Identification of Circulating Bacterial Antigens by <italic toggle="yes">In Vivo</italic> Microbial Antigen Discovery
ABSTRACT Detection of microbial antigens in clinical samples can lead to rapid diagnosis of an infection and administration of appropriate therapeutics. A major barrier in diagnostics development is determining which of the potentially hundreds or thousands of antigens produced by a microbe are actu...
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American Society for Microbiology
2011
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oai:doaj.org-article:508db936fb364d0fb580707b7420d3a52021-11-15T15:38:44ZIdentification of Circulating Bacterial Antigens by <italic toggle="yes">In Vivo</italic> Microbial Antigen Discovery10.1128/mBio.00136-112150-7511https://doaj.org/article/508db936fb364d0fb580707b7420d3a52011-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00136-11https://doaj.org/toc/2150-7511ABSTRACT Detection of microbial antigens in clinical samples can lead to rapid diagnosis of an infection and administration of appropriate therapeutics. A major barrier in diagnostics development is determining which of the potentially hundreds or thousands of antigens produced by a microbe are actually present in patient samples in detectable amounts against a background of innumerable host proteins. In this report, we describe a strategy, termed in vivo microbial antigen discovery (InMAD), that we used to identify circulating bacterial antigens. This technique starts with “InMAD serum,” which is filtered serum that has been harvested from BALB/c mice infected with a bacterial pathogen. The InMAD serum, which is free of whole bacterial cells, is used to immunize syngeneic BALB/c mice. The resulting “InMAD immune serum” contains antibodies specific for the soluble microbial antigens present in sera from the infected mice. The InMAD immune serum is then used to probe blots of bacterial lysates or bacterial proteome arrays. Bacterial antigens that are reactive with the InMAD immune serum are precisely the antigens to target in an antigen immunoassay. By employing InMAD, we identified multiple circulating antigens that are secreted or shed during infection using Burkholderia pseudomallei and Francisella tularensis as model organisms. Potential diagnostic targets identified by the InMAD approach included bacterial proteins, capsular polysaccharide, and lipopolysaccharide. The InMAD technique makes no assumptions other than immunogenicity and has the potential to be a broad discovery platform to identify diagnostic targets from microbial pathogens. IMPORTANCE Effective treatment of microbial infection is critically dependent on early diagnosis and identification of the etiological agent. One means for rapid diagnosis is immunoassay for antigens that are shed into body fluids during infection. Immunoassays can be inexpensive, rapid, and adaptable to a point-of-care format. A major impediment to immunoassay for diagnosis of infectious disease is identification of appropriate antigen targets. This report describes a strategy that can be used for identification of microbial antigens that are shed into serum during infection by the biothreats Burkholderia pseudomallei and Francisella tularensis. Termed InMAD (in vivo microbial antigen discovery), the strategy has the potential for application to a broad spectrum of microbial pathogens.Dana E. NutiReva B. CrumpFarida Dwi HandayaniNarisara ChantratitaSharon J. PeacockRichard BowenPhilip L. FelgnerD. Huw DaviesTerry WuC. Rick LyonsPaul J. BrettMary N. BurtnickThomas R. KozelDavid P. AuCoinAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 2, Iss 4 (2011) |
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Microbiology QR1-502 |
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Microbiology QR1-502 Dana E. Nuti Reva B. Crump Farida Dwi Handayani Narisara Chantratita Sharon J. Peacock Richard Bowen Philip L. Felgner D. Huw Davies Terry Wu C. Rick Lyons Paul J. Brett Mary N. Burtnick Thomas R. Kozel David P. AuCoin Identification of Circulating Bacterial Antigens by <italic toggle="yes">In Vivo</italic> Microbial Antigen Discovery |
| description |
ABSTRACT Detection of microbial antigens in clinical samples can lead to rapid diagnosis of an infection and administration of appropriate therapeutics. A major barrier in diagnostics development is determining which of the potentially hundreds or thousands of antigens produced by a microbe are actually present in patient samples in detectable amounts against a background of innumerable host proteins. In this report, we describe a strategy, termed in vivo microbial antigen discovery (InMAD), that we used to identify circulating bacterial antigens. This technique starts with “InMAD serum,” which is filtered serum that has been harvested from BALB/c mice infected with a bacterial pathogen. The InMAD serum, which is free of whole bacterial cells, is used to immunize syngeneic BALB/c mice. The resulting “InMAD immune serum” contains antibodies specific for the soluble microbial antigens present in sera from the infected mice. The InMAD immune serum is then used to probe blots of bacterial lysates or bacterial proteome arrays. Bacterial antigens that are reactive with the InMAD immune serum are precisely the antigens to target in an antigen immunoassay. By employing InMAD, we identified multiple circulating antigens that are secreted or shed during infection using Burkholderia pseudomallei and Francisella tularensis as model organisms. Potential diagnostic targets identified by the InMAD approach included bacterial proteins, capsular polysaccharide, and lipopolysaccharide. The InMAD technique makes no assumptions other than immunogenicity and has the potential to be a broad discovery platform to identify diagnostic targets from microbial pathogens. IMPORTANCE Effective treatment of microbial infection is critically dependent on early diagnosis and identification of the etiological agent. One means for rapid diagnosis is immunoassay for antigens that are shed into body fluids during infection. Immunoassays can be inexpensive, rapid, and adaptable to a point-of-care format. A major impediment to immunoassay for diagnosis of infectious disease is identification of appropriate antigen targets. This report describes a strategy that can be used for identification of microbial antigens that are shed into serum during infection by the biothreats Burkholderia pseudomallei and Francisella tularensis. Termed InMAD (in vivo microbial antigen discovery), the strategy has the potential for application to a broad spectrum of microbial pathogens. |
| format |
article |
| author |
Dana E. Nuti Reva B. Crump Farida Dwi Handayani Narisara Chantratita Sharon J. Peacock Richard Bowen Philip L. Felgner D. Huw Davies Terry Wu C. Rick Lyons Paul J. Brett Mary N. Burtnick Thomas R. Kozel David P. AuCoin |
| author_facet |
Dana E. Nuti Reva B. Crump Farida Dwi Handayani Narisara Chantratita Sharon J. Peacock Richard Bowen Philip L. Felgner D. Huw Davies Terry Wu C. Rick Lyons Paul J. Brett Mary N. Burtnick Thomas R. Kozel David P. AuCoin |
| author_sort |
Dana E. Nuti |
| title |
Identification of Circulating Bacterial Antigens by <italic toggle="yes">In Vivo</italic> Microbial Antigen Discovery |
| title_short |
Identification of Circulating Bacterial Antigens by <italic toggle="yes">In Vivo</italic> Microbial Antigen Discovery |
| title_full |
Identification of Circulating Bacterial Antigens by <italic toggle="yes">In Vivo</italic> Microbial Antigen Discovery |
| title_fullStr |
Identification of Circulating Bacterial Antigens by <italic toggle="yes">In Vivo</italic> Microbial Antigen Discovery |
| title_full_unstemmed |
Identification of Circulating Bacterial Antigens by <italic toggle="yes">In Vivo</italic> Microbial Antigen Discovery |
| title_sort |
identification of circulating bacterial antigens by <italic toggle="yes">in vivo</italic> microbial antigen discovery |
| publisher |
American Society for Microbiology |
| publishDate |
2011 |
| url |
https://doaj.org/article/508db936fb364d0fb580707b7420d3a5 |
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