Identification of a Campylobacter jejuni protein that cross-reacts with cholera toxin.
The question of whether Campylobacter jejuni produces a cholera toxin-like toxin (CTLT) has been controversial. The objective of this study was to identify the factor that cross-reacts with CT from C. jejuni. Filtrates of C. jejuni grown in four different liquid media reported to promote CTLT production were tested by Chinese hamster ovary (CHO) cell elongation assay and for reactivity with CT antibody using GM1 ganglioside enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Protein sequence was determined by matrix-assisted laser desorption ionization-time of flight (MALDI TOF-TOF). Filtrates from seven reference strains reported to produce CTLT and from 80 clinical strains were negative in the CHO cell assay, but those from three reference strains and 16 clinical strains were positive by GM1 ELISA. All strains tested, including C. jejuni NCTC 11168, which does not contain a CT gene homologue, possessed a 53-kDa protein which reacted with CT antibody by immunoblotting. This band was identified as the major outer membrane protein, PorA, of C. jejuni. CT antibody reacted by immunoblotting with a recombinant PorA, but antibody to the recombinant PorA did not react with CT. Our results indicate that C. jejuni does not produce a functional CTLT, but the reactivity of PorA with CT antibody would lead to the erroneous conclusion that C. jejuni produces a functional CTLT.
Albert
M John
MJ
Department of Microbiology, Faculty of Medicine, Kuwait University, Safat 13110, Kuwait. john@hsc.edu.kw
Haridas
Shilpa
S
Steer
David
D
Dhaunsi
Gursev S
GS
Smith
A Ian
AI
Adler
Ben
B
D000818:Animals; D001427:Bacterial Toxins / Q000302:isolation & purification / Q000378:metabolism*; D016466:CHO Cells; D016123:Campylobacter jejuni / Q000302:isolation & purification / Q000378:metabolism*; D002772:Cholera Toxin / Q000276:immunology / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003429:Cross Reactions; D003603:Cytotoxins / Q000096:biosynthesis* / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D007082:Ileum / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley
D001427:Bacterial Toxins; D003603:Cytotoxins; D002772:Cholera Toxin