کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6133859 | 1593480 | 2014 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Optimization of interferon gamma ELISPOT assay to detect human cytomegalovirus specific T-cell responses in solid organ transplants
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کلمات کلیدی
ConAConcanavalin APHAAZAELISPOTPBMCsCMIPWMMMFPKCazathioprine - آزاتیوپرین Human leukocyte antigen - آنتی ژن لوسکسی انسانHLA - آنتیژن گلبول سفید انسانیCSA - ایالات مؤتلفهٔ آمریکاCell-mediated immunity - ایمنی سلولیT-cell - سلول TCell mediated immunity - سلول های ایمنیperipheral blood mononuclear cells - سلول های تک هسته ای خون محیطیCMV - سیتومگالوویروسHuman cytomegalovirus - سیتومگالوویروس انسانیCyclosporine A - سیکلوسپورینApokeweed mitogen - ماتوژن pokeweedmycophenolic acid - مایکوفنولیک اسید، مایکوفنولاتplaque forming unit - واحد پالک تشکیل شده استProtein kinase C - پروتئین کیناز سیpfu - پفوTransplantation - پیوند
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ویروس شناسی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Assessing the CMV specific CMI in transplant subjects represents a promising strategy to determine the risk of infection on individual basis. In this study 61 adult CMV IgG seropositive solid organ transplant recipients were examined in order to improve the efficacy of CMI detection. For this purpose, pair-wise comparisons were conducted comparing positive control stimuli PWM and PMA/iono and CMV stimuli, pp65 peptide pool and whole CMV particle. Rosette pre-depletion of blood was also investigated for detecting CD4+ or CD8+ T-cell responses using the IFN-g ELISPOT assay. In the time-points 30-180 days after transplantation, PMA/iono produced statistically significant higher responses compared to PWM, probably because PMA/iono activation pathway is independent from the effect of immunosuppressive drugs. The data showed that 11% of transplant patients displayed very low or undetectable responses to pp65 peptide pool antigen while having sustained high responses to whole CMV particle. In addition, in all the subjects analyzed, CMI responses to CMV particle produced a statistically significant higher number of spots compared to pp65 peptide pool antigen. Rosette pre-depletion of whole blood proved to be effective in detecting CD4+ or CD8+ T-cell responses similarly to flow cytometry. Taken together, the following recommendations are suggested to optimize the CMV-ELISPOT for transplantation settings: (1) use PMA/iono as positive control; (2) whole virus particle should be used to avoid peptide-related false negative responses; (3) a rosette pre-depletion step may be useful to detect CD4+ or CD8+ T-cell responses.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 196, February 2014, Pages 157-162
Journal: Journal of Virological Methods - Volume 196, February 2014, Pages 157-162
نویسندگان
Davide Abate, Alda Saldan, Gabriella Forner, Daniel Tinto, Alice Bianchin, Giorgio Palù,