Abstract:
:A new strategy for rapidly selecting and testing genetic vaccines has been developed, in which a whole genome library is cloned into a bacteriophage lambda ZAP Express vector which contains both prokaryotic (P(lac)) and eukaryotic (P(CMV)) promoters upstream of the insertion site. The phage library is plated on Escherichia coli cells, immunoblotted, and probed with hyperimmune and/or convalescent-phase antiserum to rapidly identify vaccine candidates. These are then plaque purified and grown as liquid lysates, and whole bacteriophage particles are then used directly to immunize the host, following which P(CMV)-driven expression of the candidate vaccine gene occurs. In the example given here, a semirandom genome library of the bovine pathogen Mycoplasma mycoides subsp. mycoides small colony (SC) biotype was cloned into lambda ZAP Express, and two strongly immunodominant clones, lambda-A8 and lambda-B1, were identified and subsequently tested for vaccine potential against M. mycoides subsp. mycoides SC biotype-induced mycoplasmemia. Sequencing and immunoblotting indicated that clone lambda-A8 expressed an isopropyl-beta-d-thiogalactopyranoside (IPTG)-inducible M. mycoides subsp. mycoides SC biotype protein with a 28-kDa apparent molecular mass, identified as a previously uncharacterized putative lipoprotein (MSC_0397). Clone lambda-B1 contained several full-length genes from the M. mycoides subsp. mycoides SC biotype pyruvate dehydrogenase region, and two IPTG-independent polypeptides, of 29 kDa and 57 kDa, were identified on immunoblots. Following vaccination, significant anti-M. mycoides subsp. mycoides SC biotype responses were observed in mice vaccinated with clones lambda-A8 and lambda-B1. A significant stimulation index was observed following incubation of splenocytes from mice vaccinated with clone lambda-A8 with whole live M. mycoides subsp. mycoides SC biotype cells, indicating cellular proliferation. After challenge, mice vaccinated with clone lambda-A8 also exhibited a reduced level of mycoplasmemia compared to controls, suggesting that the MSC_0397 lipoprotein has a protective effect in the mouse model when delivered as a bacteriophage DNA vaccine. Bacteriophage-mediated immunoscreening using an appropriate vector system offers a rapid and simple technique for the identification and immediate testing of putative candidate vaccines from a variety of pathogens.
journal_name
Infect Immunjournal_title
Infection and immunityauthors
March JB,Jepson CD,Clark JR,Totsika M,Calcutt MJdoi
10.1128/IAI.74.1.167-174.2006keywords:
subject
Has Abstractpub_date
2006-01-01 00:00:00pages
167-74issue
1eissn
0019-9567issn
1098-5522pii
74/1/167journal_volume
74pub_type
杂志文章abstract::We identified nonhemolytic mutants of Listeria monocytogenes that were severely deficient in their ability to invade mammalian nonprofessional phagocytes. These mutants were generated spontaneously or by means of transposon Tn916 mutagenesis. In terms of their extracellular proteins, the noninvasive mutants were defic...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.58.12.3988-3995.1990
更新日期:1990-12-01 00:00:00
abstract::Herpes simplex virus (HSV) infection of mouse liver after partial hepatectomy was studied. Partial hepatectomy resulted in the rapid onset of cellular DNA synthesis and the appearance of many mitotic figures (peak, 3 days after surgery). Similar changes were not seen in control animals. After partial hepatectomy, the ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.42.1.402-408.1983
更新日期:1983-10-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.01943-06
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.65.11.4682-4689.1997
更新日期:1997-11-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.48.2.560-564.1985
更新日期:1985-05-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.73.8.4676-4683.2005
更新日期:2005-08-01 00:00:00
abstract::Gram-negative organisms incorporate hydroxy fatty acids into the lipid A moiety of lipopolysaccharide (LPS), and in the case of some members of the family Enterobacteriaceae, hydroxy fatty acids are incorporated exclusively into lipid A. However, a limited number of Bacteroides species have been shown to incorporate s...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.62.9.3753-3760.1994
更新日期:1994-09-01 00:00:00
abstract::The chlamydial inclusion occupies a unique niche within the eukaryotic cell that does not interact with endocytic compartments but instead is fusogenic with a subset of sphingomyelin-containing exocytic vesicles. The Chlamydia trachomatis inclusion acquires these distinctive properties by as early as 2 h postinfection...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/iai.71.2.973-984.2003
更新日期:2003-02-01 00:00:00
abstract::Female C57BL/6J mice were infected with Trypanosoma cruzi and subsequently given macrophages or lymphocytes from syngeneic donors which had recovered from the acute infection. Mice which received immune peritoneal macrophages, splenic lymphocytes, or lymph node lymphocytes developed lower mean parasitemias and cumulat...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.25.3.838-843.1979
更新日期:1979-09-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.57.1.41-47.1989
更新日期:1989-01-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/iai.68.1.288-293.2000
更新日期:2000-01-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.72.8.4351-4356.2004
更新日期:2004-08-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.62.5.1940-1945.1994
更新日期:1994-05-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.60.8.3238-3243.1992
更新日期:1992-08-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.21.1.310-319.1978
更新日期:1978-07-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.7.6.918-921.1973
更新日期:1973-06-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.66.8.3841-3847.1998
更新日期:1998-08-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.11.2.330-333.1975
更新日期:1975-02-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.64.8.3174-3179.1996
更新日期:1996-08-01 00:00:00
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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
pub_type: 杂志文章
doi:10.1128/IAI.69.3.1364-1372.2001
更新日期:2001-03-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
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更新日期:2006-04-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
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更新日期:2010-01-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.60.2.392-400.1992
更新日期:1992-02-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.63.2.724-728.1995
更新日期:1995-02-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
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更新日期:2008-05-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.6.3.355-363.1972
更新日期:1972-09-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.49.3.498-504.1985
更新日期:1985-09-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.40.2.629-632.1983
更新日期:1983-05-01 00:00:00