- Open Access
Cytokines derived from innate lymphoid cells assist Helicobacter hepaticus to aggravate hepatocellular tumorigenesis in viral transgenic mice
© The Author(s) 2019
- Received: 6 March 2019
- Accepted: 27 April 2019
- Published: 15 May 2019
Recently, intestinal microbiome has been involved in hepatic diseases due to the immunologic and metabolic communication between liver and intestine. Initiation of hepatocellular carcinoma (HCC) frequently attributes to conspiracy between immune cells and infectious carcinogens. Here, the hypothesis that the tumorigenesis of HCC with HBV infection will be aggravated by specific intestinal bacteria was verified in viral transgenic mouse models.
Comparative 16S rRNA sequencing was adopted to observe the intestinal enrichment of Helicobacter hepaticus in HCC. Oral administration of Helicobacter hepaticus was carried out to evaluate its hepatic carcinogenic effect in HBV transgenic mice or wildtype C57BL/6. The livers of experimental mice were collected and examined for the degree of tumorigenesis.
We found that Helicobacter hepaticus more likely colonized at lower colon of HBV-infected mice with HCC, compared with C57BL/6 and HBV-infected mice without neoplasm. Pretreatment of Helicobacter hepaticus in transgenic mice aggravated tumor formation, with higher incidence, more tumor nodule and higher serum AFP. Then, a cytokines expression patterns with inclined IFN-γ, IFN-γR1, IL-17 and IL-23 was found in HBV-infected mice with Helicobacter hepaticus. Furthermore, innate lymphoid cells, especially Th17 and NK cells which can secret IL-17 and IFN-γ respectively, might be recruited by Helicobacter hepaticus cooperated with HBV. Besides, increased expression of CD69, NKG2D and IFN-γ showed activation of cytokine production in intrahepatic NK cells. Finally, IFN-γ decreased E-cadherin expression through p-STAT1 pathway, resulting in epithelial–mesenchymal transition with inclined expression of Snail2, SIP1 and CXCR4 in vitro. p-STAT1 inhibitor was able to reverse the expression of E-cadherin and EMT resulted from IFN-γ function on HBsAg-positive hepatocytes.
Helicobacter hepaticus generate a detrimental immune microenvironment by IFN-γ/p-STAT1 axis which can promote the tumorigenesis of hepatitis B via recruiting innate lymphoid cells.
- Helicobacter hepaticus
- Hepatitis B virus
- Hepatocellular carcinoma
- Innate lymphoid cells
- IFN-γ/p-STAT1 axis
In accordance with latest statistics, hepatocellular carcinoma (HCC) is the third cause of cancer death in the world . Hepatitis B virus (HBV) infection is associated with at least 55% incidence of cirrhosis, followed by hepatic functional decompensation leading to initiation of HCC . Nowadays, progression of HCC can be significantly restrained by viral polymerase inhibition from nucleotide analogues drugs , however, hepatic cancer risk of patients with long-duration HBV infection can remain above normal after functional clearance of the virus infection [4–6]. This clinical problem indicated that some viral integration mechanisms resulting in genomic changes render hepatic carcinogenesis, including chromosomal instability [7, 8], and function regulation of human telomerase reverse transcriptase gene . Moreover, investigators believed that incontrollable inflammation and liver remodeling are highly relative to antiviral immunity [10, 11], considering HBV itself is non-cytopathic to host cells. Therefore, the immune against viral integrated-hepatocytes is vital in the pathogenesis of HBV-associated HCC.
Recently a study demonstrated that intestinal Lactobacillus gasseri triggered the production of interleukin 17 by intrahepatic γδ T cells, resulting in cholestatic hepatitis . It seems a few strains with disrupted intestinal barriers can participate in pathogenesis of hepatic disease, including hepatocellular carcinoma, by regulating innate lymphoid cells (ILCs) and their inflammatory chemokines [13–15]. Manipulating intestinal microbiome in mice induced CXCL16 expression of liver sinusoidal endothelial cells, which accumulated CXCR6+ natural killer T, reinforcing an antitumor effect by interferon-γ secretion . Evidences for the connection between intestinal microflora and HCC gradually rise, however, few study has investigated the role of specific strains in development of HCC taking viral etiology into consideration. In our study, we used HBV transgenic mice to simulate the course of human HCC, to verify the hypothesis that Helicobacter hepaticus, a specific strain highly relative to hepatitis, could facilitate the carcinogenesis of HBV infected hepatitis via regulating innate lymphoid cells.
