• IMMUNE NETWORK
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• v.9 no.2
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• pp.58-63
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• 2009

Background: T cell immunoglobulin and mucin domain containing 3 protein (Tim-3) expressed on terminally differentiated Th1 cells plays a suppressive role in Th1-mediated immune responses. Recently, it has been shown that N-glycosylation affects the binding activity of the Tim-3-Ig fusion protein to its ligand, galectin-9, but the binding properties of non-glycosylated Tim-3 on $CD4^+CD25^+$T cells has not been fully examined. In this study, we produced recombinant Tim-3-Ig fusion proteins in different cellular sources and its N-glycosylation mutant forms to evaluate their binding activities to $CD4^+CD25^+$T cells. Methods: We isolated and cloned Tim-3 cDNA from BALB/C mouse splenocytes. Then, we constructed a mammalian expression vector and a prokaryotic expression vector for the Tim-3-Ig fusion protein. Using a site directed mutagenesis method, plasmid vectors for Tim-3-Ig N-glycosylation mutant expression were produced. The recombinant protein was purified by protein A sepharose column chromatography. The binding activity of Tim-3-Ig fusion protein to $CD4^+CD25^+$T cells was analyzed using flow cytometry. Results: We found that the nonglycosylated Tim-3-Ig fusion proteins expressed in bacteria bound to $CD4^+CD25^+$T cells similarly to the glycosylated Tim-3-Ig protein produced in CHO cells. Further, three N-glycosylation mutant forms (N53Q, N100Q, N53/100Q) of Tim-3-Ig showed similar binding activities to those of wild type glycosylated Tim-3-Ig. Conclusion: Our results suggest that N-glycosylation of Tim-3 may not affect its binding activity to ligands expressed on $CD4^+CD25^+$T cells.

• Bioinformatics and Biosystems
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• v.1 no.2
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• pp.123-127
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• 2006

TIM barrel domain is widely studied since it is one of most common structure and mediates diverse function maintaining overall structure. TIM barrel domain's function is determined by local structural environment at the C-terminal end of barrel structure. We classified TIM barrel domains by local structural alignment tool, LSHEBA, to understand characteristics of TIM barrel domain's functionalvariation. TIM barrel domains classified as the same cluster share common structure, function and ligands. Over 80% of TIM barrels in clusters share exactly the same catalytic function. Comparing clustering result with that of SCOP, we found that it's important to know local structural environment of TIM barrel domains rather than overallstructure to understand specific structural detail of TIM barrel function. Non TIM barrel domains were associated to make different domain combination to form a different function. The relationship between domain combination, we suggested expected evolutional history. We finally analyzed the characteristics of amino acids around ligand interface.

• IMMUNE NETWORK
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• v.11 no.4
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• pp.203-209
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• 2011

Background: T cell immunoglobulin mucin containing molecule (TIM)-3 is expressed in differentiated Th1 cells and is involved in the suppression of the cytokine production by these cells. However, the regulation of the expression of other T cell genes by TIM-3 is unclear. Herein, we attempted to identify differentially expressed genes in cells abundantly expressing TIM-3 compared to cells with low expression of TIM-3. Methods: TIM-3 overexpressing cell clones were established by transfection of Jurkat T cells with TIM-3 expression vector. For screening of differentially expressed genes, gene fishing technology based on reverse transcription-polymerase chain reaction (RT-PCR) using an annealing control primer system was used. The selected candidate genes were validated by semi quantitative and real-time RT-PCR. Results: The transcription of TIMP-1, IFITM1, PAR3 and CCL1 was different between TIM-3 overexpressing cells and control cells. However, only CCL1 transcription was significantly different in cells transiently transfected with TIM3 expression vector compared with control cells. CCL1 transcription was increased in primary human $CD4^+$ T cells abundantly expressing TIM-3 but not in cells with low expression of TIM-3. Conclusion: CCL1 was identified as a differentially transcribed gene in TIM-3-expressing $CD4^+$ T cells.

