1.Varying expression levels of colony stimulating factor receptors in disease states and different leukocytes.
Kyo Young LEE ; Byung Gyu SUH ; Jong Wan KIM ; Won Bae LEE ; So Young KIM ; Young Yoo KIM ; Je Hoon LEE ; Ji Hyang LIM ; Myung Shin LIM ; Chang Suk KANG ; Kyung Ja HAN
Experimental & Molecular Medicine 2000;32(4):210-215
Administration of G-CSF may not always respond in rise of neutrophil counts in different patient population. In order to understand a possible inter-relationship between the G-CSF and GM-CSF induced leukocyte responses and expression levels of receptors for G-CSF (G-CSFr) and GM-CSF (GM-CSFr), the levels of each receptor and CSF were measured in patients with basophilia (8), eosinophilia (14) and bacterial infection showing neutrophilia (12) in comparison with normal healthy adults (12) and children (14). G-CSFr was expressed in neutrophils in the largest amount followed by monocytes, but GM-CSFr was expressed more in monocytes than neutrophils. Lymphocytes and basophils did not express G-CSFr or GM-CSFr. The amount of GM-CSFr in neutrophils was present less in patients with infection than normal control (P = 0.031). The neutrophils expressed more G-CSFr than GM-CSFr. The quantity of G-CSFr in eosinophil showed marked interval change, higher in acute stage. The plasma concentrations of G-CSF in patients with infection were much higher than normal adults or children (117.95 +/- 181.16 pg/ml, P < 0.05). Binding assay with excess amount of CSFs could discriminate the patient who did not show any response to G-CSF or GM-CSF administration. After incubation with excess CSFs, more receptors were blocked in children than in adults (G-CSF P = 0.024, GM-CSF P = 0.006). These results indicate that the amount of CSFr in leukocyte varies in different types of leukocyte, and changes according to the patients' condition even in the same type of leukocyte, and the CSFrs of children bind to CSFs more than those of adults.
Adult
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*Bacterial Infections
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Basophils/chemistry
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Breast Neoplasms
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Child
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Colony-Stimulating Factors/*blood
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Eosinophilia
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Human
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Leukemia, Myeloid, Chronic
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*Leukocyte Disorders
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Monocytes/chemistry
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*Neoplasms
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Neutrophils/chemistry
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Receptors, Colony-Stimulating Factor/*analysis
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Receptors, Granulocyte Colony-Stimulating Factor/analysis
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Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/analysis
2.Analysis of the GM-CSF and GM-CSF/IL-3/IL-5 receptor common beta chain in a patient with pulmonary alveolar proteinosis.
Xuanding WANG ; Fuguang LIU ; Burkhard BEWIG
Chinese Medical Journal 2002;115(1):76-80
OBJECTIVETo investigate the expression of the granulocyte-macrophage colony-stimulating factor (GM-CSF) and GM-CSF/IL-3/IL-5 receptor common beta chain (beta c receptor) in an adult patient with idiopathic pulmonary alveolar proteinosis (PAP), so as to demonstrate the possible association of the GM-CSF and beta c receptor with the pathogenesis of human PAP.
METHODSThe GM-CSF levels were measured with a commercial ELISA kit (sensitivity 5 pg/ml) and the beta c receptor expression on the cell surface was detected by flow cytometry analysis. Reverse transcription polymerase chain reaction (RT-PCR) analysis was employed to detect the expression of the GM-CSF mRNA and the beta c receptor mRNA in peripheral blood mononuclear cells and alveolar macrophages. The entire coding regions of the GM-CSF cDNA and the beta c receptor cDNA were sequenced by the Sanger dideoxy-mediated chain termination method to detect possible mutations.
RESULTSThe patient with PAP failed to release the GM-CSF protein either from circulating mononuclear cells or from alveolar macrophages. The expression of the GM-CSF mRNA was normal after the stimulation of lipopolysaccharide, whereas a point mutation at position 382 of the GM-CSF cDNA from "T" to "C" was revealed by cDNA sequencing, which caused a change in amino acid 117 of the protein from isoleucine to threonine. The beta c receptor expression on the cell surface was normal, and the beta c receptor mRNA expression and the sequence of the entire coding region of the beta c receptor were also normal.
CONCLUSIONSThe decreased GM-CSF production is associated with the pathogenesis of human PAP. A point mutation of the GM-CSF cDNA may contribute to the decreased GM-CSF production in our adult PAP patient. The mutation of the beta c receptor in some of paediatric patients with PAP may not be a common problem in adult patients.
DNA, Complementary ; chemistry ; Granulocyte-Macrophage Colony-Stimulating Factor ; analysis ; biosynthesis ; Humans ; Male ; Middle Aged ; Pulmonary Alveolar Proteinosis ; etiology ; metabolism ; RNA, Messenger ; analysis ; Receptors, Cytokine ; biosynthesis ; genetics
3.Transcriptomic microarray profiling of peripheral CD4+ T cells from asthmatic patients.
