{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T11:03:41Z","timestamp":1790420621218,"version":"4.1.0"},"reference-count":61,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2020,7,23]],"date-time":"2020-07-23T00:00:00Z","timestamp":1595462400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Anthrax toxins are produced by Bacillus anthracis throughout infection and shape the physiopathogenesis of the disease. They are produced in low quantities but are highly efficient. They have thus been long ignored, but recent biochemical methods have improved our knowledge in animal models. This article reviews the various methods that have been used and how they could be applied to clinical diagnosis.<\/jats:p>","DOI":"10.3390\/microorganisms8081103","type":"journal-article","created":{"date-parts":[[2020,7,23]],"date-time":"2020-07-23T10:14:06Z","timestamp":1595499246000},"page":"1103","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Anthrax Toxin Detection: From In Vivo Studies to Diagnostic Applications"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4542-2421","authenticated-orcid":false,"given":"Jean-Nicolas","family":"Tournier","sequence":"first","affiliation":[{"name":"Unit\u00e9 Bact\u00e9riologie Bioth\u00e9rapies Anti-infectieuses et Immunit\u00e9, Institut de Recherche Biom\u00e9dicale des Arm\u00e9es (IRBA), 1 place G\u00e9n\u00e9ral Val\u00e9rie Andr\u00e9, 91220 Br\u00e9tigny sur Orge, France"},{"name":"Centre National de R\u00e9f\u00e9rence-Laboratoire Expert Charbon, 1 place G\u00e9n\u00e9ral Val\u00e9rie Andr\u00e9, 91220 Br\u00e9tigny sur Orge, France"},{"name":"Innovative Vaccine Laboratory, Institut Pasteur, 28 rue du docteur Roux, 75015 Paris, France"},{"name":"Ecole du Val-de-Gr\u00e2ce, 1 place Alphonse Laveran, 75005 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cl\u00e9mence","family":"Rougeaux","sequence":"additional","affiliation":[{"name":"Unit\u00e9 Bact\u00e9riologie Bioth\u00e9rapies Anti-infectieuses et Immunit\u00e9, Institut de Recherche Biom\u00e9dicale des Arm\u00e9es (IRBA), 1 place G\u00e9n\u00e9ral Val\u00e9rie Andr\u00e9, 91220 Br\u00e9tigny sur Orge, France"},{"name":"Centre National de R\u00e9f\u00e9rence-Laboratoire Expert Charbon, 1 place G\u00e9n\u00e9ral Val\u00e9rie Andr\u00e9, 91220 Br\u00e9tigny sur Orge, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1146\/annurev-micro-091014-104523","article-title":"Anthrax Pathogenesis","volume":"69","author":"Moayeri","year":"2015","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"933","DOI":"10.3201\/eid0706.010604","article-title":"Bioterrorism-related inhalational anthrax: The first 10 cases reported in the United States","volume":"7","author":"Jernigan","year":"2001","journal-title":"Emerg. Infect. Dis."},{"key":"ref_3","unstructured":"WHO (2020, April 01). Anthrax in Humans and Animals. Available online: https:\/\/apps.who.int\/iris\/bitstream\/handle\/10665\/97503\/9789241547536_eng.pdf;jsessionid=3BF3DA0CD1CA21CFBCF1160F6B7FB128?sequence=1."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/j.mcp.2008.06.001","article-title":"Real-time PCR system targeting a chromosomal marker specific for Bacillus anthracis","volume":"22","author":"Antwerpen","year":"2008","journal-title":"Mol. Cell. Probes"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1016\/j.mam.2009.08.007","article-title":"Receptors of anthrax toxin and cell entry","volume":"30","author":"Young","year":"2009","journal-title":"Mol. Asp. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"15583","DOI":"10.1073\/pnas.1010145107","article-title":"Heterodimeric integrin complexes containing beta1-integrin promote internalization and lethality of anthrax toxin","volume":"107","author":"Martchenko","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"16396","DOI":"10.1016\/S0021-9258(18)42016-9","article-title":"Human furin is a calcium-dependent serine endoprotease that recognizes the sequence Arg-X-X-Arg and efficiently cleaves anthrax toxin protective antigen","volume":"267","author":"Molloy","year":"1992","journal-title":"J. Biol. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Friebe, S., van der Goot, F.G., and Burgi, J. (2016). The Ins and Outs of Anthrax Toxin. Toxins, 8.","DOI":"10.3390\/toxins8030069"},{"key":"ref_9","first-page":"706","article-title":"Anthrax lethal factor cleaves the N-terminus of MAPKKs and induces tyrosine\/threonine phosphorylation of MAPKs in cultured macrophages","volume":"248","author":"Vitale","year":"1998","journal-title":"J. Appl. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Hellmich, K.A., Levinsohn, J.L., Fattah, R., Newman, Z.L., Maier, N., Sastalla, I., Liu, S., Leppla, S.H., and Moayeri, M. (2012). Anthrax lethal factor cleaves mouse nlrp1b in both toxin-sensitive and toxin-resistant macrophages. