fa مقایسه میزان کشندگی سلول‌های CD19 CAR-CIK و CD19 CAR-T در محیط in vitro ايمونولوژي Imunology پژوهشي Research <div style="text-align: justify;"><span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><span style="font-weight:bold"><span lang="FA" style="line-height:80%"><span style="font-family:Jadid"><span style="font-weight:normal">چکید</span></span></span><span lang="AR-SA" style="line-height:80%"><span style="font-family:Jadid"><span style="font-weight:normal">ه</span></span></span><span lang="FA" style="line-height:80%"><span style="font-family:Jadid"><span style="font-weight:normal"></span></span></span></span></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><i><span lang="FA" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:Titr">سابقه و هدف </span></span></span></i></b></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:">علی‌رغم موفقیت‌های سلول‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-T</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> در درمان بدخیمی‌های خونی، استفاده از منابع آلوژنیک لنفوسیت‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">T</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> برای تولید این سلول‌ها با خطر ایجاد </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">GvHD</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> همراه است. سلول‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CIK</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> سلول‌هایی با خطر پایین </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">GvHD</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> می‌باشند؛ بنابراین سلول‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-CIK</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> می‌توانند به عنوان جایگزینی بالقوه برای سلول‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-T</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> مورد ارزیابی قرار گیرند. </span></span></span></b></span></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><i><span lang="FA" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:Titr">مواد و روش‌ها</span></span></span></i></b></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><a name="_Hlk148088372"><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:">در این مطالعه تجربی، پس از جداسازی و کشت سلول‌های تک هسته‌ای خون محیطی دو اهداکننده سالم</span></span></span></b></a><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:">، سلول‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">T</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> و </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CIK</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> با استفاده از وکتور لنتی ویروسی حاوی ژن </span></span></span></b><b><i><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CD19 CAR</span></span></span></i></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> ترانسداکت شدند. در روز شانزدهم کشت برای مقایسه کشندگی سلول‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-T</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> و </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-CIK</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> از هم‌کشتی این سلول‌ها با سلول‌های هدف </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CD19⁺ K562</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> استفاده شد و میزان مرگ در بین سلول‌های هدف با استفاده از کیت سنجش لاکتات دهیدروژناز اندازه‌گیری شد. جهت تجزیه و تحلیل آماری از نرم‌افزار &nbsp;</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">Graph Pad Prism</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> ورژن 8 و تجزیه و تحلیل&nbsp; </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">Student T-test</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> و </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">Anova</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> و پست هاک </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">Tukey</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> استفاده شد و 05/0 </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">p<</span></span></span></b><b><span style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> معنادار در نظر گرفته شد.</span></span></span></b></span></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><i><span lang="FA" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:Titr">یافته‌ها</span></span></span></i></b></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:">در هر دو گروه سلول</span></span></span></b><b> </b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:">‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-T</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> و </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-CIK</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> گیرنده </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> موجب افزایش معنادار کشندگی سلول‌ها شد؛ </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold""></span></span></span></b></span></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:">به طوری که تفاوت معناداری بین قدرت <a name="_Hlk148247629">کشندگی سلول‌های </a></span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-T</span></span></span></b><b><span style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> (7/1 </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%">&plusmn;</span></span></b><b><span style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> 8/67%) و </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-CIK</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> (8/1 </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%">&plusmn;</span></span></b><b><span style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> 1/65%) مشاهده نشد</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:">.