The Transmembrane Adaptor Protein LIME Is Essential for Chemokine-Mediated Migration of Effector T Cells to Inflammatiory Sites

박인영,Myongsun Son,안은선,Young-Woong Kim,공영윤,윤영대 한국분자세포생물학회 2020 Molecules and cells Vol.43 No.11

Lck-interacting transmembrane adaptor 1 (LIME) has been previously identified as a raft-associated transmembrane protein expressed predominantly in T and B lymphocytes. Although LIME is shown to transduce the immunoreceptor signaling and immunological synapse formation via its tyrosine phosphorylation by Lck, a Src-family kinase, the in vivo function of LIME has remained elusive in the previous studies. Here we report that LIME is preferentially expressed in effector T cells and mediates chemokine-mediated T cell migration. Interestingly, in LIME-/- mice, while T cell receptor stimulation-dependent proliferation, differentiation to effector T cells, cytotoxic T lymphocyte (CTL) function and regulatory T lymphocyte (Treg) function were normal, only T cell-mediated inflammatory response was significantly defective. The reduced inflammation was accompanied by the impaired infiltration of leukocytes and T cells to the inflammatory sites of LIME-/- mice. More specifically, the absence of LIME in effector T cells resulted in the reduced migration and defective morphological polarization in response to inflammatory chemokines such as CCL5 and CXCL10. Consistently, LIME-/- effector T cells were found to be defective in chemokine-mediated activation of Rac1 and Rap1, and dysregulated phosphorylation of Pyk2 and Cas. Taken together, the present findings show that LIME is a critical regulator of inflammatory chemokine-mediated signaling and the subsequent migration of effector T cells to inflammatory sites.

Interleukin-21 induces proliferation and modulates receptor expression and effector function in canine natural killer cells

Shin, D.J.,Lee, S.H.,Park, J.Y.,Kim, J.S.,Lee, J.J.,Suh, G.H.,Lee, Y.K.,Cho, D.,Kim, S.K. Elsevier 2015 Veterinary immunology and immunopathology Vol. No.

<P>Interleukin (IL)-21 is an important modulator of natural killer (NK) cell function. However, little is known about IL-21 function in canine NK cells because the phenotype of these cells remains undefined. In this study, we selectively expanded non-B and non-T large granular NK lymphocytes (CD3(-)CD21(-)CD5(-)CD4(-)TCR alpha beta-TCR gamma delta(-)) ex vivo from the peripheral blood mononuclear cells (PBMCs) of healthy dogs using a combination of IL-2, IL-15, and IL-21 in the presence of 100 Gy-irradiated K562 cells. We investigated the effects of varying the duration and timing of IL-21 treatment on stimulation of proliferation, expression of NK-related receptors, anti-tumor activity and production of interferon (IFN)-gamma. The expanded NK cells in each treatment group became enlarged and highly granular after 21 days in culture. NK cells proliferated rapidly in response to activation by IL-21 for 3 weeks, and IL-21 was able to induce changes in the mRNA expression of NK cell-related receptors and enhance the effector function of NK cells in perforin- and granzyme-B-dependent manners. The duration, frequency and timing of IL-21 stimulation during culture affected the rate of proliferation, patterns of receptor expression, cytokine production, and anti-tumor activity. The optimal conditions for maximizing the IL-21-induced proliferation and effector function of NK cells in the presence of IL-2 and IL-15 were seen in cells treated with IL-21 for the first 7 days of culture but without any further IL-21 stimulation other than an additional 2-day treatment prior to harvesting on day 21. The results of this study suggest that synergistic interactions of IL-21 with IL-2 and IL-15 play an important role in the proliferation, receptor expression, and effector function of canine NK cells. (C) 2015 Elsevier B.V. All rights reserved.</P>

두부 손상 정도에 따른 세포 매개성 면역의 변화에 관한 연구

이시우,손은익,이장철,김동원,임만빈,김인홍,전효진 대한신경외과학회 1995 Journal of Korean neurosurgical society Vol.24 No.10

