{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T16:32:30Z","timestamp":1777480350433,"version":"3.51.4"},"reference-count":38,"publisher":"SAGE Publications","issue":"3","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CBM"],"published-print":{"date-parts":[[2021,11,10]]},"abstract":"<jats:p>BACKGROUND: Recent studies proved that metabolic changes in malignant disorders have an impact on protein glycosylation, however, only a few attempts have been made so far to use O-GlcNAc analysis as a prognostic tool. Glucose metabolism is reported to be altered in hematological malignancies thus, we hypothesized that monitoring intracellular O-GlcNAc levels in Rai stage 0-I (Binet A) CLL patients could give deeper insights regarding subtle metabolic changes of progression which are not completely detected by the routine follow-up procedures. OBJECTIVE: In this proof of concept study we established a flow cytometric detection method for the assessment of O-GlcNAcylation as a possible prognostic marker in CLL malignancy which was supported by fluorescence microscopy. METHODS: Healthy volunteers and CLL patients were recruited for this study. Lymphocytes were isolated, fixed and permeabilised by various methods to find the optimal experimental condition for O-GlcNAc detection by flow cytometry. O-GlcNAc levels were measured and compared to lymphocyte count and various blood parameters including plasma glucose level. RESULTS: The protocol we developed includes red blood cell lysis, formalin fixation, 0.1% Tween 20 permeabilisation and employs standardized cell number per sample and unstained controls. We have found significant correlation between O-GlcNAc levels and WBC (R2= 0.8535, p&lt; 0.0029) and lymphocyte count (R2= 0.9225, p&lt; 0.0006) in CLL patients. Interestingly, there was no such correlation in healthy individuals (R2= 0.05664 for O-GlcNAc vs WBC and R2= 0.04379 for O-GlcNAc vs lymphocytes). CONCLUSION: Analyzing O-GlcNAc changes in malignant disorders, specifically in malignant hematologic diseases such as CLL, could be a useful tool to monitor the progression of the disease.<\/jats:p>","DOI":"10.3233\/cbm-203049","type":"journal-article","created":{"date-parts":[[2021,6,18]],"date-time":"2021-06-18T14:42:22Z","timestamp":1624027342000},"page":"353-362","source":"Crossref","is-referenced-by-count":1,"title":["O-GlcNAcylation in early stages of chronic lymphocytic leukemia: Protocol development for flow cytometry"],"prefix":"10.1177","volume":"32","author":[{"given":"Vikt\u00f3ria","family":"Temesf\u0151i","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine, Medical School, University of P\u00e9cs, P\u00e9cs, Hungary"},{"name":"Lab-on-a-Chip Research Group, J\u00e1nos Szent\u00e1gothai Research Center, University of P\u00e9cs, P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kinga","family":"Moln\u00e1r","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Medical School, University of P\u00e9cs, P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P\u00e9ter","family":"Kaltenecker","sequence":"additional","affiliation":[{"name":"Laboratory of Actin Cytoskeleton Regulation, Institute of Genetics, Biological Research Centre, E\u00f6tv\u00f6s Lor\u00e1nd Research Network (ELKH), Szeged, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Barbara","family":"R\u00e9ger","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Medical School, University of P\u00e9cs, P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"\u00c1rp\u00e1d","family":"Szomor","sequence":"additional","affiliation":[{"name":"Division of Hematology, 1st Department of Internal 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