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HIV vaccines: Back to basics?

With the recent linkurl:failure;http://www.the-scientist.com/blog/display/53633/ of HIV vaccine clinical trials, the HIV/AIDS research community has resolved to concentrate more on the basic science behind the disease, shifting the main focus from vaccine product development towards immunology and virology research, they said at a meeting in Maryland held today (March 25). "We need to turn the knob towards discovery," said linkurl:Anthony Fauci,;http://www.the-scientist.com/blog/display/53845/

By | March 25, 2008

With the recent linkurl:failure;http://www.the-scientist.com/blog/display/53633/ of HIV vaccine clinical trials, the HIV/AIDS research community has resolved to concentrate more on the basic science behind the disease, shifting the main focus from vaccine product development towards immunology and virology research, they said at a meeting in Maryland held today (March 25). "We need to turn the knob towards discovery," said linkurl:Anthony Fauci,;http://www.the-scientist.com/blog/display/53845/ director of NIH's National Institute of Allergies and Infectious Diseases (NIAID), which convened a linkurl:summit;http://hivsummit.dgimeetingsupport.com/ on HIV vaccine research and development, which was webcast from a Bethesda hotel. One suggestion that came from speakers and the audience, which consisted of world leaders in HIV/AIDS research: Bring new blood into HIV vaccine research by supporting young investigators. "We really do need new and novel ideas," said Carl Dieffenbach, director of NIAID's Division of Acquired Immunodeficiency Syndrome. But how? Many of the scientists who spoke at the summit called on the NIAID to raise the payline for R01 grants to fund new basic research and to encourage more participation by up-and-coming scientists. However, this would only be a temporary fix, Fauci noted. "If I could raise the payline to the 20th percentile, I would do it in a microsecond," Fauci said. "But if we don't get new money the second year, the payline will be 4 [percent]" the following year. Over the past few years, the US government has spent roughly $500 million on HIV vaccine research annually. While the summit-goers unanimously voiced a need for more resources to be poured into the HIV vaccine effort, Los Angeles-based HIV/AIDS care provider AIDS Healthcare Foundation (AHF) took advantage of the summit's lunch break to hold a teleconference suggesting the opposite. AHF president Michael Weinstein called for a linkurl:suspension;http://www.baltimoresun.com/news/opinion/oped/bal-op.aidsvaccine23mar23,0,3426636.story of government-based HIV vaccine research funding and a redirection of that money towards research into direct clinical care, prevention, and education. "After 25 years and tens of billions of dollars, we have made no progress on an AIDS vaccine," Weinstein said. "We've had a runaway train of AIDS vaccine research fueled by the best of intentions that now needs to be stopped." Back at the Maryland summit, however, no such thoughts were expressed. "This is just a pause. We will never give in," said Adel Mahmoud, chief executive of the Global HIV Vaccine Enterprise and former president of Merck's vaccine division, about the recent failure of the NIH and Merck-funded STEP and Phambili HIV vaccine clinical trials. The trials were stopped when researchers found an increased incidence of HIV infection among some trial participants who received the vaccine. Mahmoud said that the way forward will be paved by innovation from young researchers. "This is not just sloganeering," he said, "The next step is going to come from outside this room." In his closing remarks, Fauci mentioned the AHF's suggestion to stop funding HIV/AIDS vaccine research, which a panelist from South Africa had brought up earlier. "Under no circumstances will we stop funding AIDS vaccine research," Fauci said. "Not only will we not stop it, we will not cut it, and wherever possible, we will increase it."
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March 26, 2008

It has been a frustrating experience for HIV vaccine researchers. I am not a HIV vaccine researcher, yet I am involved in basic HIV biology research in an academic institution. We cannot let HIV win the war! I agree with the suggestion at the meeting that NIAID should encourage young investigators to pursue HIV vaccine research. Funding has been the major question in every academic scientists' mind today. How NIAID is going to increase the funding, when federal govt. did not increase NIH funding budget? We do need new thinking and new approach to HIV vaccine. Can someone convince the private foundations to increase the funding for HIV vaccine?
Avatar of: ROULETTE W SMITH

ROULETTE W SMITH

Posts: 10

March 26, 2008

New blood? ? Young investigators / researchers? ? Raise the payline for R01 NIH grants? ? More resources (funds) for HIV vaccine research? Adel Mahmoud said, "This is not just sloganeering, ? The next step is going to come from outside this room." An outside voice need not be young, new or a person with a long-standing NIH track record for R01 grantsmanship! Why not consider a fundamental shift in paradigms as proposed in Smith, R. Wm (2001). The Durban Declaration, The Scientist 15(2):39 (http://www.the-scientist.com/article/display/12233/)?? To wit, why not explore multivalent vaccines against nascent pathogens which would be opportunistic. This indirect approach to vaccination against lentiviruses is supported by Bjorn Sigurdsson?s 3 classic 1954 British Veterinary Journal reports on slow infections in sheep. Ethological reports of AIDS-like disease in Rhesus monkeys at the UC Davis Primate Center also are instructive and support this approach [see Lerche, N. W, Henrickson, R. V., Maul, D. H., and Gardner, M. B. (1984). Epidemiologic Aspects of an Outbreak of Acquired Immunodeficiency in Rhesus Monkeys (Macaca mulatto), Laboratory Animal Science 34(2):146-150]. At the very least, a shift in paradigms is likely to teach investigators more about how nascent (i.e., relatively uncommon) pathogens activate HIV in contrast to common pathogens.
Avatar of: Jonas Moses

