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Immunophenotyping by flow cytometry

M T Cardoso, A O Pinheiro, A S Vidane, J B Casals, V C de Oliveira, Njn Gonçalves, D S Martins, C E Ambrósio
The biosafety of innovative procedures that utilize stem cells in regenerative medicine has been addressed in several studies. Previous work has showed no tumour formation following the use of feline and human amniotic membrane-derived stem cells (AMSCs). In contrast, tumour formation was observed when canine AMSCs were utilized. These findings suggested that feline and human, but not canine, AMSCs are suitable for cell transplantation trials. This study aimed to further evaluate the feasibility of utilizing canine AMSCs for transplantation purposes as well as for felines...
October 23, 2016: Reproduction in Domestic Animals, Zuchthygiene
Alicia Martinez-Varea, Roberto Romero, Yi Xu, Derek Miller, Ahmed I Ahmed, Piya Chaemsaithong, Noppadol Chaiyasit, Lami Yeo, Majid Shaman, Kia Lannaman, Benjamin Cher, Sonia S Hassan, Nardhy Gomez-Lopez
OBJECTIVES: 1) To characterize the cellular composition of the amniotic fluid of patients diagnosed with clinical chorioamnionitis at term, as a function of the presence or absence of microorganisms determined by cultivation techniques, and 2) to characterize the cytokine production by white blood cells present in the amniotic fluid using flow cytometry-based techniques. MATERIALS AND METHODS: Amniotic fluid samples from 20 women who had the diagnosis of clinical chorioamnionitis at term were analyzed using cultivation techniques (for aerobic and anaerobic bacteria as well as genital Mycoplasmas)...
October 20, 2016: Journal of Perinatal Medicine
Annapurna Saksena, Parul Gautam, Parth Desai, Naresh Gupta, A P Dubey, Tejinder Singh
BACKGROUND & OBJECTIVES: Flow cytometry is an important tool to diagnose acute leukaemia. Attempts are being made to find the minimal number of antibodies for correctly diagnosing acute leukaemia subtypes. The present study was designed to evaluate the analysis of side scatter (SSC) versus CD45 flow dot plot to distinguish acute myeloid leukaemia (AML) from acute lymphoblastic leukaemia (ALL), with minimal immunological markers. METHODS: One hundred consecutive cases of acute leukaemia were evaluated for blast cluster on SSC versus CD45 plots...
May 2016: Indian Journal of Medical Research
Deena Samir Eissa, Eman Zaghloul Kandeel, Mohamed Ghareeb
The prognosis of acute myeloid leukemia (AML) is poor because of relapses occurring on conventional chemotherapy. The distinction between leukemic and normal stem cells relies on the expression of antigen combinations defining leukemia-associated immunophenotypes (LAIPs), which are absent or extremely infrequent in normal bone marrow. However, LAIPs are very different from patient to patient and are not necessarily stable over the course of the disease. Accordingly, we addressed the applicability of human myeloid inhibitory C-lectin (hMICL) by flow cytometry as a specific leukemic myeloid stem cell marker for the diagnosis of AML in CD34(+) and CD34(-) cases and evaluated the stability of hMICL during the course of the disease...
October 13, 2016: Hematological Oncology
Laura G Rico, Jordi Juncà, Mike D Ward, Jolene Bradford, Jordi Petriz
With the aim to detect candidate malignant primitive progenitor populations, we modified an original alkaline phosphatase (ALP) stem cell detection method based on the identification of alkaline phosphatase fluorescent cells in combination with flow cytometry immunophenotyping. Over a period of one year, we have been using this technique to study its activity in patients with leukemia and lymphoma, showing that changes in the alkaline phosphatase levels can be used to detect rare populations of highly refractory malignant cells...
October 6, 2016: Oncotarget
Zala Lužnik, Marko Hawlina, Elvira Maličev, Marina Bertolin, Andreja Nataša Kopitar, Alojz Ihan, Stefano Ferrari, Petra Schollmayer
PURPOSE: To evaluate the effect of prolonged limbal explants cultured without any scaffolds or on amniotic membrane (AM) on the viability, proliferation and differentiation potential of putative phenotypically defined cultured limbal mesenchymal (LMSC) and epithelial stem cells (LESC). METHODS: Limbal explants were cultivated on cryopreserved intact AM or plastic plates using medium supplemented with only human serum. AM was positioned with either the epithelial or stromal side up...
2016: PloS One
Prithal Gangadhar, Zulfikar Ahmed, Muktha R Pai, I Sandhya
Plasma cell leukemia (PCL) is a rare and aggressive variant of myeloma accounting for 2-3% of all plasma cell dyscrasias characterized by the presence of circulating plasma cells. The diagnosis is based on the % (≥20%) and absolute number (≥2x10 9 /L) of plasma cells in the peripheral blood. The incidence of primary PCL (pPCL) is very rare and reported to occur in <1 in a million. It is classified as either pPCL occurring at diagnosis or as secondary PCL in patients with relapsed/refractory myeloma. pPCL is a distinct clinicopathological entity with different cytogenetic and molecular findings...
