry into the cell. These glycans form a shield that blocks antibody access to specific protein epitopes, but bNAbs bypass this viral defense by targeting not just isolated self-glycans like other antibodies, but combinations of glycans and nearby proteins. Thus, they can distinguish between the virus’ glycoproteins and host cell glycoproteins.
Apart from the diversification of binding sites that bNabs target on HIV-1, one of the reasons that bNAbs are more effective than other antibodies in early infection is that they all have a higher degree of somatic hypermutation. Most mature, high-affinity antibodies have 15 to 20 somatic mutations on the VH domain, while bNAbs have 40 to 100 somatic mutations on the VH domain. This allows bNAbs to have a broad neutralizing effect on most of the HIV-1 variants. bNabs are produced later in the HIV infection, usually around a year or more after the initial exposure to the virus. This is because the immune system needs to create many strain-specific antibodies in the serum in order to reduce the virus’ options of escape before the bNAbs can pursue the HIV virus and develop a broad neutralizing effect.
Currently, there is no known way to use bNAbs as a passive vaccine for HIV. In order to produce bNAbs for a passive vaccine, the antibodies would need to undergo extensive rounds of germinal center selection in order to develop a high degree of somatic hypermutation. This can be accomplished through exposure to a variety of mutant envelope spike variants over a certain amount of time. In this case, the bNabs could be harvested from an animal that elicits the same response to HIV and antibodies as a human. These bNAbs could then be purified and isolated from the animal’s serum using centrifugation and an antibody purification method, such as size exclusion chromatography or ammonium sulfate precipitation.
Cited Work:
DeleteWest, A. P., Scharf, L., Scheid, J. F., Klein, F., Bjorkman, P. J., & Nussenzweig, M. C. (2014). Structural insights on the role of antibodies in HIV-1 vaccine and therapy. Cell, 156(4), 633-648.
Annie your research on the harvesting and production of bNAbs is very interesting. It is so interesting how the complexity of the HIV virus make it necessary for many rounds of germinal selection. However it is also interesting that the way this could possibly be accomplished is exposure to several envelope spike variants in order to be able to attack the ever changing HIV virus.
DeleteSources
ReplyDeleteBridger, H. (2010, November 5). Why is it so hard to make an HIV vaccine? Retrieved November 05, 2016, from https://www.broadinstitute.org/blog/why-it-so-hard-make-hiv-vaccine
Haynes, B. F., Moody, M. A., Liao, H.-X., Verkoczy, L., & Tomaras, G. D. (2011). B cell responses to HIV-1 infection and vaccination: pathways to preventing infection. Trends in Molecular Medicine, 17(2), 108–116. http://doi.org/10.1016/j.molmed.2010.10.008
Mascola, J. R., & Haynes, B. F. (2013). HIV-1 neutralizing antibodies: understanding nature's pathways. Immunological Reviews, 254(1), 225-244. doi:10.1111/imr.12075
Newman, E. (2015). Using Naturally-Occurring Antibodies to Treat HIV. Retrieved November 05, 2016, from http://betablog.org/using-naturally-occurring-antibodies-to-treat-hiv/
Sekaly, R.-P. (2008). The failed HIV Merck vaccine study: a step back or a launching point for future vaccine development? The Journal of Experimental Medicine, 205(1), 7–12. http://doi.org/10.1084/jem.20072681
Stoller-Conrad, J. (2015, September 11). An Antibody That Can Attack HIV in New Ways | Caltech. Retrieved November 05, 2016, from https://www.caltech.edu/news/antibody-can-attack-hiv-new-ways-47845
Determine why the vaccines developed in the past have largely been ineffective in preventing AIDS.
ReplyDeleteSince its discovery by scientists in 1983, many people have been seeking to develop some kind of cure for AIDS. This search has been unsuccessful because of several challenges that the HIV virus poses. Finding an HIV vaccination is unique because the virus is unlike small pox or polio that elicit immune responses when given at low levels that cause the production of antibodies. The HIV virus goes undetected because our immune system is blind to the virus and wont launch an effective antibody attack. (Bridger, 2010) This is because HIV integrates into the genome and forms a group of resting cells that are invisible to the immune system. HIV also is undetected because it attacks CD4 T-helper lymphocytes that play a critical role in effective immune response to pathogens by the HIV virus. (Mascola & Haynes, 2013)
Some of the vaccinations they tried to develop were designed to induce a neutralizing antibody response that defends a cell from an antigen by neutralizing any biological affect it might have. They searched here because research exhibited that transferring over neutralizing antibodies could protect primates from infection of the virus. Sadly, many of the groups test in the first trials failed because of the difficulty in inducting neutralizing antibodies.
