Richer L.M.,University of Tennessee Health Science Center |
Richer L.M.,Biopeptides, Inc. |
Richer L.M.,St Jude Childrens Research Hospital |
Brisson D.,University of Pennsylvania |
And 5 more authors.
Journal of Infectious Diseases | Year: 2014
A high prevalence of infection with Borrelia burgdorferi in ixodid ticks is correlated with a high incidence of Lyme disease. The transmission of B. burgdorferi to humans can be disrupted by targeting 2 key elements in its enzootic cycle: the reservoir host and the tick vector. In a prospective 5-year field trial, we show that oral vaccination of wild white-footed mice resulted in outer surface protein A-specific seropositivity that led to reductions of 23% and 76% in the nymphal infection prevalence in a cumulative, time-dependent manner (2 and 5 years, respectively), whereas the proportion of infected ticks recovered from control plots varied randomly over time. Significant decreases in tick infection prevalence were observed within 3 years of vaccine deployment. Implementation of such a long-term public health measure could substantially reduce the risk of human exposure to Lyme disease. © 2014 The Author 2014.
del Rio B.,UTHSC |
del Rio B.,Biopeptides, Inc. |
Fuente J.L.,UTHSC |
Fuente J.L.,Biopeptides, Inc. |
And 5 more authors.
Vaccine | Year: 2010
Consumed for centuries, lactic acid bacteria are excellent candidates for the development of safe mucosal delivery vehicles for prophylactic and therapeutic molecules. We have recently reported that the immune response to an effective OspA-expressing L. plantarum vaccine for Lyme disease is modulated by the lipid modification of the antigen. In this study, we investigated if this technology can be applied to developing vaccines for other diseases by focusing on the Class A select agent, Yersinia pestis. We used a number of biochemistry and immunology techniques to determine the localization of the immunogen in our delivery vehicle and to evaluate the mucosal as well as the systemic immune response to the immunogen. We found that only LcrV cloned downstream of the signal sequence of B. burgdorferi OspA (ssLcrV), but not wildtype LcrV (LcrV), is localized to the desired peptidoglycan layer of the delivery vehicle. In addition, only mice that received L. plantarum expressing ssLcrV produced significant titers of IgG antibody as well as IgA in distant mucosal sites such as lungs and vagina. Furthermore, only L. plantarum expressing ssLcrV induced significant amounts of pro-inflammatory cytokines TNFα, IL-12, IFNγ and IL-6 as well as anti-inflammatory IL-10 in human peripheral blood mononuclear cells derived dendritic cells, suggesting that the mechanism by which LcrV-expressing L. plantarum stimulates the immune response involves polarization to Th1 mediated immunity with some involvement of Th2. The study reported here proves that this system is a platform technology to develop oral vaccines for multiple diseases. © 2010 Elsevier Ltd.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 300.00K | Year: 2012
DESCRIPTION (provided by applicant): Lyme disease is the most common vector borne infectious disease in North America and Europe. It is a progressive disease with a wide array of clinical manifestations. Treatment of early infection is highly effective, and early diagnosis and treatment are critical to prevent disease progression. By contrast, disseminated late- phase infection is associated with debilitating, sometimes, permanent damage to the nervous and musculoskeletal systems and can be refractory to antibiotics. Currently the laboratory diagnosis of Lyme disease is based on the detection of antibodies against Borrelia burgdorferi in a two-tier serological assay. However, serological assays using native or recombinant proteins from B. burgdorferi as antigens are often insensitive for the detection of antibody present at the time many patients with early Lyme disease usually seek initial medical care and/or lack sufficient specificity as both native and recombinant B. burgdorferi protein antigens contain Bcell epitopes that cross react with antibodies against other bacterial species. As a result, it has been estimated that current IgM or IgG Lyme disease assays fail to diagnose early disease in patients ~50% of the time. The failure of serological assaysto consistently identify B. burgdorferi infection in patients with suspected Lyme disease necessitates the development of a new class of diagnostic tests. The QuantiFERON(R) assay is a cellular-based IFNg release assay for the detection of antigen-specific T cells in the blood of patients infected with Mycobacterium tuberculosis. This assay, which detects the production of the proinflammatory cytokine IFN? in response to stimulation with peptide antigens derived from M. tuberculosis, has proven to be highly sensitive and highly specific in the diagnosis of tuberculosis, supplanting ineffective serological assays and improving upon diagnosis using the tuberculin skin test. A cellular based assay would represent a new direction in the development of diagnostic assays for Lyme disease, with the potential for being significantly more specific and sensitive than current serological assays. However, the development of such an assay requires the identification of highly-specific peptide epitopes unique to B. burgdorferi. The goal of the current study is to identify unique T cell-epitopes derived from B. burgdorferi proteins for use in a QuantiFERON(R)-based cellular assay for the diagnosis of Lyme disease. PUBLIC HEALTH RELEVANCE: Lyme disease is a clinically progressive disease that can result in permanent debilitating neurological and musculoskeletal damage if the infection is allowed to persist and become disseminated. Early treatment is effective and is critical for the prevention of disseminated disease; however, the currently available diagnostic serologic tests lack sufficient specificity and sensitivity during early disease, and fail to correctly diagnose Lye disease in patients as often as 50% of the time. This application takes an entirely new approach to the design of Lyme disease diagnostics, by focusing on the detection of disease specific T cells in infected individuals using unique peptide antigens to overcome the problems of sensitivity and specificity that plague protein-based serological assays. The result will be the development of an effective diagnostic assay for Lyme disease that will improve disease outcomes in patients through early detection, allowing treatment prior to dissemination.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 600.00K | Year: 2011
