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Klebsiella Pneumoniae Infections Matthew .S Lawlor Washington University School of Medicine in St Washington University School of Medicine Digital Commons@Becker Open Access Publications 2006 Comparison of the host responses to wild-type and cpsB mutant Klebsiella pneumoniae infections Matthew .S Lawlor Washington University School of Medicine in St. Louis Scott A. Handley Washington University School of Medicine in St. Louis Virginia L. Miller Washington University School of Medicine in St. Louis Follow this and additional works at: https://digitalcommons.wustl.edu/open_access_pubs Recommended Citation Lawlor, Matthew S.; Handley, Scott A.; and Miller, Virginia L., ,"Comparison of the host responses to wild-type and cpsB mutant Klebsiella pneumoniae infections." Infection and Immunity.74,9. 5402-5407. (2006). https://digitalcommons.wustl.edu/open_access_pubs/2387 This Open Access Publication is brought to you for free and open access by Digital Commons@Becker. It has been accepted for inclusion in Open Access Publications by an authorized administrator of Digital Commons@Becker. For more information, please contact [email protected]. Comparison of the Host Responses to Wild-Type and cpsB Mutant Klebsiella pneumoniae Infections Matthew S. Lawlor, Scott A. Handley and Virginia L. Miller Infect. Immun. 2006, 74(9):5402. DOI: 10.1128/IAI.00244-06. Downloaded from Updated information and services can be found at: http://iai.asm.org/content/74/9/5402 http://iai.asm.org/ These include: REFERENCES This article cites 31 articles, 21 of which can be accessed free at: http://iai.asm.org/content/74/9/5402#ref-list-1 CONTENT ALERTS Receive: RSS Feeds, eTOCs, free email alerts (when new on March 10, 2014 by Washington University in St. Louis articles cite this article), more» Information about commercial reprint orders: http://journals.asm.org/site/misc/reprints.xhtml To subscribe to to another ASM Journal go to: http://journals.asm.org/site/subscriptions/ INFECTION AND IMMUNITY, Sept. 2006, p. 5402–5407 Vol. 74, No. 9 0019-9567/06/$08.00ϩ0 doi:10.1128/IAI.00244-06 Copyright © 2006, American Society for Microbiology. All Rights Reserved. Comparison of the Host Responses to Wild-Type and cpsB Mutant Klebsiella pneumoniae Infections Matthew S. Lawlor,1 Scott A. Handley,1 and Virginia L. Miller1,2* Departments of Molecular Microbiology1 and Pediatrics,2 Washington University School of Medicine, St. Louis, Missouri 63110 Downloaded from Received 14 February 2006/Returned for modification 28 March 2006/Accepted 14 June 2006 Previously, we established an intranasal mouse model of Klebsiella pneumoniae infection and validated its utility using a highly virulent wild-type strain and an avirulent capsular polysaccharide mutant. In the present study we compare the host responses to both infections by examining cytokine production, cellular infiltration, pulmonary histology, and intranasal immunization. Klebsiella pneumoniae has been known as an important choosing few time points for comparison, or using atypical http://iai.asm.org/ cause of gram-negative bacterial pneumonia for more than 100 inoculation routes. Two studies in particular have detailed the years. The best-characterized virulence factor of this species is difference in cytokine production between a wild-type infection capsular polysaccharide, a voluminous layer of acidic sugar and one caused by a spontaneous capsule-deficient strain and polymers responsible for a variety of K. pneumoniae pheno- examined the production of a number of cytokines during each types, including protection against complement-mediated kill- infection (30, 32). Similar levels of production of TNF-␣, IL- ing and inhibition of macrophage phagocytosis (6, 11). Capsule 1␤, and IL-6 were seen in both bronchoalveolar lavage (BAL) has also been investigated using a number of different animal and serum samples at most time points in both infected groups. models for its importance in K. pneumoniae pathogenesis. Iso- Interestingly, the level of IL-6 production was significantly on March 10, 2014 by Washington University in St. Louis genic capsule mutant strains are deficient for causing disease in higher in BAL fluid from capsule mutant infected mice than mice via intraurethral and intratracheal inoculation routes from wild-type infected mice (32). Further investigation dem- (6, 24). onstrated that the capsule-deficient strain induced an early A substantial amount of research over the last 20 years has peak of IFN-␥ production that was lacking in the wild-type focused on the importance of cytokine production in the host infected mice (30). Instead, the wild-type infection induced defense against K. pneumoniae pulmonary infection. Nearly all higher levels of IL-10 production, and it was postulated that of these studies have examined infection of a wild-type strain capsule serves to induce the production of IL-10, which helps after intratracheal inoculation and compared