α-Galcer尾根皮下注射对小鼠体内不同器官中iNKT细胞频率及亚群的影响

doi:10.3969/j.issn.1674-490X.2018.02.003

摘要/Abstract

摘要： 目的探究单次和多次尾根部皮下注射α-Galcer对小鼠胸腺、脾脏和肝脏中恒定自然杀伤T细胞(invariant nature killer T cell,iNKT)频率和亚群的影响。方法 DBA/1小鼠随机分为三组:健康对照组(尾根部皮下注射0.3 mL PBS溶液)、单次注射α-Galcer组(尾根部皮下注射,2 μg/只)、多次注射α-Galcer组(尾根部皮下注射,3次,每次间隔48 h,2 μg/只)。FACS检测iNKT细胞频率及亚群。结果(1)与健康对照组相比,单次及多次尾根部皮下注射α-Galcer后,小鼠胸腺、脾脏、肝脏中iNKT频率均显著提高(P<0.05)单次注射α-Galcer组小鼠胸腺和脾脏中的iNKT频率峰值显著高于多次注射α-Galcer组iNKT频率最大值(P<0.05)。(2)胸腺:α-Galcer组iNKT1亚群的频率在第2天已明显高于健康对照组(P<0.05),至第4天时达最大值;iNKT2亚群的频率始终显著高于健康对照组(P<0.05),第8天达最大值,之后下降。脾脏:α-Galcer组iNKT1亚群的频率以健康对照值为轴,呈现先抑后扬的的趋势,第4天降至低谷,第6天升至高峰;相反,iNKT2亚群的频率,始终高于健康对照组,在第4天上升达峰值,随后迅速下降,在第6天达最小值。肝脏:α-Galcer组iNKT1亚群的频率始终低于健康对照组,呈现逐渐增高的趋势;iNKT2亚群的频率围绕健康对照值呈现小幅升降,在第6天下降到最小值,又在第8天上升至最大值(10.13%±0.03%)。结论单次及多次尾根部皮下注射α-Galcer均能诱导体内iNKT细胞增殖,但单次注射诱导的iNKT频率更高;单次尾根部皮下注射α-Galcer可诱导iNKT亚群的变化,且各器官中的亚群变化趋势有较大差异。

关键词: α-Galcer, iNKT细胞亚群, 尾根部皮下注射

Abstract: Objective To investigate the effects of single and multiple subcutaneous injection of α-Galcer on invariant nature killer T cell(iNKT cell)frequency and subpopulation in thymus, spleen and liver of mice investigate the effects of subcutaneous injection of alpha-Galcer. Methods DBA/1 mice were randomly divided into three groups: healthy control group(subcutaneous injection of 0.3 mL PBS solution in the caudal root), single injection of α-Galcer group(subcutaneous injection at the base of the tail, 2 μg/mouse), multiple injection of α-Galcer group( subcutaneous injection at the base of the tail, 3 times, eachtime: 48 h, 2 μg/mouse), FACS detection of iNKT cell frequency and subgroup. Results(1)Compared with the healthy control group, the iNKT frequencies in the thymus, spleen and liver of mice were significantly increased(P<0.05)after single and multiple subcutaneous injection of α-Galcer at the base of tail. The peak value of iNKT in thymus and spleenof single injection of α-Galcer was significantly higher than that of multiple injection of α-Galcer(P<0.05).(2)Thymus: The frequency of iNKT1 subpopulation in α-Galcer group was significantly higher than that in healthy control group(P<0.05)on the 2nd day and reached the maximum on the 4 th day. The frequency of iNKT2 subpopulation was always significantly higher than that of healthy control Group(P<0.05)and reaching the maximum on the 8th day then decreased. Spleen: In the α-Galcer group, the frequency of iNKT1 sub-group showed a tendency of first-appending and then-descending, taking the healthy control value as the axis, dropping to the trough on the 4th day and reaching the peak on the 6th day; on the contrary, the frequency of iNKT2 subpopulation was always higher than that of the healthy control group, reaching the peak on the 4th day, then dropping rapidly and reaching the minimum on the 6th day. Liver: The frequency of iNKT1 subgroup in α-Galcer group was always lower than that of the healthy control group, showing a gradually increasing trend; The frequency of iNKT2 subpopulations showed a slight rise and fall around the healthy controls, falling to a minimum on the 6th day and reaching a maximum on the 8th day. Conclusion Both single and multiple subcutaneous injected at the base of the tail of alpha-Galcer can induce proliferation of iNKT cells in vivo, but the frequency of iNKT cells of single injected by alpha-Galcer is higher; single injectied of alpha-Galcer can induce iNKT subsets changes,and the variation trend of subgroups in each organ are quite different, which provides the basic data for the study on the treatment of diseases related to iNKT cells.

Key words: alpha-Galcer, iNKT subgroup, subcutaneous injection

中图分类号:

王园园,叶闽,张静雯,崔卓,李子琦,刘慧芳,孟明. α-Galcer尾根皮下注射对小鼠体内不同器官中iNKT细胞频率及亚群的影响[J]. 医学研究与教育, 2018, 35(2): 17-25.

