The Role of Tumor Necrosis Factor- alpha and Interferon-gamma as proinflammatory cytokines involved in Gastritis and peptic ulcers Caused by Helicobacter pylori and Evaluation of Ig G and Cag A

as Immunodiagnostic markers in Humans.

Shindy M. Sherif *; Nihal M. Al-Assaly**, Hanaa Mansour ***; Wafaa Mansour****Tropical, **Clinical Chemistry and ***Immunology departments, Theodor Bilharz Research Institute

Conference: AACC, Chicago II,USA, July 23-27, 2006; Abstract A37Accepted in Research Journal of Medicine and Medical Sciences, 2008, © 2008, INSInet Publication

AbstractHelicobacter pylori is an important cause of many abdominal and extra-abdominal

manifestations.It can induce proinflammatory cytokines and stimulate the production of free radicals. Gastritis and peptic ulcers are always accompanied with increased expression of TNF-α and IFN-γ. The aim of the present study was to evaluate the role of TNF-α and IFN-γ as proinflammatory cytokines in patients infected with H. pylori, to measure the levels of anti-H. pylori Ig G antibodies in patients sera and to estimate the correlation between Cag A positivity and gastritis and peptic ulcer diseases. The present study included 148 patients clinically screened for the diseases; 86 were males and 62 were females ; and 28 subjects as negative controls. All cases were serologically examined for Ig G and Cag A positivity using ELISA technique and TNF-α and IFN-γ were evaluated in Cag A positive sera. The results obtained showed that in patients with gastritis, anti-H. pylori Ig G positivity in males was 91.5 % and in females 81.25% while, the percentage of Cag A positivity was 87.3 % in males and 75% in females. In patients with peptic ulcers; anti-H. pylori Ig G positivity in males was 76.9 % and in females was 70 % while Cag A positivity was 69.2 % in males and 70% in females. The levels of TNF-α and IFN-γ in patients with gastritis and peptic ulcers are significantly increased in comparison to the control subjects (P< 0.05). In conclusion, it is important to study the role of cytokines in the inflammatory effect caused by H pylori infection in patients with gastritis and peptic ulcer. It is also concluded that the estimation of Ig G levels and Cag A positivity in these patients, offered a better immunodiagnostic markers before offering the endoscopy.

Introduction:

Helicobacter pylori is considered as the most common human bacterial infection. It causes a worldwide chronic diseases David and Graham, 2000). Chronic gastritis, peptic ulcers, MALT lymphoma and epidemiologically gastric carcinoma are related to this infection (Peek and Blaser, 2000; Shendy et al, 2001 and Sepulveda et al, 2002). Many studies have linked H. pylori to extragastric diseases such as migraine, skin diseases, coronary artery diseases and cholecystitis (Wedi and Kapp, 2000). Isolates of H pylori can be divided into two major distinct phenotypic groups: typeI bacteria, express the vaculating cytotoxin (VacA) and the cytotoxin-associated gene (CagA), type II bacteria do not express VacA or CagA. Such strains show different pathogenic potential: type I may induce mucosal damage by stimulating gastric epithelial cytokine response and may produce a variety of other factors that determine directly and indirectly the local and remote inflammatory response (Xiang et al., 1995). It was found that persons colonized with cagA+ strains have an increased risk of developing more severe gastric diseases such as peptic ulcer and distal (non-cardia) gastric cancer than those harboring cagA- strains (Perez-Perez et al., 1999). A correlation between CagA seropositivity and peptic ulcer disease was found in this study. Also it showed correlation between distal gastric cancer rated and CagA prevalence. Neoplasma in gastrointestinal tract was accelerated by chronic inflammation. Tsuji et al. (2003) reported that H. pylori induced proinflammatory cytokines and stimulated the production of free radicals. He

stated that TNF-α played a pivotal role in induction, growth and metastasis of neoplasma. It can induce phenotypic changes in gastric cells. Also, IFN-γ production by H. pylori causes peptic ulcer and gastric inflammation (Holk 2003). These data confirmed the relation between the increased cytokines and the inflammatory activity in gastric mucosa in H. pylori infection.

