Last data update: May 06, 2024. (Total: 46732 publications since 2009)
Records 1-14 (of 14 Records) |
Query Trace: Johnson BJ[original query] |
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Metabolic differentiation of early Lyme disease from southern tick-associated rash illness (STARI).
Molins CR , Ashton LV , Wormser GP , Andre BG , Hess AM , Delorey MJ , Pilgard MA , Johnson BJ , Webb K , Islam MN , Pegalajar-Jurado A , Molla I , Jewett MW , Belisle JT . Sci Transl Med 2017 9 (403) Lyme disease, the most commonly reported vector-borne disease in the United States, results from infection with Borrelia burgdorferi. Early clinical diagnosis of this disease is largely based on the presence of an erythematous skin lesion for individuals in high-risk regions. This, however, can be confused with other illnesses including southern tick-associated rash illness (STARI), an illness that lacks a defined etiological agent or laboratory diagnostic test, and is coprevalent with Lyme disease in portions of the eastern United States. By applying an unbiased metabolomics approach with sera retrospectively obtained from well-characterized patients, we defined biochemical and diagnostic differences between early Lyme disease and STARI. Specifically, a metabolic biosignature consisting of 261 molecular features (MFs) revealed that altered N-acyl ethanolamine and primary fatty acid amide metabolism discriminated early Lyme disease from STARI. Development of classification models with the 261-MF biosignature and testing against validation samples differentiated early Lyme disease from STARI with an accuracy of 85 to 98%. These findings revealed metabolic dissimilarity between early Lyme disease and STARI, and provide a powerful and new approach to inform patient management by objectively distinguishing early Lyme disease from an illness with nearly identical symptoms. |
Evaluation of alternative killing agents for Aedes aegypti (Diptera: Culicidae) in the Gravid Aedes Trap (GAT)
Heringer L , Johnson BJ , Fikrig K , Oliveira BA , Silva RD , Townsend M , Barrera R , Eiras AE , Ritchie SA . J Med Entomol 2016 53 (4) 873-879 The Gravid Aedes Trap (GAT) uses visual and olfactory cues to attract gravid Aedes aegypti (L.) that are then captured when knocked down by a residual pyrethroid surface spray. However, the use of surface sprays can be compromised by poor availability of the spray and pesticide resistance in the target mosquito. We investigated several "alternative" insecticide and insecticide-free killing agents for use in the GAT. This included long-lasting insecticide-impregnated nets (LLINs), vapor-active synthetic pyrethroids (metofluthrin), canola oil, and two types of dry adhesive sticky card. During bench top assays LLINs, metofluthrin, and dry sticky cards had 24-h knockdown (KD) percentages >80% (91.2 +/- 7.2%, 84.2 +/- 6.8%, and 83.4 +/- 6.1%, respectively), whereas the 24-h KD for canola oil was 70 +/- 7.7%, which improved to 90.0 +/- 3.7% over 48 h. Importantly, there were no significant differences in the number of Ae. aegypti collected per week or the number of traps positive for Ae. aegypti between the sticky card and canola oil treatments compared with the surface spray and LLIN treatments in semifield and field trials. These results demonstrate that the use of inexpensive and widely available insecticide-free agents such as those described in this study are effective alternatives to pyrethroids in regions with insecticide-resistant populations. The use of such environmentally friendly insecticide-free alternatives will also be attractive in areas where there is substantial resistance to insecticide use due to environmental and public health concerns. |
The salt-sensitive structure and zinc inhibition of Borrelia burgdorferi protease BbHtrA
Russell TM , Tang X , Goldstein JM , Bagarozzi D , Johnson BJ . Mol Microbiol 2015 99 (3) 586-96 HtrA serine proteases are highly conserved and essential ATP-independent proteases with chaperone activity. Bacteria express a variable number of HtrA homologs which contribute to the virulence and pathogenicity of bacterial pathogens. Lyme disease spirochetes possess a single HtrA protease homolog, Borrelia burgdorferi HtrA (BbHtrA). Previous studies established that, like the human orthologue HtrA1, BbHtrA is proteolytically active against numerous extracellular proteins in vitro. In this study, we utilized size exclusion chromatography and blue native polyacrylamide gel electrophoresis (BN-PAGE) to demonstrate BbHtrA oligomeric structures which were substrate-independent and salt sensitive. Examination of the influence of transition metals on the activity of BbHtrA revealed that this protease is inhibited by Zn2+ >Cu2+ >Mn2+ . Extending this analysis to two other HtrA proteases, E. coli DegP and HtrA1, revealed that all three HtrA proteases were reversibly inhibited by ZnCl2 at all micro molar concentrations examined. Commercial inhibitors for HtrA proteases are not available and physiologic HtrA inhibitors are unknown. Our observation of conserved zinc inhibition of HtrA proteases will facilitate structural and functional studies of additional members of this important class of proteases. |
Borrelia burgdorferi RevA significantly affects pathogenicity and host response in the mouse model of Lyme disease.