Helicobacter hepaticus are over-presented in feces from mice with HBV infected HCC
Helicobacter hepaticus potentiated hepatocellular tumorigenesis in HBV-transgenic mice
ILC-derived cytokines contribute to helicobacter hepaticus-associated HCC development in HBs-Tg mice
ILC-derived IFN-γ triggered epithelial–mesenchymal transition via STAT1 signaling in Helicobacter hepaticus-associated HCC
Helicobacter species contribute to several kinds of chronic inflammation and carcinoma in mice. In spite of the presence of specific helicobacter strains were proved in human liver cancer, the pathogenic role of Helicobacter species on hepatic diseases remains undefined. H. hepaticus related injury and carcinogenesis in hepatocytes by activating nuclear factor-kB-regulated networks associated with innate and T helper 1-type adaptive immunity in aflatoxin B1 induced murine model of HCC . However, few study investigated the involvement of H. hepaticus in HBV-associated HCC. In recent study, transgenic mice model with viral hepatitis mimical period were utilized to uncover the over-presented H. hepaticus was significantly related to the initiation of cirrhosis and neoplasm development in liver (Figs. 1 and 2). Moreover, our study indicated that H. hepaticus accumulation might result in the rising of the triggered anti-virus cytokines via ILCs (Fig. 3).
Hepatitis and infiltration of immune cells in H. hepaticus-infected cancer indicated that chronic inflammation may make primary contribution to the promotion for carcinogenesis . Recruited to the liver in chronic H. hepaticus infected mice, plenty of neutrophils and macrophages in microenvironment accumulated oxidized nucleoside 8-hydroxydeoxyguanosine in hepatocytes by ROS secretion, exacerbating the hepatic tumorigenesis . It seems that innate immunity against H. hepaticus is the major force attributing to this kind of inflammation . Orogastric administration of H. hepaticus in Rag2−/−ApcMin/+ mice developed breast carcinoma, an extraintestinal tumor, in the absence of lymphocytes . However, rare study has reported that NK cells can assist the malignant progress of hepatic tumors.
As we usually understood, both HCC in situ and colonic metastatic carcinoma in the liver could be restrained by phagocytosis from activated NK cells [25, 26]. In hepatitis B virus associated cancer, NK cells were guided by the cooperation between cancerous cells and virus to functionally promote HCC progression [27, 28]. This kind of role transition in molecular mechanism had been proved as increasing of inhibitory receptor on NK cells including NKG2A, Tim3 and PD-1 [29, 30]. In our study, HBV infected hepatocyte damage was initialed by reprogrammed NK cells but not NKT cells, resulting in hepatic carcinoma.
Obviously, colonization of H. hepaticus aggravate liver cirrhosis and neoplasia in the liver with HBV infection in previous reports . Our study represents the original investigation of the intestinal microbiome in hepatic carcinogenesis without over-presentation at hepatocytes (Fig. 1). It more likely involved cytokines release reacted to H. hepaticus from mesenteric lymph nodes towards microenvironment in the liver. Some studies demonstrated that H. hepaticus associated inflammation activated innate and Th1-type adaptive immunity with inclining expression of cytokines and receptors within the intestinal lymphatic system .
Our study shows that H. hepaticus conspires with HBV to impair positive immunity of ILCs to accelerate progression of HCC. Although recent findings reveal the interaction between H. hepaticus, hepatic virus and liver in the evolution of HCC, further investigations are needed for novel prevention on HBV associated cancer in humans.
Helicobacter hepaticus generate a detrimental immune microenvironment by up-regulating ILCs. ILCs-derived IFN-γ can promote the tumorigenesis of hepatitis B via E-cadherin/STAT1.
