Behring Institute Mitteilungen最新文献_第6页

Construction of a Mycoplasma penetrans expression library in E. coli using the two-phase expression system. 用两相表达系统构建支原体在大肠杆菌中的表达文库。

Behring Institute Mitteilungen Pub Date : 1997-02-01

Z X Yan, J Andreev, S Rottem, T F Meyer

引用次数: 0

A hybrid outer membrane protein antigen for vaccination against Pseudomonas aeruginosa. 一种用于铜绿假单胞菌疫苗的杂交外膜蛋白抗原。

Behring Institute Mitteilungen Pub Date : 1997-02-01

J Gabelsberger, B Knapp, S Bauersachs, U I Enz, B U von Specht, H Domdey

{"title":"A hybrid outer membrane protein antigen for vaccination against Pseudomonas aeruginosa.","authors":"J Gabelsberger, B Knapp, S Bauersachs, U I Enz, B U von Specht, H Domdey","doi":"","DOIUrl":"","url":null,"abstract":"Recently a hybrid protein containing parts of the outer membrane proteins OprF (aa 190-342) and OprI (aa 21-83) from Pseudomonas aeruginosa fused to the glutathione-S-transferase was shown to protect mice against a 975-fold 50% lethal dose of P. aeruginosa. To omit the use of the GST-protein, the hybrid protein OprF-OprI was expressed in E. coli using distinct modifications which have not to be eliminated after its expression. Using different signal peptides, the yield of the hybrid protein OprF-OprI in E. coli could be increased to 30% of the total cell protein, however, only a very small amount of the hybrid preprotein was processed and could be isolated from the periplasm of the host. A construct containing an N-terminal extension of 11 amino acids from the original OprF gene gave rise to a significantly higher expression in the cytoplasm. Purification was facilitated by the addition of a five histidine tag at the C-terminus. An even higher expression was obtained by a construct in which a six histidine tag was attached to the N-terminus of the hybrid protein. The N-terminal extended OprF-OprI as well as the N-terminal his-tagged OprF-OprI hybrid antigens were purified by immobilized-metal affinity chromatography under native and denaturing conditions and can now be tested for protectivity against P. aeruginosa in animal model systems.","PeriodicalId":8816,"journal":{"name":"Behring Institute Mitteilungen","volume":" 98","pages":"302-14"},"PeriodicalIF":0.0,"publicationDate":"1997-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20312440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The Pseudomonas aeruginosa outer membrane protein I vaccine: immunogenicity and safe administration in man. 铜绿假单胞菌外膜蛋白I疫苗:免疫原性和安全给药。

Behring Institute Mitteilungen Pub Date : 1997-02-01

B U von Specht, H C Lücking, B Blum, A Schmitt, K D Hungerer, H Domdey

引用次数: 0

Bacterial ghosts as multifunctional vaccine particles. 细菌幽灵作为多功能疫苗颗粒。

Behring Institute Mitteilungen Pub Date : 1997-02-01

M P Szostak, H Mader, M Truppe, M Kamal, F O Eko, V Huter, J Marchart, W Jechlinger, W Haidinger, E Brand, E Denner, S Resch, E Dehlin, A Katinger, B Kuen, A Haslberger, A Hensel, W Lubitz

{"title":"Bacterial ghosts as multifunctional vaccine particles.","authors":"M P Szostak, H Mader, M Truppe, M Kamal, F O Eko, V Huter, J Marchart, W Jechlinger, W Haidinger, E Brand, E Denner, S Resch, E Dehlin, A Katinger, B Kuen, A Haslberger, A Hensel, W Lubitz","doi":"","DOIUrl":"","url":null,"abstract":"Expression of cloned PhiX174 gene E in Gram-negative bacteria results in lysis of the bacteria by formation of an E-specific transmembrane tunnel structure built through the cell envelope complex. Bacterial ghosts have been produced from a variety of bacteria including Escherichia coli. Salmonella typhimurium, Salmonella enteritidis, Vibrio cholerae, Klebsiella pneumoniae, Actinobacillus pleuropneumoniae, Haemophilus influenzae, Pasteurella haemolytica, Pasteurella multocida, and Helicobacter pylori. Such ghosts are used as non-living candidate vaccines and represent an alternative to heat or chemically inactivated bacteria. In recombinant ghosts, foreign proteins can be inserted into the inner membrane prior to E-mediated lysis via specific N-, or C-, or N- and C-terminal anchor sequences. The export of proteins into the periplasmic space or the expression of recombinant S-layer proteins vastly extents the capacity of ghosts or recombinant ghosts as carriers of foreign epitopes or proteins. Oral, aerogenic or parenteral applications of (recombinant) ghosts in experimental animals induced specific humoral and cellular immune responses against bacterial and target components including protective mucosal immunity. The most relevant advantage of ghosts and recombinant bacterial ghosts as immunogens is that no inactivation procedures that denature relevant immunogenic determinants are employed in the production of ghosts used as vaccines or as carriers of relevant antigens. The inserted target antigens into the inner membrane or into S-layer proteins are not limited in size.","PeriodicalId":8816,"journal":{"name":"Behring Institute Mitteilungen","volume":" 98","pages":"191-6"},"PeriodicalIF":0.0,"publicationDate":"1997-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20311206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Immunization of cystic fibrosis patients with a Pseudomonas aeruginosa O-polysaccharide-toxin A conjugate vaccine. 用铜绿假单胞菌 O 型多糖毒素 A 结合疫苗对囊性纤维化患者进行免疫接种。