Byram R , Gaultney RA , Floden AM , Hellekson C , Stone BL , Bowman A , Stevenson B , Johnson BJ , Brissette CA . Infect Immun 2015 83 (9) 3675-83 The Lyme disease spirochete, Borrelia burgdorferi, expresses RevA and numerous outer surface lipoproteins during mammalian infection. As an adhesin that promotes bacterial interaction with fibronectin, RevA is poised to interact with the hosts' extracellular matrix. To further define the role(s) of RevA during mammalian infection, we created a mutant that is unable to produce RevA. The mutant was still infectious to mice, although it was significantly less well able to infect cardiac tissues. Complementation of the mutant with a wild-type revA gene restored heart infectivity to wild-type levels. Additionally, revA mutants had increased evidence of arthritis, with increased fibrotic collagen deposition in tibiotarsal joints. The mutants also induced increased levels of the serum chemokine CCL2, a monocyte chemoattractant, and this increase was abolished in the complemented strain. Therefore, while revA is not absolutely essential for infection, deletion of revA had distinct effects on dissemination, arthritis severity, and host response. |
Evaluation of Borrelia burgdorferi BbHtrA protease as a vaccine candidate for Lyme borreliosis in mice
Ullmann AJ , Russell TM , Dolan MC , Williams M , Hojgaard A , Weiner ZP , Johnson BJ . PLoS One 2015 10 (6) e0128868 Borrelia burgdorferi synthesizes an HtrA protease (BbHtrA) which is a surface-exposed, conserved protein within Lyme disease spirochetes with activity toward CheX and BmpD of Borrelia spp, as well as aggrecan, fibronectin and proteoglycans found in skin, joints and neural tissues of vertebrates. An antibody response against BbHtrA is observed in Lyme disease patients and in experimentally infected laboratory mice and rabbits. Given the surface location of BbHtrA on B. burgdorferi and its ability to elicit an antibody response in infected hosts, we explored recombinant BbHtrA as a potential vaccine candidate in a mouse model of tick-transmitted Lyme disease. We immunized mice with two forms of BbHtrA: the proteolytically active native form and BbHtrA ablated of activity by a serine to alanine mutation at amino acid 226 (BbHtrAS226A). Although inoculation with either BbHtrA or BbHtrAS226A produced high-titer antibody responses in C3H/HeJ mice, neither antigen was successful in protecting mice from B. burgdorferi challenge. These results indicate that the search for novel vaccine candidates against Lyme borreliosis remains a challenge. |
Lyme disease testing by large commercial laboratories in the United States
Hinckley AF , Connally NP , Meek JI , Johnson BJ , Kemperman MM , Feldman KA , White JL , Mead PS . Clin Infect Dis 2014 59 (5) 676-81 BACKGROUND: Laboratory testing is helpful when evaluating patients with suspected Lyme disease (LD). A two-tiered antibody testing approach is recommended, but single-tier and non-validated tests are also used. We conducted a survey of large commercial laboratories in the United States to assess laboratory practices. We used these data to estimate the cost of testing and number of infections among patients from whom specimens were submitted. METHODS: Large commercial laboratories were asked to report the type and volume of testing conducted nationwide in 2008, as well as the percent of positive tests for four LD endemic states. The total direct cost of testing was calculated for each test type. These data and test-specific performance parameters available in published literature were used to estimate the number of infections among source patients. RESULTS: Seven participating laboratories performed approximately 3.4 million LD tests on approximately 2.4 million specimens nationwide at an estimated cost of $492 million. Two-tiered testing accounted for at least 62% of assays performed; alternative testing accounted for less than 3% of assays. The estimated frequency of infection among patients from whom specimens were submitted ranged from 10% to 18.5%. Applied to the total numbers of specimens, this yielded an estimated 240,000 to 444,000 infected source patients in 2008. DISCUSSION: LD testing is common and costly, with most testing in accordance with diagnostic recommendations. These results highlight the importance of considering clinical and exposure history when interpreting laboratory results for diagnostic and surveillance purposes. |