• Journal of Microbiology and Biotechnology
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• v.24 no.4
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• pp.568-576
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• 2014

TIM-1 (also known as KIM-1 and HAVcr-1) is a type I transmembrane glycoprotein member of the TIM family that may play important roles in innate and adaptive immune responses. The overexpression of proteins associated with membrane proteins is a major obstacle to overcome in studies of membrane protein structures and functions. In this study, we successfully coupled the overexpression of the TIM-1 protein with a C-terminal enhanced green fluorescent protein (GFP) tag in Escherichia coli. To the best of our knowledge, this report is the first to describe the overexpression of human TIM-1 in E. coli. The purified TIM-1-EGFP fusion protein recognized and bound directly to apoptotic cells and did not to bind to viable cells. Furthermore, we confirmed that the interactions of TIM-1-EGFP with apoptotic cells were blocked by TIM-1-Fc fusion proteins. This fusion protein represents a readily obtainable source of biologically active TIM-1 that may prove useful in future studies of human TIM-1.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.22
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• pp.9945-9948
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• 2014

Background: The purpose of this study was to investigate Tim-3 expression on peripheral CD3-CD56+ natural killer (NK) cells and CD3+CD56+ natural killer T (NKT) cells in lung cancer patients. Materials and Methods: We analyzed Tim-3+CD3-CD56+ cells, Tim-3+CD3-$CD56^{dim}$ cells, Tim-3+CD3-$CD56^{bright}$ cells, and Tim-3+CD3+CD56+ cells in fresh peripheral blood from 79 lung cancer cases preoperatively and 53 healthy controls by flow cytometry. Postoperative blood samples were also analyzed from 21 members of the lung cancer patient cohort. Results: It was showed that expression of Tim-3 was significantly increased on CD3-CD56+ cells, CD3-$CD56^{dim}$ cells and CD3+CD56+ cells in lung cancer patients as compared to healthy controls (p=0.03, p=0.03 and p=0.04, respectively). When analyzing Tim-3 expression with cancer progression, results revealed more elevated Tim-3 expression in CD3-CD56+ cells, CD3-$CD56^{dim}$ cells and CD3+CD56+ cells in cases with advanced stages (III/IV) than those with stage I and II (p=0.02, p=0.04 and p=0.01, respectively). In addition, Tim-3 expression was significantly reduced on after surgical resection of the primary tumor (p<0.01). Conclusions: Tim-3 expression in natural killer cells from fresh peripheral blood may provide a useful indicator of disease progression of lung cancer. Furthermore, it was indicated that Tim-3 might be as a therapeutic target.

• Journal of the Korean Society for Precision Engineering
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• v.27 no.1
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• pp.91-98
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• 2010

The increasing of power and processing speed and miniaturization of central processor unit (CPU) used in electronics equipment requires better performing thermal management systems. A typical thermal management package consists of thermal interfaces, heat dissipaters, and external cooling systems. There have been a number of experimental techniques and procedures for estimating thermal conductivity of thin, compressible thermal interface material (TIM). The TIM performance is affected by many factors and thus TIM should be evaluated under specified application conditions. In compact packaging of electronic equipment the chip is interfaced with a thin heat spreader. As the package is made thinner, the coupling between heat flow through TIM and that in the heat spreader becomes stronger. Thus, a TIM characterization system for considering the heat spreader effect is proposed and demonstrated in detail in this paper. The TIM test apparatus developed based on ASTM D-5470 standard for thermal interface resistance measurement of high performance TIM, including the precise measurement of changes in in-situ materials thickness. Thermal impedances are measured and compared for different directions of heat dissipation. The measurement of the TIM under the practical conditions can thus be used as the thermal criteria for the TIM selection.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.6
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• pp.3897-3901
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• 2013

Background: Optimal treatment for prostate cancer remains a challenge worldwide. Recently, T cell immunoglobulin mucin-3 (TIM-3) has been implicated in tumor biology but its contribution prostate cancer remains unclear. The aim of this study was to investigate the role of TIM-3 as a prognostic marker in patients with prostate cancer. Methods: TIM-3 protein expression was determined by immunohistochemistry and Western blotting in 137 prostate cancer tumor samples and paired adjacent benign tissue. We also performed cell proliferation assays using 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl- 2H tetrazolium bromide (MTT) and cell invasion assays. The effects of small interfering RNA (siRNA)-mediated knockdown of TIM-3 (TIM-3 siRNA) in two human prostate cancer cell lines were also evaluated. Results: TIM-3 expression was higher in prostate cancer tissue than in the adjacent benign tissue (P<0.001). High TIM-3 expression was an independent predictor of both recurrence-free survival and progression-free survival. TIM-3 protein was expressed in both prostate cancer cell lines and knockdown suppressed their proliferation and invasion capacity. Conclusions: TIM-3 expression is associated with a poor prognosis in prostate cancer. Taken together, our resutlts indicate that TIM-3 is a potential prognostic marker in prostate cancer.

