人类γδT细胞识别应激分子-4型UL16结合蛋白的机制研究
摘要
UL16结合蛋白(UL16 Binding Proteins，ULBPs)是一个NK细胞活化型受体一NKG2D的配体家族，目前发现的同系物包括ULBP1～5。ULBPs可应激性表达在一些肿瘤细胞或病原体感染细胞的表面。ULBP1、2、和3之间存在55-60%同源性，而ULBP4序列则变化较大，且相关研究较少，仅限于ULBP4-NKG2D相互作用和活化NK细胞方面。
γδT细胞表面表达NKG2D，可识别MICA、ULBPs分子等配体分子。研究发现，γδT细胞表面的T细胞受体(TCR)γ和δ链异质二聚体分子也识别MICA。这种NKG2D和TCRγδ对相同配体分子双重识别的生物学意义尚不完全明了，据推测，可能与确保固有免疫细胞能及时清除表达应激分子的受损细胞有关的免疫监视功能有关。
本研究关注以下四个科学问题：
第一，回答TCRγδ是否识别ULBP4分子?这一问题至今尚无答案。事实上，ULBPs家族分子是否可与TCRγδ结合，至今无结构生物学的证据。由于ULBP4具有一定的基因表达的多样性，我们推测机体可能会启动具有受体多样性的TCRγδ对其产生识别，故以ULBP4分子为靶分子，探讨TCRγδ对其识别的可能性。在这一问题框架下，我们要研究δ1链，或δ2链对其结合的特异性，并探究δ链中与配体结合起关键作用的部位一互补决定区(CDR)3的基因序列变化的特点。
第二，澄清ULBP4反应性γδT细胞的特性，如主要亚群，细胞因子分泌谱系和肿瘤细胞毒等。
第三，利用抗体阻断实验分析ULBP4-NKG2D和ULBP4-TCRγδ双重识别的生物学意义。
第四，通过ULBP4在EB病毒感染B淋巴细胞和肿瘤细胞表面的表达和ULBP4介导的γδT细胞细胞毒分析，来探究ULBP4在免疫监视(尤其是在细胞恶变早期)中的作用。
我们希望通过针对以上四个科学问题的科学研究可基本阐明γδ-T细胞对ULBP4识别的机制，并为系统阐释ULBP4的生物学意义提供有价值的资料。
为解决上述科学问题，我们进行了以下研究：(1)利用293ET真核表达系统表达了人ULBP4分子胞外区(1～226氨基酸)的重组蛋白ULBP4_(1-226)；利用pcDNA真核表达载体系统在EL4细胞中稳定表达全长的ULBP4基因，这些系统的建立为后续的研究奠定了基础。(3)首次使用固相化ULBP4分子诱导OEC和CC中的肿瘤浸润γδT淋巴细胞的体外扩增，获得纯度为80%的Vδ2 T细胞，并开展ULBP4反应性γδT细胞TCR Vδ-CDR3区的结构生物学研究。(4)利用配体受体酶联免疫结合实验、流式细胞术和γδTCR基因转染的TCR-β链缺陷型的T细胞系J．RT3-T3．5细胞平台证实，ULBP4与TCRγ9δ2特异结合。(5)ELISA法检测ULBP4刺激γδT细胞产生细胞因子的谱系。MTT方法检测γδT细胞对表达ULBP4的肿瘤细胞的杀伤活性。(6)在抗体封闭实验中，分别分析了anti-NKG2D、anti-TCRγδ、anti-NKG2D+anti-TCRγδ单抗封闭前后，ULBP4活化的γδT细胞分泌IFN-γ水平及对ULBP4阳性的肿瘤细胞细胞毒的变化。实验结果显示，anti-NKG2D+anti-γδTCR单抗联合阻断的效果强于anti-γδTCR或anti-NKG2D单独阻断作用。(7)结合流式细胞术和组织化学方法，检测ULBP4在不同来源肿瘤细胞、组织和EB病毒转化前后B淋巴细胞表面的表达。分析ULBP4介导的γδT细胞细胞毒效应。
综上，γδT细胞可凭借表面的NKG2D和TCRγδ结合并识别与肿瘤和感染相关的应激分子ULBP4，其自身发生细胞活化，对肿瘤细胞，恶变细胞，病毒感染细胞迅速产生细胞毒效应，体现了固有免疫系统在免疫监视中所发挥的直接与迅捷的特点。以ULBP4为靶的免疫诊断与治疗策略可能为肿瘤的早期诊断与干预提供新型手段。
Ph.D.candidate:Yan Kong
Supervisor:Prof.Wei He,Prof.Lianxian Cui
ULBPs (UL16 binding proteins),is a ligand family for NKG2D activatingreceptor of human NK cells.Five members of ULBPs family have been identifiedto date and they are stressfully expressed in most epithelial tumours orpathogen-infected tissue cells.Amino acid sequence analyses indicate thatULBP1,ULBP2 and ULBP3 are 55～60% identical,while newly found ULBP4 ismore divergent,which only few reports related to,but focus on the mechanism ofNK cell activation via ULBP4 - NKG2D interaction.
MostγδT cells express NKG2D on their surface,which could recognize ULBPs,MICA and other ligands.Furthmore,many results has demonstrated that theγδTcells homodimer receptor,TCRγδalso mediated the recognition of MICA by theVδ1 subset ofγδT cells.However,it is not clear about the biological significance ofrecognition of the same ligand mediated by TCRγδand NKG2D simultaneously,which preditly benefit ed the clear of cells by innate immune cells during immunedefense
The present study focused on four scientific questions as follows:
First,Demonstrate whether ULBP4 could recognized byγδT cells viaTCRγδ.It is still enigmatic.In fact,there is no evidence in structure biology so farabout the ULBP4-TCRγδbinding.We proposed that TCRγδrespond to a diversityof ULBP4 gene due to its extremely diversity,analyzed the specificity ofδ1 orδ2 bind to ULBP4 and explored the sequence features of TCRδ-CDR3.
Second,clarify the immunological effects of ULBP4 -reactiveγδT cells,including analyzing the phentype of ULBP4 -reactiveγδT cells、measuringcytokine secreted by ULBP4-inducedγδT cells and identifing the target cells lysis.
Third,analyze biological significance of recognition of the same ligandmediated by TCRγδand NKG2D simultaneously by antibodies blockade.
Finaly,Study the roles of ULBP4 in immunosurveillance through analysis of theULBP4 expression on different tumor cells or tissue or EB virus- transformed Blymphocytes,and ULBP4-mediate target cells lysis.
To solve the scientific problems as mentioned above,we performed theinvestigations as follows.(1) we successfully expressed and purified recombinantextracellular segments (1-226aa) of ULBP41-226 protein utilizing 293ET eukaryoticexpressing system and established stable transfection of ULBP4 into EL4 cells,which make foundation for further researches.(2) The immobilized ULBP4 couldinduce marked expansion of human OEC- or CC -derivedγδTILs of Vδ2phenotype mostly in vitro andδ-CDR3 region of ULBP-reactiveγδT cells wasanalyzed (3) ULBP4 could bind to TCRγ9δ2 identified by Enzyme-linkedimmunosorbent binding assay or flow cytometry and stimulate the activation ofJ.RT3-T3.5-TCRγ9δ2 via TCRγ9δ2.(4) Cytokine secreted byγδT cells afterstimulation with ULBP4 were analyzed by ELISA and the cytotoxicity of Vδ2 T cellsto EL4/ULBP4-EL4 tumor cell lines were compared.(5) We compared the level ofIFN-γsecreted by ULBP-reactiveγδT cells and target cells lysis before or afterblockade with anti-NKG2D、anti-TCRγδ、anti-NKG2D +anti-TCRγδmAb.Our dataindicate that the percentages of blocking by combination anti-NKG2D andanti-TCRγδantibodies is stronger than anti-NKG2D mAb alone or anti-γδTCR mAbalone.(6) ULBP4 expressed on different tumor cells or tissue or EB virus-transformed B lymphocytes were analyzed by flow cytometry or immunohistochemistry.
