肝脏微环境对肝脏NK和NKT细胞发育的影响
摘要
NK细胞和NKT细胞前体来源于骨髓细胞,定向分化至自然杀伤细胞谱系,并发育成具有效应功能和异质性表型的成熟的NK细胞和NKT细胞。骨髓,胚胎胸腺,淋巴结,肝脏,脾脏和外周血中都可以检测到自然杀伤细胞前体；但是不成熟自然杀伤细胞(iNK)只能在骨髓,肝脏和脾脏中被检测到。这些发育中间体是如何离开骨髓在外周位点如肝脏、脾脏中完成发育成熟的过程仍然未知。肝脏中存在不成熟的NK细胞和NKT细胞。肝脏中不成熟的NK细胞具有特殊的表型,即持续性表达肿瘤坏死因子相关凋亡诱导配体(tumour necrosis factor-related apoptosis-inducing ligand, TRAIL),但是低水平表达其他成熟自然杀伤细胞表面标志,例如Ly49受体和CD11b.胸腺中不成熟NKT细胞不表达NK1.1标志,在外周经历发育分化成熟后表达NKl.1。肝脏是外周NKT细胞发育成熟的重要场所。在NK细胞和NKT细胞发育分化过程中,不同组织脏器中基质细胞通过分泌可溶性细胞因子等信号分子来影响其表型和功能。本研究主要使用流式细胞术探索了小鼠个体发生过程中肝脏中NK细胞的发育并且与脾脏NK细胞发育进行了比对。Y干扰素对肝脏NK细胞亚群分布以及功能的影响做了初步的研究。
我们发现,CD27-CD11b-NK细胞前体主要聚集在成年小鼠肝脏而不是脾脏中小鼠个体发生时,在肝脏中,存在更多的不成熟的NK细胞,表达高水平的NKG2A和低水平的Ly49受体。肝脏中NK细胞表面刺激性受体和粘附分子的表达也与脾脏NK细胞不同。这些结果显示了肝脏内存在特异的NK细胞发育途径,并且环境微环境在NK细胞发育分化过程中起了重要作用。我们揭示出成年Y干扰素缺陷小鼠肝脏内成熟NK细胞的比例和数目都显著减少了。成年γ干扰素缺陷小鼠肝脏内聚集了特异的NK细胞亚群和表型。然而,成年Y干扰素缺陷小鼠肝脏和骨髓内Lin-CD122+NK前体细胞比例和数目都是正常的。进一步研究发现,γ干扰素缺陷导致小鼠肝脏内NK细胞活化分子CD69和功能分子的表达减弱。这些结果揭示了γ干扰素在维持肝脏独特微环境,影响肝脏NK细胞发育中起了重要作用。另外,我们发现肝脏中存在不成熟的NKT细胞亚群。对于小鼠个体发生时肝脏中NKT细胞发育阶段及亚群变化做了初步探索。
The precursors of NK cells and NKT cells are derived from the bone marrow;they are committed to the NK cell and NKT cell lineage and develop into mature NK cells with full effector function and heterogeneous phenotypes.NK cell precursors are found in different organs, such as bone marrow, fetal thymus, lymph node (LN), liver, spleen and peripheral blood, whereas immature NK (iNK) cells are found in the bone marrow, liver and spleen. It is unknown whether these developmental intermediates leave the bone marrow to complete their differentiation elsewhere, such as the liver and spleen.Immature NKT cells of thymus do not express NK1.1.They undergo mature process in periphery and then express NK1.1.As an lymphoid organ, liver is an important site for NKT cells maturing in periphery. Stromal cells in various organs send signals through cytokines that influence the ultimate phenotypes and functions of NK cell precursors.In our study, the development of NK cells in liver was explored and compared with NK cell development in spleen during mouse ontogeny by Flow cytometry.We also explored the impact of IFN-gamma on the distribution and function of hepatic NK cells.
