شناسایی مولکولی ژن پروتئین فعال کننده نوتروفیل هلیکوباکتر پیلوری موثر در کنترل آسم آلرژیک به روش PCR
محورهای موضوعی : ایمنی شناسی
علیرضا خالقی خرمی
1
,
ساناز مهمازی
2
,
رسول شکری
3
,
سید شمس الدین اطهاری
4
1 - گروه میکروبیولوژی، دانشکده علوم پایه، واحد زنجان، دانشگاه آزاد اسلامی، زنجان، ایران
2 - گروه میکروبیولوژی، دانشکده علوم پایه، واحد زنجان، دانشگاه آزاد اسلامی، زنجان، ایران
3 - گروه میکروبیولوژی، دانشکده علوم پایه، واحد زنجان، دانشگاه آزاد اسلامی، زنجان، ایران
4 - گروه ایمونولوژی، دانشکده پزشکی، دانشگاه علوم پزشکی زنجان، زنجان، ایران
کلید واژه: هلیکوباکتر پیلوری, آسم آلرژیک, پروتئین فعال کننده نوتروفیل,
چکیده مقاله :
پروتئین فعال کننده نوتروفیل هلیکوباکتر پیلوری (HP-NAP) یکی از پروتئین های مهم هلیکوباکتر پیلوری (H. pylori) است HP-NAP.دارای خواص تعدیل کننده ایمنی است و پاسخ های آلرژیک را از Th2 به Th1 هدایت می کند. هدف از این مطالعه شناسایی مولکولی ژن HP-NAP بومی ایران است. سویه بومی هلیکوباکتر پیلوری از انستیتو پاستور خریداری شد. آزمایش اوره آز اختصاصی، رنگ‌آمیزی گرم و مشاهده میکروسکوپی انجام شد. برای شناسایی HP-NAP در باکتری های بومی از روش PCR استفاده شد. در نهایت، محصول PCR برای تشخیص ژن HP-NAP الکتروفورز شد. پس از رنگ‌آمیزی باکتریایی و نیز مشاهده میکروسکوپی، باکتری‌های گرم منفی خمیده مشاهده شدند. آزمایش اوره آز تغییر در معرف اوره (فنل قرمز) را نشان داد که نشان دهنده هلیکوباکتر پیلوری است. آزمایش PCR و الکتروفورز یک باند 260 جفت باز مربوط به ژن HP-NAP را در مقایسه با نردبان (لدر) 100 نشان داد. با توجه به مطالعه انجام شده و تایید وجود این پروتئین در ساختار باکتری های بومی و همچنین مطالعات قبلی در در مناطق مختلف جهان، با مشاهده اثرات این پروتئین به عنوان تعدیل کننده ایمنی سیستم ایمنی، می توان امیدوار بود که در آینده از این پروتئین به عنوان وسیله ای بالقوه برای کنترل و درمان بیماری های آلرژیک به ویژه آسم آلرژیک استفاده شود.
Helicobacter pylori neutrophil activating protein (HP-NAP) is one of the important proteins of Helicobacter pylori (H. pylori). HP-NAP has immunomodulatory properties and directs allergic responses from Th2 to Th1. The aim of this study is the molecular identification of the HP-NAP gene native to Iran. The native strain of H. pylori was purchased from the Pasteur Institute. A specific urease test was performed, along with gram staining and microscopic observation. To identify HP-NAP in native bacteria, the PCR method was used. Finally, the PCR product was electrophoresed to detect the HP-NAP gene. After bacterial staining and microscopic observation, bent gram-negative bacteria were observed. The urease test showed a change in the urea reagent (phenol red), indicating H. pylori. The PCR test and electrophoresis revealed a 260 bp band related to the HP-NAP gene, compared with ladder 100. According to the study conducted and the confirmation of the presence of this protein in the structure of native bacteria, as well as previous studies in different regions of the world, observing the effects of this protein as an immunomodulator of the immune system, we can hope that in the future this protein will be used as a potential means to control and treat allergic diseases, especially allergic asthma.
