Effect of Bletillae Rhizoma Polysaccharide on PI3K/Akt in Gastric Tissue of GU Model Rats / 中国实验方剂学杂志

Objective::To investigate the effect of Bletillae Rhizoma polysaccharide on the expressions of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) gene protein and its mediated cytokines interleukin-2 receptor (IL-2R) and interleukin-4 (IL-4) in gastric tissue of rats with gastric ulcer (GU). Method::Sixty SPF Wistar rats were randomly divided into blank group and model group.The GU model was replicated by direct acetic acid cauterization in model group.The GU model rats were randomly divided into five groups: model group, positive control group, and large, medium and small-dose Bletillae Rhizoma polysaccharide groups, with 10 rats in each group.Rats in blank group and GU model group were given 10 mL·kg-1·d-1 distilled water by gavage, rats in large, medium and small-dose groups were given 0.5, 0.25, 0.125 g·kg-1·d-1 Bletillae Rhizoma polysaccharide by gavage, while rats in positive control group were given 0.3 g·kg-1·d-1 ranitidine by gavage for 15 days.Serum nitric oxide (NO) content, pepsinase activity and cytokines IL-2R and IL-4 levels in rats of each group were measured by enzyme-linked immunosorbent assay (ELISA), PI3K and Akt mRNA expressions were detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR), and PI3K and Akt protein expressions were detected by Western blot. Result::Compared with the blank group, the contents and gene expressions of cytokines IL-2R and IL-4 in gastric tissue were significantly increased, and the PI3K and Akt genes and protein expressions were significantly increased, with statistical significance (P<0.01). Compared with GU model group, the content and gene expressions of IL-2R and IL-4 in large, medium and small-dose Bletillae Rhizoma polysaccharide groups were decreased significantly, and the PI3K and Akt gene and protein expressions were decreased significantly in large-dose Bletillae Rhizoma polysaccharide group, while those in large and medium-dose Bletillae Rhizoma polysaccharide groups were decreased significantly (P<0.05, P<0.01). Conclusion::Bletillae Rhizoma polysaccharide can protect gastric mucosa by down-regulating PI3K and Akt gene and protein expressions and inhibiting abnormal secretion of cytokines IL-2R and IL-4.

Changes of IL-2, TNF, TGF and IL-2R mRNA of Menopausal Rats and Effects of Preventive Acupuncture on Guanyuan Point / 中国中医药信息杂志

Objective To observe the effect on IL-2,TNF,TGF in uterus and serum,and IL-2RmRNA in spleen of menopausal rats with preventive accupuncture on Guanyuan point,and analysis the mechanism.Methods 112 ten month-aged SD female rats examined by vaginal smear were chosen and divided into seven groups.And another 16 three point five month-aged SD female rats were another group.Radio-immunity and hybridiztion in situ methods were applied to observe the effect on IL-2,TNF,TGF in uterus and serum and IL-2R mRNA in spleen of 12,14 and 16 month-aged menopausal rats with preventive accupuncture on Guanyuan point(twice a week,eight weeks continued) in 10th month.Results Compared with the normal group,IL-2,TGF in uterus and serum and TNF in uterus of menopausal rats reduced,TNF in serum came down in the 12th month and went up in 14th and 16th month,IL-2R mRNA in spleen decreased.Compared with the same-aged model group,IL-2,TGF of serum and uterus and TNF in uterus increased.TGF in serum of 14 and 16 month-aged group,IL-2 in uterus of 12 and 16 month-aged group,TNF in uterus of 12 month-aged group and TGF in uterus of 12 and 16 month-aged group rised significantly(P

IL-2 Pathway Blocking in Combination with Anti-CD154 Synergistically Establishes Mixed Macrochimerism with Limited Dose of Bone Marrow Cells and Prolongs Skin Graft Survival in Mice

