1.Role of CD47 in hematologic malignancies.
Journal of Experimental Hematology 2013;21(6):1631-1634
CD47 is a ubiquitously expressed transmembrane glycoprotein on surface of many cells. Through its interaction with integrin, signal regulatory protein alpha (SIRPα) and thrombin sensitive protein-1 (TSP-1), it plays important roles in various immunological processes including inflammatory response, immune response and tumor immunity. Recently, it has been found that CD47 interacts with SIRPα expressed on phagocytic cells, which transfers a negative signal when being activated. By the mechanisms described above, CD47-SIRPα signal complex is involved in the pathogenesis of hematological diseases and might provide some informations for the therapy of patients. This review focuses on the structure and immunoregulatory functions of CD47, the mechanism of CD47 in tumor therapy, the CD47 and hematologic malignancies including acute leukemia, B-cell lymphoma and multiple myeloma, as well as CD47 and hematopoietic stem cell transplantation.
Animals
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Antigens, Differentiation
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metabolism
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CD47 Antigen
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metabolism
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Hematologic Neoplasms
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metabolism
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Humans
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Receptors, Immunologic
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metabolism
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Signal Transduction
4.Expression of dendritic cell markers on cultured neutrophils and its modulation by anti-apoptotic and pro-apoptotic compounds.
Hae Young PARK ; Jun O JIN ; Min Gyu SONG ; Joo In PARK ; Jong Young KWAK
Experimental & Molecular Medicine 2007;39(4):439-449
Neutrophils are also known to acquire the characteristics of dendritic cells (DCs) under the appropriate conditions. In this study, neutrophils were cultivated in vitro in the presence or absence of compounds modulating their survival in an attempt to characterize the expression profile of the DC markers. Higher MHC-II, CD80, CD86, CD83, and CD40 expression levels were detected on the surface of the cultured neutrophils for 24 h than on the freshly isolated cells. The annexin V-positive cells showed a higher expression level of the DC markers than the annexin V-negative cells. The population of neutrophils double stained with annexin V and the DC markers increased after being incubated with agonistic anti-Fas Ab. LPS, the anti-apoptotic compound, decreased the CD86 and MHC-II expression levels but 50-60% of the DC marker-positive cells were detected in the annexin V-positive cells. In contrast, CD80, CD86, CD83, and HLA-DR mRNA levels increased in the GM-CSF-treated neutrophils but not in the anti-Fas Ab-treated neutrophils. T cell proliferation was inhibited by co-culturing them with anti-Fas Ab- or LPS-treated neutrophils at a high neutrophil:T cell ratio. However, the superantigen-mediated T cell proliferation was increased by the LPS-treated neutrophils but decreased by the anti-Fas Ab-treated neutrophils. There was a lower level of interferon-gamma production in the T cells co-cultured with anti-Fas Ab-treated neutrophils than with the LPS-treated neutrophils. This suggests that apoptotic neutrophils express DC markers on their surface and the differential expression of DC markers might have a detrimental effect on the immune reaction.
Antigen Presentation
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Antigens, CD/biosynthesis
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Antigens, CD95/pharmacology
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Antigens, Differentiation/*biosynthesis
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*Apoptosis
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Cells, Cultured
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Dendritic Cells/*metabolism
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Humans
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Lipopolysaccharides/pharmacology
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Lymphocyte Activation
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Neutrophils/*metabolism/physiology
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T-Lymphocytes/immunology
5.Isolation, culture and validation of CD34+ vascular wall-resident stem cells from mice.
