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Pediatric Oncology
Published in Pat Price, Karol Sikora, Treatment of Cancer, 2020
Stephen Lowis, Rachel Cox, John Moppett, Helen Rees
Anaplastic large cell lymphoma (ALCL) accounts for around 10% of childhood NHL. Patients may present with mediastinal or abdominal disease, but there is a greater likelihood of systemic symptoms and cutaneous involvement. Histologically, cells may resemble undifferentiated tumors, Hodgkin’s, and other forms of NHL. Immunophenotyping is necessary to make the diagnosis for most. Cells have T-helper phenotype (CD3 and CD4 positive) and are characteristically positive for CD30 and epithelial membrane antigen (EMA).40
The lymphoreticular system and bone marrow
Published in C. Simon Herrington, Muir's Textbook of Pathology, 2020
The diagnosis of AML requires the demonstration of more than 20% myeloid blasts in the blood or marrow. Differentiation from ALL can be difficult on morphology alone, and immunophenotyping by flow cytometry is often required. The detailed laboratory techniques used in subclassifying these tumours is beyond the scope of this textbook.
Diagnostic applications of immunology
Published in Gabriel Virella, Medical Immunology, 2019
Ajay Grover, Virginia Litwin, Gabriel Virella
Immunophenotyping is also critical in the diagnosis of leukemias and lymphomas, and to monitor therapy. Samples are analyzed using various panels of antibodies that have been established for various types of leukemia or lymphoma. The markers that are present on the cells as detected by immunophenotyping will help characterize the abnormal cells present.
Comparison of outcomes of children with acute lymphoblastic leukemia treated with BMF protocol across 2 decades
Published in Pediatric Hematology and Oncology, 2020
Evrim Çifçi Sunamak, Nihal Özdemir, Aida Koka, Leman Yantiri, Hilmi Apak, Tiraje Celkan
Patients were diagnosed as ALL if their bone marrow contained 25% or more lymphoblasts. Bone marrow and blasts were counted morphologically. Flow cytometry was used for immunophenotyping. The presence of at least five lymphoblasts per microliter in the cerebrospinal fluid or the presence of a lesion in the brain around the meninges on magnetic resonance imaging or computed tomography was accepted as central nervous system (CNS) involvement. Patients were stratified into standard, medium, and high-risk groups (SRG, MRG, and HRG) according to BFM protocol.13–15 Response to treatment was evaluated by a peripheral smear on the 8th day and bone marrow aspiration on the 15th and 33rd days. Blast counts in bone marrow aspiration of less than 5%, between 5 and 24% and more than 25% were designated M1, M2, and M3, respectively. While relapses within the first 18 months of diagnosis were defined as “very early”, relapses occurring between 18 months after diagnosis and six months after discontinuation of treatment were defined as “early” and relapses detected later than these were designated as “late” relapses. At our center, evaluation of minimal residual disease (MRD) by flow cytometry began in May 2011.
Increased Aqueous Humor CD4+/CD8+ Lymphocyte Ratio in Sarcoid Uveitis
Published in Ocular Immunology and Inflammation, 2019
Namita Dave, Priyanka Chevour, Padmamalini Mahendradas, Anitha Venkatesh, Ankush Kawali, Rohit Shetty, Arkasubhra Ghosh, Swaminathan Sethu
Aqueous humor sample (0.15 ml) was collected by anterior chamber paracentesis using a 30-G needle with tuberculin syringe under aseptic precautions and contents stored at 4°C until immunophenotyping of the cells were performed. A part of the sample obtained (0.1 ml) was sent for PCR test for herpes simplex virus 1/2, VZV, cytomegalovirus, and Mycobacterium TB as part of routine diagnostic process. The remaining 0.05 ml of aqueous humor was used for immunophenotyping of cellular infiltrates. Aqueous humor samples (0.05 ml) of non-uveitis control subjects were also collected with a 30-G needle with tuberculin syringe before cataract surgery commenced. Peripheral venous blood (2 ml) was collected by venipuncture using EDTA-coated Vacutainer® (BD, New Jersey, USA) and stored 4°C until further analysis. The cells in the samples were processed for immunophenotyping analysis immediately as described below.
Hormonal oral contraceptive influence on isolation, Characterization and cryopreservation of mesenchymal stem cells from menstrual fluid
Published in Gynecological Endocrinology, 2019
Lilian Renata Fiorelli-Arazawa, Jorge Milhem Haddad, Maria Helena Nicola, Janaína J. dos Santos Machado, Anna Carolina Coimbra, Xavier Santamaria, José Maria Soares, Edmund Chada Baracat
Characterization by immunophenotyping. Standard flow cytometry (FCM) techniques were used to determine the typical cell surface epitope profiles according to Table 2. Cell numbers were quantified with a hemocytometer and 5.0 × 105 cells were located in each tube. Antibodies (CD45, CD34, CD19, CD14, HLA, DR, CD105, CD73, CD90, CD31, CD133, CD14; all from Becton Dickinson, San Diego, CA) were added and diluted as fabricant instruction and incubated for 20 min at 4 °C protected from light. Samples were lysed with BDLyse 1X solution (BD, USA) and incubated for 15 min at 4 °C protected from light. The material was then washed in phosphate buffered saline (PBS) (LGC Biotechnology, Brazil) and centrifuged at 400 rpm for 5 min. The supernatant was discarded and the pellet resuspended in 300 µL of PBS. Image acquisition was made in FACSCanto flow cytometer (Becton Dickinson, San Diego, CA). Cell viability was measured by 7AAD (BD, San Diego, NJ) flow cytometry and it evaluated the percentage of live nucleated cells. Data analysis was conducted with the CellQuest software, and at least 50,000 events were collected and analyzed. Immunophenotype were determined through the evaluation of specific markers as described by Dominic et al. [8]: hematopoietic cells (CD45+); mesenchymal cells (CD45-, CD34-, CD19-, CD14-, HLA-, DR-, CD105+, CD73+, CD90+); and endothelial cells (CD45-, CD34+, CD31+, CD133+) (Table 2).