To facilitate the establishment of mixed chimerism with limited dose of bone marrow (BM) cells, and to achieve tolerance in skin graft model, combined blocking of costimulatory pathway and IL-2 pathway was used in minimally myeloablative model using busulfan. BM cells (2.5 x 10(7)) of BALB/c were injected into C57BL/6 mice at day 0 with full thickness skin graft after single dose injection of busulfan (25 mg/kg) on day-1. Recipients were grouped and injected the anti-CD154, CTLA4-Ig, anti-IL-2R at days 0, 2, 4, and 6 according to protocol. Mixed macrochimerism were induced in groups treated with anti-CD154+anti-CTLA4-Ig, anti-CD154+anti-IL-2R, and anti-CD154+anti-CTLA4 Ig+anti-IL-2R. Three groups having chimerism enjoyed prolonged graft survival more than 6 months. Superantigen deletion study revealed deletion of alloreactive T cells in combined blockade treated groups. In graft versus host disease model using CFSE staining, CD4+ T cell and CD8+ T cell proliferation were reduced in groups treated with CTLA4-Ig or anti-IL-2R or both in combination with anti-CD154. However, anti-IL-2R was not so strong as CTLA4-Ig in terms of inhibition of T cell proliferation. In conclusion, IL-2 pathway blocking combined with anti-CD154 can establish macrochimerism with limited dose of BM transplantation and induce specific tolerance to allograft.

IL-2 Pathway Blocking in Combination with Anti-CD154 Synergistically Establishes Mixed Macrochimerism with Limited Dose of Bone Marrow Cells and Prolongs Skin Graft Survival in Mice

To facilitate the establishment of mixed chimerism with limited dose of bone marrow (BM) cells, and to achieve tolerance in skin graft model, combined blocking of costimulatory pathway and IL-2 pathway was used in minimally myeloablative model using busulfan. BM cells (2.5 x 10(7)) of BALB/c were injected into C57BL/6 mice at day 0 with full thickness skin graft after single dose injection of busulfan (25 mg/kg) on day-1. Recipients were grouped and injected the anti-CD154, CTLA4-Ig, anti-IL-2R at days 0, 2, 4, and 6 according to protocol. Mixed macrochimerism were induced in groups treated with anti-CD154+anti-CTLA4-Ig, anti-CD154+anti-IL-2R, and anti-CD154+anti-CTLA4 Ig+anti-IL-2R. Three groups having chimerism enjoyed prolonged graft survival more than 6 months. Superantigen deletion study revealed deletion of alloreactive T cells in combined blockade treated groups. In graft versus host disease model using CFSE staining, CD4+ T cell and CD8+ T cell proliferation were reduced in groups treated with CTLA4-Ig or anti-IL-2R or both in combination with anti-CD154. However, anti-IL-2R was not so strong as CTLA4-Ig in terms of inhibition of T cell proliferation. In conclusion, IL-2 pathway blocking combined with anti-CD154 can establish macrochimerism with limited dose of BM transplantation and induce specific tolerance to allograft.

The detection of interleukin-2 receptor ?-chain gene mutation and protein expression of the tumor infiltrated Tcell in B-NHL / 中国免疫学杂志

Objective:To detect the gene mutation,protein expression of IL-2R? of TIL-T and the pr oliferation of TIL-T with the present of rIL-2 in B-NHL. Methods:The gene mutation of IL-2R? was performed on 19 TIL-T by PCR-SSCP;proliferat ion assay of 17 TIL-T with the rIL-2 was tests by MTT;IL-2R? protein express ion in cryostat section of 29 B-NHL were determined by immunohistochemical stai n. Results:SSCP showed there is no mutation happened in the cDNA of IL-2R? of TIL-T.Pro liferation test showed the intensity of response of TIL-T was decreased in 76. 5% TIL-T(13 of 17 cases).The expression of CD25 protein in 86.2%(25 of 29 c ases) of B-NHL cases were (+) or (+/-). Conclusion:No genetic mutation had been found in IL-2R? of TIL-T,but IL-2R? protein i s weakly expressed in B-NHL;It indicated that there may be abnormal in the mech anism of activation of TIL-T by cell-cell contact. [

The Levels of Interferon-gamma, Interleukin-2 Receptor, Interleukin-6 and Interleukin-10 in the patients with Malignant Pleural Effusion, Tuberculosis Effusion, Parapneumonic Effusion, and Lung Emphysema? / 결핵