Taken these data together,humanγδT cells could bind to and recognizeULBP4 stressfully expressed in tumours or pathogen-infected tissue cell via TCRand NKG2D signal,be actived,display strong cytotoxic toward tumors、transformed and virus infected cells,Which reflect the instant and direct responseof innate immune cells during immune defense.
γδT细胞表面表达NKG2D，可识别MICA、ULBPs分子等配体分子。研究发现，γδT细胞表面的T细胞受体(TCR)γ和δ链异质二聚体分子也识别MICA。这种NKG2D和TCRγδ对相同配体分子双重识别的生物学意义尚不完全明了，据推测，可能与确保固有免疫细胞能及时清除表达应激分子的受损细胞有关的免疫监视功能有关。
本研究关注以下四个科学问题：
第一，回答TCRγδ是否识别ULBP4分子?这一问题至今尚无答案。事实上，ULBPs家族分子是否可与TCRγδ结合，至今无结构生物学的证据。由于ULBP4具有一定的基因表达的多样性，我们推测机体可能会启动具有受体多样性的TCRγδ对其产生识别，故以ULBP4分子为靶分子，探讨TCRγδ对其识别的可能性。在这一问题框架下，我们要研究δ1链，或δ2链对其结合的特异性，并探究δ链中与配体结合起关键作用的部位一互补决定区(CDR)3的基因序列变化的特点。
第二，澄清ULBP4反应性γδT细胞的特性，如主要亚群，细胞因子分泌谱系和肿瘤细胞毒等。
第三，利用抗体阻断实验分析ULBP4-NKG2D和ULBP4-TCRγδ双重识别的生物学意义。
第四，通过ULBP4在EB病毒感染B淋巴细胞和肿瘤细胞表面的表达和ULBP4介导的γδT细胞细胞毒分析，来探究ULBP4在免疫监视(尤其是在细胞恶变早期)中的作用。
我们希望通过针对以上四个科学问题的科学研究可基本阐明γδ-T细胞对ULBP4识别的机制，并为系统阐释ULBP4的生物学意义提供有价值的资料。
为解决上述科学问题，我们进行了以下研究：(1)利用293ET真核表达系统表达了人ULBP4分子胞外区(1～226氨基酸)的重组蛋白ULBP4_(1-226)；利用pcDNA真核表达载体系统在EL4细胞中稳定表达全长的ULBP4基因，这些系统的建立为后续的研究奠定了基础。(3)首次使用固相化ULBP4分子诱导OEC和CC中的肿瘤浸润γδT淋巴细胞的体外扩增，获得纯度为80%的Vδ2 T细胞，并开展ULBP4反应性γδT细胞TCR Vδ-CDR3区的结构生物学研究。(4)利用配体受体酶联免疫结合实验、流式细胞术和γδTCR基因转染的TCR-β链缺陷型的T细胞系J．RT3-T3．5细胞平台证实，ULBP4与TCRγ9δ2特异结合。(5)ELISA法检测ULBP4刺激γδT细胞产生细胞因子的谱系。MTT方法检测γδT细胞对表达ULBP4的肿瘤细胞的杀伤活性。(6)在抗体封闭实验中，分别分析了anti-NKG2D、anti-TCRγδ、anti-NKG2D+anti-TCRγδ单抗封闭前后，ULBP4活化的γδT细胞分泌IFN-γ水平及对ULBP4阳性的肿瘤细胞细胞毒的变化。实验结果显示，anti-NKG2D+anti-γδTCR单抗联合阻断的效果强于anti-γδTCR或anti-NKG2D单独阻断作用。(7)结合流式细胞术和组织化学方法，检测ULBP4在不同来源肿瘤细胞、组织和EB病毒转化前后B淋巴细胞表面的表达。分析ULBP4介导的γδT细胞细胞毒效应。
综上，γδT细胞可凭借表面的NKG2D和TCRγδ结合并识别与肿瘤和感染相关的应激分子ULBP4，其自身发生细胞活化，对肿瘤细胞，恶变细胞，病毒感染细胞迅速产生细胞毒效应，体现了固有免疫系统在免疫监视中所发挥的直接与迅捷的特点。以ULBP4为靶的免疫诊断与治疗策略可能为肿瘤的早期诊断与干预提供新型手段。
Ph.D.candidate:Yan Kong
Supervisor:Prof.Wei He,Prof.Lianxian Cui
ULBPs (UL16 binding proteins),is a ligand family for NKG2D activatingreceptor of human NK cells.Five members of ULBPs family have been identifiedto date and they are stressfully expressed in most epithelial tumours orpathogen-infected tissue cells.Amino acid sequence analyses indicate thatULBP1,ULBP2 and ULBP3 are 55～60% identical,while newly found ULBP4 ismore divergent,which only few reports related to,but focus on the mechanism ofNK cell activation via ULBP4 - NKG2D interaction.
MostγδT cells express NKG2D on their surface,which could recognize ULBPs,MICA and other ligands.Furthmore,many results has demonstrated that theγδTcells homodimer receptor,TCRγδalso mediated the recognition of MICA by theVδ1 subset ofγδT cells.However,it is not clear about the biological significance ofrecognition of the same ligand mediated by TCRγδand NKG2D simultaneously,which preditly benefit ed the clear of cells by innate immune cells during immunedefense
The present study focused on four scientific questions as follows:
First,Demonstrate whether ULBP4 could recognized byγδT cells viaTCRγδ.It is still enigmatic.In fact,there is no evidence in structure biology so farabout the ULBP4-TCRγδbinding.We proposed that TCRγδrespond to a diversityof ULBP4 gene due to its extremely diversity,analyzed the specificity ofδ1 orδ2 bind to ULBP4 and explored the sequence features of TCRδ-CDR3.
Second,clarify the immunological effects of ULBP4 -reactiveγδT cells,including analyzing the phentype of ULBP4 -reactiveγδT cells、measuringcytokine secreted by ULBP4-inducedγδT cells and identifing the target cells lysis.
Third,analyze biological significance of recognition of the same ligandmediated by TCRγδand NKG2D simultaneously by antibodies blockade.
Finaly,Study the roles of ULBP4 in immunosurveillance through analysis of theULBP4 expression on different tumor cells or tissue or EB virus- transformed Blymphocytes,and ULBP4-mediate target cells lysis.
To solve the scientific problems as mentioned above,we performed theinvestigations as follows.(1) we successfully expressed and purified recombinantextracellular segments (1-226aa) of ULBP41-226 protein utilizing 293ET eukaryoticexpressing system and established stable transfection of ULBP4 into EL4 cells,which make foundation for further researches.(2) The immobilized ULBP4 couldinduce marked expansion of human OEC- or CC -derivedγδTILs of Vδ2phenotype mostly in vitro andδ-CDR3 region of ULBP-reactiveγδT cells wasanalyzed (3) ULBP4 could bind to TCRγ9δ2 identified by Enzyme-linkedimmunosorbent binding assay or flow cytometry and stimulate the activation ofJ.RT3-T3.5-TCRγ9δ2 via TCRγ9δ2.(4) Cytokine secreted byγδT cells afterstimulation with ULBP4 were analyzed by ELISA and the cytotoxicity of Vδ2 T cellsto EL4/ULBP4-EL4 tumor cell lines were compared.(5) We compared the level ofIFN-γsecreted by ULBP-reactiveγδT cells and target cells lysis before or afterblockade with anti-NKG2D、anti-TCRγδ、anti-NKG2D +anti-TCRγδmAb.Our dataindicate that the percentages of blocking by combination anti-NKG2D andanti-TCRγδantibodies is stronger than anti-NKG2D mAb alone or anti-γδTCR mAbalone.(6) ULBP4 expressed on different tumor cells or tissue or EB virus-transformed B lymphocytes were analyzed by flow cytometry or immunohistochemistry.