[1] Gonzalo JA, Lloyd CM, Kremer L, Finger E, Martinez-A C, Siegelman MH, et al. Eosinophil recruitment to the lung in a murine model of allergic inflammation. The role of T cells, chemokines, and adhesion receptors. Journal of Clinical Investigation. 1996; 98(10): 2332-45. doi:10.1172/JCI119045
. [2] Arnold IC, Dehzad N, Reuter S, Martin H, Becher B, Taube C, et al. Helicobacter pylori infection prevents allergic asthma in mouse models through the induction of regulatory T cells. Journal of Clinical Investigation. 2011; 121(8):3088-93. doi:10.1172/JCI45041
[3] Kosunen TU, Höök-Nikanne J, Salomaa A, Sarna S, Aromaa A, Haahtela T. Increase of allergen-specific immunoglobulin E antibodies from 1973 to 1994 in a Finnish population and a possible relationship to Helicobacter pylori infections. Clinical & Experimental Allergy. 2002; 32(3): 373-8. doi:10.1046/j.1365-2222.2002.01330.x
[4] Marshall BJ, Warren JR. Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet. 1984; 1(8390): 1311-5.
doi:10.1016/s0140-6736(84)91816-6
[5] Parsonnet J, Friedman GD, Vandersteen DP, Chang Y, Vogelman JH, Orentreich N, et al. Helicobacter pylori infection and the risk of gastric carcinoma. New England Journal of Medicine. 1991; 325(16): 1127-31. doi:10.1056/NEJM199110173251603
[6] Wotherspoon AC, Ortiz-Hidalgo C, Falzon MR, Isaacson PG. Helicobacter pylori-associated gastritis and primary B-cell gastric lymphoma. Lancet. 1991; 338(8776): 1175-6. doi:10.1016/0140-6736(91)92035-z
[7] Lankarani KB, Ravanbod MR, Aflaki E, Nazarinia MA, Rajaee A. High prevalence of Helicobacter pylori infection in Behcet's disease. BMC Gastroenterology. 2014; 14: 58. doi:10.1186/1471-230X-14-58
[8] Fullerton D, Britton JR, Lewis SA, Pavord ID, McKeever TM, Fogarty AW. Helicobacter pylori and lung function, asthma, atopy and allergic disease--a population-based cross-sectional study in adults. International Journal of Epidemiology. 2009; 38(2): 419-26. doi:10.1093/ije/dyn348
[9] Matricardi PM, Rosmini F, Riondino S, Fortini M, Ferrigno L, Rapicetta M, et al. Exposure to foodborne and orofecal microbes versus airborne viruses in relation to atopy and allergic asthma: epidemiological study. British Medical Journal. 2000; 320(7232): 412-7. doi:10.1136/bmj.320.7232.412
[10] Shiotani A, Miyanishi T, Kamada T, Haruma K. Helicobacter pylori infection and allergic diseases: epidemiological study in Japanese university students. Journal of Gastroenterology and Hepatology. 2008; 23(7 Pt 2): e29-33.
doi:10.1111/j.1440-1746.2007.05107.x
[11] Chen Y, Blaser MJ. Helicobacter pylori colonization is inversely associated with childhood asthma. Journal of Infectious Diseases. 2008; 198(4): 553-60. doi:10.1086/590158
[12] Wang Y, Bi Y, Zhang L, Wang C. Is Helicobacter pylori infection associated with asthma risk? A meta-analysis based on 770 cases and 785 controls. International Journal of Medical Sciences. 2012; 9(7): 603-10. doi:10.7150/ijms.4970
[13] Nyström J, Svennerholm A-M. Oral immunization with HpaA affords therapeutic protective immunity against H.pylori that is reflected by specific mucosal immune responses. Vaccine. 2007; 25(14): 2591-8.
