The presence of Helicobacter pylori in the liver depends on the Th1, Th17 and Treg cytokine profile of the patient

The hypothesis that Helicobactermight be a risk factor for human liver diseases has arisen after the detection of Helicobacter DNA in hepatic tissue of patients with hepatobiliary diseases. Nevertheless, no explanation that justifies the presence of the bacterium in the human liver has been proposed. We evaluated the presence of Helicobacterin the liver of patients with hepatic diseases of different aetiologies. We prospectively evaluated 147 patients (106 with primary hepatic diseases and 41 with hepatic metastatic tumours) and 20 liver donors as controls. Helicobacter species were investigated in the liver by culture and specific 16S rDNA nested-polymerase chain reaction followed by sequencing. Serum and hepatic levels of representative cytokines of T regulatory cell, T helper (Th)1 and Th17 cell lineages were determined using enzyme linked immunosorbent assay. The data were evaluated using logistic models. Detection of Helicobacter pylori DNA in the liver was independently associated with hepatitis B virus/hepatitis C virus, pancreatic carcinoma and a cytokine pattern characterised by high interleukin (IL)-10, low/absent interferon-γ and decreased IL-17A concentrations (p < 10(-3)). The bacterial DNA was never detected in the liver of patients with alcoholic cirrhosis and autoimmune hepatitis that are associated with Th1/Th17 polarisation. H. pylori may be observed in the liver of patients with certain hepatic and pancreatic diseases, but this might depend on the patient cytokine profile.     

Population Genetic Analyses of Helicobacter pylori Isolates from Gambian Adults and Children

The gastric pathogen Helicobacter pylori is one of the most genetically diverse of bacterial species. Much of its diversity stems from frequent mutation and recombination, preferential transmission within families and local communities, and selection during persistent gastric mucosal infection. MLST of seven housekeeping genes had identified multiple distinct H. pylori populations, including three from Africa: hpNEAfrica, hpAfrica1 and hpAfrica2, which consists of three subpopulations (hspWAfrica, hspCAfrica and hspSAfrica). Most detailed H. pylori population analyses have used strains from non-African countries, despite Africa''s high importance in the emergence and evolution of humans and their pathogens. Our concatenated sequences from seven H. pylori housekeeping genes from 44 Gambian patients (MLST) identified 42 distinct sequence types (or haplotypes), and no clustering with age or disease. STRUCTURE analysis of the sequence data indicated that Gambian H. pylori strains belong to the hspWAfrica subpopulation of hpAfrica1, in accord with Gambia''s West African location. Despite Gambia''s history of invasion and colonisation by Europeans and North Africans during the last millennium, no traces of Ancestral Europe1 (AE1) population carried by those people were found. Instead, admixture of 17% from Ancestral Europe2 (AE2) was detected in Gambian strains; this population predominates in Nilo-Saharan speakers of North-East Africa, and might have been derived from admixture of hpNEAfrica strains these people carried when they migrated across the Sahara during the Holocene humid period 6,000–9,000 years ago. Alternatively, shared AE2 ancestry might have resulted from shared ancestral polymorphisms already present in the common ancestor of sister populations hpAfrica1 and hpNEAfrica.     

Antigen-Specific Memory B-cell Responses to Enterotoxigenic Escherichia coli Infection in Bangladeshi Adults

Background

Multiple infections with diverse enterotoxigenic E. coli (ETEC) strains lead to broad spectrum protection against ETEC diarrhea. However, the precise mechanism of protection against ETEC infection is still unknown. Therefore, memory B cell responses and affinity maturation of antibodies to the specific ETEC antigens might be important to understand the mechanism of protection.

Methodology

In this study, we investigated the heat labile toxin B subunit (LTB) and colonization factor antigens (CFA/I and CS6) specific IgA and IgG memory B cell responses in Bangladeshi adults (n = 52) who were infected with ETEC. We also investigated the avidity of IgA and IgG antibodies that developed after infection to these antigens.

