About: The protective effect of recombinant Lactococcus lactis oral vaccine on a Clostridium difficile-infected animal model

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About: The protective effect of recombinant Lactococcus lactis oral vaccine on a Clostridium difficile-infected animal model Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	The protective effect of recombinant Lactococcus lactis oral vaccine on a Clostridium difficile-infected animal model
Creator	Jiang, Bo Chen, Xue-Qing Sun, Da-Yong Yang, Xiao-Qiang Zhao, Ya-Gang
Source	Medline; PMC
abstract	BACKGROUND: Oral immunization with vaccines may be an effective strategy for prevention of Clostridium difficile infection (CDI). However, application of previously developed vaccines for preventing CDI has been limited due to various reasons. Here, we developed a recombinant Lactococcus lactis oral vaccine and evaluated its effect on a C. difficile-infected animal model established in golden hamsters in attempt to provide an alternative strategy for CDI prevention. METHODS: Recombinant L. lactis vaccine was developed using the pTRKH2 plasmid, a high-copy-number Escherichia coli-L. shuttle vector: 1) L. lactis expressing secreted proteins was constructed with recombinant pTRKH2 (secreted-protein plasmid) carrying the Usp45 signal peptide (SPUsp45), nontoxic adjuvanted tetanus toxin fragment C (TETC), and 14 of the 38 C-terminal repeats (14CDTA) of nontoxic C. difficile toxin A (TcdA); and 2) L. lactis expressing secreted and membrane proteins was constructed with recombinant pTRKH2 (membrane-anchored plasmid) carrying SPUsp45, TETC, 14CDTA, and the cell wall-anchored sequence of protein M6 (cwaM6). Then, 32 male Syrian golden hamsters were randomly divided into 4 groups (n = 8 each) for gavage of normal saline (blank control) and L. lactis carrying the empty shuttle vector, secreted-protein plasmid, and membrane-anchored plasmid, respectively. After 1-week gavage of clindamycin, the animals were administered with C. difficile spore suspension. General symptoms and intestinal pathological changes of the animals were examined by naked eye and microscopy, respectively. Protein levels of anti-TcdA IgG/IgA antibodies in intestinal tissue and fluid were analyzed by enzyme-linked immunosorbent assay (ELISA). A cell culture cytotoxicity neutralization assay was done by TcdA treatment with or without anti-TcdA serum pre-incubation or treatment. Apoptosis of intestinal epithelial cells was examined by flow cytometry (FL) assay. Expression of mucosal inflammatory cytokines in the animals was detected by polymer chain reaction (PCR) assay. RESULTS: After the C. difficile challenge, the animals of control group had severe diarrhea symptoms on day 1 and all died on day 4, indicating that the CDI animal model was established in hamster. Of the 3 immunization groups, secreted-protein and membrane-anchored plasmid groups had significantly lower mortalities, body weight decreases, and pathological scores, with higher survival rate/time than the empty plasmid group (P < 0.05). The tilter of IgG antibody directed against TcdA was significantly higher in serum and intestinal fluid of secreted-protein and membrane-anchored plasmid groups than in the empty plasmid group (P < 0.05) while the corresponding titer of IgA antibody directed against TcdA had no substantial differences (P > 0.05). The anti-TcdA serum of membrane-anchored plasmid group neutralized the cytotoxicity of 200 ng/ml TcdA with the best protective effect achieved by anti-TcdA serum pre-incubation. The incidences of TcdA-induced death and apoptosis of intestinal epithelial cells were significantly reduced by cell pre-incubation or treatment with anti-TcdA serum of membrane-anchored plasmid group (P < 0.05). MCP-1, ICAM-1, IL-6, and Gro-1 mRNA expression levels were the lowest in cecum tissue of the membrane-anchored groups compared to the other groups. CONCLUSION: Recombinant L. lactis live vaccine is effective for preventing CDI in the hamster model, thus providing an alternative for immunization of C. difficile-associated diseases.
has issue date	2013-07-17(xsd:dateTime)
bibo:doi	10.1186/1471-230x-13-117
bibo:pmid	23865596
has license	cc-by
sha1sum (hex)	ec2b2544b77c203db7fb59fbcb3f2c12733685e1
schema:url	https://doi.org/10.1186/1471-230x-13-117
resource representing a document's title	The protective effect of recombinant Lactococcus lactis oral vaccine on a Clostridium difficile-infected animal model
has PubMed Central identifier	PMC3750240
has PubMed identifier	23865596
schema:publication	BMC Gastroenterol
resource representing a document's body	covid:ec2b2544b77c203db7fb59fbcb3f2c12733685e1#body_text
is schema:about of	named entity 'Here' named entity 'polymer' named entity 'fragment' named entity 'cell' named entity 'carrying' named entity 'IgG' named entity 'CDI' named entity 'application' named entity 'clindamycin' named entity 'golden' named entity 'detected' named entity 'Background' covid:arg/ec2b2544b77c203db7fb59fbcb3f2c12733685e1 named entity 'cell culture' named entity 'male' named entity 'C. difficile' named entity 'divided' named entity 'groups' named entity 'recombinant' named entity 'secreted' named entity 'assay' named entity 'plasmid' named entity 'week' named entity 'Lactococcus lactis' named entity 'chain reaction' named entity 'serum' named entity 'vaccines' named entity 'tetanus toxin' named entity 'clindamycin' named entity 'assay' named entity 'plasmid' named entity 'assay' named entity 'assay' named entity 'inflammatory cytokines' named entity 'cytotoxicity' named entity 'copy-number' named entity 'Escherichia coli' named entity 'cell culture' named entity 'CDI' named entity 'CDI' named entity 'membrane proteins' named entity 'plasmid' named entity 'normal saline' named entity 'adjuvanted' named entity 'recombinant' named entity 'protective effect' named entity 'homogenates' named entity 'control group' named entity 'C. difficile' named entity 'serum' named entity 'light microscope' named entity 'ICAM-1' named entity 'antibiotic' named entity 'C. difficile' named entity 'immunization' named entity 'SPSS' named entity 'survival time' named entity 'inflammatory cytokines' named entity 'hydrolyzate' named entity 'protein' named entity 'body weight' named entity 'C-terminal' named entity 'protein' named entity 'ANOVA' named entity 'cecum' named entity 'Guangzhou' named entity 'plasmid' named entity 'neutrophils' named entity 'Clostridium difficile' named entity 'animal models' named entity 'C. difficile' named entity 'L. lactis' named entity 'China' named entity 'Cybernetics' named entity 'fusion protein' named entity 'intestinal mucosa' named entity 'adjuvanted'

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