• Title/Summary/Keyword: interleukin 2 (IL-2)

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Expression of the Functional Recombinant Interleukin-16 in E. coli and Mammalian Cell Lines

  • Kim, Seon-Young;Lee, Chang-Hun;Kim, Kyung-Joo;Kim, Yeon-Soo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.234-241
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    • 2001
  • The C-terminal 393 bp region of the human interleukin-16 (IL-16) gene was cloned and expressed in E. coli along with mammalian cell lines. Recombinant IL-16 expressed from E. coli was 22 kDa on SDS-PAGE and showed 260% of chemoattractant activity at a concentration of $0.1\;{\mu}g/ml$. HeLa, COS, and Neuro-2a cells were transduced by recombinant retrovirus vector pLNC/IL-16/IRES/TK and the intracellular and secreted amounts of IL-16 produced by HeLa/IL-16/TK, COS/IL-16/TK, and Neuro-2a/IL-16/TK cells were determined by enzyme-linked immunosorbent assay (ELISA). HeLa/IL-16/TK $(1{\times}10^5)$ and COS/IL-16/TK $(1{\times}10^5)$ cells secreted 36.1 and 13.3 ng of IL-16 for 48 h, respectively. Forty-nine ng and 86.4 ng of IL-16 remained in the cell lysates of HeLa/IL-16/TK and COS/IL-16/TK. Intracellular and secreted amounts of IL-16 from Neuro-2a/IL-16/TK $(5{\times}10^5)$ cells during 24 h cultivation were 50 ng and 3.3 ng, respectively. Also, HeLa and COS cells wee stably transfected with mammalian expression vector pCRIII/IL-16. Both culture media and cell lysates prepared from HeLa/IL-16 cells and COS/IL-16 cells showed chemoattractant activity ranging from 190% to 460% as compared to the control experiment. Expression of the herpes simplex virus thymidine kinase (HSV0tk) gene in pLNC/IL-16/ IRES/TK bicistronic retroviral expression vector was verified by performing a genciclovir (GCV) sensitivity assay. Finally, IL-16 repressed Tat-transactivated human immunodeficiency virus type 1 long terminal repeat (HIV-1 LTR) promoter activity.

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The in Vivo Distribution of $^{99m}Tc-Phytate$ IL-2 Complex on Selective Splenic Arterial Injection (비장동맥에 선택적으로 투여한 Interleukin-2와 $^{99m}Tc-Phytate$ 혼합물의 생체내 분포)

  • Zeon, Seok-Kil;Lee, Hee-Jung;Sohn, Soo-Sang
    • The Korean Journal of Nuclear Medicine
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    • v.26 no.1
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    • pp.124-126
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    • 1992
  • Interleukin-2 (IL-2)는 많은 immunoenhancing lymphokine의 한 종류로서 lymphokine activiated killer (LAK) cell의 생성을 자극시켜 흑종의 종양세포를 죽인다고 알려져 있다. 최근 간종양에서 비장동맥 또는 간동맥으로 투여한 IL-2가 비장의 임파계를 자극하여 LAK cell을 생성하여 어느정도효과가 있음이 밝혀지면서, 여러가지의 투여 방법이 시도되고 있다. 그러나 각종의 투여 방법에서 실제로 투여한 IL-2의 인체내 분포에 관한 연구는 없다. 저자들은 비정맥과 간문맥에 이상이 없는 증례의 비동맥에 IL-2와 $^{99m}Tc-phytate$ 혼합물을 투여하고, IL-2의 생체에서의 비장과 간에 어떻게 분포하는지 알아보기 위하여 $^{99m}Tc$의 radioactivity를 계측하여 보았다. 6예의 간세포암과 3예의 위암으로부터의 전이성간암에서 동맥조영술적방법을 이용하여 초선택적 비장동맥에 투여한 IL-2와 $^{99m}Tc-phytate$ 혼합물이 비장 27%, 간73%의 분포를 보여 비장을 거쳐온 $^{99m}Tc$의 방사능이 간에 많이 침착함을 확인하였고 간과 비장이외의 부위 즉 골수, 복수 또는 폐장이나 늑막에는 전혀 방사능 분포가 없음을 알 수 있었다. 따라서 비정맥이나 간문백에 이상이 없는 증례에서 IL-2의 비장동맥 투여는 목적하는 바 IL-2의 생체내 분포를 이룩할 수 있을 것으로 사료된다.