The aim of this study is to evaluate the magnitude of H pylori infection in patients with gastritis and peptic ulcer by evaluating anti-H.pylori Ig G and Cag A positivity. Another aim is to evaluate the role of TNF-α and IFN-γ as proinflammatory cytokines affecting the response to this infection.

Patients and methods:

This work included 148 patients they were studied over 8 months at the outpatient clinic in Theodor Bilharz research Institute and Agoza Hospital. Eighty six were males and 62 were females with random age. Most of them complained from manifestations suggestive of upper gastrointestinal diseases in addition to other extragastric manifestations such as; biliary and chronic respiratory symptoms, general fatigue, bad odour of mouth, constipation and migraine.

All patients and controls were investigated for history and clinical examination, stool and urine analysis, liver function tests, kidney function tests, abdominal ultrasound and upper esophagogastroduodenoscopy. At least three gastric biopsies were taken (from anterior, posterior walls of the antrum and the body of stomach) for histological assessment and H. pylori detection (by direct microscopic examination and rapid urease test). They were also investigated for anti-H. pylori Ig G. Patients with gastritis or peptic ulcers and positive H. pylori test by any methods were investigated for Cag A positivity using ElISA technique and for TNF-α and IFN-γ levels in their sera.

1- Detection of serum anti-H pylori Ig G:Sera were collected from patient’s blood and specific anti-H pylori Ig G was assayed according to Segal et al. (1996). Cultures were diluted 1:1000 and antigen was prepared by using the sonicates from the H. pylori strain and protein content was estimated. Plates were coated with the antigen and incubated over night at room temperature, then sera applied in dilutions of 1:200, incubated for 1 hour at 370 C, then washed using PBS/T (pH 7.2), then the antihuman IgG peroxidase conjugate (Sigma) was applied in a dilution of 1:1000 and incubated 30 minutes at 370 C, then washed 5 times with PBS/T and OPD substrate (Sigma) was then added, and the reaction was stopped after 30 minutes using 8N H2SO4. optical density at 492 nm was identified.

2- Detection of Cag A:A 66-Kda Cag A antigen fragment (Boehringes Mannheim Corp.) had been cloned in Escherichia coli (PORV 220) as mentioned by Blaser et al. (1995). For the Cag A, ELISA were coated with the previously prepared Cag A antigen and incubated over night at room temperature. Plates were washed using PBS/T (pH 7.2) then patient’s sera diluted 1: 100 were added and left for 1 hour at 370 C, then after wash, anti-human specific Ig G peroxidase conjugate was added at 1:1000 dilution and left for 30 minutes at 370 C, then after wash, OPD substrate was added and the reaction was stopped after 30 minutes and optical density was measured at 492 nm. Cases with positive Cag A were proposed to TNF-α assay and IFN-γ detection assay.

3- Detection of TNF- α :Detection of TNF- α is done by sandwisch ELISA. TNF- α was assayed in patient's sera according to Damas et al. (1989), using TNF-α enzyme immunoassay kit (Immunotech, A coulter company, France). 100 μl of TNF- α conjugate was added to each well of microtitre

plate coated with anti-TNF- α standard, control and samples were added to all wells except the blank and incubated for 120 minutes at room temperature with shaking. After washing 200 μl of substrate were added to all wells including the blank well, then incubated for 30 minutes in the dark at room temperature with shaking, the 50 μl of stop solution were added to all wells and absorbance was read at 405 nm, the the concentration was calculated by interpolation from the standard curve that was performed in the same assay.4- Detection of IFN-γ by sandwisch ELISA: IFN-γ was assayed in the sera of patients according to De Maeyer et al. (1992), using IFN-γ enzyme immunoassay kit (Immunotech, A Beckman Coulter Company, France). A 96 microliter plate applied with kit coated with anti- IFN-γ monoclonal antibody, 50 μl of standard or samples were added to each well, plate was covered and incubated for 2 hours at room temperature with shaking. After washing, 50 μl of biotinylated antibody, then 100 μl of streptavidin- HRP conjugate were added to all wells, except to that for substrate blank and plate was incubated for 30 minutes at room temperature with shaking, then after washing 100 μl of TMB substrate was added to all wells. Plate was incubated for 20 minutes at room temperature in the dark with shaking, then 50 μl of stop solution were added to all wells and absorbance was read at 450 nm, then the concentration was calculated by interpolation from the standard curve that was performed in the same assay.

