{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T11:02:52Z","timestamp":1790420572898,"version":"4.1.0"},"reference-count":30,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2008,9,22]],"date-time":"2008-09-22T00:00:00Z","timestamp":1222041600000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/www.springer.com\/tdm"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Infect Dis"],"published-print":{"date-parts":[[2008,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Anthrax is caused by<jats:italic>Bacillus anthracis<\/jats:italic>that produce two exotoxins, lethal toxin and edema toxin. The lethal toxin is composed of the lethal factor (LF) complexed with the cell binding protective antigen (PA<jats:sub>83<\/jats:sub>, 83 kDa). Likewise, the edema factor (EF) binds to the PA<jats:sub>83<\/jats:sub>to form the edema toxin. Once PA83 is bound to the host cell surface, a furin-like protease cleaves the full-length, inactive protein into 63 kDa and 20 kDa antigens (PA<jats:sub>63<\/jats:sub>and PA<jats:sub>20<\/jats:sub>). PA<jats:sub>63<\/jats:sub>forms a heptamer and is internalized via receptor mediated endocytosis forming a protease-stable pore, which allows EF and LF to enter the cell and exert their toxic effects.<\/jats:p><jats:p>Both proteolytically cleaved protective antigens (PA<jats:sub>63<\/jats:sub>and PA<jats:sub>20<\/jats:sub>fragments) are found in the blood of infected animals. The 63 kDa protective antigen PA<jats:sub>63<\/jats:sub>fragment has been thoroughly studied while little is known about the PA<jats:sub>20<\/jats:sub>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>In this study we examined the role of PA<jats:sub>20<\/jats:sub>using high throughput gene expression analysis of human peripheral blood mononuclear cells (PBMC) exposed to the PA<jats:sub>20<\/jats:sub>. We constructed a PA mutant in which a Factor Xa proteolytic recognition site was genetically engineered into the protective antigen PA<jats:sub>83<\/jats:sub>to obtain PA<jats:sub>20<\/jats:sub>using limited digestion of this recombinant PA<jats:sub>83<\/jats:sub>with trypsin.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Global gene expression response studies indicated modulation of various immune functions and showed gene patterns indicative of apoptosis via the Fas pathway in a subset of the lymphoid cells. This finding was extended to include observations of increased Caspase-3 enzymatic activity and the identification of increases in the population of apoptotic, but not necrotic cells, based on differential staining methods. We identified a list of ~40 inflammatory mediators and heat-shock proteins that were altered similarly upon exposure of PBMC to either rPA<jats:sub>20<\/jats:sub>or<jats:italic>B. anthracis<\/jats:italic>spores\/vegetative cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This study shows that the PA<jats:sub>20<\/jats:sub>has an effect on human peripheral blood leukocytes and can induce apoptosis in the absence of other PA components.<\/jats:p><\/jats:sec>","DOI":"10.1186\/1471-2334-8-124","type":"journal-article","created":{"date-parts":[[2008,9,26]],"date-time":"2008-09-26T18:16:07Z","timestamp":1222452967000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Activity of the Bacillus anthracis20 kDa protective antigen component"],"prefix":"10.1186","volume":"8","author":[{"given":"Rasha","family":"Hammamieh","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wilson J","family":"Ribot","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Terry G","family":"Abshire","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marti","family":"Jett","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Ezzell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2008,9,22]]},"reference":[{"issue":"2","key":"715_CR1","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1128\/iai.49.2.291-297.1985","volume":"49","author":"BD Green","year":"1985","unstructured":"Green BD, et al: Demonstration of a capsule plasmid in Bacillus anthracis. Infect Immun. 