Borrelia burgdorferi BbHtrA degrades host ECM proteins and stimulates release of inflammatory cytokines in vitro
Russell TM , Delorey MJ , Johnson BJ . Mol Microbiol 2013 90 (2) 241-51 The Lyme disease spirochaete, Borrelia burgdorferi, causes damage to diverse host tissues and induces inflammation but the mechanisms of injury are poorly understood. We recently reported that a surface-exposed B. burgdorferi protease, which is expressed during human disease and is conserved within the major Lyme disease spirochaete species, degrades the extracellular matrix proteoglycan, aggrecan. Here we demonstrate that BbHtrA also degrades fibronectin and numerous proteoglycans found in skin, joints and neural tissues. BbHtrA degradation of fibronectin released known pro-inflammatory fibronectin fragments FnIII13-14 and Fnf-29, which may amplify the inflammatory processes triggered by the presence of the bacteria. When this hypothesis was tested directly by exposing chondrocytes to BbHtrA in vitro, inflammatory cytokines (sICAM-1 and IL-6) and chemokines (CXCL1, CCL1, CCL2 and CCL5) that are hallmarks of Lyme disease were induced. These results provide the first evidence that, by utilizing BbHtrA, B. burgdorferi may actively participate in its dissemination and in the tissue damage and inflammation observed in Lyme disease. |
Assessment of new culture method for detection of Borrelia species from serum of lyme disease patients.
Johnson BJ , Pilgard MA , Russell TM . J Clin Microbiol 2013 52 (3) 721-4 A novel method of culturing spirochetes from the serum of U.S. Lyme disease patients was recently reported by Sapi and colleagues to have 94% sensitivity and 100% specificity for Borrelia species as assessed by microscopy and DNA sequence analysis of the pyrG gene (Sapi E, Pabbati N, Datar A, Davies EM, Rattelle A, Kuo BA. 2013. Int J Med Sci 10:362-376). The majority of the spirochetes described were related by pyrG sequences to species of Borrelia previously undetected in North American patients without a reported travel history to Europe or Asia. To better understand these unexpected findings, we determined pyrG sequences of the laboratory reference strains used by the investigators for method development and testing of culture medium. Eighty percent (41/51) of the reported patient-derived pyrG sequences are identical to one of the laboratory strains and an additional 12% (6/51) differ by only a single nucleotide across a 603bp region of the pyrG gene. Thus, false positivity due to laboratory contamination of patient samples cannot be ruled out and further validation of the proposed novel culture method is required. |
The genome sequence of Lone Star virus, a highly divergent bunyavirus found in the Amblyomma americanum tick.
Swei A , Russell BJ , Naccache SN , Kabre B , Veeraraghavan N , Pilgard MA , Johnson BJ , Chiu CY . PLoS One 2013 8 (4) e62083 Viruses in the family Bunyaviridae infect a wide range of plant, insect, and animal hosts. Tick-borne bunyaviruses in the Phlebovirus genus, including Severe Fever with Thrombocytopenia Syndrome virus (SFTSV) in China, Heartland virus (HRTV) in the United States, and Bhanja virus in Eurasia and Africa have been associated with acute febrile illness in humans. Here we sought to characterize the growth characteristics and genome of Lone Star virus (LSV), an unclassified bunyavirus originally isolated from the lone star tick Amblyomma americanum. LSV was able to infect both human (HeLa) and monkey (Vero) cells. Cytopathic effects were seen within 72 h in both cell lines; vacuolization was observed in infected Vero, but not HeLa, cells. Viral culture supernatants were examined by unbiased deep sequencing and analysis using an in-house developed rapid computational pipeline for viral discovery, which definitively identified LSV as a phlebovirus. De novo assembly of the full genome revealed that LSV is highly divergent, sharing <61% overall amino acid identity with any other bunyavirus. Despite this sequence diversity, LSV was found by phylogenetic analysis to be part of a well-supported clade that includes members of the Bhanja group viruses, which are most closely related to SFSTV/HRTV. The genome sequencing of LSV is a critical first step in developing diagnostic tools to determine the risk of arbovirus transmission by A. americanum, a tick of growing importance given its expanding geographic range and competence as a disease vector. This study also underscores the power of deep sequencing analysis in rapidly identifying and sequencing the genomes of viruses of potential clinical and public health significance. |