参考文献

[1] TANIGUCHI M, NAKAYAMA T. Recognition and function of Valpha14 NKT cells[J]. Semin Immunol, 2000,12(6):543-550. DOI:10.1006/smim.2000.0270.
[2] BENDELAC A. Positive selection of mouse NK1+ T cells by CD1-expressing cortical thymocytes[J]. J Exp Med,1995,182(6):2091-2096.
[3] COLES M C, RAULET D H. NK1.1+ T cells in the liver arise in the thymus and are selected by interactions with class I molecules on CD4+CD8+ cells[J].J Immunol,2000, 164(5):2412-2418.
[4] GODFREY D I, MACDONALD H R, KRONENBERG M, et al. NKT cells: what’s in a name[J].Nat Rev Immunol,2004, 4(3):231-237. DOI:10.1038/nri1309.
[5] BENLAGHA K, KYIN T, BEAVIS A, et al. A thymic precursor to the NK T cell lineage[J].Science,2002, 296(5567):553-555.
[6] PELLICCI D G, HAMMOND K J, ULDRICH A P, et al. A natural killer T(NKT)cell developmental pathway involving a thymus-dependent NK1.1(-)CD4(+)CD1d-dependent precursor stage[J]. J Exp Med,2002,195(7):835-844. DOI:10.1084/jem.20011544.
[7] TUDHOPE S J, VON DELWIG A, FALCONER J,et al. Profound invariant natural killer T-cell deficiency in inflammatory arthritis[J]. Ann Rheum Dis, 2010,69(10):1873-1879. DOI:10.1136/ard.2009.125849.
[8] TSAIH S W, PRESA M, KHAJA S, et al. A locus on mouse chromosome 13 inversely regulates CD1d expression and the development of invariant natural killer T-cells.[J]. Genes & Immunity, 2015, 16(3):221-230. DOI:10.1038/gene.2014.81.
[9] VAN K L, PAREKH V V, WU L. Invariant natural killer T cells as sensors and managers of inflammation[J]. Trends in Immunology, 2013, 34(2):50-58. DOI:10.1016/j.it.2012.08.009.
[10] AGUIAR C F, NAFFAH-DE-SOUZA C, CASTOLDI A, et al. Administration of α-Galactosylceramide Improves Adenine-Induced Renal Injury[J]. Molecular Medicine, 2015, 21:553. DOI:10.2119/molmed.2014.00090.
[11] FICHTNER A S, PALETTA D, STARICK L, et al. Function and expression of CD1d and invariant natural killer T-cell receptor in the cotton rat(Sigmodon hispidus)[J]. Immunology, 2015, 146(4):618. DOI:10.1111/imm.12532.
[12] MASANOBU H, DAISUKE G, SEIJI S, et al. Activation of Invariant NKT Cells with Glycolipid Ligand α-Galactosylceramide Ameliorates Glucose-6-Phosphate Isomerase Peptide-Induced Arthritis[J]. Plos One, 2012, 7(12):e51215. DOI:10.1371/journal.pone.0051215.
[13] BARTHELEMY A, IVANOV S, HASSANE M, et al. Exogenous Activation of Invariant Natural Killer T Cells by α-Galactosylceramide Reduces Pneumococcal Outgrowth and Dissemination Postinfluenza[J]. Mbio, 2016, 7(6):e01440-16. DOI:10.1128/mBio.01440-16.
[14] YANG J Q, KIM P J, SSINGH R R. Brief treatment with iNKT cell ligand α-galactosylceramide confers a long-term protection against lupus[J]. Journal of Clinical Immunology, 2012, 32(1):106. DOI:10.1007/s10875-011-9590-y.
[15] TIFFANY C, VEENA K, SHUYANG YU, et al. The transcription factor lymphoid enhancer factor 1 controls invariant natural killer T cell expansion and Th2-type effector differentiation[J].Journal of Experimental Medicine, 2015, 212(5):793-80. DOI:10.1084/jem.20141849.
[16] BRENNAN P J, BRIGL M, BRENNER M B. Invariant natural killer T cells: an innate activation scheme linked to divers[J]. Nature Reviews Immunology, 2013,13(2): 101-117. DOI:10.1038/nri3369.
[17] KRIJGSMAN D, HOKLAND M, KUPPEN P J. The Role of Natural Killer T Cells in Cancer-A Phenotypical and Functional Approach[J]. Front Immunol, 2018,9:367.DOI:10.3389/fimmu.2018.00367.
[18] CORTESI F, DELFANTI G, GRILLI A, et al. Bimodal CD40/Fas-Dependent Crosstalk between iNKT Cells and Tumor-Associated Macrophages Impairs Prostate Cancer Progression[J]. Cell Rep, 2018, 22(11):3006-3020. DOI:10.1016/j.celrep.
[19] NEGISHI Y, ICHIKAWA T, TAKESHITA T, et al. Miscarriage induced by adoptive transfer of dendritic cells and invariant natural killer T cells into mice[J]. Eur J Immunol, 2018. DOI:10.1002/eji.201747162.
[20] THOMAS M S, Zúñiga-Pflücker J C. Induction of T Cell Development from Hematopoietic Progenitor Cells by Delta-like-1 In Vitro[J]. Immunity,2002,17(6):749-756.
[21] LEE Y J, WANG H, STARRETT G J, et al. Tissue-Specific Distribution of iNKT Cells Impacts Their Cytokine Response[J]. Immunity, 2015, 43(3):566. DOI:10.1016/j.immuni.2015.06.025.
[22] ONISHA P, GARTH C, DANIEL G P, et al. NKT TCR Recognition of CD1d -a- Galactosylceramide[J]. Journal of Immunology, 2011, 187: 1-10. DOI:10.4049/jimmunol.1100794.
[23] CHANDRA S,KRONENBERG M. Activation and Function of iNKT and MAIT Cells[J]. Adv Immunol,2015,127:145-201.