Jonas Moses

Posts: 26

March 26, 2008

To "The Scientist," and its readership...\n\nIn a sentence: there is to be found no hard evidence of a single retrovirus, so-called "HIV." \n\nWhy can't a vaccine be created? Simply put, you cannot vaccinate against something that does not appear to exist in nature. I propose this as a good place to begin the discussion...\n\nAs a mainstream, conservative scientist and former medical clinician, as a member of the international medical Media, as the son of world-class and highly respected scientists, I feel it is time to place all cards on the table, in questioning the veracity of all claims made about HIV and AIDS.\n\nFirst, I want to be clear that I was trained in, and raised on, the Scientific Method. This then, is the source of my concern and my reason for questioning what has been, immutably, cast in stone as The Truth: "HIV causes AIDS"\n\nFrom the recent Washington Post article:\n\n"People have been suffering immune-activating infections and getting vaccines for years, and there has never been evidence that those events\nincreased a person's risk of acquiring HIV. These vaccine trials would be odd places to first notice such a thing. Furthermore, people in the STEP\nstudy who got the vaccine did not have more activated CD4 cells than people who got placebo -- something that Merck vaccine executive Mark B.\nFeinberg called "kind of an interesting and unexplained observation.""\n\n-- Interesting and unexplained...indeed.\n\nand\n\n""There is something very, very peculiar" going on in the vaccine trials, said Anthony S. Fauci, head of the National Institute of Allergy and\nInfectious Diseases, which sponsored them."\n\n-- Very peculiar...and highly suspect.\n\nand \n\n"The multiple surprises have reminded researchers how much they still do not know about HIV's biology. It has also focused attention on questions they never asked."\n\nI find it odd that Edward Jenner (1749-1823), was able to derive a vaccine against smallpox - long considered to be the most deadly and persistent human pathogenic disease (up until the 1970's, in fact) - in a matter of weeks, with virtually no resources (read: "outside funding"), little in the way of technology, and no background in Virology. \n\nYet, with billions of dollars in funding, hundreds of laboratories and thousands of scientists, leading-edge medical and scientific technologies, highly-evolved fields of Cell/Molecular Biology and Virology, and twenty-five years of intensive study, "HIV" biology is still poorly understood and there is no vaccine or successful treatment. \n\nSomething is very wrong here.\n\nConsider the following, well-documented facts:\n\n1) The so-called retrovirus, "HIV," has never been successfully isolated. In truth, there are several organizations, world-wide, prepared to pay hefty sums of money to the first individual or group who can present unequivocal documentation of "HIV." \nThis money has not been awarded to any person or group, to date. It remains unclaimed. Why is this the case, if "HIV" is everywhere? I refer you to the World-wide Web for validation of this claim...it is quite easy to verify.\n\n2) No one has ever successfully utilized Koch's Postulates in validating "HIV" as an infectious agent. Yet, remarkably, after some 125 years, Koch's Postulates remain the "gold standard" in the determination of infectious disease agents.\n\n3) Every single one of the test kits used in "identifying" HIV infection is inferential, ergo indirect; furthermore, the manufacturers of these kits acknowledge and publish that the kits are "not to be used" in the identification of HIV infection. Why...they must know something that all of the AIDS researchers seem not to know?\n\n4) Consider the use of PCR (polymerase chain reaction) in identifying "HIV" infected blood or tissues: no less than the inventor of PCR, Nobel Laureate Kelly Mullis, has stated that PCR cannot be used in identifying "HIV." I wonder why?\nDoes he know something that the AIDS researchers do not?\n\n5)Not one of the 60-odd failed "HIV" vaccines has succeeded in evoking antibodies. Indeed, there is evidence that such vaccines may do real harm to human beings.\n\n6) Consider also, that there are no less than 15 other hypotheses (likely, many more) for what causes AIDS-like disease processes. Noting that AIDS is NOT a disease, in and of itself, but is a so-called syndrome, defined by some 40-plus existing diseases, why do so many scientists, politicians, Media professionals and government officials refer to AIDS as if it were a single disease? Why, also, are so many among the general public allowed to go along with this non-fact, blithely ignorant of the truth? Why does no one among the AIDS "community" scientists, clinicians and non-medical "experts" correct this misconception, with a high-profile public statement? Is it possible they do not want the populace to begin questioning?\n\nNote also that these so-called AIDS-defining illnesses can and do sicken and kill human beings, in the absence of any precursor retrovirus (HIV).\n\nGiven that several, if not most, of these other noteworthy AIDS hypotheses have been around for as long as the HIV=AIDS hypothesis, and that many if not all of these other hypotheses are far better at explaining the syndrome than "HIV=AIDS," why is more money not being poured into researching those avenues? Why indeed, in the face of some 25 years and billions of dollars having been spent on failed HIV=AIDS research?\n\nI am impressed with the elaborate explanations for why this virus is so elusive and difficult to treat: \n\n- capable of mutating at a furious rate, while maintaining its integrity as an infectious and disease-causing virus?\n- capable of multiplying in the very T-Cell hosts it supposedly kills off?\n- capable of laying dormant for 40 years or killing an infected host in a matter of months...and a plethora of other, intriguing-if- nonsensical claims.\n\n"Infected" (ie "tested positive") individuals may show no viral load and have completely normal T-Cell counts, may never develop any diseases, may never experience so much as a cold, and live healthy and productive lives; meanwhile, others maypresent with sky-high viral load and nearly decimated immune systems; still others may test "negative" (classed "ideopathic", and pushed into a dark corner) but develop multiple so-called AIDS-defining illnesses. \n\nSero-discordant couples may have unprotected sex for years, with no conversion of the "uninfected" partner. "HIV-positive" mothers may deliver "HIV-negative" offspring, even though they share the same blood supply...for nine months. \n\nThese are but a mere tip of the iceberg, for a disease hypothesis that abounds in bizarre and wide-ranging inconsistencies.\n\nMiracles, all. \n\nAmazing as well that there is no animal model: \n\nEven the chimpanzees who were "infected" with so-called HIV, some 25 years ago, had to be retired to a $25 million habitat, not long ago, because none of them became ill. \n\nMore amazing that many AIDS patients also happen to be chronic drug-abusers/addicts, alcoholics and smokers and/or already have hemophilia, tuberculosis, malaria, hepatitis, cancer, syphilis and a plethora of other immune-compromising diseases and disorders, and/or are immune-compromised because of starvation, tainted drinking water and lack of basic health care (think countries like Africa), and yet no one is pointing to these preexisting conditions (all of which can cause AIDS-like diseases) as causal. Amazing...and foolhardy. \n\nWhy, indeed, is it so difficult for thinking scientists and physicians to appreciate that the highly toxic drug regimens foisted upon uninformed (and often unwilling) patients are likely doing more to destroy their immune systems than any virus might? There is a growing disconnect between the practice of good science and medicine, and those who promote "HIV" as the sole and most likely cause of AIDS. \n\nPlease consider questioning this dogma - no longer even Science dogma, as it has truly become a religion for those who follow it - for, the best-informed and most balanced view is bound to be the most successful, in combating real disease, with real Science and real Medicine.\n\nIn case you are wondering, no...I am not an AIDS dissident, and do not identify myself with any such groups. I am a conservative and skeptical scientist, who has stumbled across the inconvenient and disturbing truth that the facts about HIV do not add up.\n\nThe more I read, the more convinced I am this is plainly and simply a case of practicing bad Science and bad Medicine.\n\nI would like to invite any and all medical and scientific professionals to appear as a guest on one of my weekly television programs, to discuss this matter...on the merits. If you are interested in being interviewed, regardless of how you feel about the HIV=AIDS hypothesis, I welcome a contact from you. So long as you are willing to engage in a reasoned and unemotional scientific conversation\nabout the topic, you are invited to join the discussion. My programs are peer-to-peer, worldwide, which means the conversations reach scientists, clinicians, medical ethicists, academicians, Industry leaders, biomedical innovators, Public Health/Public Policy professionals, government officials and other Media professionals in over 35 countries. I am sincerely interested in arranging a forum for discussion, and would like to include any and all perspectives. Please know that my professional credibility as moderator of such a television series rests upon my ability to remain neutral to the conversation at hand, and I will go to great lengths to protect the views and perspectives of my guests, regardless, so long as they agree to adhere to the discussion of peer-reviewed articles , from widely accepted and respected journals, and to independently reproducible (and reproduced) experimental studies. \n\nTo reach me, regarding being included in this professional on-air forum, or to be considered as a potential guest for any of my medical programs, please send me an email via the InTime TV Network: OntheEdge@intimetv.com\n\nSincerely, \n\nJonas Moses, PhD PA\n\n[Biomedical Engineering innovator of diagnostics, therapeutics and devices in the treatment of cancer and other disease processes; cancer and cell biologist and former medical clinician (in Ophthalmology), Medical/Science Liaison and TV talk show Host]
Avatar of: Mike Barr    

Mike Barr   

Posts: 2

March 27, 2008

AIDS Healthcare Foundation is widely reviled in the AIDS community for their self-serving proclamations, and this appears to be one of them.\n\nTheir statement that "rapid scale-up of HIV testing worldwide would likely prevent millions of new infections" requires better informed scrutiny. As scientists they must know that HIV testing has not been shown to prevent transmission except in the case of serodiscordant couples. Surely they must be aware of the thin ice they skate on when they claim: "Achieving universal access to treatment would make the most significant contribution to a drop... in HIV transmission rates worldwide." There is not one iota of scientific evidence that this is so!\n\nAnd there are other contentious (or flat out fictional) points in their 3/23 Baltimore Sun opinion piece:\n\n* "A vaccine against a retrovirus, the family of viruses HIV belongs to, has never been successfully developed." Not true. There is a licensed vaccine against the feline retrovirus FIV. It isn't perfect, isn't always used, but it does exist.\n\n* "If the majority of the 250,000 undiagnosed cases were diagnosed over the next 5 years, new infections could be reduced by as much as 50 percent." Perhaps. But this is pure (and rhetorically self-serving) speculation. The AHF leaders rightly point out that transmission is reduced in couples where the viral load is kept in check with drugs. The viral load is highest, however, during the first weeks of infection when conventional tests would often give a false negative result. Once a conventional test is positive, the window for most likely transmission may already be closed. Simply increasing testing without improving sensitivity to detect new infections is unlikely to produce the hoped for effect.\n\n*"Scale-up of treatment worldwide must be our highest priority." Again, highly contentious. Treatment with a combination of costly and harsh antiretroviral medicines must be maintained for a lifetime?without interruption. While early and continuous combination antiretroviral treatment represents a significant boon to commercial entities with a vested interest in keeping patients on lifelong drug regimens, long term toxicity and drug resistance may undermine this strategy. A short-term focus on drugs to the exclusion of next-generation preventative measures (including vaccines) could create an even worse situation 10-20 years down the road.\n\nThe final irony of all this is particularly rich given African representatives' reaction to the Healthcare Foundation publicity stunt:\n\n?[The call] to redirect vaccine funding into testing and treatment... stirred consternation in the AIDS community, especially in Africa, where the bulk of the pandemic's 33 million victims live and where ?AIDS is not an abstract thing,? said vaccine researcher Glenda Gray of the University of Witwatersrand, South Africa. ?Any call to halt vaccine funding is like abandoning Africa.?" (WSJ 3/26)\n\nOne cannot help wondering how in touch with the African communities they purport to serve these AIDS Healthcare Foundation people really are?and whose interests lie foremost in their minds.\n\n
Avatar of: Jesse Creel