Taken these data together,humanγδT cells could bind to and recognizeULBP4 stressfully expressed in tumours or pathogen-infected tissue cell via TCRand NKG2D signal,be actived,display strong cytotoxic toward tumors、transformed and virus infected cells,Which reflect the instant and direct responseof innate immune cells during immune defense.
引文
1. Bauer S, Groh V, Wu J, Steinle A, Phillips JH, LanierLL, Spies T. 1999. Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science. 285:727-729.
2. Andre P, Biassoni R, Colonna M, Cosman D, Lanier LL, Long EO, Lopez-Botet M, Moretta A, Moretta L, Parham P. 2001. New nomenclature for MHC receptors. Nat Immunol. 2:661.
3. Cosman, D, J. Mullberg, C. L. Sutherland, W. Chin, R. Armitage, W. Fanslow, M. Kubin, and N. J. Chalupny. 2001. ULBPs, novel MHC class I-related molecules, bind to CMV glycoprotein UL16 and stimulate NK cytotoxicity through the NKG2D receptor. Immunity. 14:123-133.
4. Sutherland, C.L, Chalupny, N.J, Schooley, K, VandenBos,T, Kubin, M, Cosman, D, 2002. UL16-binding proteins, novel MHC class I-related proteins, bind to NKG2D and activate multiple signaling pathways in primary NK cells. J. Immunol. 168:671-679.
5. Groh V, Rhinehart R, Randolph-Habecker J, Topp MS, Riddell SR, Spies T. 2001. Costimulation of CD8alphabeta T cells by NKG2D via engagement by MIC induced on virus-infected cells. Nat Immunol. 2:255-260.
6. Hayday AC. 2000. [gamma][delta] cells: a right time and a right place for a conserved third way of protection. Annu Rev Immunol. 18:975-1026.
7. Rincon-Orozco B, Kunzmann V, Wrobel P, Kabelitz D, Steinle A.Herrmann T. 2005. Activation of Vc9Vd2 T cells by NKG2D. J Immunol. 175:2144-51.
8. V. Groh, A. Steinle, S. Bauer, and T. Spies, 1998 .Recognition of stress-induced molecules by intestinal epithelium y8 T cells, Science. 279:1737-1740.
9. V. Groh, R. Rhinehart, H. Secrist, S. Bauer, K. H. Grabstein, T. Spies, 1999. Broad tumor-associated expression and recognition by tumor-derived γδ T cells of MICA and MICB, Proc. Natl. Acad. Sci. U.S.A. 96:6879-6884.
10. A. Steinle, V. Groh, T. Spies, 1998. Diversification, expression, and gamma delta T cell recognition of evolutionarily distant members of the MIC family of major histocompatibility complex class l-related molecules, Proc. Natl. Acad. Sci. U. S. A. 95:12510-12515.
11. J. Wu, V. Groh, T. Spies, 2002. T cell antigen receptor engagement and specificity in the recognition of stress-inducible MHC class l-related chains by human epithelial γδ T cells, J Immunology. 169 :1236-1240
12. Yu, W. He, J. Chen, F. Zhang, D. Ba, 1999. Expansion and immunological study of human tumor infiltrating gamma/delta lymphocytes in virto, Int. Arch. Allergy Immunol. 119:31-37.
13. Qi J, Zhang J, Zhang S, Cui L, He W. 2003. Immobilized MICA could expand human Vdelta1 gammadelta T cells in vitro that displayed major histocompatibility complex class I chain-related A-dependent cytotoxicity to human epithelial carcinomas. Scand J Immunol. 58:211-220.
14. Jianqing Zhao, Jie Huang, Hui Chen, Lianxian Cui, Wei He, Vδ1 T cell Receptor binds specifically to MHC I Chain related A: molecular and biochemical evidences
15. Jan Chalupny N, Sutherland CL, Lawrence WA, Rein-Weston A, Cosman D. 2003. ULBP4 is a novel ligand for human NKG2D. Biochem Biophys Res Commun. 305:129-135.
16. Kubin, M, Cassiano, L, Chalupny, J, Chin, W, Cosman, D,Fanslow, W, Mullberg, J, Rousseau, A.M, Ulrich, D, Armitage, R. 2001. ULBP1, 2, 3: novel MHC class I-related molecules that bind to human cytomegalovirus glycoprotein UL16, activate NK cells. Eur. J. Immunol. 31:1428-1437.
17. Cao, W, and W. He. 2004. UL16 binding proteins. Immunobiology. 209:283-290.
18. Poggi, A., C. Venturino, S. Catellani, M. Clavio, M. Miglino, M. Gobbi, A. Steinle, P. Ghia, S. Stella, F. Caligaris-Cappio, and M. R. Zocchi. 2004. Vdelta1 T lymphocytes from B-CLL patients recognize ULBP3 expressed on leukemic B cells and up-regulated by trans-retinoic acid. Cancer research. 64:9172-9179.
19. Catellani, S, A. Poggi, A. Bruzzone, P. Dadati, J. L. Ravetti, M. Gobbi, and M. R. Zocchi. 2007. Expansion of V{delta}1 T lymphocytes producing IL-4 in low-grade non-Hodgkin lymphomas expressing UL-16-binding proteins. Blood. 109:2078-2085.
20. Wei Cao, Xueyan Xi, Zhiyong Hao, Wenjing Li, Yan Kong, Lianxian Cui, Chi Ma, Wei He. 2007. RAET1E2, a soluble isoform of the UL16 binding protein RAET1E produced by tumor cells, inhibits NKG2D-mediated NK cytotoxicity. J Biol Chem.
21. Novokhatny V, Medved L, Lijnen HR . 1995. Tissue-type plasminogen activator (tPA) interacts with urokinase-type plasminogen activator (uPA) via tPA's lysine binding site. An explanation of the poor fibrin affinity of recombinant tPA/uPA chimeric molecule. J Biol Chem. 270:8680-8685.
22.Khalili-Shirazi,A,N.A.Gregson,M.Londei,L.Summers,and R.A.Hughes.1998.The distribution of CD1 molecules in inflammatory neuropathy.J.Neurol.Sci.158:154-163.
23．李伯良，胡明等．1994．人胰岛素生长因子I基因的人工合成、克隆及其 表达．生物化学与生物物理学报．26：153-159．
24.Shiping Zhang,Geoffrey Zubay,Emanuel Goldman.1991.Low-usage codons in Escherichia coli,yeast,fruit fly and primates.Gene.105:61-72.
25．许艳，万敏，张培因，卫红飞，王丽颖．2004．六聚组氨酸在重组蛋白中的位置对镍亲和层析的影响．医学分子生物学杂志．
26.Yip TT,Hutchens TVV.2004.Immobilized metal-ion affinity chromatography.Methods Mol Biol.244:179-90.
27.Porath J.1992.Immobilized metal ion affinity chromatography.Protein Expr.Purif.3:263—281
28.Sulkowski E.1989.The saga of IMAC and MIT.Bioessays.10:170—175
29.Steinle A,Li P,Morris DL,Groh V,Lanier LL,Strong RK,Spies T:Interactions of human NKG2D with its ligands MICA,MICB,and homologs of the mouse RAE-1 protein family.Immunogenetics.2001,53:279-287.
