1.A Study of EFFECT and MECHANISM of IL-2on SURVIVAL of EOSINOPHILS.
Hyo Seok KIM ; Young Mok LEE ; Young Soo CHOI ; Kyung Ho KIM ; Geon Il IM ; Sung Whan JEONG ; Seung Hyug MOON ; Hyeon Tae KIM ; Soo Taek UH ; Yong Hun KIM ; Choon Sik PARK
Tuberculosis and Respiratory Diseases 1996;43(3):348-358
BACKGROUND: Interleukin-5 (IL-5) is responsible for eosinophilia in allergic diseases. In allergic bronchial asthma, there is a correlation between the extent of eosinophil infiltration in bronchial mucosa and IL-5 concentrations. In addition, IL-2 concentration is elevated in the airways and associated with eosinophilia in symptomatic patients with bronchial asthma. In animal studies, IL-2 can induce eosinophilia by increasing the synthesis of IL-5, however, it is still unknown how IL-2 can induce eosinophila in human being. The aim of this study is to evaluation the effect and mechanism of IL-2 on prolongation of eosinophil survival. METHODS: After purifiing the eosinophils from the venous blood of allergic patients with eosinophilia, we measured the survival rates of eosinophils using trypan blue dye exclusion test, and the number of eosinophils with Randolp's solution. We compared the survival rates of eosinophils in the presence of IL-2 or IL-5. Neutralizing antibody for IL-5 was added in IL-2 treated eosinophils to reveal whether IL-2 induced prolongation of eosinophil survival was mediated by IL-5. We checked IL-5 m-RNA expression of lymphocytes in the presence of IL-2 by using Reverse transcription-Polymerase chain reaction (RT-PCR) method to revealed the effect of IL-2 on IL-5 m-RNA expression on lymphocyte. alpha and beta IL-2 receptors were measured on eosinophils and lymphocytes with flow-cytometer after stimulated with IL-2. RESULTS: 1) Eosinophil survival rates increased dose dependently on IL-5 and IL-2. 2) The eosinophil survival rates increased by IL-2 were not inhibited by the pretreatment with neutralizing antibody for IL-5. 3) IL-5 m-RNA was not expressed on lymphocytes by the treatment with IL-2 up to 96 hours. 4) IL-2 upregulate the expression of IL-2Ralpha on eosinophils, instead of no effect on the expression of IL-2Rbeta. CONCLUSION: Interleukin-2 had the enhancing effect on the survival rates of eosinophils. The mechanism behind IL-2 induced eosinophilia might be the increment of IL-2 receptors on eosinophils rather than IL-5 synthesis by lymphocytes.
Animals
;
Antibodies, Neutralizing
;
Asthma
;
Eosinophilia
;
Eosinophils*
;
Humans
;
Interleukin-2
;
Interleukin-2 Receptor alpha Subunit
;
Interleukin-5
;
Lymphocytes
;
Mucous Membrane
;
Receptors, Interleukin-2
;
Survival Rate
;
Trypan Blue
2.Essential roles for ID-1 motif of interleukin-4 receptor alpha chain in interleukin-4 signaling.
Jonghee YOUN ; Kyung Hee LEE ; Woo Youl HWANG ; Doo Jin PAIK ; Ho Sam CHUNG
Journal of Asthma, Allergy and Clinical Immunology 2003;23(2):372-384
BACKGROUND: Interleukin (IL)-4 is a pleiotropic cytokine that plays an important role in the pathogenesis of the allergic inflammation and asthma. Upon IL-4 receptor (IL-4R) engagement, a variety of signaling mediators, such as JAK kinases and STAT-6 are activated, leading to induction of IL-4 target gene expression including CD23 and germline C epsilon transcription. The function of a membrane-proximal domain of IL-4Ra, termed ID-1, remains to be characterized to date. OBJECTIVE: To assess whether the ID-1 domain mediates the induction of IL-4 target gene expression in a STAT-6-dependent manner. METHODS: The intracellular region of IL-4Ralpha was translationally fused to the extracellular region of IL-2Rbeta to provide ligand specificity to IL-2. Acidic amino acids and serine residues in the ID-1 domain of the chimeric receptor were substituted by site-directed mutagenesis. These receptor cDNAs were stably transfected to M12.4.1 murine B lymphoma cells. Following IL-2 stimulation, wild type and mutant clones for the ID-1 motif were subjected to FACS. RNA blotting and elecroporetic mobility shift assays to address the levels of CD23, germline C epsilon and STAT-6 inductions, respectively. RESULTS: ID-1 mutant clones were defective in gene induction of CD23 and germline C epsilon in response to IL-2 stimulation, as compared with wildtype clones. Moreover, IL-2-mediated STAT-6 activation was abolished in ID-1 mutant clones. CONCLUSION: These results demonstrate that the ID-1 domain of IL-4Ra is essential to induce IL-4 target gene expression through a STAT-6-dependent pathway.