Lyme disease spirochetes possess an aggrecan-binding protease with aggrecanase activity
Russell TM , Johnson BJ . Mol Microbiol 2013 90 (2) 228-40 Connective tissues are the most common area of colonization for the Lyme disease spirochete Borrelia burgdorferi. Colonization is aided by the interaction between numerous bacterial adhesins with components of the extracellular matrix (ECM). Here we describe a novel interaction between B. burgdorferi and the major ECM proteoglycan found in joints, aggrecan. Using affinity chromatography and mass spectrometry we identify two borrelial aggrecan-binding proteins: the known ECM ligand Bgp (BB0588) and an uncharacterized protease BbHtrA (BB0104). Proteinase K studies demonstrate that BbHtrA is surface exposed. Immunoblots using sera from patients with both early and late Lyme disease establish that BbHtrA is expressed during human disease, immunogenic, and conserved in the three major Lyme disease spirochete species. Consequences of the interaction between aggrecan and BbHtrA were examined by proteolysis assays. BbHtrA cleaves aggrecan at a site known to destroy aggrecan function and which has been previously observed in the synovial fluid of patients with Lyme arthritis. These data demonstrate that B. burgdorferi possess aggrecan-binding proteins which may provide the organism with additional capability to colonize connective tissues. Moreover, our studies provide the first evidence that B. burgdorferi possess proteolytic activity which may contribute to the pathogenesis of Lyme arthritis. |
Single-tier testing with the C6 peptide ELISA kit compared with two-tier testing for Lyme disease
Wormser GP , Schriefer M , Aguero-Rosenfeld ME , Levin A , Steere AC , Nadelman RB , Nowakowski J , Marques A , Johnson BJ , Dumler JS . Diagn Microbiol Infect Dis 2013 75 (1) 9-15 For the diagnosis of Lyme disease, the 2-tier serologic testing protocol for Lyme disease has a number of shortcomings including low sensitivity in early disease; increased cost, time, and labor; and subjectivity in the interpretation of immunoblots. In this study, the diagnostic accuracy of a single-tier commercial C6 ELISA kit was compared with 2-tier testing. The results showed that the C6 ELISA was significantly more sensitive than 2-tier testing with sensitivities of 66.5% (95% confidence interval [CI] 61.7-71.1) and 35.2% (95% CI 30.6-40.1), respectively (P < 0.001) in 403 sera from patients with erythema migrans. The C6 ELISA had sensitivity statistically comparable to 2-tier testing in sera from Lyme disease patients with early neurologic manifestations (88.6% versus 77.3%, P = 0.13) or arthritis (98.3% versus 95.6%, P = 0.38). The specificities of C6 ELISA and 2-tier testing in over 2200 blood donors, patients with other conditions, and Lyme disease vaccine recipients were found to be 98.9% and 99.5%, respectively (P < 0.05, 95% CI surrounding the 0.6 percentage point difference of 0.04 to 1.15). In conclusion, using a reference standard of 2-tier testing, the C6 ELISA as a single-step serodiagnostic test provided increased sensitivity in early Lyme disease with comparable sensitivity in later manifestations of Lyme disease. The C6 ELISA had slightly decreased specificity. Future studies should evaluate the performance of the C6 ELISA compared with 2-tier testing in routine clinical practice. |
Multiplex immunoassay for Lyme disease using VlsE1-IgG and pepC10-IgM antibodies: improving test performance through bioinformatics
Porwancher RB , Hagerty CG , Fan J , Landsberg L , Johnson BJ , Kopnitsky M , Steere AC , Kulas K , Wong SJ . Clin Vaccine Immunol 2011 18 (5) 851-9 The Centers for Disease Control and Prevention currently recommends a 2-tier serologic approach to Lyme disease laboratory diagnosis, comprised of an initial serum enzyme immunoassay (EIA) for antibody to Borrelia burgdorferi followed by supplementary IgG and IgM Western blotting of EIA-positive or -equivocal samples. Western blot accuracy is limited by subjective interpretation of weak-positive bands, false-positive IgM immunoblots, and low sensitivity for early disease. We developed an objective alternative second-tier immunoassay using a multiplex microsphere system that measures VlsE1-IgG and pepC10-IgM antibodies simultaneously from the same sample. Our study population was comprised of 79 patients with early acute Lyme disease, 82 with early