fa اثر ایمونومدولاتوری اگزوتوکسین A نوترکیب پسودوموناس بر سلول‌های دندریتیکی استخراج شده از طحال موش ايمونولوژي Immunology پژوهشي Research <p style="direction: rtl"><font face="Times New Roman" size="3"> </font><b><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman"">زمینه و هدف:</span></b><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""> با توجه به نقش کلیدی سلول دندریتیک در ایجاد پاسخ سیستم ایمنی بر علیه آنتی ژن‌های میکروبی و بیماری‌ها، در این مطالعه اثر پروتئین نوترکیب اگزوتوکسین </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">A</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> باکتری پسودوموناس آئروژینوزا در بلو غ و فعال‌سازی سلول‌های دندریتیک مورد بررسی قرار گرفت.</span></p><p class="MsoNormal" style="margin: 6pt 1.3pt 0pt 0cm text-align: justify line-height: normal" dir="rtl"><b><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman"">مواد و روش‌ها:</span></b><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""> با استفاده از </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">DNA</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> باکتری پسودوموناس آئروژینوزا پروتئین نوترکیب اگزوتوکسین </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">A</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> ساخته شد و اثر سیتوتوکسیک آن بر سلول دندریتیک به وسیله تست </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">MTT</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> مورد مطالعه قرار گرفت. همچنین اثر این آنتی ژن بر بیان مولکول‌های </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">CD40</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span>، </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">CD86</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> و </span><span dir="ltr"></span><span dir="ltr"></span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr"><span dir="ltr"></span><span dir="ltr"></span><font face="Times New Roman"><span style="mso-spacerun: yes"><font size="2"> </font></span>MHCΠ</font></span><span dir="rtl"></span><span dir="rtl"></span><span style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> <span lang="FA"><font face="B Nazanin" size="2">بر روی سلول‌های دندریتیک با استفاده از تکنیک فلوسایتومتری مورد بررسی قرار گرفت. علاوه بر این‌ها اثر این آنتی ژن بر تکثیر سلول </font></span></span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">T</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> به وسیله واکنش مختلط لنفوسیتی</span><span dir="ltr"></span><span dir="ltr"></span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr"><span dir="ltr"></span><span dir="ltr"></span> (MLR) </span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman"">و ترشح سایتوکاین‌های </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">IL-4</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> و </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">IFN-γ</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> بررسی شد. همچنین اثر این آنتی ژن بر تولید </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">IL-12</span><span dir="rtl"></span><span dir="rtl"></span><span style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> <span lang="FA"><font face="B Nazanin" size="2">توسط سلول‌های دندریتیک به کمک تکنیک </font></span></span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">ELISA</span><span dir="rtl"></span><span dir="rtl"></span><span style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> <span lang="FA"><font face="B Nazanin" size="2">بررسی شد. نتایج حاصل به وسیله تست آماری </font></span></span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin" mso-fareast-font-family: AdvGulliv-R" dir="ltr">one way ANOVA</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> مورد بررسی قرار گرفت.</span></p><p class="MsoNormal" style="margin: 6pt 1.3pt 0pt 0cm text-align: justify line-height: normal" dir="rtl"><b><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman"">نتایج:</span></b><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""> اگزوتوکسین </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">A</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> بقاء سلول دندریتیک را کاهش نداد و همچنین میزان بیان مولکول‌های کمک تحریکی </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">CD40</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span>، </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">CD86</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> و </span><span dir="ltr"></span><span dir="ltr"></span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr"><span dir="ltr"></span><span dir="ltr"></span><font face="Times New Roman"><span style="mso-spacerun: yes"><font size="2"> </font></span>MHCΠ</font></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman"">نسبت به کنترل منفی تغییر معنی داری پیدا نکرد. این آنتی ژن میزان تکثیر سلول </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">T</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> را کاهش داد و این کاهش تکثیر در غلظت 1/0 معنی دار بود. میزان ترشح </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">IL-12</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> توسط سلول دندریتیک تحت تاثیر این آنتی ژن افزایش یافت ولی میزان ترشح </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">IL-4</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> و </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">IFN-γ</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> در تست</span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">MLR </span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span><font size="2"><font face="B Nazanin"><span style="mso-spacerun: yes"> </span>کاهش معنی داری را نشان نداد.