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0049741"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Chavarria-Smith, J., and Vance, R.E. (2013). Direct proteolytic cleavage of NLRP1B is necessary and sufficient for inflammasome activation by anthrax lethal factor. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003452"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3162","DOI":"10.1073\/pnas.79.10.3162","article-title":"Anthrax toxin edema factor: A bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells","volume":"79","author":"Leppla","year":"1982","journal-title":"Proce. Natl. Acad. Sci. USA."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Puhar, A., Dal Molin, F., Horvath, S., Ladant, D., and Montecucco, C. (2008). Anthrax edema toxin modulates PKA- and CREB-dependent signaling in two phases. PLoS ONE, 3.","DOI":"10.1371\/annotation\/b72b80a6-ee81-49e6-a086-0292f6255d4f"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"19781","DOI":"10.1074\/jbc.M700128200","article-title":"Anthrax edema toxin inhibits endothelial cell chemotaxis via Epac and Rap1","volume":"282","author":"Hong","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1038\/173869a0","article-title":"Observations on experimental anthrax; demonstration of a specific lethal factor produced in vivo by Bacillus anthracis","volume":"173","author":"Smith","year":"1954","journal-title":"Nature"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1016\/S0140-6736(54)91881-4","article-title":"Observations on the cause of death in experimental anthrax","volume":"267","author":"Smith","year":"1954","journal-title":"Lancet"},{"key":"ref_17","first-page":"263","article-title":"The chemical basis of the virulence of Bacillus anthracis.  VII. Two components of the anthrax toxin: Their relationship to known immunising aggressins","volume":"37","author":"Smith","year":"1956","journal-title":"Br. J. Exp. Pathol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1128\/jb.76.2.192-202.1958","article-title":"Further purification studies on the protective antigen of Bacillus anthracis produced in vitro","volume":"76","author":"Strange","year":"1958","journal-title":"J. Bact."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1274","DOI":"10.1128\/jb.83.6.1274-1280.1962","article-title":"Rapid lethal effect in rats of a third component found upon fractionating the toxin of Bacillus anthracis","volume":"83","author":"Beall","year":"1962","journal-title":"J. Bacteriol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1099\/00221287-17-2-505","article-title":"An agar-diffusion method for titrating Bacillus anthracis immunizing antigen and its application to a study of antigen production","volume":"17","author":"Thorne","year":"1957","journal-title":"J. Gen. Microb."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1099\/00221287-22-1-219","article-title":"The serological relationship between purified preparations of factors I and II of the anthrax toxin produced in vivo and in vitro","volume":"22","author":"Sargeant","year":"1960","journal-title":"J. Gen. Microb."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1099\/00221287-22-1-206","article-title":"Purification of factors I and II of the anthrax toxin produced in vivo","volume":"22","author":"Stanley","year":"1960","journal-title":"J. Gen. Microbiol."},{"key":"ref_23","first-page":"32","article-title":"Serologic testing for anthrax antibodies in workers in a goat hair processing mill","volume":"72","author":"Norman","year":"1960","journal-title":"Am. J. Hyg."},{"key":"ref_24","first-page":"992","article-title":"An epidemic of inhalation anthrax, the first in the twentieth century","volume":"29","author":"Plotkin","year":"1960","journal-title":"I. Clin. Features. Am. J. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1093\/oxfordjournals.aje.a120135","article-title":"An epidemic of inhalation anthrax: The first in the twentieth century","volume":"72","author":"Brachman","year":"1960","journal-title":"Ii. Epidemiol. Am. J. Hyg."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Darlow, H.M., and Pride, N.B. (1969). Serological diagnosis of anthrax. Lancet, 2.","DOI":"10.1016\/S0140-6736(69)90130-5"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1128\/jb.95.3.919-924.1968","article-title":"In vivo-produced anthrax toxin","volume":"95","author":"Fish","year":"1968","journal-title":"J. Bacteriol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1128\/iai.52.2.356-363.1986","article-title":"Development of antibodies to protective antigen and lethal factor components of anthrax toxin in humans and guinea pigs and their relevance to protective immunity","volume":"52","author":"Turnbull","year":"1986","journal-title":"Infect. Immun."