</span></span></span></b></span></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><i><span lang="FA" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:Titr">نتیجه گیری</span></span></span></i></b></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:">سلول‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-CIK</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> در محیط </span></span></span></b><b><i><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">in vitro</span></span></span></i></b><b><span style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> کشندگی مناسبی را از خود بروز می‌دهند لذا می‌توانند به عنوان جایگزینی برای سلول‌های </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span bold="" new="" roman="" style="font-family:" times="">CAR-T</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span 2="" lotus="" style="font-family:"> مطرح باشند. با این حال مطالعه‌های بیشتری جهت مقایسه سایر ابعاد مؤثر در عملکرد این دو سلول مورد نیاز می‌باشد. </span></span></span></b></span></span></span></span></span></span></span><br> <br> &nbsp;</div> <span style="font-size:12pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="line-height:106%"><span style="font-family:Calibri,sans-serif"><span lang="FA" style="font-size:14.0pt"><span style="line-height:106%"><span style="font-family:"B Nazanin""></span></span></span></span></span></span></span></span> <div style="text-align: justify;"><span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span style="font-size:12.0pt">Abstract</span></b><b><span style="font-size:12.0pt"></span></b></span></span></span></span></span><br> <span style="font-size:10pt"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><i>Background and Objectives</i></b></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times="">Despite the success of CAR-T cells in the treatment of hematologic malignancies, the use of allogeneic sources of T lymphocytes to produce these cells is associated with risk of GvHD. Cytokine induced killer (CIK) cells are a population with a lower risk of GvHD. Therefore, CAR-CIK cells can be evaluated as a potential alternative for CAR-T cells.</span></span></span></span></span><br> <br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><i>Materials and Methods</i></b></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times="">In this experimental study, after isolation and culture of mononuclear cells isolated from the peripheral blood of two healthy donors, T and CIK cells are transduced with CD19 gene containing lentiviral vector. On the 16th day of culture, to compare the cytotoxicity of CAR-T and CAR-CIK cells, these cells were co-cultured with CD19+ K562 target cells, and the death rate among the target cells was measured using a lactate dehydrogenase release assay kit. Student T-test, Anova and post-hoc Tukey were used for statistical analysis using Graph Pad prism software 8. A p-value less than 0.05 was considered statistically significant.</span></span></span></span></span><br> <br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><i>Results</i></b></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times="">In both CAR-CIK and CAR-T cell, CAR receptor caused a significant increase in cytotoxicity. So that no significant difference was observed between the <a name="_Hlk148247586">cytotoxicity of CAR-T cells (67.8% &plusmn; 1.7) and CAR-CIK (65.1% &plusmn; 1.8)</a>.</span></span></span></span></span><br> <br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><i>Conclusions&nbsp; </i></b></span></span></span></span></span><br> <span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times="">CAR-CIK cells show a suitable <i>in vitro</i> cytotoxicity, so they can be considered as a potential alternative for CAR-T cells. However, more preclinical and clinical studies are needed to compare other effective dimensions in the function of these two cell types.</span></span></span></span></span><br> <br> <br> <br> <br> &nbsp;</div> کلمات کلیدی: Chimeric Antigen Receptors, سلول‌های T, سلول‌های کشنده القا شده با سیتوکین, بیماری پیوند علیه میزبان Key words: Chimeric Antigen Receptors, T Cells, Cytokine-Induced Killer Cells, Graft-Versus-Host Disease 298 311 http://bloodjournal.ir/browse.php?a_code=A-10-352-4&slc_lang=fa&sid=1 V. Moradi وحید مرادی 9100319475328460024672 9100319475328460024672 Yes دانشکده علوم پیراپزشکی دانشگاه علوم پزشکی تهران A. Ranjbar اعظم رنجبر 9100319475328460024673 9100319475328460024673 No مرکز تحقیقات ژن درمانی ـ پژوهشکده گوارش و کبد دانشگاه علوم پزشکی تهران A. Omidkhoda آزاده امیدخدا a-omidkhoda@tums.ac.ir 9100319475328460024674 9100319475328460024674 No دانشیار گروه هماتولوژی آزمایشگاهی و علوم انتقال خون ـ دانشکده علوم پیراپزشکی ـ دانشگاه علوم پزشکی تهران N. Ahmadbeigi ناصر احمدبیگی n-ahmadbeigi@tums.ac.ir 9100319475328460024675 9100319475328460024675 No دانشیار مرکز تحقیقات ژن درمانی پژوهشکده گوارش و کبد دانشگاه علوم پزشکی تهران