October 2016: Indian Journal of Pathology & Microbiology
Prashant R Tembhare, Sitaram Ghogale, Nisha Ghatwai, Yajamanam Badrinath, Nikesh Kunder, Nikhil V Patkar, Asma R Bibi, Gaurav Chatterjee, Brijesh Arora, Gaurav Narula, Shripad Banawali, Nilesh Deshpande, Prathibha Amare, Sumeet Gujral, Papagudi G Subramanian
BACKGROUND: Multiparametric flow cytometry (MFC) is a popular technique for minimal residual disease (MRD) analysis. However, its applicability is still limited to 90% of B-cell precursor acute lymphoblastic leukemia (BCPALL) due to two major issues, i.e. a proportion of cases do not express adequate leukemia associated immunophenotype (LAIPs) with currently used markers and drug-induced antigen-modulation. Hence, the incorporation of additional reliable markers is required for the further improvement of MFC-based MRD evaluation...
October 7, 2016: Cytometry. Part B, Clinical Cytometry
Lauren P McLaughlin, Haili Lang, Elizabeth Williams, Kaylor E Wright, Allison Powell, Conrad R Cruz, Anamaris M Colberg-Poley, Cecilia Barese, Patrick J Hanley, Catherine M Bollard, Michael D Keller
BACKGROUND AIMS: Human parainfluenza virus-3 (HPIV) is a common cause of respiratory infection in immunocompromised patients and currently has no effective therapies. Virus-specific T-cell therapy has been successful for the treatment or prevention of viral infections in immunocompromised patients but requires determination of T-cell antigens on targeted viruses. METHODS: HPIV3-specific T cells were expanded from peripheral blood of healthy donors using a rapid generation protocol targeting four HPIV3 proteins...
September 27, 2016: Cytotherapy
Oula El Atat, Diane Antonios, George Hilal, Nabil Hokayem, Joelle Abou-Ghoch, Hussein Hashim, Rim Serhal, Clara Hebbo, Mayssam Moussa, Nada Alaaeddine
The use of adipose-derived stem cells (ADSC) in regenerative medicine is rising due to their plasticity, capacity of differentiation and paracrine and trophic effects. Despite the large number of cells obtained from adipose tissue, it is usually not enough for therapeutic purposes for many diseases or cosmetic procedures. Thus, there is the need for culturing and expanding cells in-vitro for several weeks remain. Our aim is to investigate if long- term proliferation with minimal passaging will affect the stemness, paracrine secretions and carcinogenesis markers of ADSC...
2016: PloS One
Alina Nicolae, Karthik A Ganapathi, Trinh Hoc-Tran Pham, Liqiang Xi, Carlos A Torres-Cabala, Nahid M Nanaji, Hongbin D Zha, Zhen Fan, Sybil Irwin, Stefania Pittaluga, Mark Raffeld, Elaine S Jaffe
Aggressive natural killer cell leukemia (ANKL) is a systemic NK-cell neoplasm, almost always associated with Epstein-Barr virus (EBV). Rare cases of EBV-negative ANKL have been described, and some reports suggested more indolent behavior. We report the clinicopathologic, immunophenotypic, and molecular characteristics of 7 EBV-negative ANKL. All patients were adults, with a median age of 63 years (range 22 to 83 y) and an M:F ratio of 2.5:1. Five patients were White, 1 Black, and 1 Asian. All patients presented acutely, with fever (6/7), cytopenias (6/7), and splenomegaly (4/7)...
September 14, 2016: American Journal of Surgical Pathology
Edson R A Oliveira, Antônio J S Gonçalves, Simone M Costa, Adriana S Azevedo, Marcio Mantuano-Barradas, Ana Cristina M A Nogueira, Ada M B Alves
Dengue disease has emerged as a major public health issue across tropical and subtropical countries. Infections caused by dengue virus (DENV) can evolve to life-threatening forms, resulting in about 20,000 deaths every year worldwide. Several animal models have been described concerning pre-clinical stages in vaccine development against dengue, each of them presenting limitations and advantages. Among these models, a traditional approach is the inoculation of a mouse-brain adapted DENV variant in immunocompetent animals by the intracerebral (i...
2016: PloS One
Luca Aresu
Diffuse large B-cell lymphoma (DLBCL) is the most common canine aggressive B-cell lymphoma worldwide, and new recent molecular approaches have shown that DLBCL constitutes a heterogeneous tumor that cannot be unraveled by morphology and immunophenotype. DLBCL behaves aggressively, typically progressing over a short period of time, and the therapy response may be difficult to be predicted. Recently, the rate of bone marrow infiltration by flow cytometry has been demonstrated to be prognostic, but more sensible markers are needed...
2016: Frontiers in Veterinary Science
Florin Tuluc, Sergei Spitsin, Nancy B Tustin, Jennifer B Murray, Richard Tustin, Laura A Schankel, Andrew Wiznia, Sharon Nachman, Steven D Douglas
We investigated the effect of combination antiretroviral therapy (cART) on immune recovery, particularly on the percentages of PD-1-positive cells within the major leukocyte subsets. Cryopreserved peripheral blood mononuclear cells and plasma samples collected longitudinally from a subset of 13 children and adolescents (between 9.7 and 18.2 years old) who were enrolled in the International Maternal Pediatric Adolescent AIDS Clinical Trials (IMPAACT) P1066 were used for this study. Immunophenotyping by flow cytometry was performed to determine the effect of raltegravir-containing cART regimen on the distribution of leukocyte populations, on the expression of PD-1 on T cell subpopulations, and on the expression of well-established markers of T cell activation (CD38 and HLA-DR) on CD8 T cells...