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EFFECT OF INTERLEUKIM-10 ON THE BONE RESORPTION INDUCED BY INTERLEUKIN-1B (Interleukin-10 이 $interleukin-1{\beta}$로 유도되는 골흡수에 미치는 효과)

  • Yu, Yun-Jung;Kang, Yun-Sun;Lee, Syng-Ill
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.321-339
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    • 1994
  • The cytokines released by osteoblasts induce bone resorption via the differentiation of osteoclast precursors. In this process, $interleukin-1{\beta}$($IL-1{\beta}$)-induced bone resorption is mediated by granulocyte macrophage-colony stimulation factor(GM-CSF), interleukin-6 (IL-6), and tumor necrosis factor ${\alpha}$($TNF-{\alpha}$) released from osteoblasts. Since these cytokines (GM-CSF, IL-6, $TNF-{\alpha}$) are produced by not only osteoblasts but also monocytes, and interleukin-10(I1-10) inhibits the secretion of these cytokines from monocytes, it may be speculated that IL 10 could modulate the production of GM-CSF, IL-6, and $TNF-{\alpha}$ by osteoblasts, then control $IL-1{\beta}-induced$ bone resorption. Therefore, the aims of the present study were to examine the effects of IL-10 on bone resorption. The sixten or seventeen-day pregnant ICR mice were injected with $^{45}Ca$ and sacrificed one day after injection. Then fetal mouse calvaria prelabeled with $^{45}Ca$ were dissected out. In order to confirm the degree of bone resorption, mouse calvaria were treated with Lipopolysaccharide(LPS), $TNF-{\alpha}$, $IL-1{\alpha}$, IL-8, $IL-1{\beta}$, and $IL-1{\alpha}$, Then, IL-10 and $interferon-{\gamma}$ ($IFN-{\gamma}$) were added to calvarial medium, in an attempt to evaluate the effect of $IL-1{\beta}-induced$ bone resorption. In addition, osteoclasts formation in bone marrow cell cultures, and the concentration of IL-6, $TNF-{\alpha}$, and GM-CSF produced from mouse calvarial cells were investigated in response to $IL-1{\beta}$ alone and simultaneously adding f $IL-1{\beta}$ and IL-10. The degree of bone resorption was expressed as the ratio of $^{45}Ca$ release(the treated/the control). The osteoclasts in bone marrow cultures were indentified by tartrate resistant acid phosphatase(TRAP) stain and the concentration of the cytokines was quantified using enzyme linked immunosorbent method. As results of these studies, bone resorption was induced by LPS(1 ng/ml ; the ratio of $^{45}Ca$ release, $1.14{\pm}0.07$). Also $IL-1{\beta}$(1 ng/ml), $IL-1{\alpha}$(1 ng/ml), and $TNF-{\alpha}$(1 ng/ml) resulted in bone resorption(the rations of $^{45}Ca$ release, $1.61{\pm}0.26$, $1.77{\pm}0.03$, $1.20{\pm}0.15$ respectively), but IL-8 did not(the ratio of $^{45}Ca$ release, $0.93{\pm}0.21$). The ratios of $^{45}Ca$ release in response to IL-10(400 ng/ml) and $IFN-{\gamma}$(100 ng/ml) were $1.24{\pm}0.12$ and $1.08{\pm}0.04$ respectively, hence these cytokines inhibited $IL-1{\beta}$(1 ng/ml)-induced bone resorption(the ratio of $^{45}Ca$ release $1.65{\pm}0.24$). While $IL-1{\beta}$(1 ng/ml) increased the number of TRAP positive multinulcleated cells in bone marrow cultures($20{\pm}11$), simultaneously adding $IL-1{\beta}$(1 ng/ml) and IL-10(400 ng/ml) decreased the number of these cells($2{\pm}2$). Nevertheless, IL-10(400 ng/ml) did not affect the IL-6, GM-CSF, and $TNF-{\alpha}$ secretion from $IL-1{\beta}$(1 ng/ml)-activated mouse calvarial cells. From the above results, it may be suggested that IL-10 inhibites $IL-1{\beta}-induced$ osteoclast differntiation and bone resorption. However, the inhibitory effect of IL-10 on the osteoclast formation seems to be mediated not by the reduction of IL-6, GM-CSF, and $TNF-{\alpha}$ production, but by other mechanisms.