Statistical analysis: The data was represented in mean of values ± SE using ANOVA (analysis of variance).

Results:

From the screening of GIT manifestations of 148 patients, 86 males and 62 females, it was found that 71 males and 42 females suffered from different grades of gastritis as detected by gross endoscopic examination and histopathology, while 13 males and 10 females had peptic ulcers (with surrounding gastritis or duodenitis in all cases). Twenty eight subjects were considered as negative controls with negative microbiological findings (negative urease test and negative microscopic examination for H pylori and negative anti H. pylori IgG test). However, endoscopic and histopathological findings of gastritis (in 4 cases) and peptic ulcers (in one case) were all negative for H pylori by all means. They all have history of NSAID intake in the previous few weeks. Tables 1 and 2 illustrate the number and percentage positivity of the controls and patients infected with H. pylori (males and females) suffering from gastritis (gross and endoscopic) and peptic ulcers. It includes also the results of rapid urease test done on gastric biopsies taken during endoscopy. From the total patients of 148 showing microscopic gastritis, only 119 (80.4%) showed gross gastritis by endoscopic examination and 23 (15.5%) showed gross peptic ulcer (18 duodenal and 5 gastric ulcers). For males, 71 (48%) showed gross gastritis and 13 (8.8%) showed peptic ulcers (11 duodenal and 2 gastric ulcers. For females, 48 (32.4 %) showed gross gastritis and 10 (6.8%) showed peptic ulcers (7 duodenal and 3 gastric ulcers). Males showed significantly higher prevalence of microscopic gastritis than females (P<0.05) and insignificant differences for other manifestations. H. pylori organism was detected in 133 cases (89.9%), while rapid urease test was positive in (126 (85.1%) of total cases using antral biopsies. These results showed that microscopic examination of gastric biopsy is more sensitive than urease test for detection of H. pylori.

Table 3 shows the anti-H. pylori antibody and Cag A positivity in different cases and sex distribution. Antibody positivity was found in 91.5 % of male patients and in 81.25 % of female patients with gastritis. In patients with peptic ulcers, antibody positivity was found in 76.9 % in males and in 70 % in females. Cag A positivity was detected in 87.3 % of males and in 75 % of females with gastritis. It was detected in 61.5 % of males and in 60 % of females with peptic

ulcers. H pylori and Cag A positivity were significantly higher among male patients than females with gross gastritis (P<0.05).

Table 4 shows the results of TNF- α and IFN-γ levels in patients and control group. In controls, the mean TNF- α level was 301±112 Pg/ml while it was 448±257 Pg/ml for males and 352±231 Pg/ml for females with gastritis. In peptic ulcer patients, it was 498±305 Pg/ml for males and 568±341 Pg/ml for females. It was significantly higher in all types of patients than in controls (P< 0.01). The mean IFN-γ level in controls was 0.41 ± 0.23 IU/ml. It was 0.73±0.42IU/ml for males and 0..56±0.51IU/ml for females with gastritis. In peptic ulcer patients, it was 0.61 ±0.32 IU for males and 0.82 ±0.37 IU for females. It was also significantly higher in all types of patients than in controls (P< 0.01). Only TNF- α was significantly higher in males than females with gastritis (P <0.01) but not with peptic ulcer and not for IFN-γ (P>0. 05).