BACKGROUND: Cytokines are chemical mediators that control and modulate many infalmmatory processes. They work in different fashions in a variety of diseases. Discriminating between malignant effusion, tuberculous effusion, and parapneumonic effusion are crucial from the clinical view-point in Korea. In the current study, interferon-gamma(INF-γ), soluble interleukin-2 receptor(IL-2R), interleukin-6(IL-6) and interleukin-10(IL-10) were measured for this purpose. METHODS: Pleural fluids from patients with malignant disease, tuberculosis, parapneumonic effusion and lung empysema were collected and gauged using commercial ELISA kits. RESULTS: 34 patients were enrolled in this study. among these 15 cases were malignant effusions, 12 were tuberculosis pleurisy and 7 were paraneumonic effusion and lung empyema. The levels of cytokines measured in this study were as follows, in order of frequency, malignant effusion, tuberculous effusion, parapneumonic effusion and lung empyema. The levels of INF-γ were higher in tuberculous effusion than in malignant or parapneumonic effusion(295.5±585.5 vs. 16.7±50 vs. 10.0±0 pg/ml, p>0.05). The levels of IL-2R were higher in tuberculous effusion than in malignant or parapneumonic effusion(7423.5±3752.8 vs. 3247.4±1713.3 vs. 3790.2±3201.1 pg/ml, p<0.05). No significant differences were found in the levels of IL-6 between the groups(600±12.8 pg/ml in malignant effusion, 556.4±161.7 pg/ml in tuberculous effusion, 514.4±224.8 pg/ml in parapneumonic effusion). IL-10 levels were higher in parapneumonic effusion than in malignant or tuberculous effusions(98.4±141.7 vs. 28.2±55.5 vs. 11.3±11.7 pg/ml, p<0.05). CONCLUSION: These results suggest that the measurement of IL-2R levels in pleural fluids may be a useful means of differentiating between tuberculous effusion and pleural effusions of other origins, and that the measurement of IL-10 levels in pleural fluids may be useful to differentiate between parapneumonic effusion and pleural effusions of other origins.

Evaluation of Plasma and Urinary Tumor Necrosis Factor alpha Interleukin-2 Receptor and Interleukin-6 Levels in Renal Allograft Recepients / 대한이식학회지

This is the report of 98 cases in renal allograft, which were treated at Yeungnam University Hospital from January 1994 to July 1996 and compared the significance of changes of TNF alpha, IL-2R, IL-6 in blood and urine as an early diagnostic tool of acute rejection in renal allograft. The aim of this study was to investigate the value of plasma and urinary TNF alpha, IL-2R, IL-6 in patients with renal allografts. Renal allografts patients were divided into four groups (control, acute rejection, acute tubular necrosis, systemic infection) according to their postoperative diagnostic methods. Blood and urine samples in four groups were obtained: control group (2 days before transplantation, at the day of transplantation and every other day after transplantation), acute rejection group (everyday sampling from 2 days before therapy to the end of therapy), acute tubular necrosis and systemic infection group (everyday sampling from the day of diagnosis to the end of therapy). In acute rejection group, there were significant elevation of cytokines; plasma TNF alpha (68.4%, p<0.01), IL-2R (73.6%, p<0.01), and IL-6 (89.5%, p<0.01), urinary TNF-alpha (42.1%, p<0.01), IL-2R (89.5%, p<0.01) and IL-6 (94.7%, p<0.01). In systemic infection group, all cytokines except urinary TNF-alpha were significantly elevated. The results suggested that plasma and urinary TNF-alpha, IL-2R, and IL-6 may play a complementary early diagnostic tool of acute rejection in renal allograft patients although the differential diagnosis is difficult with systemic infection. Urinary TNF-alpha was not elevated in systemic infection group, so it may be used in differential diagnosis between acute rejection and systemic infection.

Evaluation of Plasma and Urinary Tumor Necrosis Factor alpha Interleukin-2 Receptor and Interleukin-6 Levels in Renal Allograft Recepients / 대한이식학회지

This is the report of 98 cases in renal allograft, which were treated at Yeungnam University Hospital from January 1994 to July 1996 and compared the significance of changes of TNF alpha, IL-2R, IL-6 in blood and urine as an early diagnostic tool of acute rejection in renal allograft. The aim of this study was to investigate the value of plasma and urinary TNF alpha, IL-2R, IL-6 in patients with renal allografts. Renal allografts patients were divided into four groups (control, acute rejection, acute tubular necrosis, systemic infection) according to their postoperative diagnostic methods. Blood and urine samples in four groups were obtained: control group (2 days before transplantation, at the day of transplantation and every other day after transplantation), acute rejection group (everyday sampling from 2 days before therapy to the end of therapy), acute tubular necrosis and systemic infection group (everyday sampling from the day of diagnosis to the end of therapy). In acute rejection group, there were significant elevation of cytokines; plasma TNF alpha (68.4%, p<0.01), IL-2R (73.6%, p<0.01), and IL-6 (89.5%, p<0.01), urinary TNF-alpha (42.1%, p<0.01), IL-2R (89.5%, p<0.01) and IL-6 (94.7%, p<0.01). In systemic infection group, all cytokines except urinary TNF-alpha were significantly elevated. The results suggested that plasma and urinary TNF-alpha, IL-2R, and IL-6 may play a complementary early diagnostic tool of acute rejection in renal allograft patients although the differential diagnosis is difficult with systemic infection. Urinary TNF-alpha was not elevated in systemic infection group, so it may be used in differential diagnosis between acute rejection and systemic infection.

