Albert M John, Haridas Shilpa, Steer David, Dhaunsi Gursev S, Smith A Ian, Adler Ben
Department of Microbiology, Faculty of Medicine, Kuwait University, Safat 13110, Kuwait.
Infect Immun. 2007 Jun;75(6):3070-3. doi: 10.1128/IAI.00139-07. Epub 2007 Apr 16.
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.
空肠弯曲菌是否产生霍乱毒素样毒素(CTLT)这一问题一直存在争议。本研究的目的是鉴定与空肠弯曲菌CT发生交叉反应的因子。使用中国仓鼠卵巢(CHO)细胞伸长试验,以及采用GM1神经节苷脂酶联免疫吸附测定(ELISA)和免疫印迹法检测空肠弯曲菌在四种据报道可促进CTLT产生的不同液体培养基中生长后的滤液与CT抗体的反应性。通过基质辅助激光解吸电离飞行时间(MALDI TOF-TOF)测定蛋白质序列。据报道能产生CTLT的7株参考菌株和80株临床菌株的滤液在CHO细胞试验中呈阴性,但3株参考菌株和16株临床菌株的滤液通过GM1 ELISA检测呈阳性。所有测试菌株,包括不含有CT基因同源物的空肠弯曲菌NCTC 11168,均拥有一种53 kDa的蛋白质,该蛋白质通过免疫印迹法与CT抗体发生反应。该条带被鉴定为空肠弯曲菌的主要外膜蛋白PorA。CT抗体通过免疫印迹法与重组PorA发生反应,但重组PorA的抗体不与CT发生反应。我们的结果表明,空肠弯曲菌不产生功能性CTLT,但PorA与CT抗体的反应性会导致得出空肠弯曲菌产生功能性CTLT这一错误结论。