Antibodies, Bacterial/immunology; Antigens, Bacterial/genetics; Bacterial Toxins/genetics; Cell Line; DNA, Bacterial/genetics; France; Humans; Immunoglobulin G/immunology; Japan; Mutation; Polymorphism, Genetic; Pore Forming Cytotoxic Proteins/genetics; Protein Transport/genetics; Pseudomonas aeruginosa/genetics/isolation & purification/metabolism; Random Amplified Polymorphic DNA Technique; Sequence Analysis, DNA; United States
Abstract :
[en] The type III secretion system of Pseudomonas aeruginosa, responsible for acute infection, is composed of over twenty proteins that facilitate cytotoxin injection directly into host cells. Integral to this process is production and secretion of PcrV. Administration of a recently developed, anti-PcrV immunoglobulin, either as a therapeutic or prophylactic has previously demonstrated efficacy against laboratory strains of P. aeruginosa in a murine model. To determine if this therapy is universally applicable to a variety of P. aeruginosa clinical isolates, genetic heterogeneity of pcrV was analyzed among strains collected from three geographically distinct regions; United States, France and Japan. Sequence analysis of PcrV demonstrated limited variation among the clinical isolates examined. Strains were grouped according to the presence of non-synonymous single nucleotide polymorphisms. Representative isolates from each mutant group were examined for the ability of anti-PcrV to bind the protein secreted by these strains. The protective effect of anti-PcrV IgG against each strain was determined using an epithelial cell line cytotoxicity assay. The majority of strains tested demonstrated reduced cytotoxicity in the presence of anti-PcrV IgG. This study provides insights into the natural sequence variability of PcrV and an initial indication of the amino acid residues that appear to be conserved across strains. It also demonstrates the protective effect of anti-PcrV immunotherapy against a multitude of P. aeruginosa strains from diverse global regions with a variety of mutations in PcrV.
Disciplines :
Anesthesia & intensive care
Author, co-author :
Lynch, Susan V.
Flanagan, Judith L.
Sawa, Teiji
Fang, Alice
Baek, Marshall S.
Rubio-Mills, Amua
Ajayi, Temitayo
Yanagihara, Katsunori
Hirakata, Yoichi
Kohno, Shigeru
Misset, Benoît ; Centre Hospitalier Universitaire de Liège - CHU > Service de Soins Intensifs
Allmond L.R., Ajayi T., Moriyama K., Wiener-Kronish J.P., Sawa T. V-antigen genotype and phenotype analyses of clinical isolates of Pseudomonas aeruginosa. J Clin Microbiol 2004, 42:3857-3860.
Almirall J., Mesalles E., Klamburg J., Parra O., Agudo A. Prognostic factors of pneumonia requiring admission to the intensive care unit. Chest 1995, 107:511-516.
Burrows T.W. An antigen determining virulence in Pasteurella pestis. Nature 1956, 177:426-427.
Burrows T.W., Bacon G.A. The basis of virulence in Pasteurella pestis: an antigen determining virulence. Br J Exp Pathol 1956, 37:481-493.
Craven D.E., Steger K.A. Nosocomial pneumonia in mechanically ventilated adult patients: epidemology and prevention in 1996. Semin Respir Infect 1996, 11:32-53.
Crouch Brewer S., Wunderink R.G., Jones C.B., Leeper K.V. Ventilator-associated pneumonia due to Pseudomonas aeruginosa. Chest 1996, 109:1019-1029.
DeBord K.L., Lee V.T., Schneewind O. Roles of LcrG and LcrV during type III targeting of effector Yops by Yersinia enterocolitica. J Bacteriol 2001, 183:4588-4598.
Duncan C.J., Scott S. What acused the black deadth?. Postgrad Med J 2005, 81:315-320.
Elvin S.J., Eyles J.E., Howard K.A., Ravichandran E., Somavarappu S., Alpar H.O., et al. Protection against bubonic and pneumonic plague with a single dose microencapsulated sub-unit vaccine. Vaccine 2005, 15:4433-4439.
Faure K., Fujimoto J., Shimabukuro D.W., Ajayi T., Shime N., Moriyama K., et al. Effects of monoclonal anti-PcrV antibody on Pseudomonas aeruginosa-induced acute lung injury in a rat model. J Immune Ther Vaccines 2003, 1:2.