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1128\/CVI.00023-06","article-title":"Detection of anthrax toxin in the serum of animals infected with Bacillus anthracis by using engineered immunoassays","volume":"13","author":"Mabry","year":"2006","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"5871","DOI":"10.1128\/IAI.00792-06","article-title":"Protective antigen as a correlative marker for anthrax in animal models","volume":"74","author":"Kobiler","year":"2006","journal-title":"Infect. Immun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5790","DOI":"10.1128\/IAI.00520-08","article-title":"Identification of a surrogate marker for infection in the African green monkey model of inhalation anthrax","volume":"76","author":"Rossi","year":"2008","journal-title":"Infect. Immun."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1016\/j.toxicon.2008.08.011","article-title":"Ratio of lethal and edema factors in rabbit systemic anthrax","volume":"52","author":"Molin","year":"2008","journal-title":"Toxicon"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1128\/CVI.00412-08","article-title":"Detection of anthrax toxin by an ultrasensitive immunoassay using europium nanoparticles","volume":"16","author":"Tang","year":"2009","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.ab.2012.02.040","article-title":"Ultra-fast pg\/mL anthrax toxin (protective antigen) detection assay based on microwave-accelerated metal-enhanced fluorescence","volume":"425","author":"Dragan","year":"2012","journal-title":"Anal. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.diagmicrobio.2013.05.006","article-title":"Detection of protective antigen, an anthrax specific toxin in human serum by using surface plasmon resonance","volume":"77","author":"Ghosh","year":"2013","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1016\/j.jim.2014.05.008","article-title":"Detection of anthrax protective antigen (PA) using europium labeled anti-PA monoclonal antibody and time-resolved fluorescence","volume":"408","author":"Stoddard","year":"2014","journal-title":"J. Immunol. Methods"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3965","DOI":"10.1007\/s00216-013-6752-1","article-title":"A novel homogeneous immunoassay for anthrax detection based on the AlphaLISA method: Detection of B. anthracis spores and protective antigen (PA) in complex samples","volume":"405","author":"Mechaly","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1007\/s10895-014-1354-7","article-title":"Rapid homogenous time-resolved fluorescence (HTRF) immunoassay for anthrax detection","volume":"24","author":"Cohen","year":"2014","journal-title":"J. Fluoresc."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2264","DOI":"10.1039\/C8AN02479K","article-title":"Accurate and selective quantification of anthrax protective antigen in plasma by immunocapture and isotope dilution mass spectrometry","volume":"144","author":"Solano","year":"2019","journal-title":"Analyst"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1321","DOI":"10.1093\/infdis\/jir543","article-title":"Lethal factor toxemia and anti-protective antigen antibody activity in naturally acquired cutaneous anthrax","volume":"204","author":"Boyer","year":"2011","journal-title":"J. Infect. Dis."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Kolesnikov, A.V., Kozyr, A.V., Ryabko, A.K., and Shemyakin, I.G. (2016). Ultrasensitive detection of protease activity of anthrax and botulinum toxins by a new PCR-based assay. Pathog. Dis., 74.","DOI":"10.1093\/femspd\/ftv112"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"8463","DOI":"10.1021\/ac701741s","article-title":"Detection and quantification of anthrax lethal factor in serum by mass spectrometry","volume":"79","author":"Boyer","year":"2007","journal-title":"Anal. Chem."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Suryadi, K., and Shine, N. (2018). Design and use of a novel substrate for simple, rapid, and specific early detection of anthrax infection. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0207084"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2391","DOI":"10.3390\/molecules16032391","article-title":"Quantitative mass spectrometry for bacterial protein toxins\u2014A sensitive, specific, high-throughput tool for detection and diagnosis","volume":"16","author":"Boyer","year":"2011","journal-title":"Molecules"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2847","DOI":"10.1007\/s00216-015-8509-5","article-title":"High-sensitivity MALDI-TOF MS quantification of anthrax lethal toxin for diagnostics and evaluation of medical countermeasures","volume":"407","author":"Boyer","year":"2015","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Rougeaux, C., Becher, F., Ezan, E., Tournier, J.N., and Goossens, P.L. (2016). In vivo dynamics of active edema and lethal factors during anthrax. Sci. Rep., 6.","DOI":"10.1038\/srep23346"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"5935","DOI":"10.1021\/ac900827s","article-title":"Femtomolar detection of the anthrax edema factor in human and animal plasma","volume":"81","author":"Duriez","year":"2009","journal-title":"Anal. Chem."