The CD27-CD11b-NK cell precursors accumulated predominantly in the adult liver and not in the spleen. In the liver, more immature NK cells were present, which express a higher level of NKG2A and lower levels of Ly49receptors. Additionally, different stimulatory receptors and adhesion molecules were expressed on NK cells in the liver and spleen during ontogeny. These results indicate that there might be a specific developmental pathway of NK cells in the liver and that the microenvironments play important roles in NK cell development and differentiation. Futhermore, we exhibited that the percentages and numbers of mature NK cells were declined significantly in the liver from adult IFN-gamma deficient mouse GKO mouse uniquely. Moreover, the NK cell subsets and phenotypes were specifically in the liver of IFN-gamma deficient mouse. However, Lin-CD122+NK progenitor cells develop normally in the liver and bone marrow of adult IFN-gamma deficient mouse. Furthermore, deficiency of IFN-gamma resulted to weak expressed of CD69and functional molecular by hepatic NK cells. All these data demonstrated that IFN-gamma may play important role to sustain the unique environment of liver and affect the development of hepatic NK cells.Futhermore, we found that there were a subset immature NKT cells in the liver, and we explored the developmental stages and subsets of hepatic NKT cells during mouse ontogeny.
我们发现,CD27-CD11b-NK细胞前体主要聚集在成年小鼠肝脏而不是脾脏中小鼠个体发生时,在肝脏中,存在更多的不成熟的NK细胞,表达高水平的NKG2A和低水平的Ly49受体。肝脏中NK细胞表面刺激性受体和粘附分子的表达也与脾脏NK细胞不同。这些结果显示了肝脏内存在特异的NK细胞发育途径,并且环境微环境在NK细胞发育分化过程中起了重要作用。我们揭示出成年Y干扰素缺陷小鼠肝脏内成熟NK细胞的比例和数目都显著减少了。成年γ干扰素缺陷小鼠肝脏内聚集了特异的NK细胞亚群和表型。然而,成年Y干扰素缺陷小鼠肝脏和骨髓内Lin-CD122+NK前体细胞比例和数目都是正常的。进一步研究发现,γ干扰素缺陷导致小鼠肝脏内NK细胞活化分子CD69和功能分子的表达减弱。这些结果揭示了γ干扰素在维持肝脏独特微环境,影响肝脏NK细胞发育中起了重要作用。另外,我们发现肝脏中存在不成熟的NKT细胞亚群。对于小鼠个体发生时肝脏中NKT细胞发育阶段及亚群变化做了初步探索。
The precursors of NK cells and NKT cells are derived from the bone marrow;they are committed to the NK cell and NKT cell lineage and develop into mature NK cells with full effector function and heterogeneous phenotypes.NK cell precursors are found in different organs, such as bone marrow, fetal thymus, lymph node (LN), liver, spleen and peripheral blood, whereas immature NK (iNK) cells are found in the bone marrow, liver and spleen. It is unknown whether these developmental intermediates leave the bone marrow to complete their differentiation elsewhere, such as the liver and spleen.Immature NKT cells of thymus do not express NK1.1.They undergo mature process in periphery and then express NK1.1.As an lymphoid organ, liver is an important site for NKT cells maturing in periphery. Stromal cells in various organs send signals through cytokines that influence the ultimate phenotypes and functions of NK cell precursors.In our study, the development of NK cells in liver was explored and compared with NK cell development in spleen during mouse ontogeny by Flow cytometry.We also explored the impact of IFN-gamma on the distribution and function of hepatic NK cells.
The CD27-CD11b-NK cell precursors accumulated predominantly in the adult liver and not in the spleen. In the liver, more immature NK cells were present, which express a higher level of NKG2A and lower levels of Ly49receptors. Additionally, different stimulatory receptors and adhesion molecules were expressed on NK cells in the liver and spleen during ontogeny. These results indicate that there might be a specific developmental pathway of NK cells in the liver and that the microenvironments play important roles in NK cell development and differentiation. Futhermore, we exhibited that the percentages and numbers of mature NK cells were declined significantly in the liver from adult IFN-gamma deficient mouse GKO mouse uniquely. Moreover, the NK cell subsets and phenotypes were specifically in the liver of IFN-gamma deficient mouse. However, Lin-CD122+NK progenitor cells develop normally in the liver and bone marrow of adult IFN-gamma deficient mouse. Furthermore, deficiency of IFN-gamma resulted to weak expressed of CD69and functional molecular by hepatic NK cells. All these data demonstrated that IFN-gamma may play important role to sustain the unique environment of liver and affect the development of hepatic NK cells.Futhermore, we found that there were a subset immature NKT cells in the liver, and we explored the developmental stages and subsets of hepatic NKT cells during mouse ontogeny.
引文
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