An Assiociation Study of Interleukin-2 Receptor beta-Chain Gene Polymorphism on Chromosome 22 in Korean Schizophrenic Patients / 신경정신의학

BACKGROUND: While a significant genetic predisposition to schizophrenia has been proposed, the mode of inheritance or nature of etiological factors is unknown. Previous reports of a genome-wide survey for schizophrenia susceptibility genes have indicated a possible region of linkage on chromosome 22. In order to test the possibility that the interleukin-2 recepto beta chain(IL-2R beta ) gene on chromosome 22 is of etiological importance in schizophrenia, a case-control association study was conducted. METHODS: Subjects were ninety-three schizophrenic patients with a diagnosis of schizophrenia by DSM- III -R criteria and ninety-seven normal controls. Schizophrenic patients were divided by clinical phenotypes such as DSM- III -R diagnostic subtypes, positive and negative symptoms, and family history so as to increase the homogeneity of schizophrenics. Genomic DNA was extracted from whole blood lymphocytes according to standard procedures. The DNA was used to study a dinucleotide repeat in the IL-2R beta gene. To reveal the dinucleotide polymorphism, genomic DNA of subjects was amplified by polymerase chain reactions(PCR). RESULTS: At the IL-2R beta gene locus, all the previously reported alleles (eight different alleles) of a dinucleotide polymorphism were identified. There was no significant difference between number of heterozygosity in schizophrenic patients and in normal controls. There was no significant difference in the distribution of frequencies of alleles between schizophrenics and normal controls. In addition, there was no significant difrfrence in the allele frequencies among subtypes of schizophrenic patients according to DSM- III -R diagnostic subtypes, positive and negative symptoms, and family history. CONCLUSIONS: The present study did not detect a difference in frequencies of alleles of a dinucleotide polymorphism at the IL-2R beta gene locus between schizophrenic patients and normal controls. These results do not support an evidence that IL-2R beta gene plays, a major role in the etiology of schizophrenia.

The Establishment of IL - 2 Dependent Cell Line from IL - 3 Dependent Mouse Hemopoietic Progenitor Cell Line and Its Analysis / 대한면역학회지

No abstract available.

Signiflcance of IL-2, IL-2R, IL-6 and TNF-alpha as a Diagnostic Test of Acute Rejection after Renal Transplantation

In order to avoid unnecessarily heavy immunosuppression and the complications due to misdiagnosing acute rejection, we checked the serial serum cytokine levels in 34 living renal transplant recipients (excluding pediatric recipients, cadaver donor grafts and one acute vascular rejection which was removed on the 7th post-transplant day) operated on between January 1994 and June 1995, and we analyzed the relation between the cytokine levels in these cases and acute rejection. Among the 34 recipients, 11 developed acute rejection within one month after transplantation, 8 of them being confirmed by renal biopsy; however, all of these recipients showed excellent initial graft function. The elevation of the cytokine levels, except IL-6, started from 1-3 days before either clinical suspicion of rejection or increase of serum creatinine levels. Postoperative levels of IL-2, IL-2R, and TNF- were significantly higher in the rejection group than in the non-rejection group, and these findings continued at least until the 15th post-transplant day. The group whose rejection episodes developed within 2 weeks after transplantation showed higher cytokine levels, except IL-6, than those of the group whose rejection episodes occurred after 2 weeks (P<0.05). These elevated cytokine levels returned to pre-rejection levels in the group who responded to anti-rejection therapy, but maintained higher levels in the no-response group (P<0.05). In summary, these results suggest that during the early post-transplant period, the serial checking of serum cytokine levels can both predict the onset of acute rejection earlier than the conventional method and tell us the status of the response to anti-rejection therapy. However, the absolute significance of the cytokine levels and the cost of daily laboratory test for cytokine should also be considered.