Behring Institute Mitteilungen Pub Date : 1997-02-01

S J Cryz, A Lang, A Rüdeberg, J Wedgwood, J U Que, E Fürer, U Schaad

{"title":"Immunization of cystic fibrosis patients with a Pseudomonas aeruginosa O-polysaccharide-toxin A conjugate vaccine.","authors":"S J Cryz, A Lang, A Rüdeberg, J Wedgwood, J U Que, E Fürer, U Schaad","doi":"","DOIUrl":"","url":null,"abstract":"Healthy, non-colonized cystic fibrosis (CF) patients (N = 26) were immunized with an octavalent Pseudomonas aeruginosa O-polysaccharide-toxin A conjugate vaccine. Vaccination was well tolerated and induced anti-lipopolysaccharide (LPS) antibodies of a high affinity capable of promoting the opsonophagocytic killing of P. aeruginosa by human peripheral lymphocytes. In contrast, anti-LPS antibodies acquired after natural infection possessed a very low affinity and were non-opsonic. To determine if immunization could prevent or delay infections due to P. aeruginosa, the infection rate among immunized patients was compared retrospectively to age and gender-matched controls. After 6 years of clinical follow-up, 15/20 (75%) of control and 8/23 (35%) of immunized subjects were classified as infected (p = 0.022). The persistence of high-affinity antibodies among immunized patients correlated with a significantly lower rate of infection after 4-6 years of observation. Infection of immunized patients was correlated with a dramatic decline in total antibody titer between year 2 and 3 of follow-up. Smooth, typeable strains of P. aeruginosa predominated among immunized patients. In contrast, rough, nontypeable strains were most frequently isolated from nonimmunized patients. Mucoid P. aeruginosa strains were isolated from 6 nonimmunized patients versus only I immunized subject.","PeriodicalId":8816,"journal":{"name":"Behring Institute Mitteilungen","volume":" 98","pages":"345-9"},"PeriodicalIF":0.0,"publicationDate":"1997-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20314417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Towards targeting strategies for oral immunization--identification of marker antigens in rat M cells. 口腔免疫的靶向策略——大鼠M细胞标记抗原的鉴定。

Behring Institute Mitteilungen Pub Date : 1997-02-01

K Rautenberg, C Cichon, M A Schmidt

{"title":"Towards targeting strategies for oral immunization--identification of marker antigens in rat M cells.","authors":"K Rautenberg, C Cichon, M A Schmidt","doi":"","DOIUrl":"","url":null,"abstract":"Specialized M cells of the follicle-associated epithelium (FAE) of Peyer's patches in the gastrointestinal and respiratory tracts play a crucial role in the immune surveillance of mucosal surfaces and are essential for the induction of mucosal immune responses. These cells transport luminal antigens to the underlying germinal centers and thereby initiate an immune response or induce tolerance. Mucosal immune responses could be significantly enhanced if oral antigens could be directly targeted to the apical surface of M cells. However, thus far M cells have not been isolated and suitable surface markers have not been described. For the characterization and identification of potential molecular markers of M cells the follicle associated epithelium of Peyer's patches from the small intestine of rats was analyzed by indirect immunofluorescence and immunogold electronmicroscopy. The expression of intermediate filament proteins in the FAE was found to be heterogeneous. Immunoreactivity of a specific monoclonal cytokeratin 8 antibody (clone 4.1.18) was selectively found in cells which were additionally characterized by the lack of staining for alkaline phosphatase in their apical membranes. In the FAE this property is an accepted criterium for the presence of M cells. The antigen recognized by the clone 4.1.18 is expressed in substantially higher amounts in M cells of Peyer's patches as compared to neighbouring epithelial cells or \"normal\" enterocytes and thus can be employed as an intracellular molecular marker for M cells of Peyer's patches in rats. It is expected, that this marker will be very helpful for the isolation of M cells.","PeriodicalId":8816,"journal":{"name":"Behring Institute Mitteilungen","volume":" 98","pages":"361-75"},"PeriodicalIF":0.0,"publicationDate":"1997-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20314419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Cloning and sequencing of the gene encoding the outer-membrane protein Tbp1 from Actinobacillus pleuropneumoniae. Expression of Tbp1 and Tbp2. 胸膜肺炎放线杆菌外膜蛋白Tbp1基因的克隆与序列分析。Tbp1和Tbp2的表达。