The other difficulty in finding a vaccination for HIV was the great genetic variability of the viral envelope proteins that allowed the virus to escape the antibodies. It is also difficult to identify immunogens and immunization platforms that will constantly neutralize the ever changing HIV virus. (Sekaly, 2008)
In addition to these challenges, no one has been fully cured of HIV. There are some medicines that keep HIV virus levels low, but no cure has been found to completely wipe out the virus from a patient.
Melia, your post on the specific difficulties in creating a vaccine from bNab's is insightful. Honestly it seems that across the board the HIV virus is in itself too complex for a "broad" or "general" antibody, but then on top of that its hard to even harvest and isolate those bNab's. Not trying to sound depressing though. we've come a long way. But this is an observation I find from reading everyone's posts not just yours!
DeleteWhy are bNAbs effective?
ReplyDeleteThe bNabs are effective because they stop the HIV virus from binding to CD4 cells by inhibiting the envelope spike proteins on the surface of the virus that usually bind CD4. These envelope spikes are very characteristic of HIV and so it is possible to find a target antibody towards it that can be effective. (Stoller-Conrad, 2015) bNAbs are also very effective because they can neutralize 195 out of 237 strains of the HIV virus (Newman, 2015).
How do bNAbs work at the molecular level at the antibody binding site?
They latch on to the surface of the HIV, specifically onto the Env protein. In addition to preventing the binding of CD4 and the invasion of host cells, it is thought that bNAbs also tag the HIV virus cells it binds to in order to alert the immune system to destroy the cell. The antibody binds to the gp120 protein at the CD4 binding site on the virus. (Haynes, Moody, Liao, Verkoczy, & Tomaras, 2011)
Why are they more effective than other antibodies in early infection?
Because they prevent the HIV virus from entering into the host cell and using the host cell to clone and spread throughout the body. If the bNAbs work early on, then the virus will not have spread and grown and will be more easily manageable.
Why are they produced later in HIV infection?
One reason the bNAbs might be produced later is the early damage of the production and function of B cells in the HIV virus (Haynes, Moody, Liao, Verkoczy, & Tomaras, 2011).
Could bNAbs be isolated and purified and used in a passive vaccine? If so how could they be harvested?
It seems like it could be possible for bNAbs to be isolated and purified in order to be used in a vaccine. Maybe obtaining serum from an HIV patient having bNAbs in their blood could be a possibility. Using the system for making monoclonal antibodies might work. They could separate out the B cells responsible for making the bNAbs and fuse them with Myeloma cells creating the hybridomas. The bNAbs monoclonal antibodies can be isolated from their hybridomas and purified and then given to HIV patients, especially in the early stages of their disease.
That's a great idea! I didn't think of fusing the B cells with myeloma cells to create more of bNAbs, I'm definitely curious to see how this would all be done, would they have to mix up the different bNAbs together or are bNAbs polyclonal antibodies. There hasn't been much success in this field but it'd be really neat to see if bNAbs are similar enough to regular antibodies to be able to be massed produced using hybridomas. This really would be a game changer.
DeleteHi Melia, I agreed with Sarah! That's a great idea that we could fuse B cells with myeloma cells to create a more stabilized bNabs. My research so far have shown that although they have not found problem isolating bNAbs, we still need more broad and potent kinds. I'm interested to see how the current research can utilize hybridomas to further the advancement of HIV vaccines. Good job!
DeleteI think the main reason vaccines developed in the past have not been effective in the prevention of AIDS is largely due to the nature of HIV. As we learned in lab, the virus is immunoevasive which poses a problem in trying to stimulate the immune system in any way. Other limitations that have been documented include unknowns regarding a truly useful animal model or whether reactivation could occur in an attenuated vaccine. As mentioned before, the HIV evasive properties are significant, but dysfunctions of B cells as a result of infection poses another limitation in creating an effective vaccine.
