Abstract
There has been a strong, positive correlation between opisthorchiasis-associated cholangiocarcinoma and infection with Helicobacter. Here a rodent model of human infection with Opisthorchis viverrini was utilized to further investigate relationships of apparent co-infections with O. viverrini and H. pylori. A total of 150 hamsters were assigned to five groups: i) Control hamsters not infected with O. viverrini; ii) O. viverrini-infected hamsters; iii) non-O. viverrini infected hamsters treated with antibiotics (ABx); iv) O. viverrini-infected hamsters treated with ABx; and v) O. viverrini-infected hamsters treated both with ABx and praziquantel (PZQ). Stomach, gallbladder, liver, colonic tissue, colorectal feces and O. viverrini worms were collected and the presence of species of Helicobacter determined by PCR-based approaches. In addition, O. viverrini worms were cultured in vitro with and without ABx for four weeks, after which the presence of Helicobacter spp. was determined. In situ localization of H. pylori and Helicobacter-like species was performed using a combination of histochemistry and immunohistochemistry. The prevalence of H. pylori infection in O. viverrini-infected hamsters was significantly higher than that of O. viverrini-uninfected hamsters (p≤0.001). Interestingly, O. viverrini-infected hamsters treated with ABx and PZQ (to remove the flukes) had a significantly lower frequency of H. pylori than either O. viverrini- infected hamsters treated only with ABx or O. viverrini-infected hamsters, respectively (p≤0.001). Quantitative RT-PCR strongly confirmed the correlation between intensity H. pylori infection and the presence of liver fluke infection. In vitro, H. pylori could be detected in the O. viverrini worms cultured with ABx over four weeks. In situ localization revealed H. pylori and other Helicobacter-like bacteria in worm gut. The findings indicate that the liver fluke O. viverrini in the biliary tree of the hamsters harbors H. pylori and Helicobacter-like bacteria. Accordingly, the association between O. viverrini and H. pylori may be an obligatory mutualism.
Highlights
Cholangiocarcinoma (CCA) is highly prevalent in Asian countries, in Thailand, China, Taiwan and Korea (Shin et al, 2010)
A total of 150 hamsters were assigned to five groups: i) Control hamsters not infected with O. viverrini; ii) O. viverrini-infected hamsters; iii) non-O. viverrini infected hamsters treated with antibiotics (ABx); iv) O. viverrini-infected hamsters treated with ABx; and v) O. viverrini-infected hamsters treated both with ABx and praziquantel (PZQ)
In situ localization of H. pylori and Helicobacter-like species was performed using a combination of histochemistry and immunohistochemistry
Summary
Cholangiocarcinoma (CCA) is highly prevalent in Asian countries, in Thailand, China, Taiwan and Korea (Shin et al, 2010). Thailand has the highest incidence of CCA worldwide due to its robust association with opisthorchiasis (Sripa and Pairojkul, 2008; Shin et al, 2010). Helicobacter pylori infection was the first bacterial infection known to be a causative factor of gastrointestinal diseases including gastric adenocarcinoma (Bouvard et al, 2009). Its virulence factors such as cagA and vacA are involved in the pathogenesis of the diseases (Cid et al, 2013; Hatakeyama, 2014). Other Helicobacter species, H. hepaticus and H. bilis are implicated in hepatobiliary disease (Zhou et al, 2013; MateosMuñoz et al, 2013; Murphy et al, 2014). We observed an association between H. pylori and hepatolithiasis or CCA in people in Northeast Thailand, a region endemic for opisthorchiasis (Boonyanugomol et al, 2012b; 2012c).
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