Amino Acids, Acidic
;
Asthma
;
Clone Cells
;
DNA, Complementary
;
Electrophoretic Mobility Shift Assay
;
Gene Expression
;
Inflammation
;
Interleukin-2
;
Interleukin-4 Receptor alpha Subunit*
;
Interleukin-4*
;
Interleukins
;
Janus Kinases
;
Lymphoma
;
Mutagenesis, Site-Directed
;
Receptors, Interleukin-4
;
RNA
;
Sensitivity and Specificity
;
Serine
3.Effects of humanized recombinant CD25 monoclonal antibody on activation and proliferation of t lymphocytes.
Journal of Experimental Hematology 2007;15(1):134-137
The study was purposed to investigate the effects of humanized recombined CD25 monoclonal antibody (rhCD25MAb) on activation and proliferation of T lymphocytes in vitro. Peripheral blood mononuclear cells (PBMNC) were incubated with phytohemagglutinin (PHA). Before or after T cell activation, the cells were cultured with or without rhCD25MAb or cyclosporine A (CsA) in vitro. After 72 hours incubation, the proliferation of lymphocytes was analyzed by MTT assay. The expression of CD3 and CD25 antigens on T lymphocytes were detected by flow cytometry. The levels of sIL-2R in the supernatants were determined with ELISA. The results showed that both rhCD25MAb and CsA could inhibit the proliferation of T lymphocytes significantly in concentration-dependent manner and CsA was more efficient than rhCD25MAb. Both rhCD25MAb and CsA could also decrease the levels of sIL-2R in the supernatant and inhibit the expression of CD25 antigen on T lymphocytes. The level of sIL-2R and the expression of CD25 on T lymphocytes decreases more profoundly in rhCD25MAb group. It is concluded that rhCD25MAb shows strong immunosuppressive activity both before and after T cell activation, suggesting that this agent may be useful in not only prophylaxis but also the treatment of acute graft-versus-host disease.
Antibodies, Monoclonal
;
pharmacology
;
CD3 Complex
;
biosynthesis
;
genetics
;
Cell Proliferation
;
Cells, Cultured
;
Cyclosporine
;
pharmacology
;
Humans
;
Immunosuppressive Agents
;
pharmacology
;
Interleukin-2 Receptor alpha Subunit
;
biosynthesis
;
genetics
;
immunology
;
Lymphocyte Activation
;
immunology
;
Receptors, Interleukin-2
;
analysis
;
Recombinant Proteins
;
immunology
;
pharmacology
;
T-Lymphocytes
;
immunology
4.Screening of short peptides binding to cell surface interleukin-2 receptor alpha chain.
Bei-yi LIU ; Ping ZHU ; Hai-bo LUO ; Ning FU
Journal of Southern Medical University 2006;26(7):971-974
OBJECTIVETo screen and characterize the short peptides which bind specifically to interleukin-2 (IL-2) receptor alpha chain (IL-2Ralpha) for acquisition of small antagonists for blocking the binding of IL-2 with IL-2Ralpha.
METHODS12-mer phage displayed peptide library was screened with the target cells of MT-2 cells which expressed IL-2Ralpha at high levels. The binding phage clones were eluted by anti-IL-2Ralpha monoclonal antibody. After 3 rounds of screening, the positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry, and the amino acid sequences of the positive clones were deduced from the DNA sequences.
RESULTSSeven positive clones were screened out of the 17 phage clones bound to MT-2 cells. The positive clone M15 could bind specifically to MT-2 cell and PHA-activated peripheral blood monouclear cells. Amino acid sequence analysis identified 6 sequences, all of which contained hydrophilic residues, and 5 of these 6 sequences included Tyr, Phe and Leu conservative residues.
