Helicobacter pylori and IFN-γ, IL-17A, IL-4 and TGF-β1 expression in patients with chronic gastritis and gastric cancer
Dinorah
N.
Martínez Carrillo1,
Rogelio
Hernández Pando2,
Miguel
Á.
Mendoza Catalán1,
Cristian
S.
Hernández Moreno1,
Josefina
Atrisco Morales1,
Salomón
Reyes Navarrete3,
Marco
A.
Jiménez López3,
Reyes
Betancourt Linares4,
Iván
Cruz Del Carmen5,
Berenice
Illades Aguiar1,
Adolfo
Román Román1 and
Gloria
Fernández Tilapa1*
-
1
Universidad Autónoma de Guerrero, Unidad Acádemica de Ciencias Químico Biológicas, Mexico
-
2
Instituto Nacional de Ciencia Médicas y Nutrición “Salvador Zubirán”, Departamento de Patología, Mexico
-
3
Instituto Estatal de Cancerología “Dr. Arturo Beltrán Ortega”, Mexico
-
4
Unidad Especializada en Gastroenterología y Endoscopía, Mexico
-
5
Hospital General “Dr. Raymundo Abarca Alarcón”, Mexico
Gastric cancer is related to the persistent infection by Helicobacter pylori (H. pylori). By utilizing various virulence factors, H. pylori targets different cellular proteins to modulate the host inflammatory response and initiate multiple pathways on the gastric mucosa, resulting in chronic gastritis and peptic ulceration. Some virulence factors of H. pylori activate or downregulate the functions of epithelial cells, monocytes, dendritic cells, neutrophils and natural killer cells. During acute H. pylori infection, neutrophils and macrophages accumulate in the gastric mucosa leading to the production of increased levels of proinflammatory cytokines and chemokines including IL-1β, TNF-α and IL-8. The molecular mechanisms by which H. pylori triggers and maintains the local immune response are complex, but there is evidence that cytokines produced by both innate and adaptive immune systems can lead to the development of ulcer disease and gastric adenocarcinoma. There are no studies that analyze the local expression of IFN-γ, IL-17A, TGF-β1 and IL-4 with vacA and cagA genotypes of H. pylori in patients with chronic gastritis and gastric cancer. The objective this study was to analyze the relationship between the expression of IFN-γ, IL-17A, TGF-β1 and IL-4 with H. pylori cagA and vacA genotypes in patients with chronic gastritis and gastric cancer. Patients attended an upper digestive endoscopy in three Hospitals of Guerrero, Mexico were selected. For each patient, three biopsies from the antrum, corpus, or tumor were taken; the first one was placed in buffer solution for H. pylori molecular diagnosis by PCR method, amplifying a fragment of the 16S rRNA gene, H. pylori positive samples were then subjected to a multiplex PCR to amplify a fragment of the genes cagA, babA2 and the s and m regions from the vacA gene. The second biopsy was fixed in absolute alcohol to verify the IFN-γ, IL-17A, TGF-β1 and IL-4 expression through immunohistochemistry with respective primary antibodies IFN-γ, IL-17A, TGF-β1 and IL-4. The data were expressed as percentage of positives cells. The last biopsy was immediately fixed in formalin for histological diagnosis. The p value was obtained with the X2 test or the Fisher exact test for qualitative variables and the Mann-Whitney and Kruskal-Wallis test to obtain the quantitative variables. A value of p<0.05 was considered statistically significant. The study was conducted with 111 patients, of which 99 were diagnosed with chronic gastritis and 12 with gastric cancer. From those histologically diagnosed with gastric cancer 58.3% (7/12) were diffuse-type, 25% (3/12) of intestinal type and 16.7% (2/12) of mixed type. The mean age of the group with chronic gastritis was 48 years with a range of 19-80 years and in the gastric cancer group the mean age was 57 years with a minimum age of 31 years and a maximum of 85 years. The prevalence of H. pylori infection was 56.8% (63/111). From those positive to H. pylori infection, the general