ReplyDeletebNAbs are effective in the way that they bind. Their ability as an antiviral is a result of interactions with the antigen binding site-Env. This interaction blocks the entry of virions and viral cell-cell transmission. Their activity is strongly dependent on the functions mediated by their Fc regions.
bNAbs work at the molecular level by targeting the CD-4 binding site. The most common presentation of HIV in the body is it’s attachment to a T cell. Until the virus binds to a cell, it is considered to be free-floating and its epitopes (spikes) are in a closed conformation. Most bNAbs neutralize the virus by binding to these closed epitopes. There is at least one bNAb that is known to be able to still bind to the open conformation (8ANC195).
I found it interesting that not only can bNAbs be more effective than other antibodies, there are specific bNAbs that are more effective than even other bNAbs. As I mentioned before, there are certain bNAbs that bind to certain conformations of the virus.
I think a major contributor to the late production of these antibodies is the fact that many B cell dysfunctions occur early in the course of HIV infection. The impact of the virus is significantly seen in the compromised nature of the body’s humoral response to HIV and other pathogens. Another possibility, presented by Dr. Haynes, is the fact that “bNAbs resemble the antibodies found in patients suffering from autoimmune diseases…the body lacks control over making antibodies that cross-react with their own cells.” The potential for cross reaction creates higher risk in creation of these Abs, which can explain their late appearance in HIV patients (he explains them as kind of a last resort).
I think it is a possibility to isolate these antibodies and something that is being researched further. Dr. Barton Haynes, director of the Duke Human Vaccine Institute explained that they are discussing the development of a preventative HIV vaccine that will be administered to prevent any infection in the first place. Other than this information, I was not able to find any other indications for future research. What is known about bNAbs has served as a template to hopefully create an effective HIV vaccine.
Sources:
Deletehttp://www.aidsmap.com/Why-is-it-so-hard-to-make-a-vaccine-against-HIV/page/1271042/
https://www.ncbi.nlm.nih.gov/pubmed/19372943
mals using Fc-mutated bNAbs http://www.nature.com/articles/ncomms10844
https://www.caltech.edu/news/antibody-can-attack-hiv-new-ways-47845
http://www.bnaber.org/?q=Home
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3964981/
I like that you found that there are bNABs that have specificity. From my understanding I thought they just neutralized whichever cells were around them.
DeleteHi Samantha, good research on the second to last question. It is interesting that bNAbs are used as the last resort because of the risk of cross reaction. It would be profitable if we can find ways to minimize cross reaction and potentially maximize the antibody ability to broad neutralize HIV particles.
DeletebNabs is really effective for suppressing viraemia but mechanisms on how it is done is still a questions. BNabs are able to exert ADCC which is antibodydependent cellula cytotoxicity in the cell. This helps with the destruction of HIV-1-infected lymphocytes through natural killer engagement. The antibodies target against the CD4-binding site. They are able to target mutiple epiptotes thus making bNabs effective. (http://www.nature.com/articles/ncomms10844#introduction)
ReplyDeleteThe antibodies target the CD4-binding site, the glycans/V3 and V1/V2 loops on gp120, or the gp41 moiety. The landscape of Env epitope exposure at the surface and the sensitivity of infected cells to ADCC vary considerably between viral strains. Efficient ADCC requires sustained cell surface binding of bNAbs to Env, and combining bNAbs allows a potent killing activity. Reactivated infected cells from HIV-positive individuals expose heterogeneous Env epitope patterns, with levels that are often but not always sufficient to trigger killing by bNAbs. (http://www.nature.com/articles/ncomms10844#abstract)
bNabs are more effective than other antibodies because they are able to target multiple epiptoes rather than just one. They can stick to more than HIV strains.
Bnabs take a while to form as they are always mutating and changing form.
There is no current vaccination through the purification of bNabs. It could be obtained by isolating the antibody and creating the serum but there are no studies that show this is effective
I appreciate the information you found about how bNAbs work on a molecular level. Great detail!
DeleteOne of the many reasons that vaccines developed over the years have not been effective against AIDS is the “enormous genetic diversity of HIV.” Despite the attack from the immune system, its ability to replicate and combat is deadly. Another dilemma is that an HIV vaccine cannot contain attenuated, actively replicating HIV that might cause reactivation. Many vaccines developed against HIV virus have targeted the immune response of CD8 or CD4. However researchers have not been able to guarantee the definite correlation between the virus and the immune response. Worst yet, CD8 cellular vaccines act at the later stage of the infection, and are only helpful in reducing viral loads. More importantly, HIV has also shown to be capable of developing immunity to the CD8 cellular response. With all the vaccines developed, there is no useful small animal model to study HIV infection.