1985, 49 (2): 291-7.","journal-title":"Infect Immun"},{"issue":"Pt 3","key":"715_CR2","doi-asserted-by":"publisher","first-page":"739","DOI":"10.1042\/bj3520739","volume":"352","author":"G Vitale","year":"2000","unstructured":"Vitale G, et al: Susceptibility of mitogen-activated protein kinase kinase family members to proteolysis by anthrax lethal factor. Biochem J. 2000, 352 (Pt 3): 739-45. 10.1042\/0264-6021:3520739.","journal-title":"Biochem J"},{"issue":"1","key":"715_CR3","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1016\/0378-1119(89)90335-1","volume":"81","author":"TS Bragg","year":"1989","unstructured":"Bragg TS, Robertson DL: Nucleotide sequence and analysis of the lethal factor gene (lef) from Bacillus anthracis. Gene. 1989, 81 (1): 45-54. 10.1016\/0378-1119(89)90335-1.","journal-title":"Gene"},{"issue":"4\u20135","key":"715_CR4","doi-asserted-by":"publisher","first-page":"421","DOI":"10.1016\/S1438-4221(00)80056-9","volume":"290","author":"R Pellizzari","year":"2000","unstructured":"Pellizzari R, et al: Lethal factor of Bacillus anthracis cleaves the N-terminus of MAPKKs: analysis of the intracellular consequences in macrophages. Int J Med Microbiol. 2000, 290 (4\u20135): 421-7.","journal-title":"Int J Med Microbiol"},{"issue":"Pt 3","key":"715_CR5","doi-asserted-by":"publisher","first-page":"739","DOI":"10.1042\/bj3520739","volume":"352","author":"G Vitale","year":"2000","unstructured":"Vitale G, et al: Susceptibility of mitogen-activated protein kinase kinase family members to proteolysis by anthrax lethal factor. Biochem J. 2000, 352 (Pt 3): 739-45. 10.1042\/0264-6021:3520739.","journal-title":"Biochem J"},{"issue":"5364","key":"715_CR6","doi-asserted-by":"publisher","first-page":"734","DOI":"10.1126\/science.280.5364.734","volume":"280","author":"NS Duesbery","year":"1998","unstructured":"Duesbery NS, et al: Proteolytic inactivation of MAP-kinase-kinase by anthrax lethal factor. Science. 1998, 280 (5364): 734-7. 10.1126\/science.280.5364.734.","journal-title":"Science"},{"issue":"1\u20132","key":"715_CR7","doi-asserted-by":"publisher","first-page":"199","DOI":"10.1016\/S0014-5793(99)01502-1","volume":"462","author":"R Pellizzari","year":"1999","unstructured":"Pellizzari R, et al: Anthrax lethal factor cleaves MKK3 in macrophages and inhibits the LPS\/IFNgamma-induced release of NO and TNFalpha. FEBS Lett. 1999, 462 (1\u20132): 199-204. 10.1016\/S0014-5793(99)01502-1.","journal-title":"FEBS Lett"},{"issue":"10","key":"715_CR8","doi-asserted-by":"publisher","first-page":"3162","DOI":"10.1073\/pnas.79.10.3162","volume":"79","author":"SH Leppla","year":"1982","unstructured":"Leppla SH: Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells. Proc Natl Acad Sci USA. 1982, 79 (10): 3162-6. 10.1073\/pnas.79.10.3162.","journal-title":"Proc Natl Acad Sci USA"},{"issue":"2","key":"715_CR9","doi-asserted-by":"publisher","first-page":"287","DOI":"10.1016\/0378-1119(88)90439-8","volume":"69","author":"SL Welkos","year":"1988","unstructured":"Welkos SL, et al: Sequence and analysis of the DNA encoding protective antigen of Bacillus anthracis. Gene. 1988, 69 (2): 287-300. 10.1016\/0378-1119(88)90439-8.","journal-title":"Gene"},{"issue":"1","key":"715_CR10","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1128\/iai.39.1.371-376.1983","volume":"39","author":"P Mikesell","year":"1983","unstructured":"Mikesell P, et al: Evidence for plasmid-mediated toxin production in Bacillus anthracis. Infect Immun. 1983, 39 (1): 371-6.","journal-title":"Infect Immun"},{"issue":"10","key":"715_CR11","doi-asserted-by":"crossref","first-page":"3381","DOI":"10.1128\/iai.59.10.3381-3386.1991","volume":"59","author":"V Escuyer","year":"1991","unstructured":"Escuyer V, Collier RJ: Anthrax protective antigen interacts with a specific receptor on the surface of CHO-K1 cells. Infect Immun. 1991, 59 (10): 3381-6.","journal-title":"Infect Immun"},{"issue":"21","key":"715_CR12","doi-asserted-by":"publisher","first-page":"10277","DOI":"10.1073\/pnas.89.21.10277","volume":"89","author":"KR Klimpel","year":"1992","unstructured":"Klimpel KR, et al: Anthrax toxin protective antigen is activated by a cell surface protease with the sequence specificity and catalytic properties of furin. Proc Natl Acad Sci USA. 1992, 89 (21): 10277-81. 