CONCLUSIONThe peptide sequences containing Tyr, Phe conservative residues identified in this study can bind to cell surface IL-2Ralpha.
Amino Acid Sequence ; Cell Line, Transformed ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunohistochemistry ; Interleukin-2 ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Peptide Library ; Peptides ; genetics ; metabolism ; Protein Binding ; Receptors, Cell Surface ; metabolism ; T-Lymphocytes ; cytology ; metabolism
5.The soluble interleukin 2 receptor levels in Kawasaki disease.
Journal of the Korean Pediatric Society 1992;35(12):1657-1666
No abstract available.
Interleukin-2*
;
Interleukins*
;
Mucocutaneous Lymph Node Syndrome*
;
Receptors, Interleukin-2*
6.Significance of soluble CD163 and soluble CD25 in diagnosis and treatment of children with hemophagocytic lymphohistiocytosis.
Yingchao WANG ; Email: YINGCHAOWANG152@163.COM. ; Dongjie LIU ; Guiying ZHU ; Chuyun YIN ; Guangyao SHENG ; Xiaoming ZHAO
Chinese Journal of Pediatrics 2015;53(11):824-829
OBJECTIVETo explore significance of serum soluble CD163(sCD163) and soluble CD25(sCD25) in diagnosis and guiding treatment of children with hemophagocytic lymphohistiocytosis (HLH).
METHODData of 42 cases of children with HLH, 32 cases of non-HLH children with infection presented to First Affiliated Hospital of Zhengzhou University pediatric clinic and ward were collected from December 2013 to December 2014. Twenty-four healthy children were enrolled into a normal control group in the same period.Peripheral venous blood specimens (3 ml) were taken from the children with HLH after fasting before treatment, two weeks after treatment and eight weeks after treatment.Peripheral venous blood specimens (3 ml) were also taken from children of non-HLH infected group and normal control group after fasting at the initial visit. Serum sCD163 and sCD25 levels in the peripheral blood in three groups were determined by ELISA. According to cause of disease, children with HLH were divided into infection-related HLH, tumor-related HLH, primary HLH and others; relationship between serum sCD163 and sCD25 level and cause of disease was analyzed.
RESULTSerum sCD163 of HLH group ((6 094 ± 2 769) µg/L) and serum sCD163 of non-HLH infection group ((2 174 ± 950) µg/L) were significantly higher than that of normal control group ((777 ± 256) µg/L), F=71.396, P<0.05), and the differences among groups were statistically significant (P<0.05); serum sCD25 of HLH group ((41 963 ± 31 821) ng/L) and serum sCD25 of non-HLH infection group ((6 700 ± 4 105) ng/L) were significantly higher than that of normal control group ((2 440 ± 1 870) ng/L, F=37.513, P<0.05).There was no statistically significant difference between the non-HLH infection group with the normal control group (P>0.05), and the difference between the remaining groups was statistically significant (P<0.05). And serum sCD163 and sCD25 level of HLH group had a positive linear correlation, and Pearson correlation coefficient r=0.742 (t=7.000, P<0.05). The difference of serum sCD163 and sCD25 level among the different cause of disease in HLH group was significant (P<0.05).Pairwise comparison showed that serum sCD163 and sCD25 level of tumor-associated HLH group significantly increased as compared with infection-associated HLH group (P<0.05), but the difference was not statistically significant between the other groups (all P>0.05). Serum sCD163 and sCD25 level of HLH group before treatment, 2 weeks and 8 weeks after treatment showed a statistically significant tendency of decrease (P<0.05). Seen from the ROC curve, when sCD163 cut-off point was 2 359.08 µg/L, the diagnostic sensitivity was 83.3%, and specificity was 83.9%.When sCD25 cut-off point was 14 901.024 ng/L, the diagnosis sensitivity was 76.2%, and specificity was 98.2%.
CONCLUSIONSerum sCD163 and sCD25 levels may be used for diagnosis of HLH.Dynamically monitoring of serum sCD163 and sCD25 level can help to determine deterioration of HLH and guide treatment.