Jesse Creel

Posts: 4

March 31, 2008

Below is a commentary of mine which I sent to all the principals of the March 25 HIV-1 Vaccine Summit in advance...Fauci, Greene, Gray, Hoxie, Watkins, DAIDS etc. It puts forth some new avenues of HIV-1 Vaccine development worthy of consideration based on my 23 year reading of the literature.\n \nThanking You In Advance\nJesse Creel\nVaccine Research Advocate\n+++++++++++++++++++++++++++++++++++++++++++++++++ \nHIV-1 Vaccine Summit: Which Direction from Here?\n \nFirst let me say I wished all success in the HIV-1 Vaccine Summit on\nMarch 25 th, 2008, hosted by DAIDS/NIAID.\n \nFollowing the failure of the Mrk Ad5 STEP trials (60) HIV-1 Vaccine\ndevelopment is searching for which path to follow. With your kind\nindulgence, below are my thoughts on this.\n \nAccording to the prevailing opinion on HIV-1 protective immunity, a\nsuccessful HIV-1 Vaccine will need to target conserved HIV-1 genome\nsequences. The question then arises, how to best target these conserved\nHIV-1 sequences?\n \nSeveral approaches are under consideration currently. At the forefront are\nvarious DNA Prime/Boost vaccine approaches involving the use of a DNA Prime\nfollowed by a recombinant microbial vector boost, e.g., MVA or Adenovirus.\nNotably are the delayed PAVE 100 DNA Prime/Adenovirus Boost (1;60) and DNA\nPrime/MVA boost (2;60). Another approach uses a DNA Prime\nfollowed by a recombinant protein boost as per ( 3 ; 59) These only being a\nsampling of the HIV-1 vaccines under serious development. (9;86)\n \nGiven this emphasis on DNA Vaccines, which offers ease of production and low\ncost logistical advantages, it seems merited to consider carefully the fact\nthat no DNA vaccine has shown efficacy in humans despite many years of\nresearch.\n \nOne possible reason for the failed efficacy of DNA Vaccines in humans may be\nthat cellular RNAi mechanisms, as spoken to in abstracts and citations below\n(4-8), may target the mRNA of a DNA Vaccine thereby curtailing or preventing\ntranslation into vaccine epitopes/antigens.\n \nOn this in email exchanges:\n \nHIV-1 Vaccine Summit Co-chair Warner C. Greene, M.D., Ph.D. Director,\nGladstone Institute of Virology and Immunology replied:\n \n"Micro RNAs, in contrast to synthetic siRNAs, block the translation of mRNAs\ninto their protein products instead of inducing mRNA degradation. However,\nfor a vaccine which depends on the production of the antigen, their net\neffect is similar. I do not know whether any of the naturally occurring\nmicro RNAs would be predicted to inhibit production of the protein products\nfrom the current spectrum of DNA vaccines that are being tested. It has\nbeen clear for some time that DNA-based vaccines work significantly less\nwell in humans than in mice. I suspect the reason for this is\nmultifactorial and not solely due to micro RNAs."\n \nBharat Ramratnam, an RNAi researcher at Brown U, said:\n \n"It would entirely depend upon the nucelic acid sequence of the\nimmunogen--if it were homologous to a miRNA, then a nil to modest to severe\nreduction in immunogen transcript level could be expected. We have been\nlooking at this, but NO conclusive data\n \nStay tuned...."\n \nMonsef Benkirane, an RNAi researcher and associate editor of the Journal\nRetrovirology, said:\n \n"Absolutely, DNA viruses often produce viral miRNA that will either target\nviral mRNA or cellular miRNA. I believe that one should try to avoid any\nmRNA structure that will serve as substrate for Drosha and Dicer in the\nprocess of making DNA viruses vaccine."\n \nA successful HIV-1 DNA Vaccine will need to target conserved sequences of\nHIV-1(10), the very same sequences of mRNA that cellular RNAi will need to\ntarget to curtail HIV-1 replication. Therefore it stands to reason that\ncellular RNAi will also target these HIV-1 DNA Vaccine mRNA sequences for\nsilencing/RNAi thereby curtailing DNA Vaccine Efficacy.\n \nIn my humble opinion this is probably the case, and along with other\nfactors, are the reason DNA vaccines have failed to show efficacy in humans\ndespite many attempts over a number of years.\n \nSpeaking to other factors as to why DNA Vaccines don't show the\nefficacy they did in proof of concept small animal trials.\n \nIn an email exchange Robert Malone, an early DNA plasmid gene therapy\npioneer, said:\n \n"Basically, the transfection efficiency in larger mammals is not good. If\nthe original hypothesis of my colleague Dennis Carson held (muscle cells as\n \nAPC), then due to absence of costimulatory molecules on muscle cells the\n"vaccination" would have evoked tolerance, not immunity! To the extent that\ndelivery to and expression in muscle is important, it appears to be due to\ncross-priming (exosomes?).\n \nRather, tissue dendritic cells are most likely the important actors."\n \n"I was well aware of the fact that the field "ran off" to prove the utility\nin humans of what was really immature technology. The rodent findings were\nmerely proof of the concept, but the delivery technology was not "enabling"\nfor larger mammals."\n \nMuscle mass in humans is much greater than in small animals therefore much\nof a DNA Vaccine is taken up by muscle cells rather than DCs, as Dr Robert\nMalone says above, this would reduce DNA Vaccine efficacy.\n \nFurther, according to my understanding, any DNA Vaccine translated peptide\nand/or protein mRNA transcript would be processed and presented as antigen\nvia the endogenous rather than the exogenous/cross-priming HLA I pathway.\nThe DNA Vaccine construct mRNA transcripts would be treated as mRNA of a\nvirus which infected the cell. This could result in the killing of some\ncells by immune system components, Th1 CTL and Th2 Antibody both I think.\n \n \nAll this doesn't even speak to safety concerns which may result in a delay\nin the FDA regulatory approval process for any DNA Vaccine, not covered here\nfor brevity.\n \nThese DNA Vaccine Efficacy and/or Safety problems bring into question the\nutility of continued investment of time, effort and money, both public and\nprivate, into a number of HIV-1 DNA Vaccines in clinical trials, spoken to\nin an article by Margaret Johnston, head of HIV-1 Vaccines at DAIDS at\nNIAID, and Anthony Fauci, head of NIAID . ( 9)\n \nThe use of recombinant viral vectors also presents us with several problems.\nPre-existing immunity, as shown in the recent failed Mrk Ad5 vaccine trials\n(11;60), is one problem. Anti-vector immune responses would curtail, even\nwhere there is no preexisting immunity, efficacy. These anti-vector immune\nresponses would also squander immune system components ---cellular and\nsoluble cytokines such as IL-2 as well as molecular building blocks, e.g., amino\nacids, peptides and proteins ---needed for protective immunity. This being\nof particular importance in populations under general immunosuppression, as per the case\nwith the HIV+ and/or HIV- malnourished populations found mainly in the poor\nnations, with limited immunologic potential.\n \nFurther, these populations are under a high state of immune activation.\n(20;65;91;93) Besides endemic gut parasite and other infections, a high state\nof immune activation occurs due to multiple insults--- generally squalid\nliving conditions, unsanitary food, un-potable water, biting insects and\nenvironmental pollution. This highlights the need to use a innocuous vaccine\nvector that avoids undue systemic immune activation in order to prevent further\nvaccine induced immunosuppressive cytokines such as IL-10 and Tregs which\nwould reduce vaccine efficacy. (65;92;94-101)\n \nSimilar would apply if a systemic adjuvant, as QS-21 adjuvant (Antigenics\nInc., Woburn,MA) (59) was used as part of the vaccine.\n \nShould we be designing vaccines that use microbial vectors, adjuvants or\nexcess HIV-1 epitopes/antigens that may induce unneeded immune activation,\nwhich may be counterproductive as regards vaccine efficacy by causing\nthe activation of T Cells unnecessarily?\n \nOne consequence of an HIV-1Vaccine that causes undo immune activation\nwould be, activated T cells are more readily infected by HIV.\n \nAnother consequence may be the induction of HIV-1 replication by latently\ninfected T cells and/or a higher rate of replication by HIV-1 infected T\ncells already replicating HIV-1 at lower levels.\n \nTaken together, an HIV-1 vaccine that induces inappropriate activation of\nT cells may lead to the higher HIV-1 viral loads characteristic of AIDS\ndisease progression, while on the other hand it is acting to curtail HIV-1\nviral loads, in total, reduced vaccine efficacy.\n \nI think we must choose HIV-1 Vaccine components carefully to insure as\nbest we can that only cells important to HIV-1 protective immune responses\nare activated. This would reduce any possible vaccine induced\ncontribution to AIDS immune hyper activation pathologies.