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Response to Bee Venom Acupuncture and Polymorphism of Interleukin 4 Receptor and Interleukin 10 in Rheumatoid Arthritis

  • Kim, Yung-Tae;Lee, Jae-Dong;Lee, Yun-Ho
    • The Journal of Korean Medicine
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    • v.25 no.2
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    • pp.207-219
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    • 2004
  • Objectives : We investigated to find the relationship between single-nucleotide polymorphism (SNP) of IL4R, IL-10 and bee venom therapy efficacy in patients with RA treated with bee venom for 8 weeks. Methods : Korean RA patients (n=114) and healthy subjects (n=109) were included in this prospective study. Korean bee venom was dissolved in saline (diluted 1:3000) and administrated into acupuncture points. Bee venom therapy was applied twice a week and continued for 8 weeks. The clinical response was evaluated using various assessments before and after treatment. Disease severity was measured by determining the number of tender joints and swollen joints. Laboratory studies included ESR, CRP, and rheumatoid factor. Genotyping for IL-4R and IL-10 polymorphism was done by pyrosequencing analysis. Results : 1. In IL4R genotypes, there was significant difference between RA ptitients tind controls group. 2. In IL4R genotypes, there was significant difference among Good, Mild and Bad responders to in RA patients, but in the frequency of alleles and carriers, there were no significant difference. 3. There was no significant difference between RA patients and controls group in IL-10 gene genotypes. 4. In IL-10 genotypes, there was no significant difference among Good, Mild and Bad responders to in RA patients. 5. There was no significant difference in the improvement of ESR, CRP and KHAQ scores after bee venom therapy in RA patients among the IL4R or IL-10 genotypes. Conclusions : In IL-4R genotypes, there was significant difference between RA patients and control group, and among Good, Mild and Bad responders in RA patients. However, in IL-10 genotypes, there was no significant difference between RA patients and controls group and among Good, Mild and Bad responders in RA patients.

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The Effect of Interleukin-10 on KC Gene Expression in Mouse Peritoneal Macrophages (케모카인 KC 유전자 발현에 대한 Interleukin-10의 억제작용)

  • Kim, Hee-Sun
    • Journal of Yeungnam Medical Science
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    • v.15 no.1
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    • pp.47-54
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    • 1998
  • Interleukin-10(IL-10) inhibits production of a wide range of cytokines in various cell types and transcriptionally inhibits lipopolysaccharide (LPS)-induced expression of proinflammatory mediators. Cytokine expression by macrophages is an important aspect to ochestrate inflammatory responses. As an approach to identify mechanistic targets of IL-10, it was examined the time course for expression of KC(murine homologue of Gro) gene in murine peritoneal macrophages stimulated with LPS with or without IL-10. The effect of IL-10 on LPS induced KC mRNA expression was delayed and only seen after 1 hour treatment. Pretreatment with IL-10 did not eliminate the delayed inhibitory response nor increase the magnitude of suppression. These effects did not depend upon time of IL-10 treatment but the time of LPS treatment. LPS-induced KC mRNA expression by inhibitory action of IL-10 was not controlled at the level of transcription. The result indicates that IL-10 acts late in the process of KC gene expression and that the prominant site of action may be mRNA stability or translation.