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Israeli, M., Rotem, S., Elia, U., Bar-Haim, E., Cohen, O., and Chitlaru, T. (2016). A Simple Luminescent Adenylate-Cyclase Functional Assay for Evaluation of Bacillus anthracis Edema Factor Activity. Toxins, 8.","DOI":"10.3390\/toxins8080243"},{"key":"ref_49","unstructured":"Boyer, A.E., Woolfitt, A.R., Candela, M., Lins, R.C., Solano, M., Lee, J., Sanford, D., Stark, G., Dreier, T., and Quinn, P. (2013, January 1\u20135). Toxin levels in organ tissues of nonhuman primates with inhalation anthrax. Proceedings of the International Conference on Bacillus anthracis, B. cereus and B. thuringiensis, Victoria, BC, Canada."},{"key":"ref_50","unstructured":"Woolfitt, A.R., Juni, B.A., Gallegos-Candela, M., Lins, R., Solano, M., Lee, J., Sanford, D., Stark, G., Dreier, T., and Barr, J. (2013, January 1\u20135). Development of anthrax toxemia in new zealand white rabbits developing systemic anthrax after exposure to low-dose ames spores. Proceedings of the International Conference on Bacillus anthracis, B. cereus, B. thuringiensis, Victoria, BC, Canada."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2493","DOI":"10.1007\/s00216-019-01730-4","article-title":"Zeptomole per milliliter detection and quantification of edema factor in plasma by LC-MS\/MS yields insights into toxemia and the progression of inhalation anthrax","volume":"411","author":"Lins","year":"2019","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1086\/512372","article-title":"A case of naturally acquired inhalation anthrax: Clinical care and analyses of anti-protective antigen immunoglobulin G and lethal factor","volume":"44","author":"Walsh","year":"2007","journal-title":"Clin. Infect. Dis."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"310","DOI":"10.3201\/eid2002.130245","article-title":"Lethal factor and anti-protective antigen IgG levels associated with inhalation anthrax, Minnesota, USA","volume":"20","author":"Sprenkle","year":"2014","journal-title":"Emerg. Infect. Dis."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Weiner, Z.P., Boyer, A.E., Gallegos-Candela, M., Cardani, A.N., Barr, J.R., and Glomski, I.J. (2012). Debridement increases survival in a mouse model of subcutaneous anthrax. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0030201"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1111\/cmi.12232","article-title":"Circulating lethal toxin decreases the ability of neutrophils to respond to Bacillus anthracis","volume":"16","author":"Weiner","year":"2014","journal-title":"Cell Microbiol"},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Marston, C.K., Ibrahim, H., Lee, P., Churchwell, G., Gumke, M., Stanek, D., Gee, J.E., Boyer, A.E., Gallegos-Candela, M., and Barr, J.R. (2016). Anthrax Toxin-Expressing Bacillus cereus Isolated from an Anthrax-Like Eschar. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0156987"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"660","DOI":"10.1093\/infdis\/jiz497","article-title":"Very Early Blood Diffusion of the Active Lethal and Edema Factors of Bacillus anthracis After Intranasal Infection","volume":"221","author":"Rougeaux","year":"2020","journal-title":"J. Infect. Dis."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"3432","DOI":"10.1128\/IAI.00346-09","article-title":"Kinetics of lethal factor and poly-D-glutamic acid antigenemia during inhalation anthrax in rhesus macaques","volume":"77","author":"Boyer","year":"2009","journal-title":"Infect. Immun."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"270","DOI":"10.7326\/0003-4819-144-4-200602210-00009","article-title":"Systematic review: A century of inhalational anthrax cases from 1900 to 2005","volume":"144","author":"Holty","year":"2006","journal-title":"Ann. Intern. Med."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.ab.2017.12.007","article-title":"Validated MALDI-TOF-MS method for anthrax lethal factor provides early diagnosis and evaluation of therapeutics","volume":"543","author":"Boyer","year":"2018","journal-title":"Anal. Biochem."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Tournier, J.N., Rougeaux, C., Biot, F.V., and Goossens, P.L. (2019). Questionable Efficacy of Therapeutic Antibodies in the Treatment of Anthrax. mSphere, 4.","DOI":"10.1128\/mSphere.00282-19"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/8\/8\/1103\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:51:08Z","timestamp":1760176268000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/8\/8\/1103"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,23]]},"references-count":61,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2020,8]]}},"alternative-id":["microorganisms8081103"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms8081103","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,23]]}}}