Table 1: endoscopic, microscopic and urease test findings in controls and patients of this study.Number Gross findings Microscopic examination. Urease Test

positivitygastritis Peptic U. gastritis H. pylori positivityControls 28 2 1 4 0 0Total patients 148 119

(80.4%) 23 148 133

(89.9%) 126 (85.1%)

Males 86 71 13 86* 81 76Females 62 48 10 48 52 50

* Males showed significantly higher prevalence of microscopic gastritis only than females (P<0.05). Table 2: summary of table 1.Patients Total Number Gastritis (gross) Peptic ulcers

Number Percentage Number Percentage Number PercentageTotal 148 100 % 119 80.4 % 23 15.5 %Males 86 58.1 % 71 48 % 13 8.8 %Females 62 41.9 % 48 32.4 % 10 6.8 %

Table 3: anti-H. pylori antibody and Cag A positivity in different cases and sex distribution

PatientsTotal number

Percentage of anti-H. pylori positivity

Percentage of Cag A positivity

Number of cases % Number of cases % gastritis Males 71 65 91.5 % * 62 87.3 %*

Females 48 39 81.25 % 36 75 %peptic ulcer

Males 13 10 76.9 % 9 61.5 %Females 10 7 70 % 6 60 %

* H pylori and Cag A positivity were significantly higher among male patients than females with gross gastritis (P<0.05).Table 4: TNF- α and IFN-γ in patients and control group

Number TNF- α IFN-γControl 28 301 ±112 Pg/ml 0.41 ± 0.23 IU/mlPatients Gastritis Peptic ulcers

No. TNF- α IFN-γ No. TNF- α IFN-γMales 65 448±257 Pg/ml 0.73±0.42IU/ml 10 498±305 Pg/ml 0.61 ±0.32 IUFemales 39 352±231 Pg/ml 0..56±0.51IU/ml 7 568±341 Pg/ml 0..82 ±0.37 IU*Both TNF- α and IFN-γ levels were also significantly higher in all types of patients than in controls (P< 0.001). Only TNF- α was significantly higher in males than females with gastritis (P <0.01) but not with peptic ulcer and not for IFN-γ (P>0. 05).

Yuhong Yuan; Ireneusz T Padol; Richard H Hunt (2006): Peptic Ulcer Disease Today Nat Clin Pract Gastroenterol Hepatol. 2006;3(2):80-89. ©2006 Nature Publishing Group

Discussion: In this study, patients with endoscopically evident gastritis and peptic ulcers were selected.

Control subjects, despite the presence of few cases of gastritis due to other causes, were negative for H. pylori by all means. Gross gastritis was detected in 119 cases of patient group (80.4%). Peptic ulcer is not as common as gastritis in our patients. It was found only in 15.5% of cases. Gastritis and peptic ulcer are often associated with H. pylori infection. MALT lymphoma and epidemiologically gastric carcinoma are also related to this infection (Peek and Blaser, 2000;Zucca et al., 2000; Shendy et al, 2001; Sepulveda et al, 2002 and Tsuji et al., 2003). Eradication of H. pylori infection is now the mainstay of treatment for peptic ulcer disease, and has resulted in very high ulcer healing rates and recurrence rates that have dropped dramatically, especially for individuals with a duodenal ulcer (Yuhong et al., 2006). Endoscopy is the best tool to diagnose H. pylori related gastric diseases either by direct visualization of gastro duodenal mucosa or by taking tissue biopsies for both histopatholological examination and microscopic detection of H pylori itself. Also biopsies can be examined immediately for H pylori using rapid urease test before the patient leaves the unit. The spectrum of the disease differs according the duration of infection. The acute phase lasts 1 to 4 weeks and is replaced gradually by a chronic, mononuclear infiltrate in the lamina propria. Active gastritis refers to the presence of neutrophils mixed with mononuclear cells in the gastric mucosa. Chronic active gastritis occurs in the majority of infected individuals and consists of surface epithelial degeneration, persistent neutrophil infiltration of the epithelium and lamina propria, and mononuclear infiltration (lymphocytes and plasma cells) of the lamina propria. Lymphoid hyperplasia in the gastric mucosa is suggestive of H pylori infection. Generally, gastritis is most prominent in the corpus and antrum, with evidence of inflammation of the cardia in most infected individuals (Sharma et al., 2000). Chronic gastritis due to H pylori infection may be separated into distinct, clinically relevant phenotypes. Non-atrophic pangastritis occurs in the majority of H pylori-infected individuals with no predisposition to peptic ulcer disease or gastric atrophy. Prominent mucosal inflammation in chronic active gastritis often is evident in the antrum (antral-predominant gastritis), predisposing to hyperacidity and duodenal ulcer disease. In contrast, multifocal atrophic pangastritis and atrophic corpus-predominant gastritis result from long-standing infection and are characterized by glandular atrophy, intestinal metaplasia, and sparse inflammatory cells (Rubin 1997 and Faller and Kirchner 2001).