Detection of Interleukin 2 Receptor and EBV Antibody in Nasopharyngeal Carcinoma / 대한이비인후과학회지

Nasopharyngeal carcinoma(NPC) is highly prevalent among southeastern Chinese,and not rare in Korea. Due to NPC usually spreads submucosally and complexity of anatomy in the nasopharynx, we have difficulty in determining the exact extent of tumor. If we know the parameters that represent tumor burden and prognosis, it is very helpful to decide planning of treatment. NPC is etiologically related to Epstein Barr Virus(EBV), and many studies have been reported dealing with EBV and its relationships to the clinical course of NPC. Interleukin 2 receptor(IL-2R) are released from activated human T lymphocyte and marked elevated levels of serum IL-2R have been reported in patients with lymphoproliferative disorders such as leukemia,lymphoma. Histologicallly NPC is associated with T lymphocyte infiltration and it was also reported that the serum levels of IL-2R represents a good marker of tumor burden in NPC. The purpose of this papers was to detect the levels of IL-2R , viral capsid antigen(VCA) IgA and early antigen(EA) IgG in the serum of 19 NPC patients and analyzed the relationships those values and clinical courses, for its usefulness in estimating the tumor burden and predicting the response to therapy and prognosis, who diagnosed as NPC in Kang Nam St. Mary Hospital from June 1993 to November 1995. The results were as follows. 1) Serum IL-2R levels of NPC was not higher than that of controls and showed no relationships with the stage of the NPC. 2) Serum IL-2R levels of recurrent group were higher than that of remission groups(p=0.019). 3) Titers of VCA(IgA), EA(IgG) were higher than that of controls(p=0.0009) and showed no relatiohships with the stage of NPC. 4) There is no differences in the titer of antibodies to EBV according to histological types of NPC. 5) The titers of EA(IgA) had relationship to clinical results(p=0.0286). Above results suggested that higher levels of IL-2R and titers of EBV antibody after therapy, detailed examination should be done for the detection of possible recurrent and remained disease with close follow up.

A Study of EFFECT and MECHANISM of IL-2on SURVIVAL of EOSINOPHILS / 결핵

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.

Immunomodulatory effects of theasinesin on human peripheral blood mononuclear cells in vitro / 中草药

Object To study the immunomodulatory effects of theasinesin (TS) on human peripheral blood mononuclear cells (PBMC) in vitro. Methods The proliferation response of lymphocytes and IL 2 production of human PBMC were detected by MTT method. IL 2R expression was determined by immunocytochemistry and indirect immunofluorescence. The intracellular cAMP level was assayed by radioimmunoassay. Results 50, 150, 500 ?g/mL TS could enhance phytagglutinin (PHA) induced human PBMC proliferation in a dose dependent manner. The IL 2 production and IL 2R expression of PHA activated PBMC were increased markedly under the presence of TS. TS also decreased cAMP level of PBMC and intensively weaken the cAMP upregulating effect of prostaglandin E 1 (PGE 1). Conclusion TS possesses a potential immunoenhancing effect which might be made by decreasing the cAMP level of PBMC, promoting IL 2 production and IL 2R expression.

Enhancement of proliferaton and tumoricidal activities of T-AK cells by plant polysaccharides / 中草药

Object To study the augmentation of the T activated killer (T AK) cell proliferating and tumor killing activities of the polysaccharide fraction from Rhodiola kirilowii (Regel) Regel, Lycium barbarum L., Hedysarum polybotrys Hand. Mazz., Glehnia littoralis F. Schmidt ex Miq., Rehmannia glutinosa (Gaert.) Libosch. ex Fisch. et Mey. and Aloe barbadensis Mill. in vitro. Methods The T AK cells were induced by anti CD 3 antibody (CD 3McAb) and rIL 2 from human peripheral blood mononuclear cells (PBMC). The effects of the above six plant polysaccharides (1～100 ?g/mL) on the proliferation of T AK cell, the cytotoxicity to Raji cells and L 1210 cells, and the IL 2 receptor (IL 2R) expression in T AK cells were determined. Results The six polysaccharides alone had no obvious effect on the proliferation of T AK cells. In the presence of rIL 2 and CD 3McAb, they could reinforce the proliferation of T AK cells and its tumor killing activities against Raji cells and L 1210 cells at a different extent, and a 21%～68% increase of IL 2R expression in T AK cells was observed. Conclusion The plant polysaccharides significantly enhance the proliferation and the tumoricidal activities of T AK cells and the enhancing actions related to the increase of IL 2R expression.

