TY - GEN A1 - George, Sandra A1 - Paulick, Silvia A1 - Knütter, Ilka A1 - Röber, Nadja A1 - Hiemann, Rico A1 - Roggenbuck, Dirk A1 - Conrad, Karsten A1 - Küpper, Jan-Heiner T1 - Stable Expression of Human Muscle-Specific Kinase in HEp-2 M4 Cells for Automatic Immunofluorescence Diagnostics of Myasthenia Gravis T2 - PLOS one Y1 - 2014 U6 - https://doi.org/10.1371/journal.pone.0083924 SN - 1932-6203 VL - 9 IS - 1 SP - e83924 ER - TY - GEN A1 - Knütter, Ilka A1 - Hiemann, Rico A1 - Brumma, Therese A1 - Büttner, Thomas A1 - Großmann, Kai A1 - Cusini, Marco A1 - Pregnolato, Francesca A1 - Borghi, Maria Orietta A1 - Anderer, Ursula A1 - Conrad, Karsten A1 - Reinhold, Dirk A1 - Roggenbuck, Dirk A1 - Csernok, Elena T1 - Automated interpretation of ANCA patterns - a new approach in the serology of ANCA-associated vasculitis T2 - Arthritis Research & Therapy KW - ANCA-associated vasculitis KW - serology KW - pattern recognition algorithms KW - discrimination and classification of cytoplasmic ANCA (C-ANCA) and perinuclear ANCA (P-ANCA) KW - automated interpretation of ANCA IIF Y1 - 2012 UR - http://arthritis-research.com/content/14/6/R271 U6 - https://doi.org/10.1186/ar4119 SN - 1465-9913 SN - 1478-6354 VL - 14 ER - TY - GEN A1 - Hiemann, Rico A1 - Roggenbuck, Dirk A1 - Sack, Ulrich A1 - Anderer, Ursula A1 - Conrad, Karsten T1 - Die HEp-2-Zelle als Target für multiparametrische Autoantikörperanalytik – Automatisierung und Standardisierung/The HEp-2 cell as target for multiparametric autoantibody analyses: automation and standardisation T2 - Journal of Laboratory Medicine N2 - Trotz Einführung von Multiplex-Assays ist die Bestimmung von Autoantikörpern (AAK) mittels indirekter Immunfluoreszenz (IIF) nach wie vor ein wesentlicher Bestandteil der Autoimmundiagnostik. Das Screening auf nichtorganspezifische AAK wird in der Routinediagnostik von autoimmunen rheumatischen Erkrankungen mittels indirekter Immunfluoreszenz an HEp-2-Zellen durchgeführt. Leider differieren die mit diesem Test erhaltenen Befunde z. T. recht erheblich auf Grund von objektiven (z. B. HEp-2-Zellfixierung) und subjektiven (z. B. Erfahrungswerte) Faktoren. Die Intra- und Interlaborvarianzen sind daher relativ hoch. Erste Ergebnisse von Evaluierungsstudien eines neu entwickelten Systems zur automatischen Bildanalyse (AKLIDES®) zeigen einen Weg in Richtung Standardisierung (bzw. Harmonisierung) des HEp-2-Zell-Assays auf, in dem zunächst subjektive Fehlinterpretationen vermieden werden können. Eine weitere Harmonisierung erfordert die Weiterentwicklung der Software-Algorithmen zur Mustererkennung sowie neue Kalibrationssysteme. KW - Autoantikörper KW - Automatisierung KW - HEp-2 KW - Immunfluoreszenz Y1 - 2011 U6 - https://doi.org/10.1515/JLM.2011.053 SN - 1439-0477 VL - 35 IS - 6 SP - 351 EP - 361 ER - TY - GEN A1 - Hiemann, Rico A1 - Hilger, Nadja A1 - Michel, Jörg A1 - Nitschke, Jörg A1 - Böhm, Alexander A1 - Anderer, Ursula A1 - Weigert, Martin A1 - Sack, Ulrich T1 - Automatic Analysis of Immunofluorescence Patterns of HEp‐2 Cells T2 - Annals of the New York Academy of Sciences N2 - The standard screening test for the recognition of autoimmune diseases is the proof of autoantibodies in serum of patients by indirect immunofluorescence (IIF) based on HEp‐2 cells. Manual evaluation of this test is very subjective, slow, and there are no objective parameters as guidelines available. Interlaboratory tests showed occasionally large deviations in the test evaluation resulting in a high variance of results. The aim of this project is fast, objective, safe, and economical automatic analysis of HEp‐2 IIF patterns. Images of IIF patterns were completely and automatically captured using an inverse motorized fluorescence microscope. Thereby, device‐specific parameters were controlled automatically, too. For fast analysis of IIF