frequencies of virulence factors were, 87.3% (55/63) carried the vacAs1m1 genotype and 77.8% (49/63) the cagA-positive genotype, whereas 73% (46/63) of the patients were H. pylori vacAs1m1/cagA-positive. The prevalence of H. pylori according to the histopathological type of gastric cancer was 71.4% (5/7) diffuse type, 33.3% (1/3) intestinal type and 50% (1/2) mixed type. The IFN-γ, IL-17A, TGF-β1 and IL-4 immunostaining was localized predominantly in the cytoplasm and less frequent in the nucleus of the infiltrating mononuclear cells. Between patients with chronic gastritis, the median of percentage of IFN-γ and TGF-β1 positive cells was significantly higher than IL-17A and IL-4 (p<0.0001). Among those who were not infected by H. pylori the median of percentage of TGF-β1 positive cells was significantly greater than IL-17A and IL-4. In H. pylori positive patients the median of percentage of TGF-β1 positive cells was significantly greater than IFN-γ, IL-17A and IL-4. No significant differences were observed between the median of percentage cells positive to the cytokines in patients infected with vacAs2m2/cagA-negative genotype. However, in patients infected with vacAs1m1/cagA-negative and vacAs1m1/cagA-positive genotype the median percentage of IFN-γ positive cells was significantly greater than TGF-β1, IL-17A and IL-4. Between patients with gastric cancer, the median of percentage of TGF-β1 positive cells was significantly greater than IFN-γ, IL-17A and IL-4. The expression patron of cytokine was similar in H. pylori-negative and H. pylori-positive patients. In patients infected with vacAs1m1/cagA-negative genotype the IL-4 expression it was not detected. In the patients infected with vacAs1m1/cagA-positive genotype the median of percentage of TGF-β1 positive cells was significantly greater than IFN-γ, IL-17A and IL-4. In this study we found a high proportion of patients with chronic gastritis infected with H. pylori vacAs1m1/cagA-positivo genotype (71.4%), in these patients the Th1 and Th17 cytokine type were the most predominant and it is likely that these patients could be at greatest risk of developing a more serious gastric pathology. In patients with gastric cancer is likely that cytokines act synergistically to maintaining inflammatory tumor microenvironment to promote tumor growth, metastasis and angiogenesis.
Acknowledgements
The authors thank the nurses and the support staff of the participants institutions who helped in obtaining the samples. We thank Dr. Victor Hugo Garzón-Barrientos and the MCB. Monica Virginia Saavedra Herrera for the facilities provided for the completion of this research. This research was supported by Universidad Autónoma de Guerrero (2013) and the Secretaría de Educación Pública through PIFI-2011 program.
Keywords:
Helicobacter pylori,
Chronic gastritis,
gastric cancer,
Cytokines,
vacA and cagA genotypes
Conference:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015.
Presentation Type:
Poster Presentation
Topic:
Infectious and parasitic diseases
Citation:
Martínez Carrillo
DN,
Hernández Pando
R,
Mendoza Catalán
MÁ,
Hernández Moreno
CS,
Atrisco Morales
J,
Reyes Navarrete
S,
Jiménez López
MA,
Betancourt Linares
R,
Cruz Del Carmen
I,
Illades Aguiar
B,
Román Román
A and
Fernández Tilapa
G
(2015). Helicobacter pylori and IFN-γ, IL-17A, IL-4 and TGF-β1 expression in patients with chronic gastritis and gastric cancer.
Front. Immunol.
Conference Abstract:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología.
doi: 10.3389/conf.fimmu.2015.05.00147
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Received:
19 May 2015;
Published Online:
14 Sep 2015.
*
Correspondence:
PhD. Gloria Fernández Tilapa, Universidad Autónoma de Guerrero, Unidad Acádemica de Ciencias Químico Biológicas, Chilpancingo, Guerrero, 39087, Mexico, gferti@hotmail.com