DESCRIPTION (provided by applicant): Caused by Borrelia burgdorferi, Lyme Disease (LD) can affect the skin, heart, joints or nervous system leading to an array of symptoms that overlap with those of other illnesses. LD is the most common vector-borne infectious disease in the United States and remains a significant public health concern. Early diagnosis and treatment is key to improving disease outcome. The laboratory diagnosis of LD depends on the demonstration of antibodies against Borrelia burgdorferi. There are few rapid point-of-care assays in the market based on lab-on-a-chip technology and none has been developed for LD. Further, current assays are poorly sensitive especially in early disease and cannot discriminate between the early and late stages of LD. New assays are needed and we plan to fill this unmet need. In this Phase I proposal our main objective is to use ultrasensitive microfluidics technology to develop and assess the performance of a rapid lab-on-a-chip, multi-antigenic, stage-specific,and quantitative Point-Of-Care (POC) assay for the serodiagnosis of Lyme disease (LD). After establishing the proof-of-concept of a lab-on-a-chip prototype first-tier Lyme disease test, we plan to move it towards commercialization in Phase II in partnership with Claros Diagnostics. This company specializes in transitioning technology from the laboratory to the physician's office or bedside and has recently received regulatory approval for a point-of-care system in Europe. A rapid, stage-specific assay for the serodiagnosis of Lyme disease is a much sought after device that will advance diagnostic testing beyond the current state of the art. PUBLIC HEALTH RELEVANCE: Lyme disease, the most common vector-borne infectious disease in the United States affects multiple organ systems. Although prompt diagnosis and treatment prevents or limits serious injury to the systems affected, current sero-diagnostics for Lyme disease lack sensitivity in early disease. Furthermore, there is no rapid point-of-care assay in the market based on lab-on-a-chip technology for Lyme disease. Our objective is to develop a novel front-line diagnostic testing device that enables simultaneous detection of multiple Borrelia burgdorferi antigens at the point of care. It is a quantitative rapid assay for serodiagnosis of Lyme disease that has the additional benefit of distinguishing between the stages of the disease. This low-cost portable device requires just one small finger-pricked volume of blood, and performs diagnostic testing thatis as accurate as laboratory testing.
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 151.78K | Year: 2016
DESCRIPTION provided by applicant Lyme disease caused by infection with Borrelia burgdorferi is the most common vector borne infectious disease in North America and Europe Early disease can be effectively cured with antibiotics however untreated late disseminated infection can result in permanent damage to the nervous and musculoskeletal systems Therefore early diagnosis is critical for ensuring good patient outcomes The laboratory diagnosis of Lyme disease is dependent upon the serological detection of antibodies against B burgdorferi However the sensitivity of current IgM and IgG Lyme disease serodiagnostic assays seldom exceeds for the detection of early disease More effective diagnostic assays are needed Serodiagnostics utilizing synthetic peptides have demonstrated significant improvements in sensitivity and specificity for the detection of early Lyme disease The focus of this application is the development of a rapid point of care POC device that utilizes an assay target consisting of a mixture of synthetic peptides that are highly specific for proteins expressed by B burgdorferi during early infection A multi peptide POC assay would offer the benefits of high specificity from the use of peptides unique to B burgdorferi elevate sensitivity from the use of peptides from multiple Bb antigens and rapid diagnosis A POC assay would also improve upon existing conventional assays by reducing the time of accurate diagnosis to a few minutes rather than several days This would improve patient outcome by reducing the likelihood of developing potentially debilitating late stage disease through early antibiotic intervention PUBLIC HEALTH RELEVANCE The CDC estimates that there are Lyme disease cases per year in the United States http www cdc gov lyme stats humanCases html Lyme disease is clinically progressive and if left untreated can result in debilitating permanent damage to the nervous and musculoskeletal systems Early diagnosis and treatment is critical to avoiding disease progression however current diagnostic assays are often unable to detect early disease The focus of this application is the development of a sensitive and specific point of care assay that will provide immediate serological detection of antibodies against Borrelia burgdorferi the causative agent of Lyme disease to aid in physician diagnosis Accurate rapid diagnosis will improve patient outcomes by reducing the likelihood of developing potentially debilitating late stage disease through early antibiotic intervention
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 176.28K | Year: 2016
