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Lack of correlation between Treg quantification assays in inflammatory bowel disease patients

World Journal of Gastroenterology 2015年第11期21卷 3325-3329页

作者： Gunnar Brandhorst darinka todorova petrova Sebastian Weigand Christoph Eberle Nicolas von Ahsen Jessica Schmitz Frank Christian Schultze Dirk Raddatz Michael Karaus Michael Oellerich Philip D Walson Department of Clinical Chemistry University Medical Center Department of Clinical Pharmacology University Medical Center Chronix Biomedical Clinics of Gastroen-terology and Endocrinology University Medical Center Department of Internal Medicine Evangelisches Krankenhaus G?ttingen--Weende

AIM:To compare the number of regulatory T-cells( Tregs) measured by flow cytometry with those obtained using a real-time quantitative PCR(q PCR) method in patients suffering from inflammatory bowel disease(IBD).METHODS:Tregs percentages obtained by both flow cytometry and q PCR methods in 35 adult IBD patients,18 out of them with Crohn′s disease(CD)and 17 with ulcerative colitis(UC)were compared to each other as well as to scores on two IBD activity questionnaires using the Harvey Bradshaw Index(HBI)for CD patients and the Simple Colitis Clinical Activity Index(SCCAI)for UC *** Treg percentages by flow cytometry were defined as CD4+CD25highCD127lowFOXP3+cells in peripheral blood mononuclear cells,whereas the Treg percentages by q PCR method were determined as FOXP3 promoter demethylation in genomic ***:We found an average of 1.56%±0.78%Tregs by using flow cytometry,compared to 1.07%±0.53%Tregs by using q PCR in adult IBD *** were no significant correlations between either the percentages of Tregs measured by flow cytometry or q PCR and the HBI or SCCAI questionnaire scores in CD or UC patients,*** addition,there was no correlation between Treg percentages measured by q PCR and those measured by flow cytometry(r=-0.06,P=0.73;Spearman Rho).These data suggest that,either Treg-related immune function or the clinical scores in these IBD patients did not accurately reflect actual disease *** the cause(s)for these differences are more clearly defined,the resultssuggest caution in interpreting studies of Tregs in various inflammatory ***:The two methods did not produce equivalent measures of the percentage of total Tregs in the IBD patients studied which is consistent with the conclusion that Tregs subtypes are not equally detected by these two assays.

关键词： Regulatory T-cells Inflammatory bowel disease Croh

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Novel MLH1 frameshift mutation in an extended hereditary nonpolyposis colorectal cancer family

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World Journal of Gastroenterology 2006年第48期12卷 7848-7851页

作者： Tanya Kirilova Kadiyska Radka petrova Kaneva Dimitar Georgiev Nedin Alexandrina Borisova Alexandrova Antonina todorova Gegova Stoyan Ganchev Lalchev Tatyana Christova Vanio Ivanov Mitev Juergen Horst Nadja Bogdanova Ivo Marinov Kremensky Laboratory of Molecular Pathology University Hospital of Obstetrics and Gynecology Bulgaria Clinic of Abdominal Surgery Queen Giovanna Hospital Bulgaria Department of Pathology Queen Giovanna Hospital Bulgaria Department of Medical Genetics Medical University Bulgaria Department of Chemistry and Biochemistry Medical University Bulgaria Institute of Human Genetics University of Munster Germany

AIM： To present novel frameshift mutation c.31delC [p.L11X] in the MLH1 gene identified in an extended Bulgarian hereditary non-polyposis colorectal cancer （HNPCC） family and to analyze the molecular and clinical findings within the pedigree concerning the proposal of adequate individual prophylactic strategy for all mutation carriers.METHODS： The pedigree of the family consists of 42 members in four generations. Search for mutations in the MLH1 and hMSH2 genes was performed in the proband. After PCR amplification of all exons including flanking intronic regions, amplicons were directly sequenced.RESULTS： The mutation was found in nine from the thirteen pedigree members who signed informed consent to participate in the study. In three adenocarcinomas, microsatellite instability and lack of the MLH1 protein expression were detected. The only one tubulovillous adenoma analyzed was microsatellite stable and the MLH1 protein showed an intact staining.CONCLUSION： The newly described mutation c.31delC is HNPCC causative. Besides the typical clinical features of the syndrome, we found a specific pathologic manifestation such as moderate to high differentiated adenocarcinomas of the colon. One of the mutation carriers developed a benign giant cell soft tissue tumor, The primary tumor Iocalizatlons were frequently extracolonic and detailed yearly gastrointestinal and gynecological examinations have been proposed to the mutation carriers. We emphasize the importance of including the HNPCC genetic counseling and testing as well in the following surveillance of all patients at risk in the services covered by the health insurance in Bulgaria.

关键词： Colon cancer Hereditary non-polyposiscolorectal cancer MLH1 Microsatellite instability

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