Ten-week-old male HBV transgenic mice C57BL/6J-TgN (AlblHBV) 44 Bri (named as HBs-Tg mice) were purchased from Department of Laboratory Animal Science of Peking University. The control C57BL/6J mice are the littermates of HBs-Tg mice. All mice were housed under specific pathogen-free conditions (22 °C, 55% humidity, and 12 h day/night rhythm). Feces, colons and livers in natural HBs-Tg mice would be collected for abundance evaluation of H. hepaticus at 24–25 months old. Besides, they were divided into four groups characterised by the presence or absence of the HBV transgene and H. hepaticus infection (shown as Fig. 1). Beginning at 4–5-month-old, mice were gavaged with 2 × 107 colony-forming units (CFU) of H. hepaticus strain or broth only every 48 h for 8 months as previously described . Mice were euthanatised at 12–13 or 24–25-month-old by CO2 inhalation. Blood and livers were harvested at 12–13-month-old, submitted for histopathology, serum test, and quantitative real-time PCR (qRT-PCR). At 24–25-month-old, survival rate would be calculated.
Helicobacter hepaticus (type strain ATCC 51448) was cultured according to previously described . Briefly, they grew under microaerobic conditions (37 °C) using trypticase soy blood agar (BD Biosciences, USA) at first, followed by inoculation in 5% fetal bovine serum-contained brucella broth on a rotary shaking incubator (Thermo Fisher Scientific, USA) for 48 h. Then, the cultured broth was centrifuged at 10,000 rpm (4 °C) for 20 min. After harvested in exponential phase using OD600 nm test, the pellet was resuspended in brucella broth containing 30% glycerol, to approximately 108 organisms/ml as confirmed by spectrophotometry (MEGATOO, Beijing, China). Oral gavage of 0.2 ml fresh culture was administrated into examed mice for three doses every 2 days, while medium alone was introduced into controls synchronously. Subculture of the inoculum and the medium on blood agar was used for maintaining the purity of the strain and the sterility of the medium.
Serum biochemical assays
Serum alanine aminotransferase (ALT), alpha-fetoprotein (AFP) was determined using ELISA kits (R&D systems, Minneapolis, MN), according to the manufacturer’s instructions.
Preparation of fecal sample and colon tissues for PCR
Based on the manufacturer’s instructions, DNA in feces was extracted using the QIAamp DNA stool Mini Kit (Qiagen, USA) with a spin column, followed by an elution in the Tris–EDTA buffer (pH 8). DNA of intestinal microbiome in situ was isolated from frozen colorectal tissues after mechanical homogenization (Tissue Lyser, Qiagen, USA), using the EZ1 DNA Tissue kit and the EZ1 BioRobot (Qiagen, USA). The parameter of the extracted DNA was measured by the NanoDrop 2000 spectrophotometer (Thermo Scientific, USA). All DNA samples were stored at − 80 °C.
All reactions were assayed in 20 μL reaction volume containing 1× final concentration TaqMan Universal Master Mix (Applied Biosystems, USA) in a 96-well optical PCR plate. Each reaction contained 5 ng of extracted faecal DNA and 5 μM of primers. Amplification and detection of DNA was performed with the Roche Lightcycler 480 Quantitative Analysis System (Applied Biosystems). the following reaction conditions would be applied: 2 min at 50 °C, 10 min at 95 °C, and 40 cycles of 15 s at 95 °C and 1 min at 60 °C. The primers for detecting H. hepaticus and total bacteria were used as previously described . The primers’ sequences were as follows:
H. hepaticus forward primer, 5′-GCAUUUGAAACUGUUACUCUG-3′;
Helicobacter hepaticus reverse primer, 5′-GGGGAGCUUGAAAACAG-3′;
Total bacterial DNA forward primer, 5′-GCAGGCCTAACACATGCAAGTC-3′;
Total bacterial DNA reverse primer, 5′-CTGCTGCCTCCCGTAGGAGT-3′.
Every sample was assayed in triplicate in same batch, and the mean of these cycle threshold (Ct) values was calculated for subsequent study. The relative abundance of H. hepaticus normalised to the total bacteria of each sample was calculated using the 2−ΔCt method (where ΔCt = the average Ct value of H. hepaticus in each sample—the average Ct value of total bacteria).