\n \nSafety issues disfavor viral vaccie vectors. They have been shown to cause\ndeath, Jesse Gelsinger's death caused by an adenovirus vector in gene\ntherapy trial (12) and disease in some cases, as per OPV actually causing\nvaccine induced polio (13;60), MVA causing heart problems(14) and worsened\nRSV disease in children upon re-exposure after inactivated RSV vaccination\n(60). Of coursethere is also a chance of reversion to virulence upon serial\npassage from one person to another over time (15).\n \nAnother approach which has fallen out of favor since Dr Donald\nFrancis/Vaxgen's AIDSVAX, which used only gp120 epitopes from a narrow range\nof HIV-1type B strains, failed to show efficacy (16), is the use of Recombinant Peptides in\nan HIV-1 vaccine. By expanding the peptide epitopes to be used in a vaccine\ntocover multiple conserved consensus HIV-1 sequences from various HIV-1\nstrains worldwide, this approach deserves further consideration I think.(10)\n \nA HepB Peptide vaccine that has shown efficacy and safety is currently being\nused in humans. (17) This should speed FDA approval of a HIV-1 Peptide\nVaccine.\n \nThe general concept of a polyvalent peptide prime followed by a\npseudoprotein boost is a viable approach according to my understanding of\nthe literature over the past 23 years. Recent human Phase I Trials by Dr\nShan Lu, UMASS Medical School, and colleagues, using a polyvalent DNA prime followed by\npseudoprotein boost regimen have shown promise.(3 ; 59) The DNA Prime\nepitopes used were from a narrow range of older HIV-1 isolates:\n \n"five plasmids each encoding a codon-optimized gp120 gene sequence from the\nfollowing primary HIV-1 envelope proteins: subtypes A (92UG037.8), B\n(92US715.6 and Bal), C (96ZM651) and E (93TH976.17) and\nthe sixth plasmid encoding a codon-optimized gag gene from\nsubtype C (96ZM651) as previously described [28]."\n \nAs regards the pseudoprotein boosters used:\n \n"The recombinant Env protein vaccine components included\nin the DP6-001 formulation contain equal amounts of five\ngp120 proteins matching that used in DNA prime components\nand were produced in CHO cell lines by Advanced BioScience\nLaboratories (ABL, Kensington, MD) using GMP compliance as\npreviously described [28]. The final protein vaccine product\nwas supplied in saline and re-formulated at the time of injec-\ntion with 50_g of QS-21 adjuvant (Antigenics Inc., Woburn,\nMA) and 30 mg of excipient cyclodextrin (Cargill Cerestar\nUSA Inc., Hammond, IN),"\n \nI think a better approach is to use Conserved Immunogenic Consensus\nSequences (ICS) defined by close computer analysis of multiple HIV\nclades/strains worldwide, bioinformatics, to point the vaccine induced\nimmune response to conserved epitopes most likely to offer protection for\nall strains of HIV-1 worldwide.(10)\n \nThe GAIA HIV-1 Vaccine abstract (10) points out a number of\nexcellent HIV-1 Conserved Immunogenic Consensus Sequences (ICS) derived from\nclose computer aided analysis, bioinformatics, of multiple HIV-1 genome\nsequences, covering various HIV-1 strains worldwide, which can be used as\nHIV-1Epitopes/Antigens in order to conduct comparative studies between DNA and\nPeptide/ Pseudoprotein formulations of a HIV-1 Vaccine, using the same ICS\nepitopes, to determine which approach is best.\n \nGAIA is a not for profit foundation\n(http://www.gaiavaccine.org/matriarch/default.asp)\ndedicated to the development of a HIV-1 Vaccine available to rich and poor\nalike.\n \nHIV-1's main portals of entry are at various mucosal surfaces and the site\nof most CD 4+ T Cell Loss early on. (21-29) It makes good vaccine sense to\ntarget a vaccine to the GALT which can result in good immunity at\nvarious mucosas as well as systemically. (30-34;48-49;85;89) This should\ncurtail or prevent infection and/or systemic dissemination of HIV-1.\n \nA Gelcap encapsulated (35-37;89)Mannosylated (38-41 ) Self-adjuvanting\n(42-47 )Freeze-dried Liposome Vector (36) which targets the GAIA HIV-1\nVaccine components to the mannose receptor on DCs (40;50-52), and DEC-205 on DCs and\nB cells (61),can orally deliver both DNA and/or Peptide/Pseudoprotein formulations\nof a HIV-1 Vaccine to DCs and B Cells of the GALT.\n \nI suggest we conduct two prong comparative trials using a GAIA HIV-1 ICS\nVaccine Polyvalent Peptide Prime, DNA ICS Prime in one prong and Plain ICS\nPeptide Prime in the other prong, and GAIA Recombinant ICS Pseudoprotein\nBoost in both prongs to determine which strategy is best.\n \nThe presence of DEC-205 on B Cells and DCs is good news. Mannosylated\nLiposomes uptake should allow B Cells (53-55 ) and DCs to be used as\nAPC(55;61).\n \nMannan-binding lectin (40) should bind the Mannosylated Liposomes vector,\nactivating the classical complement cascade(62) and inducing\nopsonization/phagocytosis and antigen presentation .\n \nMannosylated Liposome will also provide lipids which are key to proper\ntrafficking and loading of ICS epitopes onto HLA I & II. (31;43;63)\n \n"Presence of both liposomal lipids and liposomal protein in the trans-Golgi\ntherefore facilitates the entry of liposomal antigens into the MHC class I\npathway. It is also possible that liposomal lipids are presented to T cells via the\nrecently described CD1 pathway for lipid antigens. Because\nliposome-formulated vaccines have the potential to stimulate antibody as\nwell as cellular immune responses to protein and lipid components, this\napproach could prove to be extremely useful in designing vaccine\n strategies."(63)\n \nThe use of the CD1 antigen pathway is not spoken to here, however I think it\npossible that both MHC I & II and CD1 may be used with this vector strategy.\nIf so, this would greatly enhance the GAIA HIV-1 ICS Vaccine induced immune\nresponses.\n \nThis delivery of concentrated high dose multiple consensus antigens to GALT\nDCs and B cells is crucial in order to evoke mucosal and systemic immune\nresponses which are very strong in breadth and depth. This is essential\ngiven thesmall number of infectious HIV-1 vs Non-infectious HIV-1(NHIV-1), 1\nin 10,000+ HIV-1 progeny(64;66;68 ), essential to protective immunity. We have\na small target of HIV-1 in a large crowd of NHIV-1, therefore we will need\nthis very strong immune response to insure efficacy. Without infectious HIV-1 capable\nof producing both HIV-1 and NHIV-1 there will be no more AIDS.\n \nBoth DCs and B Cells have TLR-2 which can bind Mannosylated Liposome\nactivating DCs and B Cells via the Toll Pathway, adjuvanting actions\nakin to LPS/Freud's adjuvant. Causing up-regulation of co-stimulatory\nmolecules, MHC II and cytokine secretion (55-58;73-82;84) and endocytosis of\nthe GAIA ICS Liposome Vaccine Vector:\n \n"Thus, antigen linked to the TLR2 ligand can be endocytosed after binding\nTLR2, processed via the classical (exogenous) pathway of antigen\npresentation, and can enhance the stimulation of T cells. This same route\ncould be exploited to generate more efficacious vaccines." (70) (71;75)\n \nMultiple pathways of DC and B Cell phagocytosis/uptake of Mannosylated\nLiposome encapsulated GAIA ICS antigens should give a stronger vaccine\ninduced immune response by providing a reciprocal boosting of the DC and B\nCell vaccine induced immune response. (72)\n \nBest would be an oral formulation with no boost required. This would offer\nease of administration, along with several cost and safety advantages when\nyou consider the need to vaccinate several billion people living mainly in\nthe resource poor nations with limited healthcare infrastructures and public\ntransport. This is highlighted by recent reports regarding the reuse of\ndirty syringes and medical equipment occurring in the world's richest\nnation, the USA. (18) In the resource poor nations this is even more likely \nto occur owing to cost considerations as well as lack of education regarding\nproper medical hygiene and generally meager healthcare infrastructures, as\nper the case of HCV transmission by reusing unsterile syringes in Egypt. (19)\n \nTo test this one dose oral vaccination (88-90) possibility I suggest we try,\nin addition to the proposed two prong GAIA ICS Prime and Pseudoprotein Boost\nabove, the administration of peptide and protein components of the vaccine\nsimultaneously also. This would mimic reasonably well what happens in\nnatural exposure to HIV-1, where in the majority of exposures no actual\ntransmission/infection occurs.\n \nFurther there is the concern regarding Tregs commonly found in HIV-1/Aids\nas indicated by the excellent research of Gene Shearer NCI and colleagues.