Detection of intimins α, β, γ, and δ, four intimin derivatives expressed by attaching and effacing microbial pathogens

被引:211
作者
Adu-Bobie, J
Frankel, G
Bain, C
Goncalves, AG
Trabulsi, LR
Douce, G
Knutton, S
Dougan, G
机构
[1] Univ London Imperial Coll Sci Technol & Med, Dept Biochem, London SW7 2AZ, England
[2] Univ Birmingham, Inst Child Hlth, Birmingham B16 8ET, W Midlands, England
[3] Univ Sao Paulo, Inst Ciencias Biomed, Dept Microbiol, BR-05508 Sao Paulo, Brazil
基金
英国惠康基金;
关键词
D O I
10.1128/JCM.36.3.662-668.1998
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Intimins are outer membrane proteins expressed by enteric bacterial pathogens capable of inducing intestinal attachment-and-effacement lesions. A eukaryotic cell-binding domain is located within a 280-amino-acid (Int280) carboxy terminus of intimin polypeptides. Polyclonal antiserum was raised against Int280 from enteropathogenic Escherichia coli (EPEC) serotypes O127:H6 and O114:H2 (anti-Int280-H6 and anti-Int-280-H2, respectively), and Western blot analysis was used to explore the immunological relationship between the intimin polypeptides expressed by different clinical EPEC and enterohemorrhagic E. coli (EHEC) isolates, a rabbit diarrheagenic E. coli strain (RDEC-1), and Citrobacter rodentium. Anti-Int280-H6 serum reacted strongly with some EPEC serotypes, whereas anti-Int280-H2 serum reacted strongly with strains belonging to different EPEC and EHEC serotypes, RDEC-1, and C. rodentium. These observations were confirmed by using purified Int280 in an enzyme-linked immunosorbent assay and by immunogold and immunofluorescence labelling of whole bacterial cells. Some bacterial strains were recognized poorly by either antiserum (e.g., EPEC O86:H34 and EHEC O157:H7). By using PCR primers designed on the basis of the intimin-encoding eae gene sequences of serotype O127:H6, O114:H2, and O86:H34 EPEC and serotype O157:H7 EHEC, we could distinguish between different eae gene derivatives. Accordingly, the different intimin types were designated alpha, beta, delta, and gamma, respectively.
引用
收藏
页码:662 / 668
页数:7
相关论文
共 49 条
[1]   Characterization of the eaeA gene from rabbit enteropathogenic Escherichia coli strain RDEC-1 and comparison to other eaeA genes from bacteria that cause attaching-effacing lesions [J].
Agin, TS ;
Cantey, JR ;
Boedeker, EC ;
Wolf, MK .
FEMS MICROBIOLOGY LETTERS, 1996, 144 (2-3) :249-258
[2]   Identification of a family of intimins common to Escherichia coli causing attaching-effacing lesions in rabbits, humans and swine [J].
Agin, TS ;
Wolf, MK .
INFECTION AND IMMUNITY, 1997, 65 (01) :320-326
[3]   SHARING OF VIRULENCE-ASSOCIATED PROPERTIES AT THE PHENOTYPIC AND GENETIC LEVELS BETWEEN ENTEROPATHOGENIC ESCHERICHIA-COLI AND HAFNIA-ALVEI [J].
ALBERT, MJ ;
FARUQUE, SM ;
ANSARUZZAMAN, M ;
ISLAM, MM ;
HAIDER, K ;
ALAM, K ;
KABIR, I ;
ROBINSBROWNE, R .
JOURNAL OF MEDICAL MICROBIOLOGY, 1992, 37 (05) :310-314
[4]  
BARTHOLD SW, 1976, LAB ANIM SCI, V26, P889
[5]  
BEEBAKHEE G, 1992, FEMS MICROBIOL LETT, V91, P63, DOI 10.1016/0378-1097(92)90563-4
[6]  
BURNETTE WN, 1981, ANAL BIOCHEM, V112, P195, DOI 10.1016/0003-2697(81)90281-5
[7]   INHIBITION OF ENTEROPATHOGENIC ESCHERICHIA-COLI (EPEC) ADHESION TO HELA-CELLS BY HUMAN COLOSTRUM - DETECTION OF SPECIFIC SIGA RELATED TO EPEC OUTER-MEMBRANE PROTEINS [J].
CAMARA, LM ;
CARBONARE, SB ;
SILVA, MLM ;
CARNEIROSAMPAIO, MMS .
INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY, 1994, 103 (03) :307-310
[8]   DIARRHEA DUE TO ESCHERICHIA-COLI IN RABBIT - NOVEL MECHANISM [J].
CANTEY, JR ;
BLAKE, RK .
JOURNAL OF INFECTIOUS DISEASES, 1977, 135 (03) :454-462
[9]  
Carneiro-Sampaio MMS, 1996, REV MICROBIOL, V27, P120
[10]   ROLE OF THE EAEA GENE IN EXPERIMENTAL ENTEROPATHOGENIC ESCHERICHIA-COLI INFECTION [J].
DONNENBERG, MS ;
TACKET, CO ;
JAMES, SP ;
LOSONSKY, G ;
NATARO, JP ;
WASSERMAN, SS ;
KAPER, JB ;
LEVINE, MM .
JOURNAL OF CLINICAL INVESTIGATION, 1993, 92 (03) :1412-1417