30.Conejo-Garcia,J.R,F.Benencia,M.C.Courreges,E.Khang,L.Zhang,A.Mohamed-Hadley,J.M.Vinocur,R.J.Buckanovich,C.B.Thompson,B.Levine,and G.Coukos.2003.Letal,A tumor-associated NKG2D immunoreceptor ligand,induces activation and expansion of effector immune cells.Cancer biology &therapy 2:446-451.
31. Arden B, Clark S P, Kabelitz D. 1995. Human T-cell receptor variable gene segment families. Immunogenetics. 42:455-500.
32. Erin J. Adams, Yueh-Hsiu Chien, K.Christopher Garcia. 2005. Structure of a γδ T cell receptor in complex with the nonclassical MHC T22. Science. 308: 227-231.
33. Xu C, Zhang H, Hu H, He H, Wang Z, Xu Y, et al. 2007.gammadelta T cells recognizetumor cells via CDR3delta region. Mol Immunol 44(4):302-10.34.
34. Robert A, James A, Omodele Ashiru, Mark R. 2006. Regulation of NKG2D Ligand Gene Expression. Human Immunology. 67: 159 -169
1.Kaye,J,Browne,H,Stoffel,M.,Minson,T.1992.The UL16 gene of human cytomegalovirus encodes a glycoprotein that is dispensable for growth in vitro.J.Virol.66:6609-6615.
2.Cosman,D,J.Mullberg,C.L.Sutherland,W.Chin,R.Armitage,W.Fanslow,M.Kubin,and N.J.Chalupny.2001.ULBPs,novel MHC class I-related molecules,bind to CMV glycoprotein UL16 and stimulate NK cytotoxicity through the NKG2D receptor.Immunity.14:123-133.
3.Jan Chalupny N,Sutherland CL,Lawrence WA,Rein-Weston A,Cosman D.2003.ULBP4 is a novel ligand for human NKG2D.Biochem Biophys Res Commun.305:129-135.
4.Conejo-Garcia,J.R,F.Benencia,M.C.Courreges,E.Khang,L.Zhang,A.Mohamed-Hadley,J.M.Vinocur,R.J.Buckanovich,C.B.Thompson,B.Levine,and G.Coukos.2003.Letal,A tumor-associated NKG2D immunoreceptor ligand,induces activation and expansion of effector immune cells.Cancer biology & therapy 2:446-451.
5.Bacon L,Eagle RA,Meyer M,Easom N,Young NT,Trowsdale J.2004.Two human ULBP/RAET1 molecules with transmembrane regions are ligands for NKG2D.J Immunol.173:1078-1084.
6.Verneris,M.R,M.Karami,J.Baker,A.Jayaswal,and R.S.Negrin.2004.Role of NKG2D signaling in the cytotoxicity of activated and expanded CD8+ T cells.Blood.103:3065-3072
7.Wu,J,Groh,V,Spies,T.2002.T cell antigen receptor engagement and specificity in the recognition of stress-inducible MHC class I-related chains by human epithelial gamma delta T cells.J.Immunol.169:1236-1240.
8. Michael T, Nathaniel L. Harris. 2005. NKG2D Ligands are Expressed on Stressed Human Airway Epithelial Cells. Am J Physiol Lung Cell Mol Physiol. in press.
9. Isabel Y, Eddie C, Martin Rowe.2006. The switch from latent to productive infection in Epstein-Barr virus infected B cells is associated with sensitization to NK cell killing. J. Virol. in press.
10. Michael T, Nathaniel L, Scott C, Mark Vitucci, David Cosman. 2006. NKG2D Ligands are Expressed on Stressed Human Airway Epithelial Cells. Am J Physiol Lung Cell Mol Physiol. in Press
11. Robert A, James A, Omodele Ashiru, Mark R. 2006. Regulation of NKG2D Ligand Gene Expression. Human Immunology. 67: 159 -169
12. Pende, D, Rivera, P, Marcenaro, S, Chang, C.C., Biassoni,R, Conte, R, Kubin, M, Cosman, D, Ferrone, S.Moretta, L, Moretta, A. 2002. Major histocompatibility complex class I-related chain A and UL16-binding protein expression on tumor cell lines of different histo-types: analysis of tumor susceptibility to NKG2D-depen-dent natural killer cell cytotoxicity. Cancer Res. 62:6178-6186.
13. Rolle A, Mousavi-Jazi M, Eriksson M, Odeberg J, Soderberg-Naucler C, Cosman D, Karre K, and Cerboni C. 2003. Effects of human cytomegalovirus infection on ligands for the activating NKG2D receptor of NK cells:up-regulation of UL16-binding protein (ULBP)1 and ULBP2 is counteracted by the viral UL16 protein. J Immunol .171:902-908
14. Groh V, Bahram S, Bauer S, Herman A, Beauchamp M, Spies T. 1996. Cell stress-regulated human major histocompatibility complex class I gene expressed in gastrointestinal epithelium. Proc Natl Acad Sci U S A. 93:12445-12450.
15. Groh V, Steinle A, Bauer S, Spies T. 1998. Recognition of stress-induced MHC molecules by intestinal epithelial gammadelta T cells. Science. 279:1737-1740.
16. Groh V, Rhinehart R, Randolph-Habecker J, Topp MS, Riddell SR, Spies T. 2001. Costimulation of CD8alphabeta T cells by NKG2D via engagement by MIC induced on virus-infected cells. Nat Immunol. 2:255-260.
17. Cerwenka, A, Bakker, A.B, McClanahan, T, Wagner, J, Wu J, Phillips, J.H, Lanier, L.L. 2000. Retinoic acid early inducible genes define a ligand family for the activating NKG2D receptor in mice. Immunity. 12: 721-727.
18. Jinushi, M., Takehara, T, Tatsumi, T, Kanto, T, Groh, V,Spies, T., Kimura, R, Miyagi, T, Mochizuki, K., Sasaki,Y, Hayashi, N.2003. Expression and role of MICA and MICB in human hepatocellular carcinomas and their regulation by retinoic acid. Int. J. Cancer. 104: 354-361.
19. Wu, J, Chalupny, N.J, Manley, T.J, Riddell, S.R, Cosman,D, Spies, T. 2003. Intracellular retention of the MHC class l-related chain B ligand of NKG2D by the human cytomegalovirus UL16 glycoprotein. J. Immunol. 170:4196-4200.
20. Dunn, C, Chalupny, N.J., Sutherland, C.L, Dosch, S,Sivakumar, P.V, Johnson, D.C., Cosman, D. 2003. Human cytomegalovirus glycoprotein UL16 causes intra-cellular sequestration of NKG2D ligands, protecting against natural killer cell cytotoxicity. J. Exp. Med. 197:1427-1439.
21. Welte, S.A, Sinzger, C, Lutz, S.Z, Singh-Jasuja, H,Sampaio, K.L, Eknigk, U, Rammensee, H.G, Steinle,A. 2003. Selective intracellular retention of virally induced NKG2D ligands by the human cytomegalovirus UL16 glycoprotein. Eur. J. Immunol. 33:194-203.
22. 0deberg, J, Browne, H, Metkar, S., Froelich, C.J, Branden,L, Cosman, D, Soderberg-Naucler, C. 2003. The human cytomegalovirus protein UL16 mediates increased resistance to natural killer cell cytotoxicity through resistance to cytolytic proteins. J. Virol. 77: 4539-4545.