\n(67;69) which could curtail vaccine induced antigen specific CD 4+ T Helper\ncells which are crucial to the establishment of long term immunologic memory. The\nlarge number of CD 4+ T Helper cells induced by this GAIA HIV-1 ICS Vaccine\nstrategy should be able to withstand any ensuing Tregs which tend to come\ninto play later. (83) Thereby providing a excellent pool of Memory CD 4+ T\nHelper (87) able to rapidly respond to future encounters with HIV-1, be it\nfrom further exposures or bouts of HIV-1 replication in vivo in the\nchronically HIV-1 +.\n \nIn effect, it is hoped that this HIV-1 vaccine strategy will be Therapeutic\nand Preventative/Prophylactic!\n \nThanking you in advance for considering this proposed HIV-1 Vaccine\ndevelopment strategy.\n \nJesse Creel\nVaccine Research Advocate\n1104 River Valley Dr #3\nFlint, MI 48532\nEmail: JesseCreel@comcast.net\n \nA special thanks to Dr Anne De Groot and colleagues at GAIA Vaccine\nFoundation, Dr Gene Shearer at the NCI and Dr Robert Malone for the many\nhelpful insights and encouragement provided without which I could not have\never proceeded.\n \nPlease see Endnote Regarding Production of GAIA ICS Antigen\n++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++\nReferences\n1. PAVE 100\n http://www.hivpave.org/about/pave100-copy.html\n \n2. Clin Pharmacol Ther. 2007 Dec;82(6):686-93\n \n3. Springer Semin Immunopathol. 2006 Nov;28(3):255-65\n \n4. Journal of Virology, March 2008, p. 2895-2903, Vol. 82, No. 6\n \nHuman Immunodeficiency Virus Type 1 Escape Is Restricted When Conserved\nGenome Sequences Are Targeted by RNA Interference\n \nKarin Jasmijn von Eije, Olivier ter Brake, and Ben Berkhout*\nLaboratory of Experimental Virology, Department of Medical Microbiology,\nCenter for Infection and Immunity Amsterdam, Academic Medical Center of the\nUniversity of Amsterdam, K3-110, Meibergdreef 15, 1105 AZ Amsterdam, The\nNetherlands\nReceived 14 September 2007/ Accepted 5 December 2007\nRNA interference (RNAi) is a cellular mechanism in which small interfering\nRNAs (siRNAs) mediate sequence-specific gene silencing by cleaving the\ntargeted mRNA. RNAi can be used as an antiviral approach to silence the\nhuman immunodeficiency virus type 1 (HIV-1) through stable expression of\nshort-hairpin RNAs (shRNAs). We previously reported efficient HIV-1\ninhibition by an shRNA against the nonessential nef gene but also described\nviral escape by mutation or deletion of the nef target sequence. The\nobjective of this study was to obtain insight in the viral escape routes\nwhen essential and highly conserved sequences are targeted in the Gag,\nprotease, integrase, and Tat-Rev regions of HIV-1. Target sequences were\nanalyzed of more than 500 escape viruses that were selected in T cells\nexpressing individual shRNAs. Viruses acquired single point mutations,\noccasionally secondary mutations, but-in contrast to what is observed with\nnef-no deletions were detected. Mutations occurred predominantly at target\npositions 6, 8, 9, 14, and 15, whereas none were selected at positions 1, 2,\n5, 18, and 19. We also analyzed the type of mismatch in the siRNA-target RNA\nduplex, and G-U base pairs were frequently selected. These results provide\ninsight into the sequence requirements for optimal RNAi inhibition. This\nknowledge on RNAi escape may guide the design and selection of shRNAs for\nthe development of an effective RNAi therapy for HIV-1 infections.\n \n \n* Corresponding author. Mailing address: Laboratory of Experimental\nVirology, Department of Medical Microbiology, Center for Infection and\nImmunity Amsterdam, Academic Medical Center of the University of Amsterdam,\nK3-110, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Phone: 31 20\n566 4822. Fax: 31 20 691 6531. E-mail: b.berkhout@amc.uva.nl\n Published ahead of print on 12 December 2007.\n \n http://jvi.asm.org/cgi/content/abstract/82/6/2895?etoc\n \n5. Journal of Virology, March 2008, p. 2938-2951, Vol. 82, No. 6\n \nTargets of Small Interfering RNA Restriction during Human Immunodeficiency\nVirus Type 1 Replication\n \nYong Gao,1 Michael A. Lobritz,1,2 Justin Roth,3 Measho Abreha,1 Kenneth N.\nNelson,1 Immaculate Nankya,1,2 Dawn M. Moore-Dudley,1,2 Awet Abraha,1\nStanton L. Gerson,3 and Eric J. Arts1,2*\n \nDivision of Infectious Diseases, Department of Medicine,1 Department of\nMolecular Biology and Microbiology,2 Case Comprehensive Cancer Center, Case\nWestern Reserve University, 10900 Euclid Ave., Cleveland, Ohio 441063\nReceived 26 September 2007/ Accepted 4 January 2008\nSmall interfering RNAs (siRNAs) have been shown to effectively inhibit human\nimmunodeficiency virus type 1 (HIV-1) replication in vitro. The mechanism(s)\nfor this inhibition is poorly understood, as siRNAs may interact with\nmultiple HIV-1 RNA species during different steps of the retroviral life\ncycle. To define susceptible HIV-1 RNA species, siRNAs were first designed\nto specifically inhibit two divergent primary HIV-1 isolates via env and gag\ngene targets. A self-inactivating lentiviral vector harboring these target\nsequences confirmed that siRNA cannot degrade incoming genomic RNA.\nDisruption of the incoming core structure by rhesus macaque TRIM5 did,\nhowever, provide siRNA-RNA-induced silencing complex access to HIV-1 genomic\nRNA and promoted degradation. In the absence of accelerated core disruption,\nonly newly transcribed HIV-1 mRNA in the cytoplasm is sensitive to siRNA\ndegradation. Inhibitors of HIV-1 mRNA nuclear export, such as leptomycin B\nand camptothecin, blocked siRNA restriction. All HIV-1 RNA regions and\ntranscripts found 5' of the target sequence, including multiply spliced\nHIV-1 RNA, were degraded by unidirectional 3'-to-5' siRNA amplification and\nspreading. In contrast, HIV-1 RNA 3' of the target sequence was not\nsusceptible to siRNA. Even in the presence of siRNA, full-length HIV-1 RNA\nis still encapsidated into newly assembled viruses. These findings suggest\nthat siRNA can target only a relatively "naked" cytoplasmic HIV-1 RNA\ndespite the involvement of viral RNA at nearly every step in the retroviral\nlife cycle. Protection of HIV-1 RNA within the core following virus entry,\nduring encapsidation/virus assembly, or within the nucleus may reflect virus\nevolution in response to siRNA, TRIM5, or other host restriction factors.\n \n \n* Corresponding author. Mailing address: Division of Infectious Diseases,\nBRB 1034, Case Western Reserve University, 10900 Euclid Ave., Cleveland, OH\n44106. Phone: (216) 368-8904. Fax: (216) 368-2034. E-mail: eja3@case.edu\n Published ahead of print on 16 January 2008.\n \nhttp://jvi.asm.org/cgi/content/abstract/82/6/2938?etoc\n \n6. Nature Medicine 13, 1241 - 1247 (2007)\n \n7. Retrovirology 2005, 2:81doi:10.1186/1742-4690-2-81\n \n8. Nucleic Acids Research, 2007,Vol.35, No. 10 e73\n \n9. N Engl J Med 2007;356:2073-81.\n \n \n10. Vaccine 23 (2005) 2136-2148\n \nHIV vaccine development by computer assisted design: the GAIA vaccine\n \nAnne S. De Groot a,b,?, Luisa Marconc, Elizabeth A. Bishop a, Daniel Rivera\na, Michele Kutzler d, David B. Weiner d, William Martin b\n \na TB/HIV Research Laboratory, Brown University, Providence, RI 02912, USA\n \nb EpiVax Inc., Providence, RI 02903, USA\n \nc University of Padva Medical School, USA\n \nd University of Pennsylvania, Philadelphia, PA, USA\n \nAbstract\n \nThe design of epitope-driven vaccines that address the global variability of\nHIV has been significantly hampered by concerns\nabout conservation of the vaccine epitopes across clades of HIV. We\ndeveloped two computer-driven methods for improving epitope-driven HIV\nvaccines: the Epi-Assembler, which derives representative or "immunogenic\nconsensus sequence" (ICS) epitopes from multiple viral variants, and\nVaccineCAD, which reduces junctional immunogenicity when epitopes are\naligned in a\nstring-of-beads format for insertion in a DNA expression vector. In this\nstudy, we report on 20 ICS HIV-1 peptides. The core 9-mer contained in these\nconsensus peptides was conserved in 105-2250 individual HIV-1 strains.\nNineteen of the 20 ICS epitopes (95%) evaluated in this study were confirmed\nin ELISpot assays using peripheral blood monocytes obtained from 13 healthy\nHIV-1 infected subjects. Twenty-five ICS peptides (all 20 of the peptides\nevaluated in this study and 5 additional ICS epitopes) were then aligned in\na pseudoprotein string using "VaccineCAD", an epitope alignment tool that\neliminates immunogenicity created by the junctions between the epitopes.\nReordering the construct reduced the immunogenicity of the junctions between\nepitopes as measured by EpiMatrix, an epitope mapping algorithm. The\nreordered construct was also a more effective immunogen in vivo when tested\nin HLA-DR transgenic mice. These data confirm the utility of bioinformatics\ntools to design novel vaccines containing "immunogenic consensus sequence"\nTcell epitopes for a globally relevant vaccine against HIV.