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Immunological Tests for Diagnosis of Disease and Identification of Molecules
Published in Julius P. Kreier, Infection, Resistance, and Immunity, 2022
A third procedure, the sandwich test, detects receptors or antibody on or in the cytoplasm of cells. Antigen is the first reagent, then fluorescent-labeled antibody is added, and the antigen is sandwiched between the two antibodies. Cell surface markers, such as antigen binding proteins (i.e., receptors) of T lymphocytes and immunoglobulins present in and on B lymphocytes, are frequently detected by such fluorescent assays.
Regenerative Medicine in Pain Management
Published in Sahar Swidan, Matthew Bennett, Advanced Therapeutics in Pain Medicine, 2020
Sharon McQuillan, Rafael Gonzalez
Cell surface markers, including clusters of differentiation (CD), are critical for cell–cell and cell–environment interactions and are used to define mixed cell populations. Despite an enormous amount of research, there is little consensus with regard to CD characterization of the SVF. Table 13.1 shows a comparison of SVF-defined cell surface markers.15,19–25Table 13.2 provides a comparison of cell surface markers between bone marrow and adipose-derived stem cells.20
Peripheral Blood and Bone Marrow
Published in Harold R. Schumacher, William A. Rock, Sanford A. Stass, Handbook of Hematologic Pathology, 2019
Fermina Maria Mazzella, Gerardo Perrotta
Flow cytometry is virtually unique in that the laboratory may study several populations of cells within a specimen for multiple features and/or markers. The primary use of cell surface markers in hematopathology is to identify alterations in a particular group of cells that may correlate with a disease state. Flow cytometric assays, however, are by no means limited to immunophenotyping. Assays have been described for the quantitation of DNA for ploidy and proliferation studies, cell surface receptors, carbohydrates, intracellular antigens such as cytoplasmic mu and terminal deoxynucleotidyl transferase (TdT), ion fluxes, and phagocytosis.
microRNA-130b-3p delivery by mesenchymal stem cells-derived exosomes confers protection on acute lung injury
Published in Autoimmunity, 2022
Xiaoxia Wang, Jifeng Feng, Huijun Dai, Jianlan Mo, Bijun Luo, Cheng Luo, Weikang Zhang, Linghui Pan
CD105, CD29, CD44 and CD34 are commonly used to identify MSCs, among which CD105, CD29 and CD44 is a positive marker, whereas CD34 is a negative marker [36,37]. The specific function of the cells is related to its surface markers, and cell surface markers can reflect some basic characteristics of cells. MSCs are a mixed cell population, and their surface antigens are also non-specific, expressing the surface markers of mesenchymal cells, endothelial cells and epidermal cells [38]. CD29, also known as integrin β1, VLA-β chain or gpIIa, is a receptor for various extracellular matrix proteins, and CD29 acts as a fibronectin receptor involved in various cell–cell and cell–matrix interactions. CD105, also known as endoglin, is a 90-kDa type I transmembrane glycoprotein of the zona pellucida protein (ZP) family. CD44, also known as Hermes, Pgp1, H-CAM or Hutch, is an 80–95 kDa glycoprotein, which is expressed in leukocytes, endothelial cells, hepatocytes and MSCs. CD34 is a transmembrane salivary mucin that may be involved in adhesion and anti-adhesion. Therefore, we chose CD105, CD29, CD44 and CD34 to identify MSCs. Flow cytometry showed that CD105, CD29 and CD44 were positive, and CD34 was negative (Figure 1(A)). Adipogenic and osteogenic differentiation experiments uncovered that after oil red O staining, there were red lipid droplets in the cells; in the osteogenic medium, Alizarin Red stained cubes and formed mineralized nodules, indicating that the cells had multi-directional differentiation abilities (Figure 1(B)).
Triterpenoids of Rhus chinensis Supressed Colorectal Cancer Progress by Enhancing Antitumor Immunity and CD8 + T Cells Tumor Infiltration
Published in Nutrition and Cancer, 2022
Gang Wang, Yang Yu, Zi-Meng Li, Zhi-Min Zhu, Zhi-Jie Wang, Min-Fang Tao
Flow cytometry was used to analyze the characteristics of NK cells in spleen, blood, tumor NK cells, CD4 + and CD8 + T cells in spleen, blood and tumor. After 28 days of treatment, the mice in each group (n = 6) were euthanized. Then the spleen of the mice was removed, and the blood and tumor were taken for NK cells. The CD4 + T and CD8 + T cells were measured by flow cytometry. Then, a single cell suspension, namely spleen and blood suspension, was prepared by filtration with 3000 mesh screen. Tumor suspensions were prepared and modified as previously described (13). The cell suspensions were then stained at 4 degree for 30 min using the following antibodies: PE anti-mouse NKp46-PerCP, PE anti-mouse CD86, PE-cy5 anti-mouse CD3, FITC anti-mouse CD4, PE anti-mouse CD8, and the corresponding isotype control antibodies (all monoclonal antibodies were obtained from BD Bioscience). Subsequently, the intracellular staining was performed (34) by flow cytometry, and FITC labeled anti antibodies: anti- PD-L1-PE mAb and anti-PD-1-PE mAb were used for intracellular staining. The expression of cell surface markers was detected by flow cytometry and analyzed by flowjo software (FlowJo, Ashland, or, USA). Anti-mouse antibodies such as anti-IFN-γ- FITC, anti-TNF-α- FITC, anti-TGF-β-FITC, IL-10-APC mAb were used. After washing with PBS, the cells were fixed with 10% formaldehyde, and then the cell frequency was determined by BD faccibur. Finally, the samples were analyzed by FlowJo software.
Mechanisms of cellular and humoral immunity through the lens of VLP-based vaccines
Published in Expert Review of Vaccines, 2022
Hunter McFall-Boegeman, Xuefei Huang
In the last decade, a new subset of memory T cells, the tissue resident memory (TRM), has been characterized[153]. The hallmarks of such cells are not cell surface markers but rather mainly the fact that they are found in barrier tissues, such as the lungs, skin, reproductive tracts, etc., and do not circulate[154,155]. In fact, TRM cells can express different cell surface markers depending on the type of tissue they are in[156]. These cells are thought to play an important role in reactivation of the immune response to subsequent infections. VLP-based vaccines are able to elicit TRM cells[157]. For example, intranasal immunization of mice with P22 loaded with M and M2 proteins from the Respiratory Syncytial Virus (RSV), fused to the P22 scaffold protein, was able to elicit TRM cell populations in the lungs[158]. These cells were detected by flow cytometry analysis of bronchioalveolar lavage fluid up to 2 months post inoculation. Importantly, while there was a small decrease in total cell counts for both M- and M2-specific TRM cells, it was still protective as measured by lung viral titers, after re-challenge.