doi:10.1016/j.vaccine.2006.12.026
[14] Amedei A, Cappon A, Codolo G, Cabrelle A, Polenghi A, Benagiano M, et al. The neutrophil-activating protein of Helicobacter pylori promotes Th1 immune responses. Journal of Clinical Investigation. 2006; 116(4): 1092-101. doi:10.1172/JCI27177
[15] Kottakis F, Papadopoulos G, Pappa EV, Cordopatis P, Pentas S, Choli-Papadopoulou T. Helicobacter pylori neutrophil-activating protein activates neutrophils by its C-terminal Region even without dodecamer formation, which is a prerequisite for DNA protection--novel approaches against Helicobacter pylori inflammation. FEBS journal. 2008; 275(2): 302-17.
doi:10.1111/j.1742-4658.2007.06201.x
[16] Zanotti G, Papinutto E, Dundon W, Battistutta R, Seveso M, Giudice G, et al. Structure of the neutrophil-activating protein from Helicobacter pylori. Journal of Molecular Biology. 2002; 323(1): 125-30.
doi:10.1016/s0022-2836(02)00879-3
[17] Del Prete G, Chiumiento L, Amedei A, Piazza M, D'Elios MM, Codolo G, et al. Immunosuppression of TH2 responses in Trichinella spiralis infection by Helicobacter pylori neutrophil-activating protein. Journal of Allergy and Clinical Immunology. 2008; 122(5): 908-913.e5. doi:10.1016/j.jaci.2008.08.016
[18] Athari SS, Omidi R. Report of a patient with complex composites of hepatitis B virus, allergic asthma and diabetes. Asian Pacific Journal of Tropical Biomedicine. 2014; 4(Suppl1): S59-61. doi:10.12980/APJTB.4.2014C683
[19] Garcia R, Athari SS. Overview of relationship of allergic asthma with diet and food intake. Journal of food quality and hazards control. 2015; 2(1): 1-5.
[20] Codolo G, Mazzi P, Amedei A, Del Prete G, Berton G, D'Elios MM, et al. The neutrophil-activating protein of Helicobacter pylori down-modulates Th2 inflammation in ovalbumin-induced allergic asthma. Cell Microbiology. 2008; 10(11): 2355-63.
doi:10.1111/j.1462-5822.2008.01217.x
[21] Sudo N, Sawamura S, Tanaka K, Aiba Y, Kubo C, Koga Y. The requirement of intestinal bacterial flora for the development of an IgE production system fully susceptible to oral tolerance induction. Journal of Immunology. 1997; 159(4): 1739-45.
[22] Annagür A, Kendirli SG, Yilmaz M, Altintas DU, Inal A. Is there any relationship between asthma and asthma attack in children and atypical bacterial infections; Chlamydia pneumoniae, Mycoplasma pneumoniae and Helicobacter pylori. Journal of Tropical Pediatrics. 2007; 53(5): 313-8. doi:10.1093/tropej/fmm040
[23] Strachan DP. Hay fever, hygiene, and household size. British Medical Journal. 1989; 299(6710): 1259-60. doi:10.1136/bmj.299.6710.1259
[24] Herz U, Lacy P, Renz H, Erb K. The influence of infections on the development and severity of allergic disorders. Current Opinion in Immunology. 2000; 12(6): 632-40. doi:10.1016/s0952-7915(00)00155-2
[25] Engler DB, Reuter S, van Wijck Y, Urban S, Kyburz A, Maxeiner J, et al. Effective treatment of allergic airway inflammation with Helicobacter pylori immunomodulators requires BATF3-dependent dendritic cells and IL-10. Proceedings of the National Academy of Sciences. 2014; 111(32): 11810-5.
doi:10.1073/pnas.1410579111
[26] Wohlleben G, Erb KJ. Immune stimulatory strategies for the prevention and treatment of asthma. Current Pharmaceutical Design. 2006; 12(25): 3281-92. doi:10.2174/138161206778194114
[27] Papadopoulos NG, Konstantinou GN. Antimicrobial strategies: an option to treat allergy? Biomedicine & Pharmacotherapy. 2007; 61(1): 21-8.
doi:10.1016/j.biopha.2006.10.004
[28] Khamechian T, Movahedian AH, Ebrahimi Eskandari G, Heidarzadeh Arani M, Mohammadi A. Evaluation of the Correlation Between Childhood Asthma and Helicobacter pylori in Kashan. Jundishapur Journal of Microbiology. 2015; 8(6): e17842.