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THE CHANGE OF EXPRESSION OF INTERLEUKIN-6 AND -8 AFTER THE APPLICATION OF THE STATIC COMPRESSIVE PRESSURE ON THE FIBROBLAST ORIGINATED FROM THE PERIODONTAL LIGAMENTS (치주인대 기원의 섬유아 세포에 압축력을 가한 경우 Interleukin-6 및 Interleukin-8의 발현 변화에 관한 연구)

  • Lee, Yeon-Hee;Kim, Seong-Gon;Nahm, Dong-Seok
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.5
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    • pp.426-429
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    • 2006
  • The fibroblast in the periodontal ligaments received various stress. Among them, compression and tension are quite important and they are related to the remodeling of tooth and alveolar bone. We studied the change of expression of interleukin-6 (IL-6) and interleukin-8 (IL-8) in the fibroblasts of the periodontal ligaments by real-time RT-PCR and ELISA. In results, the relative activity of IL-6 mRNA in 2 hours after was 1.54${\pm}$0.08 and 1.00${\pm}$0.05 in control and test, respectively (P<0.05). Its 12 hours after was 1.23${\pm}$0.06 and 2.78${\pm}$0.14 in control and test, respectively (P<0.05). The relative activity of IL-8 mRNA in 2 hours after was 1.00${\pm}$0.05 and 0.24${\pm}$0.01 in control and test, respectively (P<0.05). Its 12 hours after was 1.23${\pm}$0.06 and 0.63${\pm}$0.03 in control and test, respectively (P<0.05). The concentration of IL-6 was 1.02${\pm}$0.16 ng/ml, 0.90${\pm}$0.14 ng/ml, and 1.32${\pm}$0.12 ng/ml (P<0.05) in control, 2, and 12 hours after, respectively. The concentration of IL-8 was 2.26${\pm}$0.17 ng/ml, 1.70${\pm}$0.26 ng/ml (P<0.05), and 0.84${\pm}$0.47 ng/ml (P<0.05) in control, 2, and 12 hours after, respectively. In conclusion, the expression of IL-6 was significantly increased after the application of the static compressive force, but IL-8 was significantly decreased. Considering their known function, their expression is quite important in tooth and bone resorption.

Effect of Induction Temperature on the $P_L$ Promoter Controlled Production of Recombinant Human Interleukin-2 in Escherichia coli

  • Lee, In-Young;Kim, Myung-Kuk;Lee, Sun-Bok
    • Journal of Microbiology and Biotechnology
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    • v.2 no.1
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    • pp.26-34
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    • 1992
  • The effect of induction temperature on fermentation parameters has been investigated extensively using Escherichia coli M5248[pNKM21], a producer of recombinant human interleukin-2 (rhIL-2). In this recombinant microorganism, the gene expression of rhIL-2 is regulated by the cI857 repressor and $P_L$ promoter system. The recombinant fermentation parameters studied in this work include the cell growth, protein synthesis, cell viability, plasmid stability, $\beta$-lactamase activity, and rhIL-2 productivity. Interrelationships of such fermentation parameters have been analyzed through a quantitative assessment of the experimental data set obtained at eight different culture conditions. While the expression of rhIL-2 gene was repressed at culture temperatures below $34^\circ{C}$ with little effect on other fermentation parameters, under the conditions of rhIL-2 production $>(36~44^\circ{C})$ the cell growth, plasmid stability, and $\beta$-lactamase activity were, as induction temperature was increased, more profoundly reduced. Although the rhIL-2 content in the insoluble protein fraction was maximum at $40^\circ{C}$, total rhIL-2 production in the culture volume was found to be highest at the induction temperature of $36^\circ{C}$. This was in contrast to the previously known optimum induction temperature of the P$_{L}$ promoter system $>(40~42^\circ{C})$.Explanations for such a discrepancy have been proposed based on a product formation kinetics, and their implications have been discussed in detail.l.