convalescent, 47 with stages II or III disease, 34 post-antibiotic treatment patients, and 794 controls. A bioinformatic technique called partial receiver-operating characteristic (ROC) regression was used to combine individual antibody levels into a single diagnostic score with a single cutoff; this technique enhances test performance when high specificity is required (e.g. ≥95%). Compared to the Western blot, the multiplex assay was equally specific (95.6%), but 20.7% more sensitive for early convalescent disease (89.0% versus 68.3%, respectively; 95% CI of difference: 12.1% to 30.9%) and 12.5% more sensitive overall (75.0% versus 62.5%, respectively; 95% CI of difference: 8.1% to 17.1%). When used as a second-tier test, a multiplex assay for VlsE1-IgG and pepC10-IgM antibodies performed as well as or better than the Western blot for Lyme disease diagnosis. Prospective validation studies appear to be warranted. |
Practice-based research network partnership with CDC to acquire clinical specimens to study the etiology of southern tick-associated rash illness (STARI)
Vaughn MF , Sloane PD , Knierim K , Varkey D , Pilgard MA , Johnson BJ . J Am Board Fam Med 2010 23 (6) 720-7 INTRODUCTION: Erythema migrans (EM) is an annular, erythematous, expanding rash that is characteristic of early Lyme disease. In the southern United States, however, many cases of EM seem to have an etiology different from that of Lyme disease. This little-understood condition is called Southern tick-associated rash illness. METHODS: With the goal of obtaining biological specimens and clinical histories from 12 to 20 STARI patients for use in etiologic research, microbiologists from the Centers for Disease Control and Prevention contacted the North Carolina Network Consortium, a statewide consortium of practice-based research networks. This article describes the methods by which the North Carolina Network Consortium successfully identified and enrolled Southern tick-associated rash illness patients into a primary care-based research protocol. RESULTS: A total of 23 patients were enrolled, with 100% attainment of the desired specimens. After an initial lack of success, the revised protocol identified and trained physicians practicing in endemic areas for the illness, used a coordinator with 24-hour availability, recruited participants using newspaper notices and medical providers, and provided regular reminders and progress updates. CONCLUSIONS: A practice-based research network can help basic scientists identify patients and collect specimens for clinically relevant research. |
2-Tiered antibody testing for early and late lyme disease using only an immunoglobulin G blot with the addition of a VlsE Band as the second-Tier test
Branda JA , Aguero-Rosenfeld ME , Ferraro MJ , Johnson BJ , Wormser GP , Steere AC . Clin Infect Dis 2009 50 (1) 20-6 BACKGROUND: Standard 2-tiered immunoglobulin G (IgG) testing has performed well in late Lyme disease (LD), but IgM testing early in the illness has been problematic. IgG VlsE antibody testing, by itself, improves early sensitivity, but may lower specificity. We studied whether elements of the 2 approaches could be combined to produce a second-tier IgG blot that performs well throughout the infection. METHODS: Separate serum sets from LD patients and control subjects were tested independently at 2 medical centers using whole-cell enzyme immunoassays and IgM and IgG immunoblots, with recombinant VlsE added to the IgG blots. The results from both centers were combined, and a new second-tier IgG algorithm was developed. RESULTS: With standard 2-tiered IgM and IgG testing, 31% of patients with active erythema migrans (stage 1), 63% of those with acute neuroborreliosis or carditis (stage 2), and 100% of those with arthritis or late neurologic involvement (stage 3) had positive results. Using new IgG criteria, in which only the VlsE band was scored as a second-tier test among patients with early LD (stage 1 or 2) and 5 of 11 IgG bands were required in those with stage 3 LD, 34% of patients with stage 1, 96% of those with stage 2, and 100% of those with stage 3 infection had positive responses. Both new and standard testing achieved 100% specificity. CONCLUSIONS: Compared with standard IgM and IgG testing, the new IgG algorithm (with VlsE band) eliminates the need for IgM testing; it provides comparable or better sensitivity, and it maintains high specificity. |
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