Principal Findings

Patients infected with ETEC expressing LT or LT+heat stable toxin (ST) and CFA/I group or CS6 colonization factors developed LTB, CFA/I or CS6 specific memory B cell responses at day 30 after infection. Similarly, these patients developed high avidity IgA and IgG antibodies to LTB, CFA/I or CS6 at day 7 that remained significantly elevated at day 30 when compared to the avidity of these specific antibodies at the acute stage of infection (day 2). The memory B cell responses, antibody avidity and other immune responses to CFA/I not only developed in patients infected with ETEC expressing CFA/I but also in those infected with ETEC expressing CFA/I cross-reacting epitopes. We also detected a significant positive correlation of LTB, CFA/I and CS6 specific memory B cell responses with the corresponding increase in antibody avidity.

Conclusion

This study demonstrates that natural infection with ETEC induces memory B cells and high avidity antibodies to LTB and colonization factor CFA/I and CS6 antigens that could mediate anamnestic responses on re-exposure to ETEC and may help in understanding the requirements to design an effective vaccination strategies.     

儿童幽门螺杆菌感染

幽门螺杆菌感染不仅能引起胃炎、消化性溃疡,诱发胃癌等胃肠道的病变,还与许多胃肠外疾病密切相关,如果不经过特殊治疗将终生带菌,严重的影响小儿的生长发育和身心健康。这些问题引起了儿科医生和儿童保健医生的共同关注。儿童期既是幽门螺杆菌感染的特殊时期,也是控制感染的关键时期。本文将从小儿幽门螺杆菌国内外的感染状况、相关疾病、诊断方法、治疗及预防等几个方面综述如下。     

Helicobacter pylori Infection in Children

The review summarizes the articles published on Helicobacter pylori in children between April 2007 and March 2008. Evidence is emerging in different populations including developing countries that the prevalence of H. pylori is declining in all age groups. The reasons for this are unclear but it is unlikely that treatment of infection or improvement in socioeconomic conditions fully explains the decline. For the first time, differences in the inflammatory response between adults and children have been well characterized in a group of adults and children from Chile with similar levels of H. pylori infection. This study suggests that the reduced inflammatory response to H. pylori at a cellular level in children could be the consequence of an enhanced Treg cell response, which in turn down-regulates H. pylori -induced inflammation. The publication of the Paediatric European Register for Treatment of Helicobacter pylori study (PERTH) is important as it demonstrates the advantages of different centers working in collaboration for the benefit of children. It also highlights the fact that while bismuth-based treatment is more effective than proton pump inhibitor-based treatment in children, bismuth preparations are not widely available for use in children.     

Stromal Cells Induce Th17 during Helicobacter pylori Infection and in the Gastric Tumor Microenvironment

Gastric cancer is associated with chronic inflammation and Helicobacter pylori infection. Th17 cells are CD4⁺ T cells associated with infections and inflammation; but their role and mechanism of induction during carcinogenesis is not understood. Gastric myofibroblasts/fibroblasts (GMF) are abundant class II MHC expressing cells that act as novel antigen presenting cells. Here we have demonstrated the accumulation of Th17 in H. pylori-infected human tissues and in the gastric tumor microenvironment. GMF isolated from human gastric cancer and H. pylori infected tissues co-cultured with CD4⁺ T cells induced substantially higher levels of Th17 than GMF from normal tissues in an IL-6, TGF-β, and IL-21 dependent manner. Th17 required interaction with class II MHC on GMF for activation and proliferation. These studies suggest that Th17 are induced during both H. pylori infection and gastric cancer in the inflammatory milieu of gastric stroma and may be an important link between inflammation and carcinogenesis.     

Human dendritic cells respond to Helicobacter pylori, promoting NK cell and Th1-effector responses in vitro

Helicobacter pylori infection leads to chronic gastric inflammation. The current study determined the response of human APCs, NK cells, and T cells toward the bacteria in vitro. Human monocyte-derived dendritic cells (DC) were incubated with bacteria for 48 h. Intact H. pylori at a multitude of infection 5 stimulated the expression of MHC class II (4- to 7-fold), CD80, and CD86 B7 molecules (10- to 12-fold) and the CD83 costimulatory molecule (>30-fold) as well as IL-12 secretion (>50-fold) in DCs, and thereby, strongly induced their maturation and activation. CD56(+)/CD4(-) NK cells, as well as CD4(+)/CD45RA(+) naive T cells, were isolated and incubated with DCs pulsed with intact bacteria or different cellular fractions. Coculture of H. pylori-pulsed DCs with NK cells strongly potentiated the secretion of TNF-alpha and IFN-gamma. Coculture of naive T cells with H. pylori-pulsed DCs significantly enhanced TNF-alpha, IFN-gamma, and IL-2 secretion as well as T-bet mRNA levels, while GATA-3 mRNA was lowered. However, the effect appeared attenuated compared with coculture with Escherichia coli. A greater stimulation was seen with naive T cells and DCs pulsed with H. pylori membrane preparations. Intact H. pylori potently induced the maturation and activation of human monocyte-derived DC and thereby promote NK and Th1 effector responses. The strong activation of NK cells may be important for the innate immune response. Th1-polarized T cells were induced especially by incubation with membrane preparations of H. pylori, suggesting that membrane proteins may account for the specific adaptive immune response.     