A New Approach to Enhance the Induction and Activation of PHA-LAK Cells / 中国肿瘤生物治疗杂志

A new type of killer cells, named PHA-?CD3LAK, was induced by means of costimulating the peripheral blood mononuclear cells (PBMC) with anti-CD3McAb (?CD3) and rIL-2 after PHA-priming for 48 hours. Some biological characteristics of PHA-?CD3LAK, PHA-LAK and CD3AK were compared. The results showed that PHA-?CD3LAK exhibited some advantages over CD3AK and PHA-LAK in proliferation, cytotoxicity, the expression level of mIL-2R, as well as the utilizing of IL-2, suggesting the synergistic enhancing role of PHA, ?CD3 and IL-2. All three groups of effector cells were heterogeneous populations, predominantly CD3 + CD8 + T cells. The CD8 ~(+) cell percentage of PHA-?CD3LAK was higher than that of the other two groups. The application of PHA-?CD3LAK might open a new prospect to clinical therapeutic approach.

Study on the construction and expression of anti-IL2R? genetically engineering an-tibody Fab / 中国免疫学杂志

Abstract Objective: To construct and express anti-IL-2R? genetically engineering antibody Fab and identify its biological activity.Methods: PCR were used to construct cloning vector 5G1-pA22 of anti-IL-2R? antibody Fab,by digestion of vector 5G1-pA22, anti-IL-2R? an-tibody Fab gene fragment was obtained and then this fragment was coupled with digesting product of pET28b plasmid by EcoRI and HindIII toget a recombinant expression vector 5G1-pET28b, it was highly expressed in the E. coli BL21(DE3) in the form of inclusion bodies, compoing20% of the total protein. After the renaturation of antibody Fab, its binding activity was identified by ELISA and antibody competitive inhibitionassay in vitro. Results: Anti-IL-2R? antibody Fab was successfully constructed and highly expressed in the form of inclusion bodies, its yield wasup to 1 mg/ml.When applied to ELISA, antibody Fab exhibited good spectfic reactivity with rIL-2R, antibody competitive inhibition assay invitro indicated that antibody Fab can compete with IL-2 for binding IL-2R in activated T cell surface.As a result,the activation and proliferationof T cells were inhibited and inhibition percentage exceeded 90%. Conclusion: Had successfully constructed and expressed anti-IL-2R? geneti-cally engineering antibody. Fab,it had good biological ativity.

The preparation of human IL-2R cDNA probe and its application in the detection of mRNA levels / 中国免疫学杂志

We obtained human IL-2R cDNA 673 bp fragments from plasmid pJSB15 and prepared ?-32 p-IL-2 R cDNA probe. Dot hybridizations of RNA extracted from human lymphocytes were done by using the probe. The results showed that PHA or TPA could induce IL-2 R mRNA, respectively; and the synergistic effects were seen when used them together. IL-2R mRNA could be expressed spontaneously in many kinds of leukemic lymphocytes isolated freshly from PBL of leukemic patients.

Expression of human IL-2 receptor chain cDNA in vitro transcripts / 中国免疫学杂志

Recombinat plasmid pJSB13 has an insert of 121-937 bp of human IL-2R_? cDNA in multiple cloning site of pGEM3 vector.This plasmid was digested with EcoR I to linearize it and then in vitro transcripted with SP6 RNA polymerase and capped with cap structure analog 7mG (5') G.The capped transcripts were translated in rabbit reticutocyte lysate or Xenopus Iaevis oocytes respectively. The results demonstrated that they can be efficiently translated as functional mRNA in both systems. Moreover, the translation product, IL-2R_?,can be secreted into oocvte culture medium.The program we had used enables one to produce and identify mRNA and conse quently protein from any cDNA clone. Furthermore this research can be of much helpful to the researches on the relationship between structure, function and expression of IL-2R_?.

Transforming growth factor type beta(TGF—?1 )can increase IL—2R expression and suppress the proliferation of immature(CD~(4-)CD~(8-))thymocytes / 中国免疫学杂志

Transforming growth factor type ?1(TGF—?1)belong to a family of polypeptides with regulato-ry effects on growth and differention of a variety of cells types.TGF—?1 play an important rolein regulation of immune response by acting as a negative modulate the cell proliferation throughstill unknow mechanisms.We investigated the effect of TGF—?1 on the IL—2R expression andproliferation activited DN cells and we observed that TGF—?1 was able to increase the expres-sion of IL—2R in DN cells and to suppress the proliferation of DN cells and we have discussedabout these results.