It was found in this study that microscopic examination of gastric biopsy is the best diagnostic method for detection of H. pylori. It is well known that more than 90% of duodenal ulcers are associated with H pylori, which is present in highest concentrations in the gastric antrum. A proximal-distal gradient of increasing organism densities exists along the corpus and antrum in duodenal ulcer disease and extends toward the transitional zone and gastroduodenal junction. Virtually all patients with duodenal ulcer disease have chronic, active, antral-predominant gastritis. With respect to duodenal ulcer disease, endoscopic visualization of the ulcer may be sufficient for diagnosis (Greenberg et al., 1996). Diagnostic confirmation of the presence of H pylori necessitates biopsy sampling of the gastric corpus and antrum. In contrast, the diagnostic evaluation of gastric ulcers requires biopsy specimens of the ulcer base and areas adjacent to gross ulceration to assess the histologic features for the presence of atrophic or neoplastic changes (Kuipers, 1997). Adjacent mucosa is evaluated directly for the presence of concomitant atrophy, dysplasia, intestinal metaplasia, or gastric adenocarcinoma (Arista-Nasr et al., 2001).

Atrophic gastritis is characterized by glandular atrophy and likely represents a precursor state for gastric adenocarcinoma (Correa 1992). Nonneoplastic mucosa adjacent to gastric carcinomas and lymphomas often displays atrophy, dysplasia, and intestinal metaplasia (Arista-Nasr et al., 2001). H. pylori organisms rarely are observed in association with regions of atrophic gastritis or gastric adenocarcinoma. A second form of gastric neoplasia, gastric MALT lymphoma, may be evaluated by histopathologic studies. Gastric marginal zone B-cell lymphomas (or MALT lymphomas) are highlighted histologically by the presence of lymphoepithelial lesions, prominent lymphoid follicles, and extensive zones of marginal B lymphocytes infiltrating the mucosa (Isaacson 1994). H pylori organisms may be visualized adjacent to areas of low-grade gastric MALT lymphomas but are observed less commonly in association with diffuse large B-cell lymphomas (high-grade MALT lymphomas).

In the present study, type 1 bacteria with Cag A positivity were significantly higher (P < 0.05) in patients with both diseases of H. pylori. It was detected in 87.3% of male patients and in 75% of female patients with gastritis and in 61.5% of males and in 60% of females with peptic ulcers. These results were in accordance with that of Gasbarrini et al., (1997). It was found that H pylori type 1 caused more severe illness and other associated diseases (Sozzi et al., 1998; Rudi et al., 2000 and Kato et al., 2000). Gasbarrini et al., (1997) studied and discussed the association of Cag A positivity with abdominal and extra-abdominal manifestations. He found that 11 out of 15 patients with peptic ulcers, were Cag A positive which is statistically significant.

In our study, anti-H pylori Ig G antibody was detected in 91.5% of males and in 81.25% of females with gastritis while in patients with peptic ulcers, it was detected in 76.9% in males and in 70% in females. A previous study showed that antibodies to H. pylori infection were present in 64.5 % of infected patients (Durazzo, 2002). In another study, Gasbarrini et al., (1999) found a prevalence of anti H. pylori antibodies of 77 % in patients with cirrhosis due to HCV infection and in 59 % in matched blood donors.