최근 6개월간의 계명대학교 신경외과에 입원한 심한 전신 손상을 동반하지 않은 두부 손상 환자와 비교군에서 CMI multitest kit를 이용한 DTH 피부검사와 환자의 말초 혈액에서 T세포 및 그 아형과 B세포의 백분율을 조사함으로써 외상정도와 감염과의 상관 관계를 연구한 바 세포 매개성 면역 저하의 지표로 effecter T 임파구인 helper T세포(T4)와 cytotoxic T세포(T8)의 감소 DTH 피부검사에서 반응정도의 감소가 이용될 수 있음을 관찰 하였으며 이러한 현상은 종래의 심한 두부 손상 후 세포 매개성 면역 기능의 저하에 대한 보고와는 달리 비교적 경미한 두부 손상에 의해서도 유발될 수 있음을 알았다. 또한 DTH 피부검사의 Anergy 반응은 감염 가능성을 예측하는데 도움을 주는 지표로 이용 될 수 있음도 보여 주었으며, 통계적 유의성은 떨어지나 effector T 임파구의 백분율 검사는 외상 정도와 비례해 감소하는 경향을 보여 주었다. 그러나 두부 손상 정도가 심할수록 감염율이 증가하는 소견과 연관하여 설명하기에는 DTH 피부검사의 외상정도에 따른 차이성이나, effector T 세포 백분율 측정의 감염군과 비감염군간의 비교에서 각각 특이성이 떨어짐을 보이므로 앞으로 이에 대한 보완이 요구되며, 아울러 세포 매개성 면역이외의 감염에 중요하게 관여하는 다른 요인도 고려한다면, 두 검사 모두 두부 외상 환자에서 세포 매개성 면역성 조사나 환자의 감염에 대한 예측과 치료에 필요한 유용한 정보를 얻는데 비교적 감수성 높은 검사로 사료 되었다. Severe head injury results in the suppression of cellular immunity associated with dysfunctioning of effector lymphocytes, such as helper T cells(CD4) (and cytotoxic T cells(CD8). Despite progress in the management of increased intracranial pressure following head injury, infection remains the most common complication and the primary cause of prolonged hospitalization and death. This study attempts to assess the cellular immune function following head injury according to the degree of severity, and to establish the clinically available parameters of cell mediated immune(CMI) function, which can then be used for coherent prediction of infection risk. Eighteen head injury patients without severe systemic injury, who divided into three subgroups depending on the severity of head injury, were estimated with the use of CMI multitest kit(Merieux Institute, France) to test delayed-type hypersensitivity(DTH) and enumerated the circulating lymphocyte subpopulation(pan T-cell marker CD3, helper T cell marker CD4, cytotoxic T cell marker CD8 and B-cell marker CDl9) on the 1st, 7th, and 21th day of injury. Patients were monitored for evidence of infection for this period. Fourteen patients had no reaction to any antigens of the DTH skin test(anergy) and the remaining four patients had also some degree of anergy. Seven patients became infected and all of them were anergic. There were significant decrease of circulating effector T Iymphocytes, both CD4-positive and CD8-positive cells, within 24 hours of injury in the mild as well as the moderate and severe head injury group. CD4-positive cells were nearly completely recovered by the 7th day of injury. CD8-positive cells had sustained significant decrease even after 3 weeks of injury. There was no significant change in pan T-cells(CD3-positive cells) and B-cells(CDl9-positive cells). The results suggest that DTH skin test and effector T cell enumeration are both relatively simple and highly sensitive parameters for monitoring CMI function. Especially, anergy of DTH skin test can be used for indicator to predict risk of infection. Mild as well as moderate and severe head injuries may result in the suppresion of cellular immunity associated with the dysfunctioning of effector T cell.

Sex-Based Selectivity of PPARγ Regulation in Th1, Th2, and Th17 Differentiation

Park, Hong-Jai,Park, Hyeon-Soo,Lee, Jae-Ung,Bothwell, Alfred L. M.,Choi, Je-Min MDPI 2016 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.17 No.8

<P>Peroxisome proliferator-activated receptor gamma (PPARγ) has recently been recognized to regulate adaptive immunity through Th17 differentiation, Treg functions, and T<SUB>FH</SUB> responses. However, its role in adaptive immunity and autoimmune disease is still not clear, possibly due to sexual differences. Here, we investigated in vitro treatment study with the PPARγ agonist pioglitazone to compare Th1, Th2, and Th17 differentiation in male and female mouse splenic T cells. Pioglitazone treatment significantly inhibited various effector T cell differentiations including Th1, Th2, and Th17 cells from female naïve T cells, but it selectively reduced IL-17 production in male Th17 differentiation. Interestingly, pioglitazone and estradiol (E2) co-treatment of T cells in males inhibited differentiation of Th1, Th2, and Th17 cells, suggesting a mechanism for the greater sensitivity of PPARγ to ligand treatment in the regulation of effector T cell differentiation in females. Collectively, these results demonstrate that PPARγ selectively inhibits Th17 differentiation only in male T cells and modulates Th1, Th2, and Th17 differentiation in female T cells based on different level of estrogen exposure. Accordingly, PPARγ could be an important immune regulator of sexual differences in adaptive immunity.</P>

