دانلود مقاله ترجمه شده کاهش مخصوص هتروزایگوسیتی لوسمی MHC در بیمار CLL – مجله الزویر

فهرست مطالب:

چکیده
۱ مقدمه
۲ مطالعه موردی
۳ روش ها
۳ ۱سازگاری بافتی سلولی
۳ ۲ تجزیه تحلیل و آرایه VNTR و aCGH SNP
۴ نتایج
۵ بحث

بخشی از ترجمه:

 لوسمی لیمفوسیتی مزمن  با طیف وسیعی از   عملیات بالینی و پزشکی مورد بررسی قرار می گیرند. این   لوسمی را می توان  توسط  طیف وسیعی  از ناهنجاری  سیتوژنتیکی مکرر که توسط فلروسانس و باند گذاری G در هیبریداسیون سلولی شناسایی می شوند  مشخص کرد که خود یک روش  تشخیصی بوده و برای تعیین روش ها و رویکرد های درمانی استفاده می شود. با این حال بسیاری از  موتاسیون های ژنومی  دارای   اندازه و تفکیک پذیری پایین تری از باند گذاری G  بوده و آنالیز های FISH قادر به ارزیابی  ناهنجاری های  ژنتیکی نمی باشد. آنالیزهای  ژنومی با استفاده از  میکروآرایه  اطلاعات جامعی در  خصوص مکانیسم های  ژنتیکی ترانسفورماسیون در اختیار می گذارند. پتیفر (۲)  به آزمایش۷۰  CLL متوالی با استفاده از   آرایه های  ۱۰ و ۵۰ K پرداخت. آن ها یک ناتوازنی کروموزومی را در  ۶۵ و ۸۰%  موارد مشاهده کردند. در  ۱۴ مورد،  کاهش  هتروزاگوسیتی رونوشتی خنثی در مناطق  بزرگ تر از  ۱۰ Mb  که توسط FISH شناسایی نشده بودند   دیده شد. آفلبل ،  آرایش کاریوتیپی  آرایه بنیان SNP را در  آنمی ماژور در برابر سندرم  میلدوپلاستیک فوق سلولی  در ۹۳ بیمار گزارش کرد.هدف ایشان بهبود  دقت تشخیصی  با شناسایی برخی نواقص بود.

بخشی از مقاله انگلیسی:

۱٫ Introduction Chromosomal abnormalities are common among patients with hematologic malignancies and may be useful in tailoring therapy. In chronic lymphocytic leukemia several, such as del 13q14, del 17p and trisomy 12 are of prognostic significance [1]. Fluorescent in situ hybridization (FISH) testing is used routinely to identify such aberrations, but detection is limited to the probes used in the panel [2]. Thus other, potentially important genetic changes may be undetected. We describe a CLL case with undetected genetic aberration including loss of heterozygosity of MHC genes which impacted recipient-donor matching for hematopoietic stem cell transplant. 2. Case The patient is a 63 year old Caucasian male diagnosed with CLL in 1985 when he presented with generalized lymphadenopathy. Lymph node biopsy and bone marrow aspirate and biopsy were consistent with diagnosis of chronic lymphocytic leukemia. He was observed through July 2009 when his white blood cell count was found to be 83,000 and 79% lymphocytes. He received 2 cycles Fludarabine and Rituxan, but within 3 weeks presented with fever and was found to be pancytopenic. He was subsequently referred to our institution for consultation. At time of evaluation his WBC was 87,100 with 99% lymphocytes, hematocrit was 34% and platelets count was 27,000. Peripheral blood flow cytometry revealed a kappa-restricted CD10, CD20+, CD5+, CD23+, CD19+, FMC7 CD38 clonal population of B cells. A bone marrow aspirate and biopsy demonstrated diffuse involvement of the marrow by CLL. FISH on the marrow revealed 13q14.3 (D13S319) deletion detected in 200/200 cells as well as a 17p13 (p53) deletion detected in 200/ 200 cells. Routine G-banding was not performed. He underwent HLA typing for consideration of allogeneic transplant using a peripheral blood sample obtained at the time of evaluation. In addition, he consented to participate in an IRB approved protocol in which peripheral blood and marrow samples are prospectively collected in patients with hematologic malignancies to be used in research studies. Following a cycle of Fludarabine, Cyclophosphamide and Rituxan, he was switched to Campath chemotherapy for two cycles but developed pancytopenia with transfusion dependence. His neutropenia was refractory to filgrastim. 3. Methods 3.1. Histocompatibility Recipient HLA typing was performed by molecular method. DNA was extracted from white blood cells using a manual blood kit (Qiagen). Low resolution typing for HLA-A, B, C and DQ and high resolution typing for HLA-DRB1 was performed in a bead based assay using reverse sequence specific oligonucleotide probes (rSSO), Labtype (One Lambda, Canoga Park CA). High resolution typing for HLA-A, B, C, and DQB1 used sequence specific primers (SSP) and gel electrophoresis (Unitray; Invitrogen). Related donors were screened by serological HLA typing (Gentrak and Transtype); those that are potential matches proceed to molecular typing. T and B subsets are obtained from peripheral blood by Dynal immunomagnetic bead method for CD8+ and CD19+ cells respectively (Invitrogen: Carlsbad, CA). 3.2. VNTR and aCGH-SNP array and analysis The fluorescently-labeled PCR products targeting 8 loci (D11S554, D12S391, D16S539, FGA, Penta E, SE33, THO, VWF-B) were resolved using capillary electrophoresis with the relative quantity of each product measured by its fluorescence. Microarray CGH experiment was carried out on custom-designed Agilent’s SurePrint G3 CGH-SNP 4 180 K cancer targeted platform (Agilent Technologies. Santa Clara, CA, USA) containing approximately 120,000 CGH probes covering over 500 cancer genes and more than 130 cancer-associated genomic regions and 60,000 SNP probes. Genotypes on this array are measured using one SNP probe per SNP, providing ۵–۱۰ Mb resolution for LOH/UPD detection across the entire genome. Test DNA was referenced against same-sex control DNA. Patient DNA was isolated from peripheral blood and the control DNA (NA12891) was obtained from the Coriell Cell Repository (Camden, NJ, USA). Briefly, patient’s DNA and control DNA (1.0 lg) were digested with restriction enzymes, Alu1 and Rsa1, and enzymatically labeled with dyes Cyanine- 5dUTP and Cyanine-3dUTP respectively by using Agilent’s Enzymatic Labeling Kit (Cat #5190–۰۴۴۹) as per the manufacturer’s recommendations. The labeled DNA was hybridized as per the Agilent’s recommendations at 65 C for 40 h. Following washing, the slides were scanned in Agilent’s high resolution scanner (Model #G2505C) at 3 lm resolution. The output files were processed using Feature Extraction software v10.10 and analyzed by Genomic Workbench Software v6.5 (Agilent Technologies. Santa Clara, CA, USA), using an ADM-1 algorithm for aberration analysis and visualization, with the threshold set at 6.7. To control small variations appearing in the data analysis, we used an extra aberration filter, defining the minimum number of probes that should be present in an aberrant region as 3 (gain or loss), with the minimum absolute level of average log2 ratio of 0.25. The centralization was on with threshold and bin size set at 6.0 and 10, respectively, and GC correction and fuzzy zero were also included in the analysis. For the detection of SNP and LOH, 95% confidence level and thresholds 6.0 were set respectively.

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