Antigens, CD ; blood ; Antigens, Differentiation, Myelomonocytic ; blood ; Case-Control Studies ; Child ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interleukin-2 Receptor alpha Subunit ; blood ; Lymphohistiocytosis, Hemophagocytic ; blood ; diagnosis ; therapy ; ROC Curve ; Receptors, Cell Surface ; blood ; Sensitivity and Specificity
7.Levels of Soluble Interleukin-2 Receptors in Serum of Patients with Behçet's Disease.
Won HUR ; Seung Hun LEE ; Dongsik BANG ; Sungnack LEE ; Jung Chul KIM ; Tai Ho CHUNG
Annals of Dermatology 1993;5(1):13-16
BACKGROUND: Interleukin-2 receptor (IL-2) is expressed and released predominantly activated T lymphocyte. Increased serum levels of soluble IL-2R have been noted in a variety of autoimmune diseases and in conditions associated with T lymphocyte activation. OBJECTIVE: We aimed to examine whether the T lymphocyte activation has any association with the pathogenesis of Behçet's disease. METHOD: We have measured the serum level of soluble IL-2R in serum samples obtained from 67 patients with Behçet's disease and 30 healthy people as a control group, using a double-antibody sandwich enzyme-linked immunosorbent assay technique. RESULTS: Serum soluble IL-2R levels were found to be significantly elevated in the group of Behçet's disease as compared with the control group. No significant differences were found within clinical subtypes of Behçet's disease. CONCLUSION: These findings suggest the presence of an ongoing T lymphocyte activation in this disease process.
Autoimmune Diseases
;
Enzyme-Linked Immunosorbent Assay
;
Humans
;
Interleukin-2*
;
Lymphocyte Activation
;
Lymphocytes
;
Methods
;
Receptors, Interleukin-2*
8.Levels of Soluble Interleukin-2 Receptors in Serum of Patients with Behçet's Disease.
Won HUR ; Seung Hun LEE ; Dongsik BANG ; Sungnack LEE ; Jung Chul KIM ; Tai Ho CHUNG
Annals of Dermatology 1993;5(1):13-16
BACKGROUND: Interleukin-2 receptor (IL-2) is expressed and released predominantly activated T lymphocyte. Increased serum levels of soluble IL-2R have been noted in a variety of autoimmune diseases and in conditions associated with T lymphocyte activation. OBJECTIVE: We aimed to examine whether the T lymphocyte activation has any association with the pathogenesis of Behçet's disease. METHOD: We have measured the serum level of soluble IL-2R in serum samples obtained from 67 patients with Behçet's disease and 30 healthy people as a control group, using a double-antibody sandwich enzyme-linked immunosorbent assay technique. RESULTS: Serum soluble IL-2R levels were found to be significantly elevated in the group of Behçet's disease as compared with the control group. No significant differences were found within clinical subtypes of Behçet's disease. CONCLUSION: These findings suggest the presence of an ongoing T lymphocyte activation in this disease process.
Autoimmune Diseases
;
Enzyme-Linked Immunosorbent Assay
;
Humans
;
Interleukin-2*
;
Lymphocyte Activation
;
Lymphocytes
;
Methods
;
Receptors, Interleukin-2*
9.The Enhancing IL-2R alpha mRNA Expression induces A Marked T Cel Proliferation with Interleukin-2 and Anti-CD3 mAb.
Hwa Jung KIM ; Eun Kyeong JO ; Jeong Kyu PARK ; Jong Kun KIM
Korean Journal of Immunology 1998;20(4):427-434
Culture of human peripheral T lymphocytes with irnmobilized anti-CD3 rnAb plus IL-2 resulted in a marked proliferation and the enhancing IL-2Ra mRNA expression. The process of the T cell activation involves a series of biochemical events which ultimately lead to the proliferation and IL-2Ra mRNA expression. Although the above results have been observed, the celluar signal mechanisms between the proliferative response and the IL-2Ra mRNA expression through T cell receptor and IL-2 receptor remains unresolved. In the present study, We have used genistein (the selective PTK inhibitor) or chronic PMA treatment (depletion of intracelluar PKC activity), to investigate the role of PTK or PKC both in a synergistic proliferation and in the enhancing IL-2Ra mRNA expression by IL-2/anti-CD3. Genistein (30 ug/ml) completely blocked IL-2 induced T cell proliferation, and inhibited anti-CD3 induced T cell proliferation (93.4%). But genistein downregulated the IL-2Ra mRNA expression by IL-2, anti-CD3 and IL-2/anti-CD3. The chronic PMA treatment failed to inhibit the proliferation and the IL-2R#u mRNA expression by IL-2 alone. But PKC depleted T cells stimulated with anti-CD3 mAb showed the decrease of the proliferation (68.6%) and IL-2Ra mRNA expression. In activated with IL-2/anti- CD3, the proliferative response showed a half of reduction, but the IL-2Ra mRNA expression were not regulated. These results demonstrate that proliferative response to IL-2 appears to be dependent on PTKs activity and independent of PKC involvement, but the IL-2Ra mRNA expression may be required another signals. PTKs and PKC activity may be important in TCR/CD3 signaling. But IL-2/anti-CD3 are coupled up different signal transduction pathways responsible for the synergistic T cell proliferation and the enhancing IL-2Ru mRNA expression.