All primers of cytokines in livers were synthesized by Invitrogen (Thermo Fisher Scientific, USA), as shown in Additional file 1: Table S1. Results were analyzed using the ΔΔCt method and β-actin as the reference.
Primary mouse hepatocyte isolation and culture
Details are shown in supplementary materials and methods. The isolation and culture of primary mouse hepatocytes were performed as described . IFN-γ (PeproTech, USA) treatment was using 2 ng/ml. STAT1 activity inhibition was done by incubation with 20 μM Fludarabine (Selleckchem, USA) for 24 h, followed by IFN-γ treatment for another 24 h.
Details are shown in supplementary materials and methods. Liver tissues were lysed as described . After SDS-PAGE, proteins were transferred onto PVDF membranes (Millipore Corporation, USA), and incubated with primary Abs over-night at 4 °C. Membranes were washed with 0.1% (vol/vol) Tween 20 in TBS (pH 7.6) and incubated with a 1:2, 500 dilution of horseradish peroxidase-conjugated secondary Abs for 60 min at room temperature. Protein bands were visualized by ECL reaction (Pierce Biotechnology, Rockford, IL).
Isolation of mononuclear cells in hepatic tumor
Hepatic mononuclear cells were prepared as previously described . Briefly, liver of mice was removed and washed with Ca and Mg free Dulbecco’s Phosphate Buffered Saline (DPBS). After pressing through a 200-gauge stainless steel mesh, the cell mixture was resuspended in 40% Percoll solution (General Electric Company, USA), followed by overlaying gently onto 70% Percoll solution. Then, this cell mixture was centrifuged at 1260×g for 30 min at room temperature. The interface cells between the percoll solutions were aspirated and washed twice with PBS medium. Single cell suspensions were resuspended in cell staining solution (PBS with 2% FCS) for flow cytometry.
Multicolor flow cytometry analysis
After Fc receptors blocking (BD Biosciences, USA), an appropriate concentration of the fluorescence-labeled antibody was used for the staining of surface antigens at 4 °C for 30 min in dark place. Fluorochrome-conjugated monoclonal antibodies of cellular markers: PercpCy5.5-anti-CD3, FITC-anti-IL-17A, PE-anti-CD4, PE-Cy7-anti-NK1.1 (BD Bioscience, USA). FITC-anti-CD69, FITC-anti-IFN-γ; PE-anti-FasL, PE-anti-TRAIL, PE-anti-CD107a, and APC-anti-NKG2D (eBioscience, San Diego, CA). The intracellular cytokine staining, including INF-γ and IL-17A, was using Mouse Intracellular Cytokine Staining Starter Kit (BD Biosciences, USA), according to the kit’s instructions. Samples were measured by a BD Accuri C6 plus flow cytometer (BD Biosciences, USA), and the data were managed using BD Accuri C6 plus analysis (BD Biosciences, USA).
All statistics were performed using GraphPad Prism 5.0 software (La Jolla, CA). Results of inflammatory genes, tumor number, ALT activity, AFP activity, cellular analysis, survival analysis and richness of bacteria were analyzed using Mann–Whitney U test, Log-rank (Mantel–Cox) test as appropriate. Data are expressed as the medians with interquartile range. P < 0.05 was considered significant in comparison of medians.
The research and manuscript preparation are funded by Dr. Junqing Han.
XH performed the animal experiments and immunoassays, and was a major contributor in writing the manuscript. TH helped XH to analyzed the data of chemokines from livers. JH funded and directed all these researches and reviewed this manuscript. All authors read and approved the final manuscript.
Ethics approval and consent to participate
Consent for publication
The authors declare that they have no competing interests.