patterns new algorithms of image processing were developed. Artifacts were recognized and excluded from analysis by the developed software. Analysis of more than 80,000 images clearly demonstrated full automatization and fast processing of IIF patterns. Additionally serum‐specific fluorescence could be easily distinguished from background. Even very weak but positive patterns can be recognized and used for diagnosis. A detailed separation into different basic patterns is possible. Objective, fast, and disease‐related economical analysis of HEp‐2 immunofluorescence patterns is feasible. The implemented software algorithms allowed a mathematical way of describing IIF patterns and can therefore be a useful tool for the needed standardization process. KW - Autoimmune diagnostics KW - HEp-2 cell KW - Immunofluorescence KW - Automatic analysis KW - Screening Y1 - 2007 U6 - https://doi.org/10.1196/annals.1398.042 SN - 0077-8923 VL - 1109 IS - 1 SP - 358 EP - 371 ER - TY - GEN A1 - Schierack, Peter A1 - Rödiger, Stefan A1 - Kuhl, Christoph A1 - Hiemann, Rico A1 - Roggenbuck, Dirk A1 - Li, Ganwu A1 - Weinreich, Jörg A1 - Berger, Enrico A1 - Nolan, Lisa K. A1 - Nicholson, Bryon A1 - Römer, Antje A1 - Frömmel, Ulrike A1 - Wieler, Lothar H. A1 - Schröder, Christian T1 - Porcine E. coli: Virulence-Associated Genes, Resistance Genes and Adhesion and Probiotic Activity Tested by a New Screening Method T2 - PLoS One N2 - We established an automated screening method to characterize adhesion of Escherichia coli to intestinal porcine epithelial cells (IPEC-J2) and their probiotic activity against infection by enteropathogenic E. coli (EPEC). 104 intestinal E. coli isolates from domestic pigs were tested by PCR for the occurrence of virulence-associated genes, genes coding for resistances to antimicrobial agents and metals, and for phylogenetic origin by PCR. Adhesion rates and probiotic activity were examined for correlation with the presence of these genes. Finally, data were compared with those from 93 E. coli isolates from wild boars. Isolates from domestic pigs carried a broad variety of all tested genes and showed great diversity in gene patterns. Adhesions varied with a maximum of 18.3 or 24.2 mean bacteria adherence per epithelial cell after 2 or 6 hours respectively. Most isolates from domestic pigs and wild boars showed low adherence, with no correlation between adhesion/probiotic activity and E. coli genes or gene clusters. The gene sfa/foc, encoding for a subunit of F1C fimbriae did show a positive correlative association with adherence and probiotic activity; however E. coli isolates from wild boars with the sfa/foc gene showed less adhesion and probiotic activity than E. coli with the sfa/foc gene isolated from domestic pigs after 6 hour incubation. In conclusion, screening porcine E. coli for virulence associated genes genes, adhesion to intestinal epithelial cells, and probiotic activity revealed a single important adhesion factor, several probiotic candidates, and showed important differences between E. coli of domestic pigs and wild boars. Y1 - 2013 U6 - https://doi.org/10.1371/journal.pone.0059242 VL - 8 IS - 4 ER - TY - GEN A1 - Schierack, Peter A1 - Rödiger, Stefan A1 - Kolenda, Rafał A1 - Hiemann, Rico A1 - Berger, Enrico A1 - Grzymajlo, Krzysztof A1 - Swidsinski, Alexander A1 - Juretzek, Thomas A1 - Meissner, Dirk A1 - Mydlak, Karsten A1 - Reinhold, Dirk A1 - Nolan, Lisa K. A1 - Roggenbuck, Dirk T1 - Species-specific and pathotype-specific binding of bacteria to zymogen granule membrane glycoprotein 2 (GP2) T2 - Gut Y1 - 2015 U6 - https://doi.org/10.1136/gutjnl-2014-307854 SN - 1468-3288 SN - 0017-5749 VL - 64 IS - 3 SP - 517 EP - 519 ER - TY - GEN A1 - Scholz, Juliane A1 - Großmann, Kai A1 - Knütter, Ilka A1 - Hiemann, Rico A1 - Sowa, Mandy A1 - Röber, Nadja A1 - Rödiger, Stefan