\n \nKeywords: Epitope; Immunoinformatics; T cell; Major histocompatibility\ncomplex\n \n? Corresponding author. Tel.: +1 401 863 6083; fax: +1 401 863 6087.\n \nE-mail addresses: annied@epivax.com, annied@brown.edu,\nanne degroot@brown.edu (A.S. De Groot).\n \n11. Aids Vaccine Advocacy Coalition\nMrk Ad5 HIV-1 Vaccine Trial Failure\nhttp://www.avac.org/ANRS_mtng_summary.htm\n \n12. Nature Medicine 6, 6 (2000) doi:10.1038/71545\n \n13. OPV As Cause of Polio\nhttp://www.polioeradication.org/vaccines.asp\n \n14. MVA and Heart Problems\nhttp://www.bt.cdc.gov/agent/smallpox/vaccination/cardiacrecentvaccinees.asp\n \n15. Serial Human Passage of Simian Immunodeficiency Virus by Unsterile\nInjections and the Emergence of Epidemic Human Immunodeficiency Virus in\nAfrica\nPreston A. Marx, Phillip G. Alcabes, Ernest Drucker\nPhilosophical Transactions: Biological Sciences, Vol. 356, No. 1410, Origins\nof HIV and the AIDS Epidemic (Jun. 29, 2001), pp. 911-920\n \n16. Vaccine has no impact\nAIDSVAX's failure a blow to treatment\nDavid R. Baker, SF Chronicle Staff Writer\nThursday, November 13, 2003\n \n17. ENGERIX-B®\n[Hepatitis B Vaccine (Recombinant)]\nDESCRIPTION\nENGERIX-B [Hepatitis B Vaccine (Recombinant)] is a noninfectious recombinant\npeptide vaccine\nhttp://us.gsk.com/products/assets/us_engerixb.pdf\n \n18. CDC Head: Problems at Nevada Clinic Could Be 'Tip of Iceberg'"\nAssociated Press , (03.04.2008)\n \n19. Dirty needles lead to hepatitis C outbreak - Medical News From Around\nThe World - Brief Article\nNutrition Health Review, Fall, 2002\nEGYPT -- Up to 20 percent of people living in Egypt have tested positive for\nhepatitis C, a disease that can cause liver failure, and dirty needles are\nthought to be the reason, according to an article in The Lancet (March 11,\n2000).\nhttp://findarticles.com/p/articles/mi_m0876/is_2002_Fall/ai_95147890\n \n20. J Acquir Immune Defic Syndr. 2001 Dec 15;28(5):429-36.\nEvaluation of immune activation in HIV-infected and uninfected African\nindividuals by single-cell analysis of cytokine production.\n \n21. Retrovirology. 2007 Dec 4;4:87.\nCompartmentalization of the gut viral reservoir in HIV-1 infected patients.\n \n22. J Immunol. 2007 Sep 1;179(5):3035-46.\nAcute loss of intestinal CD4+ T cells is not predictive of simian\nimmunodeficiency virus virulence.\n \n23. J Virol. 2007 Jan;81(2):599-612.\nMechanisms of gastrointestinal CD4+ T-cell depletion during acute and early\nhuman immunodeficiency virus type 1 infection.\n \n24. J Virol. 2003 Nov;77(21):11708-17.\nSevere CD4+ T-cell depletion in gut lymphoid tissue during primary human\nimmunodeficiency virus type 1 infection and substantial delay in restoration\nfollowing highly active antiretroviral therapy.\n \n25. J Virol. 2003 Nov;77(21):11708-17.\nAntiviral Therapy During Primary SIV Infection Fails to Prevent Acute CD4+\nT-cell Loss In Gut Mucosa But Enhances Their Rapid Restoration Through\nCentral Memory T-cells\n \n26. J Infect Dis. 2008 Feb 8\nPersistence of HIV in Gut-Associated Lymphoid Tissue despite Long-Term\nAntiretroviral Therapy.\n \n27. J Clin Microbiol. 2008 Feb;46(2):757-8.\nEarly impairment of gut function and gut flora supporting a role for\nalteration of gastrointestinal mucosa in human immunodeficiency virus\npathogenesis.\n \n28. the prn notebook® |volume 12 | www.prn.org\nThe Gastrointestinal Tract in HIV-1 Infection: Questions, Answers, and More\nQuestions!\nSaurabh Mehandru, MD\n \n \n \n29. Journal of Clinical Microbiology, February 2008, p. 757-758, Vol. 46,\nNo. 2\nEarly Impairment of Gut Function and Gut Flora Supporting a Role for\nAlteration of Gastrointestinal Mucosa in Human Immunodeficiency Virus\nPathogenesis\n \n30. Expert Rev Vaccines. 2007 Apr;6(2):203-12.\nClarification of how HIV-1 DNA and protein immunizations may be better used\nto obtain HIV-1-specific mucosal and systemic immunity\n \n \n \n31. Eur J Immunol. 2002 Aug;32(8):2274-81.\nSystemic immune responses induced by mucosal administration of lipopeptides\nwithout adjuvant\n \n32. Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1709-14.\nMucosal immunization with HIV-1 peptide vaccine induces mucosal and systemic\ncytotoxic T lymphocytes and protective immunity in mice against intrarectal\nrecombinant HIV-vaccinia challenge.\n \n33. J Virol. 2004 Jan;78(2):1020-5.\nMucosal and systemic immune responses to a human immunodeficiency virus type\n1 epitope induced upon vaginal infection with a recombinant influenza A\nvirus.\n \n34. Nature Medicine 11, S45 - S53 (2005)\nMucosal immunity and vaccines\n \n35. United States Patent 6726924\nOral liposomal delivery system\nUS Patent Issued on April 27, 2004\nhttp://www.patentstorm.us/patents/6726924-description.html\n \n36. NanoSorb Gelcaps\nPlease note towards the end of this slide presentation Nanocaps, dedydrated\nliposomes encapsulated in gel caps to protect from degradation in the\nstomach and delivery to the gut lumen for release, rehydration and uptake by\nDC of GALT\nhttp://www.biopharmasci.com/downloads/nanosorb.ppt\n \n37. BioZone Laboratories Inc Drug Delivery Platforms Available\nHyperSorbT - Oral delivery in gel caps of liposomes for improved\nbioavailability http://www.biozonelabs.com/html/TechnologyLicensing.htm\n \n38. J Control Release. 2005 Nov 28;108(2-3):484-95. Epub 2005 Sep 19.\nThe role of dioleoylphosphatidylethanolamine (DOPE) in targeted gene\ndelivery with mannosylated cationic liposomes via intravenous route.\n \n39. Gene Ther. 2000 Feb;7(4):292-9.\nMannose receptor-mediated gene transfer into macrophages using novel\nmannosylated cationic liposomes.\n \n40. Biochim Biophys Acta. 2001 Mar 9;1511(1):134-45.\nInvolvement of serum mannan binding proteins and mannose receptors in uptake\nof mannosylated liposomes by macrophages.\n \n41. J Control Release. 2008 Jan 22;125(2):121-30. Epub 2007 Oct 22.\nEfficient targeting to alveolar macrophages by intratracheal administration\nof mannosylated liposomes in rats.\n \n42. J Immunol. 2001 Feb 1;166(3):1885-93.\nThe potent adjuvant activity of archaeosomes correlates to the recruitment\nand activation of macrophages and dendritic cells in vivo.\n \n43. Vaccine. 2001 May 14;19(25-26):3509-17.\nImmunization of mice with lipopeptide antigens encapsulated in novel\nliposomes prepared from the polar lipids of various Archaeobacteria elicits\nrapid and prolonged specific protective immunity against infection with the\nfacultative intracellular pathogen, Listeria monocytogenes.\n \n44. Curr Drug Deliv. 2005 Oct;2(4):407-21.\nArchaeosome immunostimulatory vaccine delivery system.\nPatel GB, Chen W.\n \n45. Infect Immun. 2000 Jan;68(1):54-63. Links\nArchaeosome vaccine adjuvants induce strong humoral, cell-mediated, and\nmemory responses: comparison to conventional liposomes and alum.\n \n46. J Drug Target. 2003;11(8-10):515-24.\nArchaeosomes as self-adjuvanting delivery systems for cancer vaccines.\n \n47. Vaccine. 2007 Dec 12;25(51):8622-36. Epub 2007 Oct 5.\nMucosal and systemic immune responses by intranasal immunization using\narchaeal lipid-adjuvanted vaccines.\n \n48. Yakugaku Zasshi. 2007 Feb;127(2):319-26.\n[Uniqueness of the mucosal immune system for the development of prospective\nmucosal vaccine\n \n49. Med Sci (Paris). 2007 Apr;23(4):371-8.\n[Mucosal immunity and vaccine development]\n[Article in French]\n \n50. Annu Rev Immunol. 2007;25:381-418.\nMucosal dendritic cells.\n \n51. J Allergy Clin Immunol. 2008\n4. Gastrointestinal mucosal immunity\n \n52. Biologicals. 2001 Sep-Dec;29(3-4):183-8.\nChemoselective ligation and antigen vectorization.\n \n53. J Immunol. 2007 Mar 1;178(5):2803-12.\nStaphylococcus aureus protein A triggers T cell-independent B cell\nproliferation by sensitizing B cells for TLR2 ligands\n \n54. Eur J Immunol. 1994 Oct;24(10):2506-14.\nRole of antigen-presenting cells in the polarized development of helper T\ncell subsets: evidence for differential cytokine production by Th0 cells in\nresponse to antigen presentation by B cells and macrophages\n \n55. Eur J Immunol. 2007 Nov;37(11):3040-53.\nTLR-mediated stimulation of APC: Distinct cytokine responses of B cells and\ndendritic cells.\n \n56. J Exp Med. 2008 Jan 21;205(1):169-81. Epub 2008 Jan 7.\nMannose-binding lectin enhances Toll-like receptors 2 and 6 signaling from\nthe phagosome.\n \n57. Nature. 1999 Oct 21;401(6755):811-5.\nComment in:\nNature. 1999 Oct 21;401(6755):755-6.\nThe Toll-like receptor 2 is recruited to macrophage phagosomes and\ndiscriminates between pathogens\n \n58. . Immunobiology Part I. An Introduction to Immunobiology and Innate\nImmunity 2. Innate Immunity--Receptors of the innate immune system.\nhttp://www.ncbi.nlm.nih.gov/books/bv.fcgi?highlight=toll,pathway&rid=imm.section.193#198\n \n \n59. Vaccine. 