October 18, 2016: AIDS Research and Human Retroviruses
Alessandra Stacchini, Anna Demurtas, Sabrina Aliberti, Antonella Barreca, Domenico Novero, Donatella Pacchioni
OBJECTIVES: Flow cytometry (FC) has become a useful support for cytomorphologic evaluation (CM) of fine-needle aspirates (FNA) and serous cavity effusions (SCE) in cases of suspected non-Hodgkin lymphoma (NHL). FC results may be hampered by the scarce viability and low cellularity of the specimens. STUDY DESIGN: We developed a single-tube FC assay (STA) that included 10 antibodies cocktailed in 8-color labeling, a cell viability dye, and a logical gating strategy to detect NHL in hypocellular samples...
2016: Acta Cytologica
R M Ribeiro Itaborahy, S Freitas de Medeiros
OBJECTIVE: To evaluate the impact of estrogen therapy on cellular and humoral immune markers in postmenopausal women. METHODS: This prospective, controlled cohort study included 30 patients who used oral estradiol (1 mg) for 14-17 weeks and 28 patients who served as controls. Total leukocytes and leukocyte subtypes were counted and immunophenotyped by flow cytometry. The concentrations of immunoglobulins and pro- and anti-inflammatory cytokines were also measured in the peripheral blood before and after estrogen therapy...
October 2016: Climacteric: the Journal of the International Menopause Society
Vinodh Pillai, David M Dorfman
Many epithelial neoplasms can be analyzed by flow cytometry (FC), particularly from serous cavity effusion samples, using EpCAM, a cell adhesion molecule expressed on most normal epithelial cells and expressed at a higher level in most epithelial neoplasms. A simple 3-color flow cytometric panel can provide a high sensitivity and specificity compared to cytomorphology. FC provides more rapid immunophenotyping than conventional immunohistochemical staining, can identify rare malignant cells that could be missed by a cytological exam alone, and can be utilized to evaluate limited samples such as cerebrospinal fluid or fine-needle aspiration samples...
2016: Acta Cytologica
Bianca Mendes Souza, Tatiane Melo Preisser, Vanessa Bastos Pereira, Meritxell Zurita-Turk, Camila Prósperi de Castro, Vanessa Pecini da Cunha, Rafael Pires de Oliveira, Ana Cristina Gomes-Santos, Ana Maria Caetano de Faria, Denise Carmona Cara Machado, Jean-Marc Chatel, Vasco Ariston de Carvalho Azevedo, Philippe Langella, Anderson Miyoshi
BACKGROUND: Inflammatory bowel diseases are characterized by chronic intestinal inflammation that leads to severe destruction of the intestinal mucosa. Therefore, the understanding of their aetiology as well as the development of new medicines is an important step for the treatment of such diseases. Consequently, the development of Lactococcus lactis strains capable of delivering a eukaryotic expression vector encoding the interleukin 4 (IL-4) of Mus musculus would represent a new strategy for the elaboration of a more effective alternative therapy against Crohn's disease...
2016: Microbial Cell Factories
Anna Lucia Peluso, Antonio Ieni, Chiara Mignogna, Pio Zeppa
Lymph node (LN) fine-needle cytology (FNC) coupled with flow cytometry immunophenotyping provides relevant information for the diagnosis of non-Hodgkin lymphoma (NHL). Numerous studies have shown FNC samples to be suitable for different molecular procedures; in this review, some of the molecular procedures most commonly employed for NHL are briefly described and evaluated in this perspective. Fluorescence in situ hybridization and chromogenic in situ hybridization are briefly described. Polymerase chain reaction (PCR)-based assays are used to identify and quantify mutations and translocations, namely immunoglobulin (IGH) and T-cell receptor rearrangements by clonality testing and IGVH somatic hypermutations either by Sanger sequencing, single-strand conformational polymorphisms or RT-PCR strategies...
2016: Acta Cytologica
Hui-Fang Li, Wen-Tong Meng, Yong-Qian Jia, Neng-Gang Jiang, Ting-Ting Zeng, Yong-Mei Jin, Qiao-Rong Huang, Xue Li, Hong Xu, Xian-Ming Mo
B cell acute lymphoblastic leukemia (B-ALL) exhibits phenotypes reminiscent of normal stages of B-cell development. As demonstrated by flow cytometry, the immunophenotypes are able to determine the stages of B cell development. Multicolor flow cytometry (MFC) is more accurate at identifying cell populations. In this study, 9-color panels, including CD10, CD19, CD20, CD22, CD34, CD79a, CD179a, and IgM, which are sequentially expressed during B cell development, were designed to detect the leukemia cell subpopulations in adult B-ALL patients...
August 2016: Medicine (Baltimore)
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