23. Vales-Gomez, M., Browne, H., Reyburn, H.T. 2003. Expression of the UL16 glycoprotein of Human Cytomegalovirus protects the virus-infected cell from attack by natural killer cells. BMC Immunol. 4, 4.
24. Lodoen MB, Abenes G, Umamoto S, Houchins JP, Liu F, Lanier LL. 2004. The cytomegalovirus m155 gene product subverts natural killer cell antiviral protection by disruption of H60-NKG2D interactions. J Exp Med. 200:1075-1081
25. Kubin, M, Cassiano, L, Chalupny, J, Chin, W, Cosman, D,Fanslow, W, Mullberg, J, Rousseau, A.M, Ulrich, D,Armitage, R. 2001. ULBP1, 2, 3: novel MHC class l-related molecules that bind to human cytomegalovirus glycoprotein UL16, activate NK cells. Eur. J. Immunol. 31:1428-1437.
26. Bauer, S, Groh, V, Wu, J, Steinle, A, Phillips, J.H, Lanier,L.L, Spies, T.1999. Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science. 285:727-729.
27. Braud VM, Allan DS, O'Callaghan CA, Soderstrom K, D'Andrea A, Ogg GS, Lazetic S, Young NT, Bell Jl, Phillips JH. 1998. HLA-E binds to natural killer cell receptors CD94/NKG2A, B and C. Natur. 391:795-799.
28. Braud VM, Allan DS, McMichael AJ. 1999. Functions of nonclassical MHC and non-MHC-encoded class I molecules. Curr Opin Immunol.11:100-108.
29. Andre P, Biassoni R, Colonna M, Cosman D, Lanier LL, Long EO, Lopez-Botet M, Moretta A, Moretta L, Parham P, et al. 2001. New nomenclature for MHC receptors. Nat Immunol. 2:661.
30. Diefenbach A, Tomasello E, Lucas M, Jamieson AM, Hsia JK, Vivier E, Raulet DH. 2002. Selective associations with signaling proteins determine stimulatory versus costimulatory activity of NKG2D. Nat Immunol. 3:1142-1149.
31. Wu J, Song Y, Bakker AB .1999. An activating immunoreceptor complex formed by NKG2D and DAP10. Science. 285 : 730-2.
32. Sutherland, C.L, Chalupny, N.J, Schooley, K, VandenBos,T, Kubin, M, Cosman, D, 2002. UL16-binding proteins, novel MHC class l-related proteins, bind to NKG2D and activate multiple signaling pathways in primary NK cells. J. Immunol. 168:671-679.
33. Hayday AC. 2000. [gamma][delta] cells: a right time and a right place for a conserved third way of protection. Annu Rev Immunol. 18:975-1026.
34. Li JQ, Cui LX, He W. 2005. Distinct Pattern of Human Vdelta1 gammadelta T Cells Recognizing MICA. Cell Mol Immuno. 2:253-258.
35. Groh V, Rhinehart R, Secrist H, Bauer S, Grabstein KH, Spies T. 1999. Broad tumor-associated expression and recognition by tumor-derived gamma delta T cells of MICA and MICB. Proc Natl Acad Sci U S A. 96:6879-6884.
36. Wu J, Groh V, Spies T. 2002. T cell antigen receptor engagement and specificity in the recognition of stress-inducible MHC class l-related chains by human epithelial gamma delta T cells. J Immunol. 169:1236-1240.
37. Poggi, A., C. Venturino, S. Catellani, M. Clavio, M. Miglino, M. Gobbi, A. Steinle, P. Ghia, S. Stella, F. Caligaris-Cappio, and M. R. Zocchi. 2004. Vdelta1 T lymphocytes from B-CLL patients recognize ULBP3 expressed on leukemic B cells and up-regulated by trans-retinoic acid. Cancer research. 64:9172-9179.
38. Catellani, S, A. Poggi, A. Bruzzone, P. Dadati, J. L. Ravetti, M. Gobbi, and M. R. Zocchi. 2007. Expansion of V{delta}1 T lymphocytes producing IL-4 in low-grade non-Hodgkin lymphomas expressing UL-16-binding proteins. Blood. 109:2078-2085.
39. P. Wrobel, H. ShojaeiB. Schittek F, Gieselera, B. Wollenberg? H. Kalthoff-, D. Kabelitz & D. Wesch. 2007. Lysis of a Broad Range of Epithelial Tumour Cells by Human cd T Cells: Involvement of NKG2D ligands and T-cell Receptor- versus NKG2D-dependent Recognition. Scandinavian Journal of Immunology. 66:20-328
40. Jamieson AM, Diefenbach A, McMahon CW, Xiong N, Carlyle JR, Raulet DH. 2002. The role of the NKG2D immunoreceptor in immune cell activation and natural killing. Immunity. 17:19-29.
41. Pende D, Rivera P, Marcenaro S, Chang CC, Biassoni R, Conte R, Kubin M, Cosman D, Ferrone S, Moretta L, et al. 2002. Major histocompatibility complex class l-related chain A and UL16-binding protein expression on tumor cell lines of different histotypes: analysis of tumor susceptibility to NKG2D-dependent natural killer cell cytotoxicity. Cancer Res.62:6178-6186.
42. Diefenbach A, Jensen ER, Jamieson AM, Raulet DH. 2001. Rae1 and H60 ligands of the NKG2D receptor stimulate tumour immunity. Nature. 413:165-171.
43. Kulski JK, Dunn DS, Hui J, Martinez P, Romphruk AV, Leelayuwat C, Tay GK, Oka A, Inoko H: 2002. Alu polymorphism within the MICB gene and association with HLA-B alleles. Immunogenetics. 53:975-979.
44.Spreu, J, T. Stehle, and A. Steinle. 2006. Human cytomegalovirus-encoded UL16 discriminates MIC molecules by their alpha2 domains. J Immunol 177:3143-3149.
45. Waldhauer, I, and A. Steinle. 2006. Proteolytic release of soluble UL16-binding protein 2 from tumor cells. Cancer research. 66:2520-2526.
46.Wei Cao, Xueyan Xi, Zhiyong Hao, Wenjing Li, Yan Kong, Lianxian Cui, Chi Ma, Wei He. 2007. RAET1E2, a soluble isoform of the UL16 binding protein RAET1E produced by tumor cells, inhibits NKG2D-mediated NK cytotoxicity. J Biol Chem.
47. Eisele, G., J. Wischhusen, M. Mittelbronn, R. Meyermann, I. Waldhauer, A. Steinle, M. Weller, and M. A. Friese. 2006. TGF-beta and metalloproteinases differentially suppress NKG2D ligand surface expression on malignant glioma cells. Brain. 129:2416-2425.
48. Zhou H, Luo Y, Lo JF, Kaplan CD, Mizutani M, Mizutani N, Lee JD, Primus FJ, Becker JC, Xiang R, et al. 2005. DNA-based vaccines activate innate and adaptive antitumor immunity by engaging the NKG2D receptor. Proc Natl Acad Sci U S A. 102:10846-10851.
49. Zhou H, Luo Y, Kaplan CD, Kruger JA, Lee SH, Xiang R, Reisfeld RA. 2006. A DNA-based cancer vaccine enhances lymphocyte cross talk by engaging the NKG2D receptor. Blood . 107:3251-3257.
50. Conejo-Garcia, J.R, Benencia, F, Courreges, M.C, Khang,E, Zhang, L, Mohamed-Hadley, A, Vinocur, J.M, Buckanovich, R.J, Thompson, C.B, Levine, B., Coukos,G. 2003. Letal, a tumor-associated NKG2D immunor-eceptor ligand, induces activation and expansion of effector immune cells. Cancer Biol. Ther. 2: 446-451.