10.1073\/pnas.89.21.10277.","journal-title":"Proc Natl Acad Sci USA"},{"issue":"23","key":"715_CR13","doi-asserted-by":"crossref","first-page":"16396","DOI":"10.1016\/S0021-9258(18)42016-9","volume":"267","author":"SS Molloy","year":"1992","unstructured":"Molloy SS, et al: Human furin is a calcium-dependent serine endoprotease that recognizes the sequence Arg-X-X-Arg and efficiently cleaves anthrax toxin protective antigen. J Biol Chem. 1992, 267 (23): 16396-402.","journal-title":"J Biol Chem"},{"issue":"21","key":"715_CR14","doi-asserted-by":"publisher","first-page":"10198","DOI":"10.1073\/pnas.90.21.10198","volume":"90","author":"PC Hanna","year":"1993","unstructured":"Hanna PC, Acosta D, Collier RJ: On the role of macrophages in anthrax. Proc Natl Acad Sci USA. 1993, 90 (21): 10198-201. 10.1073\/pnas.90.21.10198.","journal-title":"Proc Natl Acad Sci USA"},{"issue":"10","key":"715_CR15","doi-asserted-by":"publisher","first-page":"3162","DOI":"10.1073\/pnas.79.10.3162","volume":"79","author":"SH Leppla","year":"1982","unstructured":"Leppla SH: Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells. Proc Natl Acad Sci USA. 1982, 79 (10): 3162-6. 10.1073\/pnas.79.10.3162.","journal-title":"Proc Natl Acad Sci USA"},{"issue":"3","key":"715_CR16","doi-asserted-by":"publisher","first-page":"543","DOI":"10.1099\/00221287-138-3-543","volume":"138","author":"JW Ezzell Jr","year":"1992","unstructured":"Ezzell JW, Abshire TG: Serum protease cleavage of Bacillus anthracis protective antigen. J Gen Microbiol. 1992, 138 (3): 543-9.","journal-title":"J Gen Microbiol"},{"key":"715_CR17","first-page":"2","volume-title":"Proceedings of the International Workshop on Anthrax","author":"JW Ezzell","year":"1990","unstructured":"Ezzell JW, Abshire TG, Brown C: Analyses of Bacillus anthracis vegetative cell surface antigens and serum protease cleavage of protective antigen. Proceedings of the International Workshop on Anthrax. Edited by: Turnbull. 1990, Wiltshire: Salisbury Printing Co, 68: 2-"},{"issue":"6","key":"715_CR18","doi-asserted-by":"crossref","first-page":"1606","DOI":"10.1128\/iai.58.6.1606-1613.1990","volume":"58","author":"SF Little","year":"1990","unstructured":"Little SF, Leppla SH, Friedlander AM: Production and characterization of monoclonal antibodies against the lethal factor component of Bacillus anthracis lethal toxin. Infect Immun. 1990, 58 (6): 1606-13.","journal-title":"Infect Immun"},{"issue":"32","key":"715_CR19","doi-asserted-by":"crossref","first-page":"20607","DOI":"10.1016\/S0021-9258(17)32036-7","volume":"269","author":"JC Milne","year":"1994","unstructured":"Milne JC, et al: Anthrax protective antigen forms oligomers during intoxication of mammalian cells. J Biol Chem. 1994, 269 (32): 20607-12.","journal-title":"J Biol Chem"},{"issue":"3","key":"715_CR20","doi-asserted-by":"publisher","first-page":"647","DOI":"10.1111\/j.1365-2958.1993.tb00936.x","volume":"10","author":"JC Milne","year":"1993","unstructured":"Milne JC, Collier RJ: pH-dependent permeabilization of the plasma membrane of mammalian cells by anthrax protective antigen. Mol Microbiol. 1993, 10 (3): 647-53. 10.1111\/j.1365-2958.1993.tb00936.x.","journal-title":"Mol Microbiol"},{"issue":"3","key":"715_CR21","doi-asserted-by":"publisher","first-page":"1047","DOI":"10.1021\/bi047791s","volume":"44","author":"KA Christensen","year":"2005","unstructured":"Christensen KA, et al: Interaction of the 20 kDa and 63 kDa fragments of anthrax protective antigen: kinetics and thermodynamics. Biochemistry. 2005, 44 (3): 1047-53. 10.1021\/bi047791s.","journal-title":"Biochemistry"},{"issue":"32","key":"715_CR22","doi-asserted-by":"crossref","first-page":"19103","DOI":"10.1016\/S0021-9258(19)47273-6","volume":"264","author":"Y Singh","year":"1989","unstructured":"Singh Y, Chaudhary VK, Leppla SH: A deleted variant of Bacillus anthracis protective antigen is non-toxic and blocks anthrax toxin action in vivo. J Biol Chem. 1989, 264 (32): 19103-7.","journal-title":"J Biol Chem"},{"issue":"5","key":"715_CR23","doi-asserted-by":"publisher","first-page":"741","DOI":"10.1046\/j.1365-2672.1998.00405.x","volume":"84","author":"L Baillie","year":"1998","unstructured":"Baillie L, Moir A, Manchee R: The expression of the protective antigen of Bacillus anthracis in Bacillus subtilis. J Appl Microbiol. 