Most patients infected with H pylori produce a measurable systemic immune response, composed primarily of IgG (Rathbone et al.,1986 and Perez-Perez et al., 1988). Serum IgA may be detected in fewer than half (range, 39%-82%) of infected patients, and serum IgM is found rarely. Since few patients have been evaluated during the acute phase of infection, data about the nature of the acute antibody response and seroconversion from IgM to IgG are sparse. Seroconversion to IgG was demonstrated between 22 and 33 days after infection in one volunteer study (Morris and Nicholson 1987). In naturally acquired infection, an initial serum IgM response was observed, and seroconversion of IgM to IgA was documented. Circulating anti-H pylori IgG antibodies persist at constant levels for years during infection. Levels of IgG1, IgG2, and IgG4 subclasses typically are elevated, whereas IgG3 antibodies (associated with acute infections) are not detected (Sobala et al., 1991). Autoimmune and corpus-predominant atrophic gastritis have many overlapping features. Antigastric antibodies are generated in a subset of infected individuals and include antiluminal and anticanalicular antibodies. A principal target of H pylori-associated autoantibodies is the parietal cell proton pump, H+,K+-ATPase (Claeys et al., 1998).

The key pathophysiologic event in H. pylori infection is the initiation of an inflammatory response (Israel et al., 2001). This response is most probably triggered by the bacterium's lipopolysaccharide, urease, and/or cytotoxins and is mediated by cytokines. The cytokine repertoire comprises a multitude of pro- and anti-inflammatory mediators whose function is to co-ordinate an effective immune/inflammatory response against invading pathogens without causing undue damage to the host. In addition to their pro- or anti-inflammatory properties, some H. pylori-induced cytokines have direct effects on gastric epithelial cells that have a profound effect

on gastric physiology. For example, the pro-inflammatory cytokine interleukin-1 beta (IL-1beta ) is the most potent of known agents that are gastric cytoprotective, antiulcer, antisecretory, and inhibitors of gastric emptying (Robert et al., 1991 and Wolfe and Nompleggi, 1992). Another important pro-inflammatory cytokine that is up-regulated by H. pylori infection is tumor necrosis factor alpha (TNF-alpha ), which also inhibits gastric acid secretion, but to a lesser extent than IL-1beta (Beales and Calam, 1998). Cytokines appear to regulate the migration of Th 1 and Th 2 lymphocytes to inflamed tissues during the immune response. A dual effect of H. pylori on the Th 1 response was verified. A stimulation response was verified by increase of IFN-γ and TNF-α (berribi et al., 2003 and Holk,2003). These studies confirmed the connection between increase of IFN-γ and TNF-α and the inflammatory activity in gastric mucosa due H. pylori infection. A significant increase of IFN-γ and TNF-α levels in the sera of patients infected with H. pylori has been reported by Tsuji et al (2003), Konturk et al. (2003) and Slomiany (2003). In the present study, a highly significant increase of TNF-α levels (P<0.01) was detected in both males and females with gastritis and peptic ulcer. Also a significant increase of IFN-γ levels (P<0.01) was detected in both males and females with gastritis and peptic ulcer due to H. pylori infection. These results are in accordance with the results obtained by Holk (2003), Tsuji (2003), Konturk et al. (2003) and Slomiany (2003). In one study, it was found that the severity of gastritis associated with H pylori infection was correlated with mucosal expression of the tumor necrosis factor alpha subunit and IFN-gamma (Lehmann , et al. 2002)

In this study, classification of patients according sex showed some differences in the manifestations and blood tests among both males and females. Males showed significantly higher prevalence of microscopic gastritis than females (P<0.05). There was no difference among both sexes as regards to peptic ulcer which might be due to small number of cases. H pylori and Cag A positivity were significantly higher among male patients than females with gross gastritis (P<0.05). Only TNF- α was significantly higher in males than females with gastritis (P <0.01) but not with peptic ulcer and not for IFN-γ (P>0. 05). Previous studies showed also that the prevalence of peptic ulcer was shifted from being a disease predominant in males to one with a nearly comparable prevalence in both sexes and nearly at the same age (Kurata et al.,1986 and Turkdoan et al., 1999). However, the incidence and male to female ratio still increased with advancing age (Mowaten et al., 1975).

From the present study, it was concluded that H. pylori is the main cause of gastritis and peptic ulcer and the disease was caused mainly by type 1 bacterium as obtained by the detection of Cag A positivity. Also, it seems from this study that Cag A positivity can be used as an additive diagnostic marker to other methods of diagnosis. It could be also concluded that evaluation of IFN-γ and TNF-α levels might play a role in the diagnosis and prognosis of H. pylori infection. In this study, there were sex predilection of some manifestations, particularly microscopic gastritis, and Cag A positivity which were more in males than females.