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
- Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68(6):394–424. https://doi.org/10.3322/caac.21492.View ArticlePubMedGoogle Scholar
- Maucort-Boulch D, de Martel C, Franceschi S, Plummer M. Fraction and incidence of liver cancer attributable to hepatitis B and C viruses worldwide. Int J Cancer. 2018;142(12):2471–7. https://doi.org/10.1002/ijc.31280.View ArticlePubMedGoogle Scholar
- Singal AK, Salameh H, Kuo YF, Fontana RJ. Meta-analysis: the impact of oral anti-viral agents on the incidence of hepatocellular carcinoma in chronic hepatitis B. Aliment Pharm Ther. 2013;38(2):98–106. https://doi.org/10.1111/apt.12344.View ArticleGoogle Scholar
- Wong GLH, Tse YK, Yip TCF, Chan HLY, Tsoi KKF, Wong VWS. Long-term use of oral nucleos(t)ide analogues for chronic hepatitis B does not increase cancer risk—a cohort study of 44 494 subjects. Aliment Pharm Ther. 2017;45(9):1213–24. https://doi.org/10.1111/apt.14015.View ArticleGoogle Scholar
- Papatheodoridis GV, Idilman R, Dalekos GN, Buti M, Chi H, van Boemmel F, et al. The risk of hepatocellular carcinoma decreases after the first 5 years of entecavir or tenofovir in Caucasians with chronic hepatitis B. Hepatology. 2017;66(5):1444–53. https://doi.org/10.1002/hep.29320.View ArticlePubMedGoogle Scholar
- Yip TCF, Chan HLY, Wong VWS, Tse YK, Lam KLY, Wong GLH. Impact of age and gender on risk of hepatocellular carcinoma after hepatitis B surface antigen seroclearance. J Hepatol. 2017;67(5):902–8. https://doi.org/10.1016/j.jhep.2017.06.019.View ArticlePubMedGoogle Scholar
- Sung WK, Zheng HC, Li SY, Chen RH, Liu X, Li YR, et al. Genome-wide survey of recurrent HBV integration in hepatocellular carcinoma. Nat Genet. 2012;44(7):765. https://doi.org/10.1038/ng.2295.View ArticlePubMedGoogle Scholar
- Jiang ZS, Jhunjhunwala S, Liu JF, Haverty PM, Kennemer MI, Guan YH, et al. The effects of hepatitis B virus integration into the genomes of hepatocellular carcinoma patients. Genome Res. 2012;22(4):593–601. https://doi.org/10.1101/gr.133926.111.View ArticlePubMedPubMed CentralGoogle Scholar
- Schluter V, Meyer M, Hofschneider PH, Koshy R, Caselmann WH. Integrated hepatitis-B virus-X and 3′ truncated pres/S-sequences derived from human hepatomas encode functionally active transactivators. Oncogene. 1994;9(11):3335–44.PubMedGoogle Scholar
- Tu T, Buhler S, Bartenschlager R. Chronic viral hepatitis and its association with liver cancer. Biol Chem. 2017;398(8):817–37. https://doi.org/10.1515/hsz-2017-0118.View ArticlePubMedGoogle Scholar
- Tu T, Budzinska MA, Shackel NA, Jilbert AR. Conceptual models for the initiation of hepatitis B virus-associated hepatocellular carcinoma. Liver Int. 2015;35(7):1786–800. https://doi.org/10.1111/liv.12773.View ArticlePubMedGoogle Scholar
- Tedesco D, Thapa M, Chin CY, Ge Y, Gong MH, Li J, et al. Alterations in intestinal microbiota lead to production of interleukin 17 by intrahepatic gamma delta T-cell receptor-positive cells and pathogenesis of cholestatic liver disease. Gastroenterology. 2018;154(8):2178–93. https://doi.org/10.1053/j.gastro.2018.02.019.View ArticlePubMedGoogle Scholar
- Sandler NG, Koh C, Roque A, Eccleston JL, Siegel RB, Demino M, et al. Host response to translocated microbial products predicts outcomes of patients with HBV or HCV infection. Gastroenterology. 2011;141(4):1220. https://doi.org/10.1053/j.gastro.2011.06.063.View ArticlePubMedPubMed CentralGoogle Scholar