Th1-Mediated Immunity against Helicobacter pylori Can Compensate for Lack of Th17 Cells and Can Protect Mice in the Absence of Immunization

Helicobacter pylori (H. pylori) infection can be significantly reduced by immunization in mice. Th17 cells play an essential role in the protective immune response. Th1 immunity has also been demonstrated to play a role in the protective immune response and can compensate in the absence of IL-17. To further address the potential of Th1 immunity, we investigated the efficacy of immunization in mice deficient in IL-23p19, a cytokine that promotes Th17 cell development. We also examined the course of Helicobacter infection in unimmunized mice treated with Th1 promoting cytokine IL-12. C57BL/6, IL-12 p35 KO, and IL-23 p19 KO mice were immunized and challenged with H. pylori. Protective immunity was evaluated by CFU determination and QPCR on gastric biopsies. Gastric and splenic IL-17 and IFNγ levels were determined by PCR or by ELISA. Balb/c mice were infected with H. felis and treated with IL-12 therapy and the resulting gastric bacterial load and inflammatory response were assessed by histologic evaluation. Vaccine induced reductions in bacterial load that were comparable to wild type mice were observed in both IL-12 p35 and IL-23 p19 KO mice. In the absence of IL-23 p19, IL-17 levels remained low but IFNγ levels increased significantly in both immunized challenged and unimmunized/challenged mice. Additionally, treatment of H. felis-infected Balb/c mice with IL-12 resulted in increased gastric inflammation and the eradication of bacteria in most mice. These data suggest that Th1 immunity can compensate for the lack of IL-23 mediated Th17 responses, and that protective Th1 immunity can be induced in the absence of immunization through cytokine therapy of the infected host.     

幽门螺杆菌感染者Th1/Th2细胞免疫应答变化

目的观察幽门螺杆菌(H.pylori)相关性胃病患者血清Th1/Th2细胞因子(干扰素-γ,IFN-γ、白细胞介素-4,IL-4)水平变化,以探讨其在发病中的可能免疫致病机制。方法采用酶联免疫吸附测定法(ELISA)测定17例慢性浅表性胃炎、15例胃癌前病变和20例胃癌患者血清IFN-γ及IL-4的含量。比较H.pylori阳性3组患者之间、H.pylori阳性与阴性各相应组患者之间血清2种细胞因子的差异。结果 H.pylori阳性的浅表性胃炎组、胃癌前病变组及胃癌组血清IL-4含量随病变的进展有逐渐升高的趋势,但3组之间差异无统计学意义(P>0.05);H.pylori阳性的3组血清IFN-γ含量差异无统计学意义(P>0.05);H.pylori阳性与阴性的各相应组血清IFN-γ含量差异无统计学意义(P>0.05);H.pylori阳性的胃癌前病变组和胃癌组与H.pylori阴性的相应组血清IL-4含量差异无统计学意义(P>0.05);H.pylori阳性的浅表性胃炎组血清IL-4含量较H.pylori阴性的浅表性胃炎组明显降低(P<0.05)。结论 H.pylori感染可能抑制Th2型免疫应答,导致H.pylori感染持续存在;H.pylori感染相关胃部病变进展过程中,可能存在Th1型应答向Th2型应答漂移,与胃癌的发生可能有一定的相关性。     

Cutting edge: cyclooxygenase-2 activation suppresses Th1 polarization in response to Helicobacter pylori