doi:10.5812/jjm.8(6)2015.17842
[29] Reibman J, Marmor M, Filner J, Fernandez-Beros ME, Rogers L, Perez-Perez GI, Blaser MJ. Asthma is inversely associated with Helicobacter pylori status in an urban population. PLoS One. 2008; 3(12): e4060.
doi:10.1371/journal.pone.0004060
[30] Blaser, Martin J et al. “Does Helicobacter pylori protect against asthma and allergy?” Gut. 2008; 57(5): 561-7. doi:10.1136/gut.2007.133462
[31] Taube C, Muller A. The role of Helicobacter pylori infection in the development of allergic asthma. Expert Review of Respiratory Medicine. 2012; 6(4): 441–9. doi:10.1586/ers.12.40
[32] Blaser MJ. The Jeremiah Metzger Lecture: global warming redux: the disappearing microbiota and epidemic obesity. Transactions of the American Clinical and Climatological Association. 2012; 123: 230-8.
[33] Blaser MJ. The theory of disappearing microbiota and the epidemics of chronic diseases. Nature Reviews Immunology. 2017; 17(8): 461-3. doi:10.1038/nri.2017.77
[34] O’Connor A, O’Morain CA, Ford AC. Population screening and treatment of Helicobacter pylori infection. Nature Reviews Gastroenterology & Hepatology. 2017; 14(4): 230-40. doi:10.1038/nrgastro.2016.195
[35] Yap TW, Leow AH, Azmi AN, Callahan DL, Perez-Perez GI, Loke MF, et al. Global fecal and plasma metabolic dynamics related to Helicobacter pylori eradication. Frontiers in Microbiology. 2017; 8: 536.
doi:10.3389/fmicb.2017.00536
[36] Abadi AT, Kusters JG. Management of Helicobacter pylori infections. BMC Gastroenterology. 2016; 16(1): 94.
[37] Lankarani KB, Honarvar B, Athari SS. The mechanisms underlying Helicobacter pylori-mediated protection against allergic asthma. Tanaffos. 2017; 16(4): 251.
[38] van Wijck Y, de Kleijn S, John-Schuster G, Mertens TCJ, Hiemstra PS, Muller A, et al. Therapeutic application of an extract of Helicobacter pylori ameliorates the development of allergic airway disease. Journal of Immunology. 2018; 200(5): 1570-9.
doi:10.4049/jimmunol.1700987
[39] Arnold IC, Hitzler I, Muller A. The immunomodulatory properties of Helicobacter pylori confer protection against allergic and chronic inflammatory disorders. Frontiers in Cellular and Infection Microbiology. 2012; 2:10. doi:10.3389/fcimb.2012.00010
[40] Sehrawat A, Sinha S, Saxena A. Helicobacter pylori neutrophil-activating protein: a potential Treg modulator suppressing allergic asthma? Frontiers in Microbiology. 2015; 6: 493. doi:10.3389/fmicb.2015.00493
[41] Permin H, Andersen LP. Inflammation, immunity, and vaccines for Helicobacter pylori. Helicobacter. 2009; 14(Suppl 1): 21-8. doi:10.1111/j.1523-5378.2005.00337.x
[42] Wijck YV, John-Schuster G, van Schadewijk A, van den Oever RL, Obieglo K, Hiemstra PS, et al. Extract of Helicobacter pylori ameliorates parameters of airway inflammation and goblet cell hyperplasia following repeated allergen exposure. International Archives of Allergy and Immunology. 2019; 180(1): 1-9.
doi:10.1159/000500598
[43] Kyburz A, Fallegger A, Zhang X, Altobelli A, Artola-Boran M, Borbet T, et al. Transmaternal Helicobacter pylori exposure reduces allergic airway inflammation in offspring through regulatory T cells. Journal of Allergy and Clinical Immunology. 2019; 143(4): 1496-e11.
doi:10.1016/j.jaci.2018.07.046