Frank D.W., Vallis A., Wiener-Kronish J.P., Roy-Burman A., Spack E.G., Mullaney B.P., et al. Generation and characterization of a protective monoclonal antibody to Pseudomonas aeruginosa PcrV. J Infect Dis 2002, 186:64-73.
Hill J., Leary S.E., Griffin K.F., Williamson E.D., Titball R.W. Regions of Yersinia pestis V antigen that contribute to protection against plague identified by passive and active immunization. Infect Immun 1997, 65:4476-4482.
Holder I.A. Pseudomonas immunotherapy: a historical overview. Vaccine 2004, 22:831-839.
Holder I.A., Neely A.N., Frank D.W. PcrV immunization enhances survival of burned Pseudomonas aeruginosa-infected mice. Infect Immun 2001, 69:5908-5910.
Hueck C.J. Type III protein secretion systems in bacterial pathogens of plants and animals. Microbiol Mol Biol Rev 1998, 62:379-433.
Kurahashi K., Kajikawa O., Sawa T., Ohara M., Gropper M.A., Frank D.W., et al. Pathogenesis of septic shock in Pseudomonas aeruginosa pneumonia. J Clin Invest 1999, 104:743-750.
Matson J.S., Nilles M.L. LcrG-LcrV interaction is required for control of Yops secretion in Yersinia pestis. J. Bacteriol. 2001, 183(17):5082-5091.
Nanao M., Ricard-Blum S., Di Guilmi A.M., Lemaire D., Lascoux D., Chabert J., et al. Type III secretion proteins PcrV and PcrG from Pseudomonas aeruginosa form a 1:1 complex through high affinity interactions. BMC Microbiol 2003, 3:21.
Neely A.N., Holder I.A., Wiener-Kronish J.P., Sawa T. Passive anti-PcrV treatment protects burned mice against Pseudomonas aeruginosa challenge. Burns 2005, 31:153-158.
Nicas T.I., Iglewski B.H. Isolation and characterization of transposon-induced mutants of Pseudomonas aeruginosa deficient in production of exoenzyme S. Infect Immun 1984, 45:470-474.
Roggenkamp A., Geiger A.M., Leitritz L., Kessler A., Heesemann J. Passive immunity to infection with Yersinia spp. mediated by anti-recombinant V antigen is dependent on polymorphism of V antigen. Infect Immun 1997, 65:446-451.
Roy-Burman A., Savel R.H., Racine S., Swanson B.L., Revadigar N.S., Fujimoto J., et al. Type III protein secretion is associated with death in lower respiratory and systemic Pseudomonas aeruginosa infections. J Infect Dis 2001, 183:1767-1774.
Rumbaugh K.P., Sawa T., Wiener-Kronish J.P. New perspectives on prevention and management of Pseudomonas aeruginosa infections. In Severe infections caused by Pseudomonas aeruginosa 2003, Kluwer Academic Publishers, Boston. A.R. Hauser (Ed.).
Sato H., Feix J.B., Hillard C.J., Frank D.W. Characterization of phospholipase activity of the Pseudomonas aeruginosa type III cytotoxin, ExoU. J Bacteriol 2005, 187:1192-1195.
Sato H., Frank D.W. ExoU is a potent intracellular phospholipase. Mol Microbiol 2004, 53:1279-1290.
Sawa T., Wiener-Kronish J.P. A therapeutic strategy against the shared virulence mechanism utilized by both Yersinia pestis and Pseudomonas aeruginosa. Anesthesiol Clin North Am 2004, 22:591-606.
Sawa T., Yahr T.L., Ohara M., Kurahashi K., Gropper M.A., Wiener-Kronish J.P., et al. Active and passive immunization with the Pseudomonas V antigen protects against type III intoxication and lung injury. Nat Med 1999, 5:392-398.
Schmidt A., Schaffelhofer S., Muller K., Rollinghoff M., Beuscher H.U. Analysis of the Yersinia enterocolitica 0:8 V antigen for cross protectivity. Microb Pathog 1999, 26:221-233.
Shime N., Sawa T., Fujimoto J., Faure K., Allmond L.R., Karaca T., et al. Therapeutic administration of anti-PcrV F(ab′)(2) in sepsis associated with Pseudomonas aeruginosa. J Immunol 2001, 167:5880-5886.
Une T., Brubaker R.R. Roles of V antigen in promoting virulence and immunity in yersiniae. J Immunol 1984, 133:2226-2230.