- Rahman K, Desai C, Iyer SS, Thorn NE, Kumar P, Liu YS, et al. Loss of junctional adhesion molecule a promotes severe steatohepatitis in mice on a diet high in saturated fat, fructose, and cholesterol. Gastroenterology. 2016;151(4):733. https://doi.org/10.1053/j.gastro.2016.06.022.View ArticlePubMedPubMed CentralGoogle Scholar
- Singh V, Yeoh BS, Chassaing B, Xiao X, Saha P, Olvera RA, et al. Dysregulated microbial fermentation of soluble fiber induces cholestatic liver cancer. Cell. 2018;175(3):679. https://doi.org/10.1016/j.cell.2018.09.004.View ArticlePubMedGoogle Scholar
- Ma C, Han MJ, Heinrich B, Fu Q, Zhang QF, Sandhu M, et al. MICROBIOME Gut microbiome-mediated bile acid metabolism regulates liver cancer via NKT cells. Science. 2018;360(6391):876. https://doi.org/10.1126/science.aan5931.View ArticleGoogle Scholar
- Daniel SG, Ball CL, Besselsen DG, Doetschman T, Hurwitz BL. Functional changes in the gut microbiome contribute to transforming growth factor beta-deficient colon cancer. Msystems. 2017;2(5):e00065-17. https://doi.org/10.1128/msystems.00065-17.View ArticlePubMedPubMed CentralGoogle Scholar
- Ge ZM, Feng Y, Ge LL, Parry N, Muthupalani S, Fox JG. Helicobacter hepaticus cytolethal distending toxin promotes intestinal carcinogenesis in 129Rag2-deficient mice. Cell Microbiol. 2017;19(7):e12728. https://doi.org/10.1111/cmi.12728.View ArticleGoogle Scholar
- Sunami Y, Ringelhan M, Kokai E, Lu M, O’Connor T, Lorentzen A, et al. Canonical NF-kappa B signaling in hepatocytes acts as a tumor-suppressor in hepatitis B virus surface antigen-driven hepatocellular carcinoma by controlling the unfolded protein response. Hepatology. 2016;63(5):1592–607. https://doi.org/10.1002/hep.28435.View ArticlePubMedGoogle Scholar
- Chen YY, Sun R, Jiang W, Wei HM, Tian ZG. Liver-specific HBsAg transgenic mice are over-sensitive to Poly(I: C)-induced liver injury in NK cell- and IFN-gamma-dependent manner. J Hepatol. 2007;47(2):183–90. https://doi.org/10.1016/j.jhep.2007.02.020.View ArticlePubMedGoogle Scholar
- Li WY, Xiao J, Zhou X, Xu M, Hu CB, Xu XY, et al. STK4 regulates TLR pathways and protects against chronic inflammation-related hepatocellular carcinoma. J Clin Invest. 2015;125(11):4239–54. https://doi.org/10.1172/Jci81203.View ArticlePubMedPubMed CentralGoogle Scholar
- Sipowicz MA, Chomarat P, Diwan BA, Anver MA, Awasthi YC, Ward JM, et al. Increased oxidative DNA damage and hepatocyte overexpression of specific cytochrome P450 isoforms in hepatitis of mice infected with Helicobacter hepaticus. Am J Pathol. 1997;151(4):933–41.PubMedPubMed CentralGoogle Scholar
- Mandell L, Moran AP, Cocchiarella A, Houghton J, Taylor N, Fox JG, et al. Intact gram-negative Helicobacter pylori, Helicobacter felis, and Helicobacter hepaticus bacteria activate innate immunity via toll-like receptor 2 but not toll-like receptor 4. Infect Immun. 2004;72(11):6446–54. https://doi.org/10.1128/Iai.72.11.6446.6454.2004.View ArticlePubMedPubMed CentralGoogle Scholar
- Rao VP, Poutahidis T, Ge ZM, Nambiar PR, Horwitz BH, Fox JG, et al. Proinflammatory CD4(+)CD45RB(hi) lymphocytes promote mammary and intestinal carcinogenesis in Apc(Min/+) mice. Cancer Res. 2006;66(1):57–61. https://doi.org/10.1158/0008-5472.Can-05-3445.View ArticlePubMedGoogle Scholar