ReplyDeletebNAbs are effective because they act against different sub-types of HIV that circulate worldwide, hence the term “broad”. Its ability to neutralize so many strains of HIV is crucial because HIV is so diversified. They act by attaching themselves to surface protein “Env”, in turn, preventing HIV from invading host cells. Env protein helps HIV enter and infect host cells, such as CD4 cells. If bNAbs latch on to Env, the HIV is blocked and hindered from entering CD4 target cells. It was also revealed that once bNAbs bind to the surface of HIV, these antibodies tag the virus cell for immune system, alerting natural killer cells to destroy the virus cell. bNAbs are more effective than other antibodies in early infection because they can protect a patient’s healthy cells by recognize the Env protein, present on the surface of all HIV strains and neutralizing the effects of the virus.
HIV can continually and rapidly evolve to escape neutralization. Thus the bNAb responses lag behind the virus escape and produced later in the infection. bNAbs can be isolated, purified and used in a passive vaccine. First bNAbs need to be screened from HIV infected patients to identify those who exhibit broad and potent neutralizing serum activity. Secondly the activity of these antibodies from memory B cells culture need to be tested for its binding to Env. Antibody genes can be obtained from the B cell cultures and be reproduced in tissues.
Sally, upon reading your post I agree with how such a complex infection cannot be wholly tackled by any general or "broad" antibody, though they still help a lot. As you said, HIV even "rapidly evolve to escape neutralization.
DeleteBroadly neutralizing antibodies (bNAB) are used as templates for HIV vaccine design, can effectively act as therapeutics, and help in AIDs care. BNabs mainly workand are effective by prevention early in the infection stage.
ReplyDeleteHow do bNAbs work at the molecular level at the antibody binding site?
Essentially the bNab binds to a target antigen and signals white blood cells to destroy the antigen. Nab's attach to the lipids on viral membranes and can prevent HIV from penetrating host cells. A concrete explanation for the lag in Nab production is elusive. But mostly, researchers have found Nab’s to be associated with Viral escape and often lag behind virus changes.
Only a few antibodies have been successfully isolated. One of them being b12, which was found to neutralize 40% of HIV-1 isolates. Attempts to isolate more broad monoclonal antibodies from human B-cells have been currently unsuccessful seeing that the specific proteins used for isolation (the gp120 or gp140 proteins) were too reactive with the HIV-1 antibodies. There are few method to harvesting these bNabs, one of them being Fluorescence-activated cell sorting after using antigen-specific B-cell memory.
Hi Abby, I like how you found other uses for bNAbs (i.e. therapeutics and AIDS care). I also found it interesting how you found fluorescence-activated cell sorting to be a method for harvesting bNAbs. I had a hard time finding possible methods, so it's interesting to read what you found in your research.
DeleteVaccines in the past have largely been ineffective in preventing AIDS because of the diversity of HIV-1 through mutation and its ability to escape host immune responses. Because certain regions on the HIV-1 viral envelope are often poorly immunogenic, antibody response to the virus is weak. However, in the minority of patients with HIV-1, the viral envelope will elicit broadly neutralizing antibody responses (bNAbs) after several years of infection. Over time, the humoral immune system learns to target epitopes on the native viral spike. BNAbs are the third line of neutralizing antibodies produced and are generally not found until more than two years of infection. Unlike normal antibodies, bNAbs are able to defend against mutated strains. They achieve this by recognition of conserved epitopes on the HIV-1 virus – aiding in the immune response. Molecularly, bNAbs target highly conserved epitopes, bind to those epitopes, and render the virus neutral (or inactive). The lag in production of bNAbs can be explained by the polyreactivity of some HIV-1 strains, and BNAbs can elicit tolerance from the host’s immune system or the host can develop immunoregulatory controls – limiting the frequent production of these antibodies.
ReplyDeleteThere are several issues to tackle when attempting to develop a vaccine for HIV, many of which deal with the ever-changing viral envelope and possibility of host tolerance. Because of these problems, a possible approach to create a vaccine should be structure-based. This means that a fairly large pool of donors should be used in order to collect as many different bNAbs as possible. Passive immunization is still in the experimental/hypothetical stage, but I believe it may be possible if the bNAbs are isolated (via centrifugation and possibly size exclusion chromatography) and purified. To harvest these antibodies, blood serum samples of many HIV positive/bNAb positive donors must be collected.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3738265/#R129
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ReplyDelete