1998, 84 (5): 741-6. 10.1046\/j.1365-2672.1998.00405.x.","journal-title":"J Appl Microbiol"},{"issue":"3","key":"715_CR24","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1128\/AEM.64.3.982-991.1998","volume":"64","author":"JW Farchaus","year":"1998","unstructured":"Farchaus JW, et al: Fermentation, purification, and characterization of protective antigen from a recombinant, avirulent strain of Bacillus anthracis. Appl Environ Microbiol. 1998, 64 (3): 982-91.","journal-title":"Appl Environ Microbiol"},{"issue":"2","key":"715_CR25","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1089\/omi.2007.4322","volume":"11","author":"R Hammamieh","year":"2007","unstructured":"Hammamieh R, et al: GeneCite: a stand-alone open source tool for high-throughput literature and pathway mining. Omics. 2007, 11 (2): 143-51. 10.1089\/omi.2007.4322.","journal-title":"Omics"},{"issue":"11","key":"715_CR26","doi-asserted-by":"publisher","first-page":"6382","DOI":"10.1128\/IAI.72.11.6382-6389.2004","volume":"72","author":"AK Pickering","year":"2004","unstructured":"Pickering AK, et al: Cytokine response to infection with Bacillus anthracis spores. Infect Immun. 2004, 72 (11): 6382-9. 10.1128\/IAI.72.11.6382-6389.2004.","journal-title":"Infect Immun"},{"issue":"7","key":"715_CR27","doi-asserted-by":"publisher","first-page":"335","DOI":"10.1016\/j.tibs.2004.05.002","volume":"29","author":"DJ Rigden","year":"2004","unstructured":"Rigden DJ, Mello LV, Galperin MY: The PA14 domain, a conserved all-beta domain in bacterial toxins, enzymes, adhesins and signaling molecules. Trends Biochem Sci. 2004, 29 (7): 335-9. 10.1016\/j.tibs.2004.05.002.","journal-title":"Trends Biochem Sci"},{"issue":"1","key":"715_CR28","doi-asserted-by":"crossref","first-page":"395","DOI":"10.4049\/jimmunol.160.1.395","volume":"160","author":"S Deaglio","year":"1998","unstructured":"Deaglio S, et al: Human CD38 (ADP-ribosyl cyclase) is a counter-receptor of CD31, an Ig superfamily member. J Immunol. 1998, 160 (1): 395-402.","journal-title":"J Immunol"},{"issue":"11","key":"715_CR29","doi-asserted-by":"crossref","first-page":"3864","DOI":"10.1182\/blood.V94.11.3864","volume":"94","author":"G Sconocchia","year":"1999","unstructured":"Sconocchia G, et al: CD38 triggers cytotoxic responses in activated human natural killer cells. Blood. 1999, 94 (11): 3864-71.","journal-title":"Blood"},{"issue":"3","key":"715_CR30","doi-asserted-by":"publisher","first-page":"690","DOI":"10.1016\/j.bbrc.2007.06.124","volume":"360","author":"I Chvyrkova","year":"2007","unstructured":"Chvyrkova I, Zhang XC, Terzyan S: Lethal factor of anthrax toxin binds monomeric form of protective antigen. Biochem Biophys Res Commun. 2007, 360 (3): 690-5. 10.1016\/j.bbrc.2007.06.124.","journal-title":"Biochem Biophys Res Commun"}],"container-title":["BMC Infectious Diseases"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/1471-2334-8-124.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/article\/10.1186\/1471-2334-8-124\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/1471-2334-8-124","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1471-2334-8-124.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,5,20]],"date-time":"2023-05-20T04:34:17Z","timestamp":1684557257000},"score":1,"resource":{"primary":{"URL":"https:\/\/bmcinfectdis.biomedcentral.com\/articles\/10.1186\/1471-2334-8-124"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,9,22]]},"references-count":30,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2008,12]]}},"alternative-id":["715"],"URL":"https:\/\/doi.org\/10.1186\/1471-2334-8-124","relation":{},"ISSN":["1471-2334"],"issn-type":[{"value":"1471-2334","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,9,22]]},"assertion":[{"value":"31 December 2007","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 September 2008","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 September 2008","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"124"}}