Cell Proliferation
;
Genistein
;
Humans
;
Interleukin-2*
;
Receptors, Antigen, T-Cell
;
Receptors, Interleukin-2
;
RNA, Messenger*
;
Signal Transduction
;
T-Lymphocytes
10.IL-12-STAT4-IFN-gamma axis is a key downstream pathway in the development of IL-13-mediated asthma phenotypes in a Th2 type asthma model.
You Sun KIM ; Seng Jin CHOI ; Jun Pyo CHOI ; Seong Gyu JEON ; Sun Young OH ; Byung Jae LEE ; Yong Song GHO ; Chun Geun LEE ; Zhou ZHU ; Jack A ELIAS ; Yoon Keun KIM
Experimental & Molecular Medicine 2010;42(8):533-546
IL-4 and IL-13 are closely related cytokines that are produced by Th2 cells. However, IL-4 and IL-13 have different effects on the development of asthma phenotypes. Here, we evaluated downstream molecular mechanisms involved in the development of Th2 type asthma phenotypes. A murine model of Th2 asthma was used that involved intraperitoneal sensitization with an allergen (ovalbumin) plus alum and then challenge with ovalbumin alone. Asthma phenotypes, including airway-hyperresponsiveness (AHR), lung inflammation, and immunologic parameters were evaluated after allergen challenge in mice deficient in candidate genes. The present study showed that methacholine AHR and lung inflammation developed in allergen-challenged IL-4-deficient mice but not in allergen-challenged IL-13-deficient mice. In addition, the production of OVA-specific IgG2a and IFN-gamma-inducible protein (IP)-10 was also impaired in the absence of IL-13, but not of IL-4. Lung-targeted IFN-gamma over-expression in the airways enhanced methacholine AHR and non-eosinophilic inflammation; in addition, these asthma phenotypes were impaired in allergen-challenged IFN-gamma-deficient mice. Moreover, AHR, non-eosinophilic inflammation, and IFN-gamma expression were impaired in allergen-challenged IL-12Rbeta2- and STAT4-deficient mice; however, AHR and non-eosinophilic inflammation were not impaired in allergen-challenged IL-4Ralpha-deficient mice, and these phenomena were accompanied by the enhanced expression of IL-12 and IFN-gamma. The present data suggest that IL-13-mediated asthma phenotypes, such as AHR and non-eosinophilic inflammation, in the Th2 type asthma are dependent on the IL-12-STAT4-IFN-gamma axis, and that these asthma phenotypes are independent of IL-4Ralpha-mediated signaling.
Allergens/immunology
;
Animals
;
Asthma/complications/*immunology/pathology/physiopathology
;
Bronchial Hyperreactivity/complications/immunology/pathology
;
Disease Models, Animal
;
Interferon-gamma/*immunology
;
Interleukin-12/*immunology
;
Interleukin-12 Receptor beta 2 Subunit/metabolism
;
Interleukin-13/deficiency/*immunology
;
Interleukin-4/deficiency
;
Methacholine Chloride
;
Mice
;
Mice, Transgenic
;
Models, Immunological
;
Organ Specificity
;
Pneumonia/complications/immunology/pathology
;
Receptors, Cell Surface/metabolism
;
STAT4 Transcription Factor/*metabolism
;
Signal Transduction/*immunology
;
Th2 Cells/*immunology