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 - 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 - 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 - 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 - 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 - 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 - 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 -