Helicobacter pylori infection causes a Th1-driven mucosal immune response. Cyclooxygenase (COX)-2 is up-regulated in lamina propria mononuclear cells in H. pylori gastritis. Because COX-2 can modulate Th1/Th2 balance, we determined whether H. pylori activates COX-2 in human PBMCs, and the effect on cytokine and proliferative responses. There was significant up-regulation of COX-2 mRNA and PGE(2) release in response to H. pylori preparations. Addition of COX-2 inhibitors or an anti-PGE(2) Ab resulted in a marked increase in H. pylori-stimulated IL-12 and IFN-gamma production, and a decrease in IL-10 levels. Addition of PGE(2) or cAMP, the second messenger activated by PGE(2), had the opposite effect. Similarly, stimulated cell proliferation was increased by COX-2 inhibitors or anti-PGE(2) Ab, and was decreased by PGE(2). Our findings indicate that COX-2 has an immunosuppressive role in H. pylori gastritis, which may protect the mucosa from severe injury, but may also contribute to the persistence of the infection.     

Human serum antibody response to Helicobacter pylori whole cell antigen in an institutionalized Bangladeshi population

J. NESSA, H. CHART, R.J. OWEN AND B. DRASAR. 2001 .
Aims: To use a commercial ELISA kit and an immunoblot assay to investigate the antibody levels of selected members of the Bangladeshi population to Helicobacter pylori protein antigens.
Methods and Results: Using immunoblotting, high seroprevalence rates were observed in all age groups, although the subjects within the 1–9 years age group had the highest seroprevalence of antibodies to H. pylori antigens. By ELISA, the highest level of seroprevalence was observed in those over the age of 20 years.
Conclusions: On the basis of these results the overall prevalence rate of H. pylori infection for the whole population was 77·4%; 77·9% for orphan boys and 76% for carers. CagA antibodies were detected in 86% of those with high levels of antibodies to H. pylori antigens.
Significance and Impact of the Study: A combination of immunoblotting and ELISA was the most efficient means of detecting serum antibodies to H. pylori antigens and could be applied to the screening of human sera for H. pylori -specific antibodies.     

Difference in Th1 and Th17 lymphocyte adhesion to endothelium

T cell subset-specific migration to inflammatory sites is tightly regulated and involves interaction of the T cells with the endothelium. Th17 cells often appear at different inflammatory sites than Th1 cells, or both subsets appear at the same sites but at different times. Differences in T cell subset adhesion to endothelium may contribute to subset-specific migratory behavior, but this possibility has not been well studied. We examined the adhesion of mouse Th17 cells to endothelial adhesion molecules and endothelium under flow in vitro and to microvessels in vivo and we characterized their migratory phenotype by flow cytometry and quantitative RT-PCR. More Th17 than Th1 cells interacted with E-selectin. Fewer Th17 than Th1 cells bound to TNF-α-activated E-selectin-deficient endothelial cells, and intravital microscopy studies demonstrated that Th17 cells engage in more rolling interactions with TNF-α-treated microvessels than Th1 cells in wild-type mice but not in E-selectin-deficient mice. Th17 adhesion to ICAM-1 was dependent on integrin activation by CCL20, the ligand for CCR6, which is highly expressed by Th17 cells. In an air pouch model of inflammation, CCL20 triggered recruitment of Th17 but not Th1 cells. These data provide evidence that E-selectin- and ICAM-1-dependent adhesion of Th17 and Th1 cells with endothelium are quantitatively different.     

Human serum antibody response to Helicobacter pylori whole cell antigen in an institutionalized Bangladeshi population

AIMS: To use a commercial ELISA kit and an immunoblot assay to investigate the antibody levels of selected members of the Bangladeshi population to Helicobacter pylori protein antigens. METHODS AND RESULTS: Using immunoblotting, high seroprevalence rates were observed in all age groups, although the subjects within the 1-9 years age group had the highest seroprevalence of antibodies to H. pylori antigens. By ELISA, the highest level of seroprevalence was observed in those over the age of 20 years. CONCLUSION: On the basis of these results the overall prevalence rate of H. pylori infection for the whole population was 77.4%; 77.9% for orphan boys and 76% for carers. CagA antibodies were detected in 86% of those with high levels of antibodies to H. pylori antigens. SIGNIFICANCE AND IMPACT OF THE STUDY: A combination of immunoblotting and ELISA was the most efficient means of detecting serum antibodies to H. pylori antigens and could be applied to the screening of human sera for H. pylori-specific antibodies.