Immunofluorescent labeling of CD20 tumor marker with quantum dots for rapid and quantitative detection of diffuse large B‐cell non‐Hodgkin's lymphoma

• Published in: Journal of cellular biochemistry Vol. 120; no. 3; pp. 4564 - 4572
• Main Authors: Shariatifar, Hanifeh, Hakhamaneshi, Mohammad‐Saeed, Abolhasani, Maryam, Ahmadi, Fahimeh Haji, Roshani, Daem, Nikkhoo, Bahram, Abdi, Mohammad, Ahmadvand, Davoud
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.03.2019
• Subjects: Adult; Aged; Aged, 80 and over; Antigens, CD20 - metabolism; Biomarkers; Biomarkers, Tumor - metabolism; CD20 antigen; CD20 tumor marker; Diagnostic systems; diagnostic value; diffuse large B‐cell lymphoma; Female; Fluorescence; Fluorescence microscopy; Humans; Immunotherapy; K562 Cells; Lymphocytes B; Lymphoma; Lymphoma, Large B-Cell, Diffuse - metabolism; Lymphoma, Large B-Cell, Diffuse - pathology; Male; Markers; Medical imaging; Middle Aged; Monoclonal antibodies; Nanocrystals; Patients; quantum dot; Quantum dots; Quantum Dots - chemistry; Rituximab; Rituximab - chemistry; Sensitivity; Staining and Labeling; Targeted cancer therapy; Tumor markers; Tumors; quantum dot; CD20 tumor marker; diffuse large B-cell lymphoma; diagnostic value; rituximab
• ISSN: 0730-2312; 1097-4644; 1097-4644
• DOI: 10.1002/jcb.27745

Fluorescent semiconductor quantum dots (QDs) are newfound nanocrystal probes which have been used in bioimaging filed in recent years. The purpose of this study is to evaluate the diagnostic value of specific QDs coupled to rituximab monoclonal antibody against CD20 tumor markers for patients with diffuse large B‐cell lymphoma (DLBCL). In current study rituximab‐conjugated quantum dots (QDs‐rituximab) were prepared against CD20 tumor markers for detection of CD20‐positive cells (human Raji cell line) using flowcytometry. A total of 27 tumor tissue samples were collected from patients with DLBCL and 27 subjects with negative pathological tests as healthy ones, which stained by QD‐rituximab. The detection signals were obtained from QDs using fluorescence microscopy. The flowcytometry results demonstrated a remarkable difference in fluorescent intensity and FL2‐H + (CD20‐positive cells percentage) between two groups. Both factors were significantly higher in Raji in comparison with K562 cell line (P < 0.05). Lot of green fluorescence signals was observed due to the selectively binding of QD‐rituximab to CD20 tumor markers which overexpressed in tumor tissues and a few signals observed on the defined healthy ones. Based on these observations the cut‐off point was 46.8 dots and the sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 89.5%, 91.3%, and 100%, respectively (LR+, 9.52; LR−, 0). The QD ‐rituximab could be beneficial as a bioimaging tool with high sensitivity to provide an accurate molecular imaging technique for identifying CD20 tumor markers for early diagnosis of the patients with DLBCL. This manuscript is discussing an innovating method, using stable nano‐immunofluorescent probes (QDs coupled to rituximab mAb) for detecting low numbers of CD20 tumor marker for early in vitro cancer diagnosis. Our research is related to fluorescent imaging of CD20 tumor marker targeted by immunofluorescent QD nanoparticles on tissue sections and CD20‐positive cell line (Raji), quantified by flowcytometry.