Abstract:
:Newly recognized Fic family virulence proteins may be important in many bacterial pathogens. To relate cellular mechanisms to pathogenesis and immune protection, we studied the cytotoxicity of the Histophilus somni immunoglobulin-binding protein A (IbpA) direct repeat 2 Fic domain (DR2/Fic) for natural host target cells. Live virulent IbpA-producing H. somni strain 2336, a cell-free culture supernatant (CCS) of this strain, or recombinant DR2/Fic (rDR2/Fic) caused dramatic retraction and rounding of bovine alveolar type 2 (BAT2) epithelial cells. IbpA-deficient H. somni strain 129Pt and a Fic motif His(298)Ala mutant rDR2/Fic protein were not cytotoxic. The cellular mechanism of DR2/Fic cytotoxicity was demonstrated by incubation of BAT2 cell lysates with strain 2336 CCS or rDR2/Fic in the presence of [alpha-(32)P]ATP, which resulted in adenylylation of Rho GTPases and cytoskeletal disruption. Since IbpA is not secreted by type III or type IV secretion systems, we determined whether DR2/Fic entered the host cytoplasm to access its Rho GTPase targets. Although H. somni did not invade BAT2 cells, DR2/Fic was internalized by cells treated with H. somni, CCS, or the rDR2/Fic protein, as shown by confocal immunomicroscopy. Transwell bacterial migration assays showed that large numbers of strain 2336 bacteria migrated between retracted BAT2 cells, but IbpA-deficient strain 129Pt did not cross a monolayer unless the monolayer was pretreated with strain 2336 CCS or rDR2/Fic protein. Antibody to rDR2/Fic or passively protective convalescent-phase serum blocked IbpA-mediated cytotoxicity and inhibited H. somni transmigration across BAT2 monolayers, confirming the role of DR2/Fic in pathogenesis and corresponding to the results for in vivo protection in previous animal studies.
journal_name
Infect Immunjournal_title
Infection and immunityauthors
Zekarias B,Mattoo S,Worby C,Lehmann J,Rosenbusch RF,Corbeil LBdoi
10.1128/IAI.01277-09subject
Has Abstractpub_date
2010-05-01 00:00:00pages
1850-8issue
5eissn
0019-9567issn
1098-5522pii
IAI.01277-09journal_volume
78pub_type
杂志文章abstract::Lack of efficacious vaccines against Mycobacterium tuberculosis (MTB) infection is a limiting factor in the prevention and control of tuberculosis (TB), the leading cause of death from an infectious agent. Improvement or replacement of the BCG vaccine with one that reliably protects all age-groups is urgent. Concerns ...
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journal_title:Infection and immunity
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doi:10.1128/IAI.63.6.2352-2355.1995
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journal_title:Infection and immunity
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doi:10.1128/IAI.11.1.14-22.1975
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journal_title:Infection and immunity
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journal_title:Infection and immunity
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doi:10.1128/IAI.22.2.480-485.1978
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journal_title:Infection and immunity
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doi:10.1128/IAI.63.1.301-308.1995
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journal_title:Infection and immunity
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journal_title:Infection and immunity
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journal_title:Infection and immunity
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doi:10.1128/IAI.62.11.5075-5084.1994
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journal_title:Infection and immunity
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doi:10.1128/IAI.2.5.686-688.1970
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journal_title:Infection and immunity
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doi:10.1128/IAI.32.2.415-419.1981
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journal_title:Infection and immunity
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更新日期:2005-05-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.64.6.2295-2299.1996
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journal_title:Infection and immunity
pub_type: 杂志文章
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journal_title:Infection and immunity
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.06140-11
更新日期:2012-05-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.65.4.1258-1266.1997
更新日期:1997-04-01 00:00:00