51. Groh V, Bruhl A, El-Gabalawy H, Nelson JL, Spies T. 2003. Stimulation of T cell autoreactivity by anomalous expression of NKG2D and its MIC ligands in rheumatoid arthritis. Proc Natl Acad Sci U S A. 100:9452-9457.
52. Ogasawara K, Hamerman JA, Ehrlich LR, Bour-Jordan H, Santamaria P, Bluestone JA, Lanier LL. 2004. NKG2D blockade prevents autoimmune diabetes in NOD mice. Immunity. 20:757-767.
2. Andre P, Biassoni R, Colonna M, Cosman D, Lanier LL, Long EO, Lopez-Botet M, Moretta A, Moretta L, Parham P. 2001. New nomenclature for MHC receptors. Nat Immunol. 2:661.
3. Cosman, D, J. Mullberg, C. L. Sutherland, W. Chin, R. Armitage, W. Fanslow, M. Kubin, and N. J. Chalupny. 2001. ULBPs, novel MHC class I-related molecules, bind to CMV glycoprotein UL16 and stimulate NK cytotoxicity through the NKG2D receptor. Immunity. 14:123-133.
4. Sutherland, C.L, Chalupny, N.J, Schooley, K, VandenBos,T, Kubin, M, Cosman, D, 2002. UL16-binding proteins, novel MHC class I-related proteins, bind to NKG2D and activate multiple signaling pathways in primary NK cells. J. Immunol. 168:671-679.
5. Groh V, Rhinehart R, Randolph-Habecker J, Topp MS, Riddell SR, Spies T. 2001. Costimulation of CD8alphabeta T cells by NKG2D via engagement by MIC induced on virus-infected cells. Nat Immunol. 2:255-260.
6. Hayday AC. 2000. [gamma][delta] cells: a right time and a right place for a conserved third way of protection. Annu Rev Immunol. 18:975-1026.
7. Rincon-Orozco B, Kunzmann V, Wrobel P, Kabelitz D, Steinle A.Herrmann T. 2005. Activation of Vc9Vd2 T cells by NKG2D. J Immunol. 175:2144-51.
8. V. Groh, A. Steinle, S. Bauer, and T. Spies, 1998 .Recognition of stress-induced molecules by intestinal epithelium y8 T cells, Science. 279:1737-1740.
9. V. Groh, R. Rhinehart, H. Secrist, S. Bauer, K. H. Grabstein, T. Spies, 1999. Broad tumor-associated expression and recognition by tumor-derived γδ T cells of MICA and MICB, Proc. Natl. Acad. Sci. U.S.A. 96:6879-6884.
10. A. Steinle, V. Groh, T. Spies, 1998. Diversification, expression, and gamma delta T cell recognition of evolutionarily distant members of the MIC family of major histocompatibility complex class l-related molecules, Proc. Natl. Acad. Sci. U. S. A. 95:12510-12515.
11. J. Wu, V. Groh, T. Spies, 2002. T cell antigen receptor engagement and specificity in the recognition of stress-inducible MHC class l-related chains by human epithelial γδ T cells, J Immunology. 169 :1236-1240
12. Yu, W. He, J. Chen, F. Zhang, D. Ba, 1999. Expansion and immunological study of human tumor infiltrating gamma/delta lymphocytes in virto, Int. Arch. Allergy Immunol. 119:31-37.
13. Qi J, Zhang J, Zhang S, Cui L, He W. 2003. Immobilized MICA could expand human Vdelta1 gammadelta T cells in vitro that displayed major histocompatibility complex class I chain-related A-dependent cytotoxicity to human epithelial carcinomas. Scand J Immunol. 58:211-220.
14. Jianqing Zhao, Jie Huang, Hui Chen, Lianxian Cui, Wei He, Vδ1 T cell Receptor binds specifically to MHC I Chain related A: molecular and biochemical evidences
15. Jan Chalupny N, Sutherland CL, Lawrence WA, Rein-Weston A, Cosman D. 2003. ULBP4 is a novel ligand for human NKG2D. Biochem Biophys Res Commun. 305:129-135.
16. Kubin, M, Cassiano, L, Chalupny, J, Chin, W, Cosman, D,Fanslow, W, Mullberg, J, Rousseau, A.M, Ulrich, D, Armitage, R. 2001. ULBP1, 2, 3: novel MHC class I-related molecules that bind to human cytomegalovirus glycoprotein UL16, activate NK cells. Eur. J. Immunol. 31:1428-1437.
17. Cao, W, and W. He. 2004. UL16 binding proteins. Immunobiology. 209:283-290.
18. Poggi, A., C. Venturino, S. Catellani, M. Clavio, M. Miglino, M. Gobbi, A. Steinle, P. Ghia, S. Stella, F. Caligaris-Cappio, and M. R. Zocchi. 2004. Vdelta1 T lymphocytes from B-CLL patients recognize ULBP3 expressed on leukemic B cells and up-regulated by trans-retinoic acid. Cancer research. 64:9172-9179.
19. Catellani, S, A. Poggi, A. Bruzzone, P. Dadati, J. L. Ravetti, M. Gobbi, and M. R. Zocchi. 2007. Expansion of V{delta}1 T lymphocytes producing IL-4 in low-grade non-Hodgkin lymphomas expressing UL-16-binding proteins. Blood. 109:2078-2085.
20. Wei Cao, Xueyan Xi, Zhiyong Hao, Wenjing Li, Yan Kong, Lianxian Cui, Chi Ma, Wei He. 2007. RAET1E2, a soluble isoform of the UL16 binding protein RAET1E produced by tumor cells, inhibits NKG2D-mediated NK cytotoxicity. J Biol Chem.
21. Novokhatny V, Medved L, Lijnen HR . 1995. Tissue-type plasminogen activator (tPA) interacts with urokinase-type plasminogen activator (uPA) via tPA's lysine binding site. An explanation of the poor fibrin affinity of recombinant tPA/uPA chimeric molecule. J Biol Chem. 270:8680-8685.
22.Khalili-Shirazi,A,N.A.Gregson,M.Londei,L.Summers,and R.A.Hughes.1998.The distribution of CD1 molecules in inflammatory neuropathy.J.Neurol.Sci.158:154-163.
23．李伯良，胡明等．1994．人胰岛素生长因子I基因的人工合成、克隆及其 表达．生物化学与生物物理学报．26：153-159．
24.Shiping Zhang,Geoffrey Zubay,Emanuel Goldman.1991.Low-usage codons in Escherichia coli,yeast,fruit fly and primates.Gene.105:61-72.
25．许艳，万敏，张培因，卫红飞，王丽颖．2004．六聚组氨酸在重组蛋白中的位置对镍亲和层析的影响．医学分子生物学杂志．
26.Yip TT,Hutchens TVV.2004.Immobilized metal-ion affinity chromatography.Methods Mol Biol.244:179-90.
27.Porath J.1992.Immobilized metal ion affinity chromatography.Protein Expr.Purif.3:263—281
28.Sulkowski E.1989.The saga of IMAC and MIT.Bioessays.10:170—175
29.Steinle A,Li P,Morris DL,Groh V,Lanier LL,Strong RK,Spies T:Interactions of human NKG2D with its ligands MICA,MICB,and homologs of the mouse RAE-1 protein family.Immunogenetics.2001,53:279-287.
30.Conejo-Garcia,J.R,F.Benencia,M.C.Courreges,E.Khang,L.Zhang,A.Mohamed-Hadley,J.M.Vinocur,R.J.Buckanovich,C.B.Thompson,B.Levine,and G.Coukos.2003.Letal,A tumor-associated NKG2D immunoreceptor ligand,induces activation and expansion of effector immune cells.Cancer biology &therapy 2:446-451.