outcomes in both to suppress the host inflammatory response and allow the wild-type and immunodeficient mice (23, 26). Among the host bacteria to grow in a more permissive environment. factors that have been investigated include tumor necrosis fac- Our initial experiments in the establishment of this intrana- ␣ tor alpha (TNF- ), interleukin-12 (IL-12), IL-17, and macro- sal model demonstrated that a cpsB mutant of K. pneumoniae ␣ ␣ phage inflammatory protein 1 (MIP-1 ) (12, 29). These ex- was severely attenuated in its ability to cause disease (18). This periments have helped to reinforce the importance of the cpsB mutant (VK20) appears to be wild type in its production inflammatory response in clearing K. pneumoniae infection and of lipopolysaccharide (LPS) and enterobacterial common an- have provided substantial evidence for the protective role of a tigen, two other surface polysaccharides of K. pneumoniae (18). T 1-mediated response. Any interference with a rapid host H All infected mice survived intranasal challenge with as many as response (e.g., suppression of the proinflammatory cytokines) 9 ϫ 108 CFU of the cpsB mutant strain. Furthermore, when leads to a more severe disease process (16). Conversely, aug- inoculated at a dose equivalent to the wild-type 50% lethal menting the immune response with exogenous inflammatory dose (LD ), 104 CFU, the cpsB mutant bacteria were cleared mediators decreases the morbidity and mortality associated 50 from all tissues by 48 h. All future infections were performed with infection (23). with doses that would result in similar concentrations of bac- Despite the importance of capsular polysaccharide for the teria in the lungs of infected mice at the 12-h time point, pathogenesis of K. pneumoniae, few studies have compared the host responses against wild-type and well-defined capsule mu- allowing for a more relevant comparison between infections ϳ 7 ϳ 4 tant strains. In examining the previous literature for in vivo ( 10 CFU of the cpsB mutant strain or 10 CFU of the studies of capsule mutant infections, only a few of these have wild-type KPPR1 strain) (18). However, bacterial concentra- included immunological assays as part of their comparison tions in the lungs increasingly diverge after the 12-h time point, (Table 1). Many of these studies feature one or more deficien- with the wild-type infection peaking at 72 h postinfection with cies, including the use of spontaneous capsule mutant strains, ensuing lethality. In contrast, the cpsB mutant is unable to grow in either the trachea or lungs and fails to disseminate to the spleen. However, small numbers of this attenuated strain can persist in the lungs for several days (18). * Corresponding author. Mailing address: Department of Molecular Despite past achievements in elucidating the host response Microbiology, Campus Box 8230, 660 S. Euclid Ave., St. Louis, MO 63110. Phone: (314) 286-2891. Fax: (314) 286-2896. E-mail: virginia to K. pneumoniae infection, we believe that our comprehensive @borcim.wustl.edu. model can be used in concert with our defined capsule mutant 5402 VOL. 74, 2006 NOTES 5403 TABLE 1. Comparison of in vivo studies with capsule mutants Type of cps Capsule Infection Tissue(s); time point(s)d LD e Immunological assay(s)f Reference mutationa productionb routec 50 UV, HS Precipitin i.p. None Y None 1 Sp Cell vol i.p. None Y None 25 Sp India ink i.t. Lung; 1–28 days Y Lung histology 10 Sp None Burn Skin, blood, liver; 12 h–6 days Y None 8 Sp Swelling s.c. Skin abscess; 24 h N None 22 UV ELISA i.u. Bladder, urine, kidney; 5 days N None 2 Tn Immunoblot i.p. None Y None 3 Downloaded from In Uronic acid i.g. Intestine; 1–20 days N Intestine histology 11 Sp India ink i.n. Lung, serum; 6–72 h Y Lung histology, BAL, and 32 serum cytokines Sp India ink i.n. None N BAL and serum cytokines 31 In ELISA i.t. Lung; 7 days N Lung histology 6 Sp Hydrophobicity i.t. Lung; 4–14 days N Lung histology 4 Sp Swelling i.g., i.u. Intestine, bladder; 1–10 days N None 24 Tn Uronic acid i.n. Trachea, lung, spleen; 12–72 h Y Lung histology 18 a The type of capsule mutation used in the study. UV, induced by UV light; Sp, spontaneous mutation; Tn, transposon insertion mutant; In, insertional mutation; http://iai.asm.org/ HS, induced by high salt concentrations. b The method used to monitor capsule production. ELISA, enzyme-linked immunosorbent assay. c The route of inoculation for the animal studies. i.p., intraperitoneal; s.c., subcutaneous; i.u., intraurethral; i.g., intragastric; i.n., intranasal; i.t., intratracheal. d The types of tissues examined and the time frame of the study. e Whether the LD50 was either determined in the study. Y, yes; N, no. f The type of immunological assay(s) (if any) performed in the study. to bring further insight to this interesting area of research. In tissues with the highest bacterial counts also had the most on March 10, 2014 by Washington University in St.
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