Behring Institute Mitteilungen Pub Date : 1997-02-01

A Medrano, E Querol, M Daban

引用次数: 0

Pilin-based anti-Pseudomonas vaccines: latest developments and perspectives. 基于匹林的抗假单胞菌疫苗:最新发展和前景。

Behring Institute Mitteilungen Pub Date : 1997-02-01

H Hahn, P M Lane-Bell, L M Glasier, J F Nomellini, W H Bingle, W Paranchych, J Smit

{"title":"Pilin-based anti-Pseudomonas vaccines: latest developments and perspectives.","authors":"H Hahn, P M Lane-Bell, L M Glasier, J F Nomellini, W H Bingle, W Paranchych, J Smit","doi":"","DOIUrl":"","url":null,"abstract":"Among the several adhesins produced by Pseudomonas aeruginosa (Pa), the type-4 pilus promotes the majority of the adherence capability of the bacterium to epithelial cells and it is a major virulence factor in an AB.Y/SnJ mouse infection model. Vaccines targeting the disulfide loop (DSL) adherence binding domain of the pilin protein should therefore provide an effective protection against initial colonization and infection with Pa. To selectively elicit adherence blocking antibodies, the pilin DSL domain was chosen as peptide antigen for the construction of recombinant protein and live vaccines. While synthetic peptide-carrier protein conjugates provided some strain-specific protection, chimeric proteins with N- or C-terminally fused pilin DSL peptides did not engender protective IgG titers mice. Integral fusions of the pilin DSL peptide with the minor coat protein of filamentous phage or surface exposed regions of an outer membrane protein resulted in a display of the peptide on the surface of the phage particles and bacterial cells respectively. However, in immunization studies neither of these live vaccines were effective immunogens. The paracrystalline S-layer of Caulobacter crescentus combines several advantages of an effective antigen surface display system. Recombinant S-layer proteins with singlecopy insertions of a pilin peptide did not engender significant IgG titers, whereas multiple tandem insertions of the same peptide increased the serum IgG response in mice a thousand times. Multiple insertions of DSL peptides from different frequent pilin prototypes may be an interesting alternative for a recombinant cross-protective anti-Pseudomonas vaccine.","PeriodicalId":8816,"journal":{"name":"Behring Institute Mitteilungen","volume":" 98","pages":"315-25"},"PeriodicalIF":0.0,"publicationDate":"1997-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20314413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Bacterial ADP-ribosylating toxins: form, function, and recombinant vaccine development. 细菌adp核糖基化毒素:形式、功能和重组疫苗开发。

Behring Institute Mitteilungen Pub Date : 1997-02-01

W N Burnette

{"title":"Bacterial ADP-ribosylating toxins: form, function, and recombinant vaccine development.","authors":"W N Burnette","doi":"","DOIUrl":"","url":null,"abstract":"No products of the biotechnology revolution will likely have a greater legacy than recombinant vaccines. Clinical efficacy trials of new acellular pertussis vaccines have recently been completed; among them, a vaccine containing a genetically modified pertussis toxin showed superior effectiveness in protection against disease caused by Bordetella pertussis. The foundations for this vaccine derive from the work of many investigators, but most notably: Japanese researchers who demonstrated the potential for subcomponents of B. pertussis, and particularly pertussis toxin, to confer protective immunity; research teams in Italy and the United States who cloned and sequenced the pertussis toxin operon; and our own group who molecularly dissected the toxin molecule to produce recombinant analogs of this heterohexameric protein that retained protective immunogenicity yet lacked the intrinsic enzyme activity that results in the adverse reactogenic effects of immunization. Another result of the research leading to this new pertussis vaccine is an intimate understanding of the relationship between form and function in the ADP-ribosylating toxins with AB5 architecture, including the structure of their catalytic domains their immunologic and adjuvant properties, characteristics and possible pathologic consequences of host cell receptor recognition, and the assembly and subunit interactions of these complex multimeric proteins.","PeriodicalId":8816,"journal":{"name":"Behring Institute Mitteilungen","volume":" 98","pages":"434-41"},"PeriodicalIF":0.0,"publicationDate":"1997-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20311588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Microbiological background for anti-Pseudomonas aeruginosa vaccination in cystic fibrosis. 囊性纤维化中抗铜绿假单胞菌疫苗的微生物学背景。

Behring Institute Mitteilungen Pub Date : 1997-02-01

A Bauernfeind, B Przyklenk

引用次数: 0