2008 Feb 20;26(8):1098-110. Epub 2008 Jan 10.\nCross-subtype antibody and cellular immune responses induced by a polyvalent\nDNA prime-protein boost HIV-1 vaccine in healthy human volunteers.\n \n \n \n60. Expert Rev Vaccines. 2008 Mar;7(2):151-3.\n \nHuman versus HIV: round 2 defeat in AIDS vaccine development. Lu S. PMID:\n18324884\n \n \n \n61. Int Immunol. 2006 Jun;18(6):857-69. Epub 2006 Mar 31.\nExpression of human DEC-205 (CD205) multilectin receptor on leukocytes.\n \n \n \n62. Immunobiology Part I. An Introduction to Immunobiology and Innate\nImmunity 2. Innate Immunity\nThe complement system and innate immunity.\n \nhttp://www.ncbi.nlm.nih.gov/books/bv.fcgi?highlight=mannan%20binding%20lectin&rid=imm.section.161#170\n \n63. Adv Drug Deliv Rev. 2000 Mar 30;41(2):171-88.\nDelivery of lipids and liposomal proteins to the cytoplasm and Golgi of\nantigen-presenting cells. mangala.rao@na.amedd.army.mil.\nRao M, Alving CR.\nDepartment of Membrane Biochemistry, Bldg. 40, Walter Reed Army Institute of\nResearch, Washington, DC 20307-5100, USA.\n \nLiposomes have the well-known ability to channel protein and peptide\nantigens into the MHC class II pathway of phagocytic antigen-presenting\ncells (APCs) and thereby enhance the induction of antibodies and\nantigen-specific T cell proliferative responses. Liposomes also serve as an\nefficient delivery system for entry of exogenous protein and peptide\nantigens into the MHC class I pathway and thus are very efficient inducers\nof cytotoxic T cell responses. Soluble antigens that are rendered\nparticulate by encapsulation in liposomes are localized both in vacuoles and\nin the cytoplasm of bone marrow-derived macrophages. Utilizing\nfluorophore-labeled proteins encapsulated in liposomes we have addressed the\nquestion of how liposomal antigens enter the MHC class I pathway. After\nphagocytosis of the liposomes, the fluorescent liposomal protein and\nliposomal lipids enter the cytoplasm where they are processed by the\nproteasome complex. The processed liposomal protein is then transported via\nthe TAP complex into the endoplasmic reticulum and the Golgi complex. Both\nthe liposomal lipids and the liposomal proteins appear to follow the same\nintracellular route and they are processed as a protein-lipid unit. In the\nabsence of a protein antigen (empty liposomes), there is no\norganelle-specific localization of the liposomal lipids. In contrast, when a\nprotein is encapsulated in these liposomes, the distribution of the\nliposomal lipids is dramatically affected and the liposomal lipids localize\nto the trans-Golgi area. Localization of the protein in the trans-Golgi area\nrequires liposomal lipids. Similarly, for the localization of liposomal\nlipids in the trans-Golgi area, there is an obligatory requirement for\nprotein. Therefore, the intracellular trafficking patterns of liposomal\nlipids and liposomal protein are reciprocally regulated. Presence of both\nliposomal lipids and liposomal protein in the trans-Golgi therefore\nfacilitates the entry of liposomal antigens into the MHC class I pathway. It\nis also possible that liposomal lipids are presented to T cells via the\nrecently described CD1 pathway for lipid antigens. Because\nliposome-formulated vaccines have the potential to stimulate antibody as\nwell as cellular immune responses to protein and lipid components, this\napproach could prove to be extremely useful in designing vaccine strategies.\n \nPMID: 10699313 [PubMed - indexed for MEDLINE]\n \n \nRelated Links\n a.. Trafficking of liposomal antigen to the trans-Golgi of murine\nmacrophages requires both liposomal lipid and liposomal protein. [Exp Cell\nRes. 1999]\n b.. Visualization of peptides derived from liposome-encapsulated proteins\nin the trans-Golgi area of macrophages. [Immunol Lett. 1997]\n c.. Liposomes containing lipid A serve as an adjuvant for induction of\nantibody and cytotoxic T-cell responses against RTS,S malaria antigen.\n[Infect Immun. 1998]\n d.. Human dendritic cells and macrophages exhibit different intracellular\nprocessing pathways for soluble and liposome-encapsulated antigens.\n[Immunobiology. 2005]\n e.. Cytotoxic T lymphocytes induced by liposomal antigens: mechanisms of\nimmunological presentation. [AIDS Res Hum Retroviruses. 1994]\n \nAll Related Articles Link:\n \n \n \nhttp://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&DbFrom=pubmed&Cmd=Link&LinkName=pubmed_pubmed&LinkReadableName=Related%20Articles&IdsFromResult=10699313&ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DiscoveryPanel.Pubmed_Discovery_RA\n \n \n \n64. Program Abstr Conf Retrovir Oppor Infect 11th 2004 San Franc Calif. 2004\nFeb 8-11; 11: abstract no. 450.\nCD4+ T-cell Depletion in AIDS: Synergy between Non-infectious HIV-1 and\nOther Viruses Induces Selective Apoptosis via a TRAIL/TRAIL\nReceptor-dependent Mechanism\n \n65. Clin Immunol. 2008 Mar;126(3):235-42. Epub 2007 Oct 3. Chronic innate\nimmune activation as a cause of HIV-1 immunopathogenesis.\n \n66. Clin Immunol. 2007 May;123(2):121-8. Epub 2006 Nov 16. HIV-1\nimmunopathogenesis: how good interferon turns bad.\n \n67. Blood. 2006 Dec 1;108(12):3808-17. Epub 2006 Aug 10. HIV-1-driven\nregulatory T-cell accumulation in lymphoid tissues is associated with\ndisease progression in HIV/AIDS.\n \n68. Blood. 2005 Nov 15;106(10):3524-31. Epub 2005 Jul 26. CD4+ T-cell death\ninduced by infectious and noninfectious HIV-1: role of type 1\ninterferon-dependent, TRAIL/DR5-mediated apoptosis.\n \n69. J Immunol. 2005 Mar 15;174(6):3143-7. The prevalence of regulatory T\ncells in lymphoid tissue is correlated with viral load in HIV-infected\npatients.\n \n70. Eurekah Bioscience Collection Signal Transduction The Function of\nToll-Like Receptors--TLR Ligand Linked Antigen Presentation in Immature DCs\n \nHuman immature DCs derived from bone marrow, pulsed with antagonistic TLR2\nspecific mAbs containing ? light chains, could stimulate a C? specific CD4+\nT cell clone in the absence of maturation effects on iDCs (Fig. 7C). An\nisotype/light-chain matched control antibody produced a two to three orders\nof magnitude lower response, indicating enhanced antigen presentation via\nTLR2. Stimulation was TLR2 specific, as antibodies against other surface\nmolecules such as CD62 and CXCR1 were not stimulatory. Inhibitors of\nlysosomal degradation, processing and MHC class II presentation like\nchloroquine, leupeptin or brefeldin A almost completely abolished T cell\nstimulation. Furthermore, an anti-TLR2 mAb was directly shown to reside in\nendosomal vesicles in pulsed iDCs. 31 Thus, antigen linked to the TLR2\nligand can be endocytosed after binding TLR2, processed via the classical\n(exogenous) pathway of antigen presentation, and can enhance the stimulation\nof T cells. This same route could be exploited to generate more efficacious\nvaccines.\n \nhttp://www.ncbi.nlm.nih.gov/books/bv.fcgi?highlight=tlr,maturation,antibody&rid=eurekah.section.26308\n \n71.J Biol Chem. 2007 Jul 20;282(29):21145-59. Epub 2007 Apr 26. Distinct\nuptake mechanisms but similar intracellular processing of two different\ntoll-like receptor ligand-peptide conjugates in dendritic cells.\n \n72. Blood. 2006 Jul 15;108(2):544-50. Epub 2006 Mar 14. Synergistic\nactivation of dendritic cells by combined Toll-like receptor ligation\ninduces superior CTL responses in vivo.\n \n73. J Immunol. 2004 Apr 15;172(8):4733-43. A Toll-like receptor 2 ligand\nstimulates Th2 responses in vivo, via induction of extracellular\nsignal-regulated kinase mitogen-activated protein kinase and c-Fos in\ndendritic cells.\n \n74. J Virol. 2003 Oct;77(19):10250-9. Yeast-derived human immunodeficiency\nvirus type 1 p55(gag) virus-like particles activate dendritic cells (DCs)\nand induce perforin expression in Gag-specific CD8(+) T cells by\ncross-presentation of DCs.\n \n75. J Immunol. 2003 Jul 1;171(1):32-6.\n \nCutting edge: link between innate and adaptive immunity: Toll-like receptor\n2 internalizes antigen for presentation to CD4+ T cells and could be an\nefficient vaccine target.Schjetne KW, Thompson KM, Nilsen N, Flo TH,\nFleckenstein B, Iversen JG, Espevik T, Bogen B.\n \nInstitute of Immunology, University of Oslo, Rikshospitalet, Oslo, Norway.\nk.w.schjetne@labmed.uio.no\n \nAn ideal vaccine for induction of CD4(+) T cell responses should induce\nlocal inflammation, maturation of APC, and peptide loading of MHC class II\nmolecules. Ligation of Toll-like receptor (TLR) 2 provides the first two of\nthese three criteria. We have studied whether targeting of TLR2 results in\nloading of MHC class II molecules and enhancement of CD4(+) T cell\nresponses. To dissociate MHC class II presentation from APC maturation, we\nhave used an antagonistic, mouse anti-human TLR2 mAb (TL2.1) as ligand and\nmeasured proliferation of a mouse Ckappa-specific human CD4(+) T cell clone.