Min ZHU ; Min HE ; Yarong HE ; Yulin JI
Chinese Journal of Medical Genetics 2018;35(6):828-831
OBJECTIVE:
To identify differentially expressed genes in peripheral blood mononuclear cells between patients with continuous mild-to-moderate asthma and healthy controls using mRNA microarray in order to explore the underlying signaling pathways and clarify the roles of CD4+ T cells in the pathogenesis of asthma.
METHODS:
Global transcriptomic profiles of the CD4+ T cells were defined by using Agilent Sure Print G3 Human GE 8×60K microarray. Enrichment pathways were analyzed with Ingenuity Pathway Analysis (IPA) software.
RESULTS:
Compared with controls, 805 genes were up-regulated, 192 were down-regulated in asthma patients. Among these, the expression of 38 annotated genes have varied by 4 times or more. Expression of CD300A was inversely proportional to the absolute value of eosinophils (r=-0.89, P=0.02) as well as the proportion of eosinophils (r=-0.94, P=0.004), while CSF1R was inversely proportional to PD20 (r=-0.83, P=0.04) and AQLQ (r=-0.88, P=0.02) by correlation analysis.
CONCLUSION
Numerous pathophysiological pathways may be involved in the pathogenesis of asthma. Above findings have provided a basis for the delineation the pathogenesis of asthma.
Antigens, CD
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genetics
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Asthma
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immunology
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CD4-Positive T-Lymphocytes
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cytology
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Case-Control Studies
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Eosinophils
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Gene Expression Profiling
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Humans
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Leukocytes, Mononuclear
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Oligonucleotide Array Sequence Analysis
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Receptors, Granulocyte-Macrophage Colony-Stimulating Factor
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genetics
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Receptors, Immunologic
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genetics
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Transcriptome
4.Single-cell Analysis of CAR-T Cell Activation Reveals A Mixed T1/T2 Response Independent of Differentiation.
Iva XHANGOLLI ; Burak DURA ; GeeHee LEE ; Dongjoo KIM ; Yang XIAO ; Rong FAN
Genomics, Proteomics & Bioinformatics 2019;17(2):129-139
The activation mechanism of chimeric antigen receptor (CAR)-engineered T cells may differ substantially from T cells carrying native T cell receptor, but this difference remains poorly understood. We present the first comprehensive portrait of single-cell level transcriptional and cytokine signatures of anti-CD19/4-1BB/CD28/CD3ζ CAR-T cells upon antigen-specific stimulation. Both CD4 helper T (T) cells and CD8 cytotoxic CAR-T cells are equally effective in directly killing target tumor cells and their cytotoxic activity is associated with the elevation of a range of T1 and T2 signature cytokines, e.g., interferon γ, tumor necrotic factor α, interleukin 5 (IL5), and IL13, as confirmed by the expression of master transcription factor genes TBX21 and GATA3. However, rather than conforming to stringent T1 or T2 subtypes, single-cell analysis reveals that the predominant response is a highly mixed T1/T2 function in the same cell. The regulatory T cell activity, although observed in a small fraction of activated cells, emerges from this hybrid T1/T2 population. Granulocyte-macrophage colony stimulating factor (GM-CSF) is produced from the majority of cells regardless of the polarization states, further contrasting CAR-T to classic T cells. Surprisingly, the cytokine response is minimally associated with differentiation status, although all major differentiation subsets such as naïve, central memory, effector memory, and effector are detected. All these suggest that the activation of CAR-engineered T cells is a canonical process that leads to a highly mixed response combining both type 1 and type 2 cytokines together with GM-CSF, supporting the notion that polyfunctional CAR-T cells correlate with objective response of patients in clinical trials. This work provides new insights into the mechanism of CAR activation and implies the necessity for cellular function assays to characterize the quality of CAR-T infusion products and monitor therapeutic responses in patients.
Antigens
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metabolism
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CTLA-4 Antigen
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metabolism
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Cell Differentiation
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drug effects
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Cell Line
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Cytokines
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metabolism
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Cytotoxicity, Immunologic
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drug effects
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Granulocyte-Macrophage Colony-Stimulating Factor
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pharmacology
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Humans
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Lymphocyte Activation
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drug effects
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immunology
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Lymphocyte Subsets
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drug effects
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metabolism
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Phenotype
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Proteomics
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Receptors, Chimeric Antigen
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metabolism
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Single-Cell Analysis
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methods
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T-Lymphocytes, Regulatory
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drug effects
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metabolism
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Th1 Cells
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cytology
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drug effects
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Th2 Cells
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cytology
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drug effects
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Transcription, Genetic
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drug effects
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Up-Regulation
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drug effects