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Serum levels of interleukin-6 and interleukin-8 as diagnostic markers of acute pyelonephritis in children

  • Mahyar, Abolfazl;Ayazi, Parviz;Maleki, Mohammad Reza;Daneshi-Kohan, Mohammad Mahdi;Sarokhani, Hamid Reza;Hashemi, Hassan Jahani;Talebi-Bakhshayesh, Mousa
    • Clinical and Experimental Pediatrics
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    • v.56 no.5
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    • pp.218-223
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    • 2013
  • Purpose: Early diagnosis and treatment of acute pyelonephritis in children is of special importance in order to prevent serious complications. This study was conducted to determine the diagnostic value of serum interleukin (IL)-6 and IL-8 in children with acute pyelonephritis. Methods: Eighty-seven patients between 1 month to 12 years old with urinary tract infection (UTI) were divided into 2 groups based on the result of 99m-technetium dimercapto-succinic acid renal scan: acute pyelonephritis (n=37) and lower UTI (n=50) groups. White blood cell (WBC) count, neutrophil (Neutl) count, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) concentration, platelet count, and serum IL-6 and IL-8 concentrations of both groups were measured and compared. Results: There was a significant difference between two groups regarding WBC count, Neutl count, ESR, and CRP concentration (P<0.05). In addition, the difference between the two groups regarding serum IL-6 and IL-8 concentrations was not significant (IL-6, 60 and 35.4 pg/mL and IL-8, 404 and 617 pg/mL, respectively). The sensitivity and specificity of serum IL-6 and IL-8 for diagnosis of acute pyelonephritis were 73%, 42% and 78%, 32%, respectively. Sensitivity, specificity, negative and positive predictive values of serum IL-6 and IL-8 were less than those of acute phase serum reactants such as CRP. Conclusion: This study showed that there was no significant difference between acute pyelonephritis and lower UTI groups regarding serum IL-6 and IL-8 levels. Therefore, despite confirming results of previous studies, it seems that IL-6 and IL-8 are not suitable markers for differentiating between acute pyelonephritis and lower UTI.

Simple Purification of Escherichia coli-Derived Recombinant Human Interleukin-2 Expressed with N-terminus Fusion of Glucagon

  • Won Hye-Soon;Lee Jeewon;Kim In-Ho;Park Young-Hoon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.13-16
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    • 2000
  • Simple procedures have been devised for purifying recombinant human interleukin-2 (hIL-2), which was expressed in Escberichia coli using sequences of glucagon molecules and enterokinase cleavage site as an N-terminus fusion partner. The insoluble aggregates of recombinant fusion protein produced in E. coli cytoplasm were easily dissolved by simple alkaline pH shift $(8\rightarrow12\rightarrow8)$. Following enterokinase cleavage, the recombinant hIL-2 was finally purified by one-step reversed-phase HPLC with high purity. The ease and high efficiency of this simple purification process seem to mainly result from the role of used glucagon fusion partner, which could be applied to the production of other therapeutically important proteins.

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Effects of Cardiovascular Training on Interleukin-6 in Stroke Patients: a Systematic Review and Meta-analysis of Randomized Controlled Trials

  • Jeong Keun Song;Ju Hui Moon
    • Physical Therapy Rehabilitation Science
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    • v.12 no.2
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    • pp.115-122
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    • 2023
  • Objective: A previous study reported that cardiovascular training (CT) decreased interleukin-6 (IL-6), a pro-inflammatory cytokine with bidirectional effects. However, because of conflicting results of increasing and decreasing IL-6 levels in stroke patients, it is essential to clarify the effects of CT on IL-6 levels in this population. Therefore, this review aimed to investigate the effects of CT on IL-6 levels in stroke patients through a meta-analysis of randomized controlled trials (RCTs), synthesizing and analyzing the effects qualitatively and quantitatively. Design: A systematic review and meta-analysis of randomized controlled trials. Methods: In this review, conducted in April 2023, electronic databases (Web of Science, CINAHL, Embase, MEDLINE, Google Scholar) were searched to ascertain the effects of CT on IL-6 levels in stroke patients. For qualitative evaluation, ReVMan, provided by the Cochrane Group, was used, and for quantitative evaluation, a random-effects model and SMD (Standardized Mean Difference) were used. Results: Three RCTs measured IL-6 in 117 patients with stroke. The experimental group to which CT was applied showed no significant change compared to the control group.The result of analysis using the random effect model is SMD=-0.23; 95% confidence interval, -0.66 to 0.20. Conclusions: CT does not affect IL-6 levels in stroke patients. These results suggest that CT can be applied regardless of its positive or negative effect on IL-6 levels in stroke patients.