31. Arden B, Clark S P, Kabelitz D. 1995. Human T-cell receptor variable gene segment families. Immunogenetics. 42:455-500.
32. Erin J. Adams, Yueh-Hsiu Chien, K.Christopher Garcia. 2005. Structure of a γδ T cell receptor in complex with the nonclassical MHC T22. Science. 308: 227-231.
33. Xu C, Zhang H, Hu H, He H, Wang Z, Xu Y, et al. 2007.gammadelta T cells recognizetumor cells via CDR3delta region. Mol Immunol 44(4):302-10.34.
34. Robert A, James A, Omodele Ashiru, Mark R. 2006. Regulation of NKG2D Ligand Gene Expression. Human Immunology. 67: 159 -169
1.Kaye,J,Browne,H,Stoffel,M.,Minson,T.1992.The UL16 gene of human cytomegalovirus encodes a glycoprotein that is dispensable for growth in vitro.J.Virol.66:6609-6615.
2.Cosman,D,J.Mullberg,C.L.Sutherland,W.Chin,R.Armitage,W.Fanslow,M.Kubin,and N.J.Chalupny.2001.ULBPs,novel MHC class I-related molecules,bind to CMV glycoprotein UL16 and stimulate NK cytotoxicity through the NKG2D receptor.Immunity.14:123-133.
3.Jan Chalupny N,Sutherland CL,Lawrence WA,Rein-Weston A,Cosman D.2003.ULBP4 is a novel ligand for human NKG2D.Biochem Biophys Res Commun.305:129-135.
4.Conejo-Garcia,J.R,F.Benencia,M.C.Courreges,E.Khang,L.Zhang,A.Mohamed-Hadley,J.M.Vinocur,R.J.Buckanovich,C.B.Thompson,B.Levine,and G.Coukos.2003.Letal,A tumor-associated NKG2D immunoreceptor ligand,induces activation and expansion of effector immune cells.Cancer biology & therapy 2:446-451.
5.Bacon L,Eagle RA,Meyer M,Easom N,Young NT,Trowsdale J.2004.Two human ULBP/RAET1 molecules with transmembrane regions are ligands for NKG2D.J Immunol.173:1078-1084.
6.Verneris,M.R,M.Karami,J.Baker,A.Jayaswal,and R.S.Negrin.2004.Role of NKG2D signaling in the cytotoxicity of activated and expanded CD8+ T cells.Blood.103:3065-3072
7.Wu,J,Groh,V,Spies,T.2002.T cell antigen receptor engagement and specificity in the recognition of stress-inducible MHC class I-related chains by human epithelial gamma delta T cells.J.Immunol.169:1236-1240.
8. Michael T, Nathaniel L. Harris. 2005. NKG2D Ligands are Expressed on Stressed Human Airway Epithelial Cells. Am J Physiol Lung Cell Mol Physiol. in press.
9. Isabel Y, Eddie C, Martin Rowe.2006. The switch from latent to productive infection in Epstein-Barr virus infected B cells is associated with sensitization to NK cell killing. J. Virol. in press.
10. Michael T, Nathaniel L, Scott C, Mark Vitucci, David Cosman. 2006. NKG2D Ligands are Expressed on Stressed Human Airway Epithelial Cells. Am J Physiol Lung Cell Mol Physiol. in Press
11. Robert A, James A, Omodele Ashiru, Mark R. 2006. Regulation of NKG2D Ligand Gene Expression. Human Immunology. 67: 159 -169
12. Pende, D, Rivera, P, Marcenaro, S, Chang, C.C., Biassoni,R, Conte, R, Kubin, M, Cosman, D, Ferrone, S.Moretta, L, Moretta, A. 2002. Major histocompatibility complex class I-related chain A and UL16-binding protein expression on tumor cell lines of different histo-types: analysis of tumor susceptibility to NKG2D-depen-dent natural killer cell cytotoxicity. Cancer Res. 62:6178-6186.
13. Rolle A, Mousavi-Jazi M, Eriksson M, Odeberg J, Soderberg-Naucler C, Cosman D, Karre K, and Cerboni C. 2003. Effects of human cytomegalovirus infection on ligands for the activating NKG2D receptor of NK cells:up-regulation of UL16-binding protein (ULBP)1 and ULBP2 is counteracted by the viral UL16 protein. J Immunol .171:902-908
14. Groh V, Bahram S, Bauer S, Herman A, Beauchamp M, Spies T. 1996. Cell stress-regulated human major histocompatibility complex class I gene expressed in gastrointestinal epithelium. Proc Natl Acad Sci U S A. 93:12445-12450.
15. Groh V, Steinle A, Bauer S, Spies T. 1998. Recognition of stress-induced MHC molecules by intestinal epithelial gammadelta T cells. Science. 279:1737-1740.
16. Groh V, Rhinehart R, Randolph-Habecker J, Topp MS, Riddell SR, Spies T. 2001. Costimulation of CD8alphabeta T cells by NKG2D via engagement by MIC induced on virus-infected cells. Nat Immunol. 2:255-260.
17. Cerwenka, A, Bakker, A.B, McClanahan, T, Wagner, J, Wu J, Phillips, J.H, Lanier, L.L. 2000. Retinoic acid early inducible genes define a ligand family for the activating NKG2D receptor in mice. Immunity. 12: 721-727.
18. Jinushi, M., Takehara, T, Tatsumi, T, Kanto, T, Groh, V,Spies, T., Kimura, R, Miyagi, T, Mochizuki, K., Sasaki,Y, Hayashi, N.2003. Expression and role of MICA and MICB in human hepatocellular carcinomas and their regulation by retinoic acid. Int. J. Cancer. 104: 354-361.
19. Wu, J, Chalupny, N.J, Manley, T.J, Riddell, S.R, Cosman,D, Spies, T. 2003. Intracellular retention of the MHC class l-related chain B ligand of NKG2D by the human cytomegalovirus UL16 glycoprotein. J. Immunol. 170:4196-4200.
20. Dunn, C, Chalupny, N.J., Sutherland, C.L, Dosch, S,Sivakumar, P.V, Johnson, D.C., Cosman, D. 2003. Human cytomegalovirus glycoprotein UL16 causes intra-cellular sequestration of NKG2D ligands, protecting against natural killer cell cytotoxicity. J. Exp. Med. 197:1427-1439.
21. Welte, S.A, Sinzger, C, Lutz, S.Z, Singh-Jasuja, H,Sampaio, K.L, Eknigk, U, Rammensee, H.G, Steinle,A. 2003. Selective intracellular retention of virally induced NKG2D ligands by the human cytomegalovirus UL16 glycoprotein. Eur. J. Immunol. 33:194-203.
22. 0deberg, J, Browne, H, Metkar, S., Froelich, C.J, Branden,L, Cosman, D, Soderberg-Naucler, C. 2003. The human cytomegalovirus protein UL16 mediates increased resistance to natural killer cell cytotoxicity through resistance to cytolytic proteins. J. Virol. 77: 4539-4545.
23. Vales-Gomez, M., Browne, H., Reyburn, H.T. 2003. Expression of the UL16 glycoprotein of Human Cytomegalovirus protects the virus-infected cell from attack by natural killer cells. BMC Immunol. 4, 4.
24. Lodoen MB, Abenes G, Umamoto S, Houchins JP, Liu F, Lanier LL. 2004. The cytomegalovirus m155 gene product subverts natural killer cell antiviral protection by disruption of H60-NKG2D interactions. J Exp Med. 200:1075-1081
25. Kubin, M, Cassiano, L, Chalupny, J, Chin, W, Cosman, D,Fanslow, W, Mullberg, J, Rousseau, A.M, Ulrich, D,Armitage, R. 2001. ULBP1, 2, 3: novel MHC class l-related molecules that bind to human cytomegalovirus glycoprotein UL16, activate NK cells. Eur. J. Immunol. 31:1428-1437.