</font></font></span></p><p class="MsoNormal" style="margin: 6pt 1.3pt 0pt 0cm text-align: justify line-height: normal" dir="rtl"><b><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman"">نتیجه‌گیری:</span></b><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""> این آنتی ژن باعث تغییر در ترشح سایتوکاین‌ها توسط سلول‌های ایمنی شده و تکثیر سلول‌های </span><span style="font-family: "Times New Roman",serif font-size: 8pt mso-bidi-font-size: 10.0pt mso-bidi-font-family: "B Nazanin"" dir="ltr">T</span><span dir="rtl"></span><span dir="rtl"></span><span lang="FA" style="font-family: "B Nazanin" font-size: 10pt mso-ansi-font-size: 8.0pt mso-ascii-font-family: "Times New Roman" mso-hansi-font-family: "Times New Roman""><span dir="rtl"></span><span dir="rtl"></span> را کاهش می‌دهد.<o:p /></span></p><p><font face="Times New Roman" size="3"> </font></p><p><font face="Times New Roman" size="3"> </font></p> <font face="Times New Roman" size="3"> </font><p class="MsoNormal" style="margin: 6pt 0cm 0pt 1.3pt text-align: justify line-height: normal unicode-bidi: embed direction: ltr"><b><i><span style="color: black font-family: "Times New Roman",serif font-size: 10pt">Background & Objective:</span></i></b><span style="color: black font-family: "Times New Roman",serif font-size: 10pt"> Dendritic cell (DC) is as a key cell in activation of immune response against microbes and disease. Therefore, the effect of recombinant exotoxin A of Pseudomonas aeruginosa on the maturity and the activation of DCs was evaluated in this study.<span lang="FA" dir="rtl"><o:p /></span></span></p><font face="Times New Roman" size="3"> </font><p class="MsoNormal" style="margin: 6pt 0cm 0pt 1.3pt text-align: justify line-height: normal unicode-bidi: embed direction: ltr"><b><i><span style="color: black font-family: "Times New Roman",serif font-size: 10pt">Materials & Methods:</span></i></b><span style="color: black font-family: "Times New Roman",serif font-size: 10pt"> Recombinant exotoxin A was produced from Pseudomonas aeruginosa DNA. MTT assay was used to evaluate the cytotoxicity of this protein on DCs. The expression of co-stimulatory molecules CD40, CD86, and MHCΠ was evaluated by flow cytometry. Moreover, the effect of this antigen (Ag) on T-cell proliferation was evaluated using Mixed Lymphocyte Reaction (MLR) assay and the secretion of IL-4 and IFN- γ. Secretion of IL-12 by DCs was measured with </span><span class="st"><span style="font-family: "Times New Roman",serif font-size: 10pt">Enzyme-Linked Immunosorbent </span></span><em><span style="font-size: 10pt"><font face="Calibri">Assay</font></span></em><i style="mso-bidi-font-style: normal"><span style="color: black font-family: "Times New Roman",serif font-size: 10pt"> </span></i><span style="color: black font-family: "Times New Roman",serif font-size: 10pt mso-bidi-font-style: italic">(</span><span style="color: black font-family: "Times New Roman",serif font-size: 10pt">ELISA) method. The data were collected and analyzed with </span><span style="color: black font-family: "Times New Roman",serif font-size: 10pt mso-fareast-font-family: AdvGulliv-R">one way ANOVA test.</span><span style="color: black font-family: "Times New Roman",serif font-size: 10pt"><o:p /></span></p><font face="Times New Roman" size="3"> </font><p class="MsoNormal" style="margin: 6pt 0cm 0pt 1.3pt text-align: justify line-height: normal unicode-bidi: embed direction: ltr"><b><i><span style="color: black font-family: "Times New Roman",serif font-size: 10pt">Results</span></i></b><b><span style="color: black font-family: "Times New Roman",serif font-size: 10pt">:</span></b><span style="color: black font-family: "Times New Roman",serif font-size: 10pt"> Recombinant exotoxin A had no effect on DCs viability. In addition, expression of CD40, CD86, and MHCΠ did not change significantly compared to the negative control cells. Moreover, T-cells proliferation was decreased significantly at the concentration of 0.1µg/ml of this Ag. The secretion of IL-12 was increased by DCs, in contrast the secretion of IL-4 and IFN-γ in MLR supernatant did not decrease significantly.<o:p /></span></p><font face="Times New Roman" size="3"> </font><p class="MsoNormal" style="margin: 6pt 0cm 0pt 1.3pt text-align: justify line-height: normal unicode-bidi: embed direction: ltr"><b><i><span style="color: black font-family: "Times New Roman",serif font-size: 10pt">Conclusio</span></i></b><b><span style="color: black font-family: "Times New Roman",serif font-size: 10pt">n:</span></b><span style="color: black font-family: "Times New Roman",serif font-size: 10pt"> Exotoxin A decreases the proliferation of T-cells and also leads to a change in the pattern of cytokine secretion of immune cells.<span lang="FA" dir="rtl"><o:p /></span></span></p><font face="Times New Roman" size="3"> </font><p class="MsoNormal" style="margin: 6pt 0cm 0pt 1.3pt text-align: justify line-height: normal unicode-bidi: embed direction: ltr"><span style="color: black font-family: "Times New Roman",serif font-size: 10pt"><!--stripped--><font size="2"></font><!--stripped--></span></p><font face="Times New Roman" size="3"> </font> اگزوتوکسین A, سلول دندریتیک, اثرات ایمونومدولاتوری Exotoxin A, Dendritic cell, Immunomodulatory effects 436 445 http://jabs.fums.ac.ir/browse.php?a_code=A-10-581-1&slc_lang=fa&sid=1 Mohammad Hossein Karimi محمد حسین کریمی karimi70@yahoo.com 2298252107 10031947532846006245 No Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. مرکز تحقیقات پیوند و ترمیم اعضاء، دانشگاه علوم پزشکی شیراز، شیراز، ایران. Aziz Japoni عزیز ژاپونی Rasouliman@yahoo.com 2298252107 10031947532846006246 No Department of Immunology, Professor Alborzi Clinical Microbiology Research Center, Namazi Hospital, Shiraz, Iran. مرکز تحقیقات میکروب شناسی بالینی پروفسور البرزی، دانشگاه علوم پزشکی شیراز، شیراز، ایران. Manouchehr Rasouli منوچهر رسولی rasouliman@yahoo.com 2298252107 10031947532846006247 Yes Department of Immunology, Professor Alborzi Clinical Microbiology Research Center, Namazi Hospital, Shiraz, Iran. مرکز تحقیقات میکروب شناسی بالینی پروفسور البرزی، دانشگاه علوم پزشکی شیراز، شیراز، ایران. Salimeh Ebrahimnezhad سلیمه ابراهیم نژاد karimi70@yahoo.com 2298252107 10031947532846006248 No Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. مرکز تحقیقات پیوند و ترمیم اعضاء، دانشگاه علوم پزشکی شیراز، شیراز، ایران.