دور عامل التأكل الورمى ألفا والتنترفيرون جاما كسيتوكينا ت مسببة لللتهاب فى حال ت التهاب المعععدة والقرح الهضمية الناتجععة ععن الاصععابة بالبكتريععا الحلزوتنيععة البوابيععة وتقييععم الجسععام المضععادة ج وال

Cag A.كوسائل تشخيصية مناعية فى التنسان شندى محمد شندى*، تنهال العسالى**، هناء منصور*** و وفاء منصور***

معهد تيودور بلهارس للبحاث، قسم الجهععاز الهضعمى والكبعد والمعراض المتوطنعة *و قسععم الكيميععاء الكلينيكيعة و**هناء منصور، شندى محمد شندى، وفاء منصور و تنهال العسالى و*** قسم المناعة

ان البكتريععا الحلزوتنيععة البوابيععة هععى سععبب مهععم للكععثير مععن العععراض الهضععمية والغيععر هضععمية وإتنهععا تحفععز تكععوين سيتوكينا ت المسببة لللتهابا ت وتكوين الجذور الحرة. كم ان التهاب المعدة والقرح الهضمية غالبا ما يصاحبها ارتفععاعال

فى التعبير عن عامل التأكل الورمى ألفا والتنترفيرون جاما. وكان الهدف من هعذه الدراسعة هعو تقييعم دور عامعل التأكعل العورمى ألفعا والتنعترفيرون جامعا كمسعببا ت لللتهابعا ت فعى المرضى المصابين بالبكتريا الحلزوتنية البوابية وقياس الجسام المضادة ج فى دم المرضى وعمععل ربععط بيععن ايجابيععة ال

Cag A. وبين التهاب المعدة والقرح الهضمية حالة سليمة للمقارتنة. وتم عمل فحص شامل لجميععع۲٨ أتنثى) و٦۲ رجل و٨٦ مريضا (١٤٨وقد شملت هذه الدراسة

Cagالمرضى وعمل الجسام المضادة وال A بطريقة ELISAكما تم قياس عامل التأكل الورمى ألفععا والتنععترفيرون الجسععام المضععادة ايجابيععة التهاب المعععدة فععان. و قد أظهر ت النتائج أتنه فى حالةCag Aجاما فى المرضى ايجابي ال

% أما فى٧٥% وفى التناث ٣,٨٧ فى الذكور Cag A% وايجابية ال ۲٥,٨١% وفى التناث٥,٩١كاتنت فى الذكور Cag% وايجابيععة ال ٧٠% وفعى التنعاث ٩,٧٦الجسام المضادة فى العذكور ايجابية حال ت القرحة الهضمية فقد كاتنت

A وقد كان مستوى عامل التأكل الورمى ألفا والتنعترفيرون جامعا أعلعى علعوا٧٠% وفى التناث ۲,٦٩ فى الذكور .% ذو دللة فى المرضى عنهما فى حال ت المقارتنة.

يستنتج من هذا البحث أتنه من المهم دراسة السيتوكينا ت ودورها فى اللتهابا ت الناتجة عن الاصابة بالبكتريعا الحلزوتنيعة فععىCag Aالبوابية فى مرضى التهاب المعدة والقرح الهضمية، كما يستنتج أيضا أن قيععاس الجسععام المضععادة ج وال

هؤلء المرضى أعطى دلل ت مناعية تشخيصية جيدة قبل عمل المنظار المعدى.

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total patients gastritis peptic ulcer0

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Figure 1: Number of patients with gastritis and peptic ulcer (according sex).

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Figure 2: anti-H. pylori antibody and Cag A positivity in male and female patients with gastritis

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Figure 3: anti-H. pylori antibody and Cag A positivity in male and female patients with p. ulcer.

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Figure 4: TNF-α in controls and patients with gastritis and peptic ulcers.

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Figure 5: IFN-γ in controls and patients with gastritis and peptic ulcers.