신경교종의 세포고사체로 감작시킨 수지상세포에 의한 세포성 면역반응의 유도

김인수,김종태,조현일,박영섭,김문찬,정동섭 대한신경외과학회 2003 Journal of Korean neurosurgical society Vol.34 No.4

Post-Translational Modifications in Transcription Factors that Determine T Helper Cell Differentiation

Kim, Hyo Kyeong,Jeong, Mi Gyeong,Hwang, Eun Sook Korean Society for Molecular and Cellular Biology 2021 Molecules and cells Vol.44 No.5

CD4+ T helper (Th) cells play a crucial role in the modulation of innate and adaptive immune responses through the differentiation of Th precursor cells into several subsets, including Th1, Th2, Th17, and regulatory T (Treg) cells. Effector Th and Treg cells are distinguished by the production of signature cytokines and are important for eliminating intracellular and extracellular pathogens and maintaining immune homeostasis. Stimulation of naive Th cells by T cell receptor and specific cytokines activates master transcription factors and induces lineage specification during the differentiation of Th cells. The master transcription factors directly activate the transcription of signature cytokine genes and also undergo post-translational modifications to fine-tune cytokine production and maintain immune balance through cross-regulation with each other. This review highlights the post-translational modifications of master transcription factors that control the differentiation of effector Th and Treg cells and provides additional insights on the immune regulation mediated by protein argininemodifying enzymes in effector Th cells.

An Intelligent Robotic Biological Cell Injection System

심재홍(Jae-Hong Shim),조영임(Young-Im Cho),김종형(Jong-Hyeong Kim) 한국지능시스템학회 2004 한국지능시스템학회논문지 Vol.14 No.4

최근 바이오 관련산업의 발전과 함께 바이오 장비 및 장치들에 대한 연구 및 개발이 활발하게 진행되고 있다. 특히 바이오 세포 조작관련 연구들이 많이 진행되어 오고 있다. 일반적으로 바이오 세포들에 대해 기계적인 엔드 이펙터들이 조작을 위해 접촉될 때 과도한 힘이 발생될 경우가 발생하며 이런 힘들에 의해 세포막이나 조직들이 피해를 입을 수 있다. 본 논문에서는 상기 문제들을 극복하기 위해 바이오 세포 조작을 위한 새로운 시스템을 제안하였다. 제안된 시스템은 내장된 힘 센서를 이용하여 바이오 세포와 엔드 이펙터간의 발생 힘을 측정할 수 있다. 또한, 비전기술을 이용하여 엔드 이펙터의 피펫 팁을 바이오 세포막까지 정확하게 가이드 할 수 있다. 결과적으로 제안된 시스템은 바이오 세포에 피해를 주지 않고 안전하게 조작이 가능하다. 제안된 기술을 이용하여 실제 시작품을 제작하여 다양한 실험을 수행한 결과 향후 DNA 조작과 같은 바이오 세포 조작용 정밀 인젝션 시스템으로의 사용 가능성을 보여 주었다. Recently, instruments and systems related on biological technology have been enormously developed. Particularly, many researches for biological cell injection have been carried out. Usually, excessive contact force occurring when the end-effector and a biological cell contact might make a damage on the cell. Unfortunately, the excessive force could easily destroy the membrane and tissue of the cell. In order to overcome the problem, we proposed a new injection system for biological cell manipulation. The proposed injection system can measure the contact force between a pipette and a cell by using a force sensor. Also, we used vision technology to correctly guide he tip of the pipette to the cell. Consequently, the proposed injection system could safely manipulate the biological cells without any damage. This paper presents the introduction of our new injection system and design concepts of the new micro end-effector. Through a series of experiments the proposed injection system shows the possibility of application for precision biological cell manipulation such as DNA operation.