A1 - Schierack, Peter A1 - Reinhold, Dirk A1 - Bogdanos, Dimitrios Petrou A1 - Meroni, Pier Luigi A1 - Radice, Antonella A1 - Conrad, Karsten A1 - Roggenbuck, Dirk T1 - Second generation analysis of antinuclear antibody (ANA) by combination of screening and confirmatory testing T2 - Clinical Chemistry and Laboratory Medicine Y1 - 2015 U6 - https://doi.org/10.1515/cclm-2015-0083 SN - 1437-4331 VL - 53 IS - 12 SP - 1991 EP - 2002 ER - TY - GEN A1 - Willitzki, Annika A1 - Lorenz, Sebastian A1 - Hiemann, Rico A1 - Guttek, Karina A1 - Goihl, Alexander A1 - Hartig, Roland A1 - Conrad, Karsten A1 - Feist, Eugen A1 - Sack, Ulrich A1 - Schierack, Peter A1 - Heiserich, Lisa A1 - Eberle, Caroline A1 - Peters, Vanessa A1 - Roggenbuck, Dirk A1 - Reinhold, Dirk T1 - Fully automated analysis of chemically induced γH2AX foci in human peripheral blood mononuclear cells by indirect immunofluorescence T2 - Cytometry Part A Y1 - 2013 U6 - https://doi.org/10.1002/cyto.a.22350 SN - 1552-4930 SN - 1552-4922 VL - 83 IS - 11 SP - 1017 EP - 1026 ER - TY - GEN A1 - Reddig, Annika A1 - Lorenz, Sebastian A1 - Hiemann, Rico A1 - Guttek, Karina A1 - Hartig, Roland A1 - Heiserich, Lisa A1 - Peter, Vanessa A1 - Schierack, Peter A1 - Sack, Ulrich A1 - Roggenbuck, Dirk A1 - Reinhold, Dirk A1 - Eberle, Caroline T1 - Cytostatic drug resistance monitored by γH2AX T2 - Cytometry Part A Y1 - 2015 U6 - https://doi.org/10.1002/cyto.a.22667 SN - 1552-4930 SN - 1552-4922 VL - 87 IS - 8 SP - 724 EP - 732 ER - TY - GEN A1 - Sowa, Mandy A1 - Großmann, Kai A1 - Knütter, Ilka A1 - Hiemann, Rico A1 - Röber, Nadja A1 - Anderer, Ursula A1 - Csernok, Elena A1 - Bogdanos, Dimitrios Petrou A1 - Borghi, Maria Orietta A1 - Meroni, Pier Luigi A1 - Schierack, Peter A1 - Reinhold, Dirk A1 - Conrad, Karsten A1 - Roggenbuck, Dirk T1 - Simultaneous automated screening and confirmatory testing for vasculitis-specific ANCA T2 - PLoS ONE KW - ANCA KW - vasculitis KW - screening KW - confirmatory testing KW - multiplexing KW - CytoBead technology Y1 - 2014 U6 - https://doi.org/10.1371/journal.pone.0107743 SN - 1932-6203 VL - 9 IS - 9 ER - TY - GEN A1 - Roggenbuck, Dirk A1 - Hiemann, Rico A1 - Schierack, Peter A1 - Reinhold, Dirk A1 - Conrad, Karsten T1 - Digital immunofluorescence enables automated detection of antinuclear antibody endpoint titers avoiding serial dilution T2 - Clinical Chemistry and Laboratory Medicine Y1 - 2013 U6 - https://doi.org/10.1515/cclm-2013-0543 SN - 1437-4331 SN - 1434-6621 VL - 52 IS - 2 SP - e9 EP - e11 ER - TY - GEN A1 - Großmann, Kai A1 - Röber, Nadja A1 - Hiemann, Rico A1 - Rödiger, Stefan A1 - Schierack, Peter A1 - Reinhold, Dirk A1 - Laass, Martin W. A1 - Conrad, Karsten A1 - Roggenbuck, Dirk T1 - Simultaneous detection of celiac disease-specific IgA antibodies and total IgA T2 - Autoimmunity Highlights Y1 - 2016 U6 - https://doi.org/10.1007/s13317-016-0073-2 SN - 2038-0305 SN - 2038-3274 VL - 7 IS - 2 ER - TY - GEN A1 - Schierack, Peter A1 - Kleta, Sylvia A1 - Tedin, Karsten A1 - Babila, T. B. A1 - Oswald, S. A1 - Oelschlaeger, T. A. A1 - Hiemann, Rico A1 - Paetzold, S. A1 - Wieler, Lothar H. T1 - E. coli Nissle 1917 affects Salmonella adhesion to porcine intestinal epithelial cells T2 - PLoS One Y1 - 2011 U6 - https://doi.org/10.1371/journal.pone.0014712 VL - 6 IS - 2 ER - TY - GEN A1 - Willitzki, Annika A1 - Hiemann, Rico A1 - Peters, Vanessa A1 - Sack, Ulrich A1 - Schierack, Peter A1 - Rödiger, Stefan A1 - Anderer, Ursula A1 - Conrad, Karsten A1 - Bogdanos, Dimitrios Petrou A1 - Reinhold, Dirk A1 - Roggenbuck, Dirk T1 - New PlatformTechnology for Comprehensive Serological Diagnostics of Autoimmune Diseases T2 - Clinical and Developmental Immunology KW - systemic autoimmune rheumatic disease (SARD) KW - automated IIF methods KW - platform for automated interpretation of cell-based IIF testing KW - microbead immunoassays KW - multiplexing Y1 - 2012 U6 - https://doi.org/10.1155/2012/284740 SN - 1740-2530 SN - 1740-2522 SP - 1 EP - 8 ER - TY - GEN A1 - Sowa, Mandy A1 - Großmann, Kai A1 - Scholz, Juliane A1 - Röber, Nadja A1 - Rödiger, Stefan A1 - Schierack, Peter A1 - Conrad, Karsten A1 - Roggenbuck, Dirk A1 - Hiemann, Rico T1 - Der CytoBead-Assay – Eine neue Möglichkeit der multiparametrischen Autoantikörperanalytik bei systemischen Autoimmunerkrankungen T2 - Journal of Laboratory Medicine Y1 - 2014 U6 - https://doi.org/10.1515/labmed-2014-0041 VL - 38 IS - 6 SP - 309 EP - 317 ER - TY - GEN A1 - Sowa, Mandy A1 - Trezzi, Barbara A1 - Hiemann, Rico A1 - Schierack, Peter A1 - Großmann, Kai A1 - Scholz, Juliane A1 - Somma, Valentina A1 - Sinico, Renato Alberto A1 - Roggenbuck, Dirk A1 - Radice, Antonella T1 - Simultaneous comprehensive multiplex autoantibody analysis for rapidly progressive glomerulonephritis T2 - Medicine Y1 - 2016 U6 - https://doi.org/10.1097/MD.0000000000005225 SN - 0025-7974 VL - 95 IS - 44 SP - 5225 ER - TY - GEN A1 - Sowa, Mandy A1 - Hiemann, Rico A1 - Schierack, Peter A1 - Reinhold, Dirk A1 - Conrad, Karsten A1 - Roggenbuck, Dirk T1 - Next-Generation Autoantibody Testing by Combination of Screening and Confirmation-the CytoBead® Technology T2 - Clinical Reviews in Allergy & Immunology Y1 - 2017 U6 - https://doi.org/10.1007/s12016-016-8574-3 SN - 1559-0267 SN - 1080-0549 VL - 53 IS - 1 SP - 87 EP - 104 ER - TY - GEN A1 - Liedtke, Victoria A1 - Rose, Laura A1 - Hiemann, Rico A1 - Nasser, Abdullah A1 - Rödiger, Stefan A1 - Bonaventura, Alena A1 - Winkler, Laura A1 - Sowa, Mandy A1 - Stöckle, Michael A1 - Schierack, Peter A1 - Junker, Kerstin A1 - Roggenbuck, Dirk T1 - Over-Expression of LEDGF/p75 in HEp-2 Cells Enhances Autoimmune IgG Response in Patients with Benign Prostatic Hyperplasia—A Novel Diagnostic Approach with Therapeutic Consequence? T2 - International Journal of Molecular Sciences N2 - Lens epithelium-derived growth factor splice variant of 75 kDa (LEDGF/p75) is an autoantigen over-expressed in solid tumors and acts as a stress-related transcriptional co-activator. Participation of autoimmune responses in the pathophysiology of benign prostatic hyperplasia (PBH) and a corresponding immunosuppressive therapy by TNFalpha antagonists has been recently suggested. Thus, autoAb testing could aid in the diagnosis of BPH patients profiting from such therapy. We generated CRISPR/Cas9 modified HEp-2 LEDGF knock-out (KO) and HEp-2 LEDGF/p75 over-expressing (OE) cells and examined IgG autoantibody reactivity to LEDGF/p75 in patients with prostate cancer (PCa, n = 89), bladder cancer (BCa, n = 116), benign prostatic hyperplasia (BPH, n = 103), and blood donors (BD, n = 60) by indirect immunofluorescence assay (IFA). Surprisingly, we could not detect elevated binding of autoAbs against LEDGF/p75 in cancer patients, but autoAb reactivity to LEDGF/p75 OE cells in about 50% of patients with BPH was unexpectedly significantly increased. Furthermore, a line immunoassay enabling the detection of 18 different autoAbs revealed a significantly increased occurrence of anti-dsDNA autoAbs in 34% of BPH patients in contrast to tumor patients and BD. This finding was confirmed by anti-mitochondrial (mDNA) autoAb detection with the Crithidia luciliae immunofluorescence test, which also showed a significantly higher prevalence (34%) of anti-mDNA autoAbs in BPH. In summary, our study provided further evidence for the occurrence of autoimmune responses in BPH. Furthermore, LEDGF/p75 over-expression renders HEp-2 cells more autoantigenic and an ideal target for autoAb analysis in BPH with a potential therapy consequence. KW - LEDGF/p75 KW - autoimmunity KW - CRISPR/Cas9 KW - dsDNA KW - mDNA Y1 - 2023 U6 - https://doi.org/10.3390/ijms24076166 SN - 1422-0067 VL - 24 IS - 7 ER -