Li-Ju YANG ; Ying MA ; Yuan LI ; Qing-Ya DANG ; Jun CHENG ; Yan YANG ; Peng-Yun LI
Acta Physiologica Sinica 2023;75(2):205-215
Vascular wall-resident stem cells (VW-SCs) play a critical role in maintaining normal vascular function and regulating vascular repair. Understanding the basic functional characteristics of the VW-SCs will facilitate the study of their regulation and potential therapeutic applications. The aim of this study was to establish a stable method for the isolation, culture, and validation of the CD34+ VW-SCs from mice, and to provide abundant and reliable cell sources for further study of the mechanisms involved in proliferation, migration and differentiation of the VW-SCs under various physiological and pathological conditions. The vascular wall cells of mouse aortic adventitia and mesenteric artery were obtained by the method of tissue block attachment and purified by magnetic microbead sorting and flow cytometry to obtain the CD34+ VW-SCs. Cell immunofluorescence staining was performed to detect the stem cell markers (CD34, Flk-1, c-kit, Sca-1), smooth muscle markers (SM22, SM MHC), endothelial marker (CD31), and intranuclear division proliferation-related protein (Ki-67). To verify the multipotency of the isolated CD34+ VW-SCs, endothelial differentiation medium EBM-2 and fibroblast differentiation medium FM-2 were used. After culture for 7 days and 3 days respectively, endothelial cell markers and fibroblast markers of the differentiated cells were evaluated by immunofluorescence staining and q-PCR. Furthermore, the intracellular Ca2+ release and extracellular Ca2+ entry signaling were evaluated by TILLvisION system in Fura-2/AM loaded cells. The results showed that: (1) High purity (more than 90%) CD34+ VW-SCs from aortic adventitia and mesenteric artery of mice were harvested by means of tissue block attachment method and magnetic microbead sorting; (2) CD34+ VW-SCs were able to differentiate into endothelial cells and fibroblasts in vitro; (3) Caffeine and ATP significantly activated intracellular Ca2+ release from endoplasmic reticulum of CD34+ VW-SCs. Store-operated Ca2+ entry (SOCE) was activated by using thapsigargin (TG) applied in Ca2+-free/Ca2+ reintroduction protocol. This study successfully established a stable and efficient method for isolation, culture and validation of the CD34+ VW-SCs from mice, which provides an ideal VW-SCs sources for the further study of cardiovascular diseases.
Mice
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Animals
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Endothelial Cells
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Cell Differentiation/physiology*
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Stem Cells
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Adventitia
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Fibroblasts
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Cells, Cultured
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Antigens, CD34/metabolism*
7.Atrophic dermatofibrosarcoma protuberans: report of a case.
Xiao-yu HAN ; Hong-quan WEI ; Qing PAN ; Jun LIU
Chinese Journal of Pathology 2013;42(1):52-53
Adult
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Antigens, CD
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metabolism
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Antigens, CD34
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metabolism
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Antigens, Differentiation, Myelomonocytic
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metabolism
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Dermatofibrosarcoma
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metabolism
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pathology
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surgery
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Diagnosis, Differential
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Female
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Fibroma
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metabolism
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pathology
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Humans
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Lipoma
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pathology
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Neurofibroma
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metabolism
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pathology
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Receptors, Cell Surface
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metabolism
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Skin Neoplasms
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metabolism
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pathology
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surgery
8.A preliminary study of markers for human hair follicle melanin stem cell.
Xing-Yu MEI ; Zhou-Wei WU ; Cheng-Zhong ZHANG ; Yue SUN ; Wei-Min SHI
Chinese Medical Journal 2019;132(9):1117-1119
Antigens, CD34
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metabolism
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Biomarkers
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metabolism
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Cell Differentiation
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physiology
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Hair Follicle
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cytology
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Humans
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Intramolecular Oxidoreductases
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metabolism
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Keratinocytes
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metabolism
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Melanins
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metabolism
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Melanocytes
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metabolism
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PAX3 Transcription Factor
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metabolism
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Stem Cells
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metabolism
9.WT1 gene expression and its isoform ratio in different cell subsets of normal human bone marrow.