\nTL2.1 mAb was 100-1000 times more efficiently presented by APC compared with\nisotype-matched control mAb. Moreover, TL2.1 mAb was internalized into\nendosomes and processed by the conventional MHC class II pathway. This novel\nfunction of TLR2 represents a link between innate and adaptive immunity and\nindicates that TLR2 could be a promising target for vaccines\n \n76. J Immunol. 2004 Sep 15;173(6):3916-24. Microglia initiate central\nnervous system innate and adaptive immune responses through multiple TLRs\n \n77. Nippon Ishinkin Gakkai Zasshi. 2002;43(3):133-6. Receptor-mediated\nrecognition of Cryptococcus neoformans.\n \n78. Curr Mol Med. 2005 Jun;5(4):413-20. The cellular responses induced by\nthe capsular polysaccharide of Cryptococcus neoformans differ depending on\nthe presence or absence of specific protective antibodies.Vecchiarelli A.\n \n79. J Immunol. 2001 Sep 15;167(6):3316-23. Predominant role of toll-like\nreceptor 2 versus 4 in Chlamydia pneumoniae-induced activation of dendritic\ncells.\n \n80, J Immunol. 2003 Aug 1;171(3):1441-6. Heat-killed Brucella abortus\ninduces TNF and IL-12p40 by distinct MyD88-dependent pathways: TNF, unlike\nIL-12p40 secretion, is Toll-like receptor 2 dependent.\n \n81. Nat Immunol. 2000 Dec;1(6):502-9. OmpA targets dendritic cells, induces\ntheir maturation and delivers antigen into the MHC class I presentation\npathway\n \n82. J Clin Invest. 2005 Nov;115(11):3265-75. Epub 2005 Oct 13. Endocytosis\nof HIV-1 activates plasmacytoid dendritic cells via Toll-like receptor-viral\nRNA interactions.\n \n83. The Journal of Immunology, 2008, 180: 1405-1413. Suppression of Mature\nDendritic Cell Function by Regulatory T Cells In Vivo Is Abrogated by CD40\nLicensing1\n \n84. Mucosal Immunology (2008) 1, 156-168 The contribution of PARs to\ninflammation and immunity to fungi\n \nhttp://www.nature.com/mi/journal/v1/n2/full/mi200713a.html\n \n85. Yakugaku Zasshi. 2007 Feb;127(2):319-26.\n[Uniqueness of the mucosal immune system for the development of prospective\nmucosal vaccine]\n \n86. The Maturing Immune System: Implications for Development and Testing\nHIV-1 Vaccines for Children and Adolescents\n \nhttp://www.medscape.com/viewarticle/524225_print\n \n87. J Immunol. 2004 May 1;172(9):5240-8. Antigen-specific T cell repertoire\nmodification of CD4+CD25+ regulatory T cells.\n \n88. Expert Opin Drug Deliv. 2007 Jul;4(4):323-40. Oral vaccination: where we\nare?\n \n89. Methods. 2006 Feb;38(2):150-7. Mucosal immunization using recombinant\nplant-based oral vaccines.\n \n90. Immunol Cell Biol. 2005 Jun;83(3):257-62. Oral hepatitis B vaccine\ncandidates produced and delivered in plant material.\n \n91. AIDS 1998, 12: 2387-2396 Immune activation in HIV-infected African\nindividuals\n \n92. J. Exp. Med., Volume 188, Number 12, December 21, 1998 2205-2213 Viral\nImmune Evasion Due to Persistence of Activated T Cells Without Effector\nFunction\n \n93. The Journal of Infectious Diseases 1999; 179: 859 -870 Shorter Survival\nin Advanced Human Immunodeficiency Virus Type 1 Infection Is More Closely\nAssociated with T Lymphocyte Activation than with Plasma Virus Burden or\nVirus Chemokine Coreceptor Usage\n \n94. Clin Cancer Res. 2007 Aug 1;13(15 Pt 1):4345-54. A unique subset of\nCD4+CD25highFoxp3+ T cells secreting interleukin-10 and transforming growth\nfactor-beta1 mediates suppression in the tumor microenvironment.\n \n95. Clin Vaccine Immunol. 2007 Sep;14(9):1127-37. Epub 2007 Jul 18.\nDendritic cell function during chronic hepatitis C virus and human\nimmunodeficiency virus type 1 infection.\n \n96.Eur J Immunol. 2007 Jul;37(7):1887-904. Impairment of dendritic cell\nfunction by excretory-secretory products: a potential mechanism for\nnematode-induced immunosuppression.\n \n97. Nature Dec 5;420:502-507. Belkaid Y, Piccirillo CA, Mendez S, Shevach\nEM, Sacks DL. CD4+CD25+regulatory T cells control Leishmania major\npersistence and immunity.\n \n98. Current Opinion in Immunology Published online 2/7/04 Development and\nfunction of CD25+CD4+ regulatory T cells\n \n99. Current Opinion in Immunology Published online 2/11/04 Immunoregulatory\nT cells in tumor immunity\n \n100. Cancer Res. 2003 Aug 1;63(15):4516-20. Human CD4(+) CD25(+) Regulatory\nT Cells Suppress NKT Cell Functions.\n \n101. J Virol. 2002 Aug;76(15):7528-34. Selective loss of innate CD4(+) V\nalpha 24 natural killer T cells in human immunodeficiency virus infection.\n \n+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++\n \nEndnote Regarding Production of GAIA ICS Antigen\n \nGMP Production of GAIA ICS cell-derived Peptide and Peusdoprotien antigen\nfor R & D and clinical trials. Followed up with full scale production in\ntransgenic plants should result in high yield, low cost, high quality and\nstable for many years at room temperatures GAIA ICS Peptide and\nPeusdoprotein antigen to be used in a Global HIV-1 Vaccination Campaign.\n \n \n \nJ Drug Target. 2003;11(8-10):539-45. Advantageous features of plant-based\nsystems for the development of HIV vaccines.\n \nJournal of the American College of Nutrition, Vol. 21, No. 90003, 212S-217S\n(2002) Foods as Production and Delivery Vehicles for Human Vaccines\n \nInfluenza and Other Respiratory Viruses, Volume 1, Number 1, January 2007 ,\npp. 19-25(7) A launch vector for the production of vaccine antigens in\nplants\n \nMethods. 2006 Feb;38(2):150-7. Mucosal immunization using recombinant\nplant-based oral vaccines.\n \nImmunol Cell Biol. 2005 Jun;83(3):257-62. Oral hepatitis B vaccine\ncandidates produced and delivered in plant material.\n \nExpert Opin Drug Deliv. 2005 Jul;2(4):719-28. Delivery of plant-derived\nvaccines.\n \nInt J Parasitol. 2003 May;33(5-6):479-93. Plant-based vaccines.\n \nVaccine Volume 24, Issue 5 , 30 January 2006, Pages 691-695 Oral\nimmunogenicity of a plant-made, subunit, tuberculosis vaccine\n \nJ Biotechnol. 2005 Oct 17;120(1):121-34. Epub 2005 Jul 18. Plants as\nbioreactors: a comparative study suggests that Medicago truncatula is a\npromising production system.\n \nTransgenic Res. 2007 Jun;16(3):315-32. Epub 2007 Apr 14.Production of\nvaccines and therapeutic antibodies for veterinary applications in\ntransgenic plants: an overview.\n \nProc Natl Acad Sci U S A. 2007 Apr 17;104(16):6864-9. Epub 2007 Apr 11.\nSmallpox subunit vaccine produced in Planta confers protection in mice.\n \nAnn N Y Acad Sci. 2007 Apr;1102:121-34. Bioproduction of therapeutic\nproteins in the 21st century and the role of plants and plant cells as\nproduction platforms.\n \nBMC Biotechnol. 2007 Feb 26;7:12. Expression, intracellular targeting and\npurification of HIV Nef variants in tobacco cells.\n \nJ Biotechnol. 2007 Feb 20;128(3):512-8. Epub 2006 Nov 16. Multimerization of\npeptide antigens for production of stable immunogens in transgenic plants.\n \nExpert Rev Vaccines. 2006 Apr;5(2):249-60. Plant-derived vaccines: a look\nback at the highlights and a view to the challenges on the road\nahead.Thanavala Y, Huang Z, Mason HS.\n \nRoswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263,\nUSA. yasmin.thanavala@roswellpark.org\n \nThe sobering reality is that each year, 33 million children remain\nunvaccinated for vaccine-preventable diseases. Universal childhood\nvaccination would have profound effects on leveling the health inequities in\nmany parts of the world. As an alternative to administration of vaccines by\nneedle and syringe, oral vaccines offer significant logistical advantages,\nas the polio eradication campaign has demonstrated. Over the past decade,\nthe expression of subunit vaccine antigens in plants has emerged as a\nconvenient, safe and potentially economical platform technology, with the\npotential to provide a novel biotechnological solution to vaccine production\nand delivery. As this technology has come of age, many improvements have\nbeen made on several fronts, as a growing number of research groups\nworldwide have extensively investigated plants as factories for vaccine\nproduction. This review attempts to highlight some of the achievements over\nthe past 15 years, identify some of the potential problems and discuss the\npromises that this technology could fulfill. PMID: 16608424\n \n================================================================\nThe Hunger Site: A Click a day sends FREE FOOD to fight World Hunger and Malnutrition Diseases\n \nhttp://www.thehungersite.com/clickToGive/home.faces?siteId=1\n================================================================\nThe Hunger Site: A Click a day sends FREE FOOD to fight World Hunger and Malnutrition Diseases\n \nhttp://www.thehungersite.com/clickToGive/home.faces?siteId=1

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