- Miyagi T, Takehara T, Tatsumi T, Kanto T, Suzuki T, Jinushi M, et al. CD1d-mediated stimulation of natural killer T cells selectively activates hepatic natural killer cells to eliminate experimentally disseminated hepatoma cells in murine liver. Int J Cancer. 2003;106(1):81–9. https://doi.org/10.1002/ijc.11163.View ArticleGoogle Scholar
- Dupaul-Chicoine J, Arabzadeh A, Dagenais M, Douglas T, Champagne C, Morizot A, et al. The Nlrp3 inflammasome suppresses colorectal cancer metastatic growth in the liver by promoting natural killer cell tumoricidal activity. Immunity. 2015;43(4):751–63. https://doi.org/10.1016/j.immuni.2015.08.013.View ArticlePubMedPubMed CentralGoogle Scholar
- Zhang QF, Yin WW, Xia Y, Yi YY, He QF, Wang X, et al. Liver-infiltrating CD11b(−)CD27(−) NK subsets account for NK-cell dysfunction in patients with hepatocellular carcinoma and are associated with tumor progression. Cell Mol Immunol. 2017;14(10):819–29. https://doi.org/10.1038/cmi.2016.28.View ArticlePubMedGoogle Scholar
- Yang YL, Han QJ, Hou ZH, Zhang C, Tian ZG, Zhang J. Exosomes mediate hepatitis B virus (HBV) transmission and NK-cell dysfunction. Cell Mol Immunol. 2017;14(5):465–75. https://doi.org/10.1038/cmi.2016.24.View ArticlePubMedGoogle Scholar
- Wolf MJ, Adili A, Piotrowitz K, Abdullah Z, Boege Y, Stemmer K, et al. Metabolic activation of lntrahepatic CD8(+) T cells and NKT Cells causes nonalcoholic steatohepatitis and liver cancer via cross-talk with hepatocytes. Cancer Cell. 2014;26(4):549–64. https://doi.org/10.1016/j.ccell.2014.09.003.View ArticleGoogle Scholar
- Wang J, Zhao WX, Cheng LA, Guo MZ, Li DL, Li XZ, et al. CD137-mediated pathogenesis from chronic hepatitis to hepatocellular carcinoma in hepatitis B virus-transgenic mice. J Immunol. 2010;185(12):7654–62. https://doi.org/10.4049/jimmunol.1000927.View ArticlePubMedPubMed CentralGoogle Scholar
- Dore MP, Realdi G, Mura D, Graham DY, Sepulveda AR. Helicobacter infection in patients with HCV-related chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Digest Dis Sci. 2002;47(7):1638–43. https://doi.org/10.1023/A:1015848009444.View ArticlePubMedGoogle Scholar
- Myles MH, Dieckgraefe BK, Criley JM, Franklin CL. Characterization of cecal gene expression in a differentially susceptible mouse model of bacterial-induced inflammatory bowel disease. Inflamm Bowel Dis. 2007;13(7):822–36. https://doi.org/10.1002/ibd.20138.View ArticlePubMedGoogle Scholar
- Whary MT, Morgan TJ, Dangler CA, Gaudes KJ, Taylor NS, Fox JG. Chronic active hepatitis induced by Helicobacter hepaticus in the A/JCr mouse is associated with a Th1 cell-mediated immune response. Infect Immun. 1998;66(7):3142–8.PubMedPubMed CentralGoogle Scholar
- Shames B, Fox JG, Dewhurst F, Yan LL, Shen ZL, Taylor NS. Identification of widespread helicobacter-Hepaticus infection in feces in commercial mouse colonies by culture and PCR assay. J Clin Microbiol. 1995;33(11):2968–72.PubMedPubMed CentralGoogle Scholar
- Chen YY, Sun R, Wu XY, Cheng M, Wei HM, Tian ZG. CD4+CD25+ regulatory T cells inhibit natural killer cell hepatocytotoxicity of hepatitis B virus transgenic mice via membrane-bound TGF-beta and OX40. J Innate Immun. 2016;8(1):30–42. https://doi.org/10.1159/000431150.View ArticlePubMedPubMed CentralGoogle Scholar