Jing XU ; Hong-Wei WANG ; Xiao-Hong LI ; Lei ZHU ; Li ZHANG ; Fan ZHANG ; Yan-Hong TAN ; Tao YANG
Journal of Experimental Hematology 2007;15(3):603-606
The Wilms' tumor gene (WT1) is a transcription factor involved in tumorigenesis, especially in leukemogenesis. However, the role of WT1 expression in nonmalignant hematopoietic cells remains unclear. Furthermore, due to alternative splicing at two sites: 17 amino acid residues of exon 5 (+17AA) and 3 amino acid residues (+KTS) between exons 9 and 10, WT1 gene has four main isoforms (17AA+/KTS+, 17AA+/KTS-, 17AA-/KTS+, 17AA-/KTS-, abbreviation: +/+, +/-, -/+, -/-). The isoforms probably existed in hematopoietic cells, which make the research more complex. The aim of study was to elucidate the expression and its isoforms of WT1 gene in different cell subsets of healthy bone marrow donors. The fluorescence RT-PCR detection system was established to measure the expressions of full-length WT1, WT1 (+17AA) and WT1 (+KTS) in CD34(+)CD38(-) (stem cell), CD34(+)CD38(+) (progenitor cell), CD15(+)CD11b(+) (granulocyte), CD33(+)CD14(+) (monocyte), CD20(+)CD5(-) (B-lymphocyte) and CD20(-)CD5(+) (T-lymphocyte) subsets from 18 normal human bone marrow samples. The results showed that WT1 expressed in CD34(+)CD38(-), CD34(+)CD38(+), CD15(+)CD11b(+) and CD33(+)CD14(+), but not in CD20(+)CD5(-) and CD20(-)CD5(+) subsets. The highest expression was in CD34(+)CD38(-), but decreased gradually in CD15(+)CD11b(+) and CD33(+)CD14(+) subsets. WT1 (+17AA), WT1 (+KTS) and WT1 (+/+) isoforms were predominant in CD34(+)CD38(-) and CD34(+)CD38(+) primitive subsets, while in CD15(+)CD11b(+) and CD33(+)CD14(+) the dominant isoforms were WT1 (-17AA), WT1 (-KTS) and WT1 (-/-). It is concluded that the expression of WT1 in normal bone marrow decreases gradually with cell differentiation. Hematopoietic cells may adjust the ratios of WT1 isoforms to inhibit or promote cell differentiation.
Antigens, CD34
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analysis
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Bone Marrow Cells
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metabolism
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Cell Differentiation
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Gene Expression Regulation
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Humans
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Protein Isoforms
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metabolism
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WT1 Proteins
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metabolism
10.DNAM-1 regulates the proliferation and function of T regulatory type 1 cells via the IL-2/STAT5 pathway.
Ning WANG ; Yi Han WANG ; Peng Tao JIANG ; Ming Hua LÜ ; Zhi Fang HU ; Xi XU
Journal of Southern Medical University 2022;42(9):1288-1295
OBJECTIVE:
To explore the role of DNAM-1 in the activation, proliferation and function of type Ⅰ regulatory T cells (Tr1 cells).
METHODS:
Anti-CD3/CD28 antibodies were used to stimulate mouse T cells derived from the spleen of wild-type (WT) mice, and the expression level of DNAM-1 in resting and activated Tr1 cells was evaluated with flow cytometry. Na?ve CD4+ T cells isolated by magnetic cell sorting from the spleens of WT mice and DNAM-1 knockout (KO) mice were cultured in Tr1 polarizing conditions for 3 days, after which CD25 and CD69 expressions were measured using flow cytometry. The induced Tr1 cells were labelled with CFSE and cultured in the presence of anti-CD/CD28 antibodies for 3 days, and their proliferative activity was analyzed. The expressions of IL-10 and p-STAT5 in DNAM-1-deficient Tr1 cells were detected before and after IL-2 stimulation.
RESULTS:
The expression level of DNAM-1 was significantly upregulated in CD4+ T cells and Tr1 cells after stimulation with anti-CD3/CD28 antibodies (P < 0.05). DNAM-1 knockout did not cause significant changes in the number or proportion of Tr1 cells, but but significantly increased the expression levels of the activation markers CD69 and CD25 (P < 0.05). Compared with WT Tr1 cells, DNAM-1-deficient Tr1 cells exhibited reduced proliferative activity in vitro (P < 0.05) with downregulated IL-10 production (P < 0.05) and decreased expressions of Il-10 and Gzmb mRNA (P < 0.05). In DNAM-1-deficient Tr1 cells, IL-2 stimulation significantly reduced IL-10 secretion level and the expression of p-STAT5 as compared with WT Tr1 cells.
CONCLUSION
DNAM-1 participate in the activation and proliferation of Tr1 cells and affect the biological functions of Tr1 cells through the IL-2/STAT5 pathway.
Animals
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Antigens, Differentiation, T-Lymphocyte
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CD28 Antigens/metabolism*
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Cell Proliferation
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Cells, Cultured
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Interleukin-10
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Interleukin-2/metabolism*
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Mice
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RNA, Messenger
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STAT5 Transcription Factor/metabolism*
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T-Lymphocytes, Regulatory