26. Bauer, S, Groh, V, Wu, J, Steinle, A, Phillips, J.H, Lanier,L.L, Spies, T.1999. Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science. 285:727-729.
27. Braud VM, Allan DS, O'Callaghan CA, Soderstrom K, D'Andrea A, Ogg GS, Lazetic S, Young NT, Bell Jl, Phillips JH. 1998. HLA-E binds to natural killer cell receptors CD94/NKG2A, B and C. Natur. 391:795-799.
28. Braud VM, Allan DS, McMichael AJ. 1999. Functions of nonclassical MHC and non-MHC-encoded class I molecules. Curr Opin Immunol.11:100-108.
29. Andre P, Biassoni R, Colonna M, Cosman D, Lanier LL, Long EO, Lopez-Botet M, Moretta A, Moretta L, Parham P, et al. 2001. New nomenclature for MHC receptors. Nat Immunol. 2:661.
30. Diefenbach A, Tomasello E, Lucas M, Jamieson AM, Hsia JK, Vivier E, Raulet DH. 2002. Selective associations with signaling proteins determine stimulatory versus costimulatory activity of NKG2D. Nat Immunol. 3:1142-1149.
31. Wu J, Song Y, Bakker AB .1999. An activating immunoreceptor complex formed by NKG2D and DAP10. Science. 285 : 730-2.
32. Sutherland, C.L, Chalupny, N.J, Schooley, K, VandenBos,T, Kubin, M, Cosman, D, 2002. UL16-binding proteins, novel MHC class l-related proteins, bind to NKG2D and activate multiple signaling pathways in primary NK cells. J. Immunol. 168:671-679.
33. Hayday AC. 2000. [gamma][delta] cells: a right time and a right place for a conserved third way of protection. Annu Rev Immunol. 18:975-1026.
34. Li JQ, Cui LX, He W. 2005. Distinct Pattern of Human Vdelta1 gammadelta T Cells Recognizing MICA. Cell Mol Immuno. 2:253-258.
35. Groh V, Rhinehart R, Secrist H, Bauer S, Grabstein KH, Spies T. 1999. Broad tumor-associated expression and recognition by tumor-derived gamma delta T cells of MICA and MICB. Proc Natl Acad Sci U S A. 96:6879-6884.
36. Wu J, Groh V, Spies T. 2002. T cell antigen receptor engagement and specificity in the recognition of stress-inducible MHC class l-related chains by human epithelial gamma delta T cells. J Immunol. 169:1236-1240.
37. Poggi, A., C. Venturino, S. Catellani, M. Clavio, M. Miglino, M. Gobbi, A. Steinle, P. Ghia, S. Stella, F. Caligaris-Cappio, and M. R. Zocchi. 2004. Vdelta1 T lymphocytes from B-CLL patients recognize ULBP3 expressed on leukemic B cells and up-regulated by trans-retinoic acid. Cancer research. 64:9172-9179.
38. Catellani, S, A. Poggi, A. Bruzzone, P. Dadati, J. L. Ravetti, M. Gobbi, and M. R. Zocchi. 2007. Expansion of V{delta}1 T lymphocytes producing IL-4 in low-grade non-Hodgkin lymphomas expressing UL-16-binding proteins. Blood. 109:2078-2085.
39. P. Wrobel, H. ShojaeiB. Schittek F, Gieselera, B. Wollenberg? H. Kalthoff-, D. Kabelitz & D. Wesch. 2007. Lysis of a Broad Range of Epithelial Tumour Cells by Human cd T Cells: Involvement of NKG2D ligands and T-cell Receptor- versus NKG2D-dependent Recognition. Scandinavian Journal of Immunology. 66:20-328
40. Jamieson AM, Diefenbach A, McMahon CW, Xiong N, Carlyle JR, Raulet DH. 2002. The role of the NKG2D immunoreceptor in immune cell activation and natural killing. Immunity. 17:19-29.
41. Pende D, Rivera P, Marcenaro S, Chang CC, Biassoni R, Conte R, Kubin M, Cosman D, Ferrone S, Moretta L, et al. 2002. Major histocompatibility complex class l-related chain A and UL16-binding protein expression on tumor cell lines of different histotypes: analysis of tumor susceptibility to NKG2D-dependent natural killer cell cytotoxicity. Cancer Res.62:6178-6186.
42. Diefenbach A, Jensen ER, Jamieson AM, Raulet DH. 2001. Rae1 and H60 ligands of the NKG2D receptor stimulate tumour immunity. Nature. 413:165-171.
43. Kulski JK, Dunn DS, Hui J, Martinez P, Romphruk AV, Leelayuwat C, Tay GK, Oka A, Inoko H: 2002. Alu polymorphism within the MICB gene and association with HLA-B alleles. Immunogenetics. 53:975-979.
44.Spreu, J, T. Stehle, and A. Steinle. 2006. Human cytomegalovirus-encoded UL16 discriminates MIC molecules by their alpha2 domains. J Immunol 177:3143-3149.
45. Waldhauer, I, and A. Steinle. 2006. Proteolytic release of soluble UL16-binding protein 2 from tumor cells. Cancer research. 66:2520-2526.
46.Wei Cao, Xueyan Xi, Zhiyong Hao, Wenjing Li, Yan Kong, Lianxian Cui, Chi Ma, Wei He. 2007. RAET1E2, a soluble isoform of the UL16 binding protein RAET1E produced by tumor cells, inhibits NKG2D-mediated NK cytotoxicity. J Biol Chem.
47. Eisele, G., J. Wischhusen, M. Mittelbronn, R. Meyermann, I. Waldhauer, A. Steinle, M. Weller, and M. A. Friese. 2006. TGF-beta and metalloproteinases differentially suppress NKG2D ligand surface expression on malignant glioma cells. Brain. 129:2416-2425.
48. Zhou H, Luo Y, Lo JF, Kaplan CD, Mizutani M, Mizutani N, Lee JD, Primus FJ, Becker JC, Xiang R, et al. 2005. DNA-based vaccines activate innate and adaptive antitumor immunity by engaging the NKG2D receptor. Proc Natl Acad Sci U S A. 102:10846-10851.
49. Zhou H, Luo Y, Kaplan CD, Kruger JA, Lee SH, Xiang R, Reisfeld RA. 2006. A DNA-based cancer vaccine enhances lymphocyte cross talk by engaging the NKG2D receptor. Blood . 107:3251-3257.
50. Conejo-Garcia, J.R, Benencia, F, Courreges, M.C, Khang,E, Zhang, L, Mohamed-Hadley, A, Vinocur, J.M, Buckanovich, R.J, Thompson, C.B, Levine, B., Coukos,G. 2003. Letal, a tumor-associated NKG2D immunor-eceptor ligand, induces activation and expansion of effector immune cells. Cancer Biol. Ther. 2: 446-451.
51. Groh V, Bruhl A, El-Gabalawy H, Nelson JL, Spies T. 2003. Stimulation of T cell autoreactivity by anomalous expression of NKG2D and its MIC ligands in rheumatoid arthritis. Proc Natl Acad Sci U S A. 100:9452-9457.
52. Ogasawara K, Hamerman JA, Ehrlich LR, Bour-Jordan H, Santamaria P, Bluestone JA, Lanier LL. 2004. NKG2D blockade prevents autoimmune diabetes in NOD mice. Immunity. 20:757-767.