DESCRIPTION provided by applicant We propose to develop a more sensitive specific multiplex serologic assay for the diagnosis of Lyme disease LD LD is the most common vector borne infectious disease in the United States and as such it is a significant public health concern The disease affects multiple organ systems including musculoskeletal skin and nervous system and is included in the differential diagnosis of multiple diseases In the absence of Erythema migrans the classic skin lesion of early LD the diagnosis is established by the detection of an antibody response to Borrelia burgdorferi Bb in patients with objective findings suggestive of the disease Prompt diagnosis is important because early treatment of LD limits or prevents serious damage to the systems affected Current serodiagnostics lack sensitivity in early disease We laid the ground work for a new generation of seroassays by mapping and defining the specific linear B cell epitopes of key Bb antigens expressed in early infection We identified specific epitopes from of antigens that are suited for use in multi peptide based assays In collaboration with Bio Rad Laboratories we will develop a highly sensitive and specific Luminex r LD serodiagnostic utilizing multiple peptides containing specific linear epitopes key Bb antigens Luminex r X Map is becoming a standard technology in most large clinical diagnostic labs Bio Radandapos s BioPlex is currently used at over locations in the US including commercial clinical labs such as Quest Laboratories and LabCorp large medical groups such as the Mayo and Cleveland Clinics and many University based medical centers Our novel and highly innovative approach will fill the void in LD diagnostics especially in early LD and will provide superior specificity and sensitivity compared to current assays in all phases of LD In addition our collaboration with Bio Rad Laboratories offers a clear cost effective path to the commercialization of this much needed technology PUBLIC HEALTH RELEVANCE Lyme disease the most common vector borne infectious disease in the United States has a wide array of clinical manifestations that are similar to those of other rheumatologic and neurologic illnesses Current laboratory tests are based on twentieth century technology Despite a great deal of effort to improve Lyme disease laboratory tests over the past decades there have been only incremental improvements Laboratory tests measure antibodies produced against spirochetal proteins These proteins can also bind antibodies produced against other bacteria Tests using whole proteins to detect antibodies are not specific We determined the places where antibodies bind to on the different proteins of the Lyme disease spirochete and identified sites that are not similar to those of other bacteria Using this knowledge we are constructing a new test that will limit the problem of nonspecific antibodies This test will be better able to detect antibody produced against the Lyme disease spirochete and less likely to detect antibodies produced against other bacteria
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase II | Award Amount: 605.37K | Year: 2016
Project Summary Lyme disease is a progressive disease that in later stages can lead to debilitating permanent damage to the nervous and musculoskeletal systems Early treatment with an appropriate course of antibiotics is highly effective at clearing infection and preventing disease progression Therefore early diagnosis is critical for ensuring patient outcome However most current IgM and IgG serological assays are not sensitive enough to detect disease at the time most patients seek medical attention More effective alternatives are necessary A T cell based cytokine release assay offers a unique and innovative alternative to serological assays Unlike antibody responses which may remain elevated for years after pathogen clearance the T cell response rapidly wanes with the cessation of effector function and a collapse in T cell numbers Thus monitoring the T cell activity may provide a more sensitive measure of active disease and pathogen clearance than serum antibody responses In phase I we identified unique peptides derived from several antigens expressed by Borrelia burgdorferi during the course of human disease Using a mixture of peptides from four of these antigens we generated a prototype assay that induced detectable secretion of IFN following overnight stimulation of whole blood obtained from patients with well characterized early Lyme disease and observed that IFN secretion was significantly decreased in the same patients days after antibiotic therapy indicating successful clearance of the bacteria These data demonstrate concrete proof of principle that a cytokine release assay for the diagnosis of early Lyme disease can be created and can be successful In this phase II application we will optimize this assay by selecting additional peptide epitopes that stimulate release of IFN from T cells activated in response to infection with Borrelia spp confirming specificity by evaluating samples from patients with potential cross reactive illnesses and evaluating the performance of a QuantiFERON like test before and after treatment of all stages of Lyme disease This will result in the creation of a finalized cytokine release assay for Lyme disease for FDA submission Project Narrative Lyme disease is a progressive disease that can result in debilitating permanent damage to the nervous and musculoskeletal systems if appropriate treatment is not administered at the early stage Unfortunately current laboratory tests for Lyme disease are not very sensitive during early disease and cannot determine if antibiotic treatment was effective We are developing an innovative diagnostic assay that measures the response of T lymphocytes specific for the bacteria that causes Lyme disease Borrelia burgdorferi rather than measuring antibody levels This test is both highly sensitive and highly specific for the detection of disease and can also distinguish between individuals that currently have Lyme disease vs those that had it previously and can determine if treatment was successful
Biopeptides, Inc. | Date: 2015-08-06
The present invention relates to vaccines for control of Borrelia infections in animal and human populations. In particular, the present invention provides compositions and methods comprising recombinant bacteria engineered to express one or more Borrelia burgdorferi antigens for use as Lyme disease vaccines. In some embodiments, the recombinant bacteria are freeze-dried.