• IMMUNE NETWORK
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• v.12 no.5
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• pp.207-212
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• 2012

T cell immunoglobulin mucin domain (TIM)-3 is an immunomodulatory molecule and upregulated in T cells by several cytokines. TIM-3 also influences mast cell function but its transcriptional regulation in mast cells has not been clarified. Therefore, we examined the transcript level and the promoter activity of TIM-3 in mast cells. The TIM-3 transcript level was assessed by real-time RT-PCR and promoter activity by luciferase reporter assay. TIM-3 mRNA levels were increased in HMC-1, a human mast cell line by TGF-${\beta}1$ stimulation but not by stimulation with interferon (IFN)-${\alpha}$, IFN-${\lambda}$, TNF-${\alpha}$, or IL-10. TIM-3 promoter -349~+144 bp region relative to the transcription start site was crucial for the basal and TGF-${\beta}1$-induced TIM-3 promoter activities in HMC-1 cells. TIM-3 promoter activity was increased by over-expression of Smad2 and Smad4, downstream molecules of TGF-${\beta}1$ signaling. Our results localize TIM-3 promoter activity to the region spanning -349 to ＋144 bp in resting and TGF-${\beta}1$ stimulated mast cells.

• The Southeast Asian review
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• v.22 no.2
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• pp.133-170
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• 2012

This thesis aims to analyze the political, social, and cultural activities of the Acehnese ethnic group living in Jakarta, Indonesia. Based on analysis, this thesis examines how their ethnicity and national identity have been formed and expressed. For this purpose, this study deals with Taman Iskandar Muda (hereinafter referred to as TIM), a group of Acehnese transmigrants living in Jakarta. The immigration of the Acehnese to Jakarta started in the 1950s and the number of Acehnese people living in Jakarta persently amounts to 100,000. TIM, which was organized by the first generational of immigrants, functions to group Acehnese immigrants of various generations and class. Forum Keprihatinan Untuk Aceh(hereinafter referred to as Forka), an organization designed to solve the political problems of TIM, undertook various activities to maintain the peace of Aceh as the representative of TIM. Through those activities, TIM and Forka were able to confirm the feeling of homogeneity among the Acehnese who were living in their hometown and also strengthen their identity within the organizations. However, the fact that TIM and Forka put their focus on humanitarian activities paradoxically shows the political limitations that they sustain. TIM and Forka take care not to make their humanitarian activities seem as if they intend to openly strengthen their Acehnese identity and deny their Indonesian one. These political characteristics of Forka's identity are commonly found in groups that practice long-distance nationalism, as transmigrants in diaspora circumstances do. In the organization of TIM, there exists the menasah, which is a space where discussions of the ethnicity and the nation are practiced. As it is the space for local exchange, menasah reveals the identity of TIM through educational/social activities and public services. Menasah functions as the public arena where people practice ethnic identity on the basis of national integration. As a minority ethnic group living in Jakarta and its neighborhood, they are accustomed to double and selective political activities, social activities, and cultural practices. In order to adapt themselves to the double circumstance that they are faced with, they should live extemporaneously, and this life may be the fate that minority ethnic and transmigrants should endure.

• Journal of Korea Technology Innovation Society
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• v.14 no.4
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• pp.813-832
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• 2011

This is an analysis of three hypotheses in the fields of Technology, Innovation, and Management (TIM) analyzing the journal of Jishu Jingji (Technology Economics) which is published by the Chinese Society of Techno-economics. The Jishu Jingji journal is similar to other Chinese journals in TIM as follows: First, it's short in length. Second, there are a few references and foreign references can be counted with fingers of one hand. Third, the journal's scope is similar to others in TIM such as technolog-firm-industry. The differences are as follows: First, they have more general topics on the firm and the economy than other journals in TIM. Second, technologies covered are those of existing industries and infrastructure rather than emerging technologies and industries. Third, editors rarely submit papers to the journal. Finally, the expertise of the editors is diverse which is similar to the scope of the journal. First proposed hypothesis on journals in TIM is that journals represent country specific characteristics over field specificities, and this view is widely accepted in China. Second hypothesis is that editors are representatives of authors, but this hypothesis is rejected, at least in this journal. Third hypothesis is that the composition of editors reflects the scope of journals, and this hypothesis is generally accepted.