Induction of MAP kinase phosphatase 3 through Erk/MAP kinase activation in three oncogenic Ras (H-, K- and N-Ras)-expressing NIH/3T3 mouse embryonic fibroblast cell lines

( Jaehyung Koo ),( Sen Wang ),( Nana Kang ),( Sun Jin Hur ),( Young Yil Bahk ) 생화학분자생물학회(구 한국생화학분자생물학회) 2016 BMB Reports Vol.49 No.7

Ras oncoproteins are small molecular weight GTPases known for their involvement in oncogenesis, which operate in a complex signaling network with multiple effectors. Approximately 25% of human tumors possess mutations in a member of this family. The Raf1/MEK/Erk1/2 pathway is one of the most intensively studied signaling mechanisms. Different levels of regulation account for the inactivation of MAP kinases by MAPK phosphatases in a cell type- and stimuli-dependent manner. In the present study, using three inducible Ras-expressing NIH/3T3 cell lines, we demonstrated that MKP3 upregulation requires the activation of the Erk1/2 pathway, which correlates with the shutdown of this pathway. We also demonstrated, by applying pharmacological inhibitors and effector mutants of Ras, that induction of MKP3 at the protein level is positively regulated by the oncogenic Ras/Raf/MEK/Erk1/2 signaling pathway. [BMB Reports 2016; 49(7): 370-375]

The proteins encoded by the Drosophila Planar Polarity Effector genes inturned, fuzzy and fritz interact physically and can re-pattern the accumulation of ''upstream'' Planar Cell Polarity proteins

Wang, Y.,Yan, J.,Lee, H.,Lu, Q.,Adler, P.N. Academic Press 2014 Developmental Biology Vol.394 No.1

The frizzled/starry night pathway regulates planar cell polarity in a wide variety of tissues in many types of animals. It was discovered and has been most intensively studied in the Drosophila wing where it controls the formation of the array of distally pointing hairs that cover the wing. The pathway does this by restricting the activation of the cytoskeleton to the distal edge of wing cells. This results in hairs initiating at the distal edge and growing in the distal direction. All of the proteins encoded by genes in the pathway accumulate asymmetrically in wing cells. The pathway is a hierarchy with the Planar Cell Polarity (PCP) genes (aka the core genes) functioning as a group upstream of the Planar Polarity Effector (PPE) genes which in turn function as a group upstream of multiple wing hairs. Upstream proteins, such as Frizzled accumulate on either the distal and/or proximal edges of wing cells. Downstream PPE proteins accumulate on the proximal edge under the instruction of the upstream proteins. A variety of types of data support this hierarchy, however, we have found that when over expressed the PPE proteins can alter both the subcellular location and level of accumulation of the upstream proteins. Thus, the epistatic relationship is context dependent. We further show that the PPE proteins interact physically and can modulate the accumulation of each other in wing cells. We also find that over expression of Frtz results in a marked delay in hair initiation suggesting that it has a separate role/activity in regulating the cytoskeleton that is not shared by other members of the group.

Rpi-blb2-Mediated Hypersensitive Cell Death Caused by Phytophthora infestans AVRblb2 Requires SGT1, but not EDS1, NDR1, Salicylic Acid-, Jasmonic Acid-, or Ethylene-Mediated Signaling

오상근,권석윤,최도일 한국식물병리학회 2014 Plant Pathology Journal Vol.30 No.3

Potato Rpi-blb2 encodes a protein with a coiled-coil-nucleotidebinding site and leucine-rich repeat (CC-NBS-LRR) motif that recognizes the Phytophthora infestans AVRblb2 effector and triggers hypersensitive cell death (HCD). To better understand the components required for Rpi-blb2-mediated HCD in plants, we used virus-induced gene silencing to repress candidate genes in Rpi-blb2-transgenic Nicotiana benthamiana plants and assayed the plants for AVRblb2 effector. Rpi-blb2 triggersHCD through NbSGT1-mediated pathways, but not NbEDS1- or NbNDR1-mediated pathways. In addition,the role of salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) in Rpi-blb2-mediated HCD were analyzed by monitoring of the responses of NbICS1-, NbCOI1-, or NbEIN2-silenced or Rpi-blb2::NahG-transgenic plants. Rpi-blb2-mediated HCD in response to AVRblb2 was not associated with SA accumulation. Thus, SA affects Rpi-blb2-mediated resistance against P. infestans, but not Rpi-blb2-mediated HCD in responseto AVRblb2. Additionally, JA and ET signaling were not required for Rpi-blb2-mediated HCD in N. benthamiana. Taken together, these findings suggest that NbSGT1 is a unique positive regulator of Rpi-blb2-mediated HCD in response to AVRblb2, but EDS1, NDR1, SA, JA, and ET are not required.