刺参体腔细胞表面抗原分类及体壁组织中免疫细胞的研究
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摘要
为检测刺参Apostichopus japonicus体腔细胞表面抗原的分类,确定其体壁组织中免疫细胞的定位,选取体质量为(160±10)g的健康刺参,采用流式细胞术(Flow cytometry, FCM)技术对刺参体腔细胞表面抗原CD类似物类型(CD3、CD4、CD7、CD8、CD11b、CD14、CD38)进行了检测,并使用免疫组化法对刺参体壁组织中的免疫细胞进行初步定位研究。结果表明:直径大于和小于5μm的体腔细胞,表面抗原CD7和CD14类似物均呈阳性,刺参体壁结缔组织经CD7、CD14类似物免疫组化染色后,可见分布在结缔组织细胞间隙中褐色阳性细胞,认为该两种表面抗原呈阳性。研究表明,在刺参体腔和体壁细胞中均能检测到CD7及CD14类似物,提示CD7和CD14类似物在免疫机能上具有T细胞和NK细胞的免疫学功能。
CD analog types(CD3, CD4, CD7, CD8, CD11 b, CD14, and CD38) on surface antigens on coelomocytes were investigated by flow cytometry(FCM) and localization was immunohistochemistrically carried out on immune cells in body walls in sea cucumber Apostichopus japonicus with body weight of(160±10)g in order to detect the classification of surface antigens on coelomocytes and localization of immune cells in body walls. It was found that CD7 analogues and CD14 analogues on coelomocytes with diameter of > and < 5 μm were concerned as positive surface antigens, and the positive cells CD7 and CD14 stained brown distributed in the connective tissue were observed. This finding indicated that CD7 and CD14 analogs were explored in coelomocytes and somatic tissue cells, and had immunological functions of T cells and NK cells, which provides theoretical supports for immune evolution among organisms.
CD analog types(CD3, CD4, CD7, CD8, CD11 b, CD14, and CD38) on surface antigens on coelomocytes were investigated by flow cytometry(FCM) and localization was immunohistochemistrically carried out on immune cells in body walls in sea cucumber Apostichopus japonicus with body weight of(160±10)g in order to detect the classification of surface antigens on coelomocytes and localization of immune cells in body walls. It was found that CD7 analogues and CD14 analogues on coelomocytes with diameter of > and < 5 μm were concerned as positive surface antigens, and the positive cells CD7 and CD14 stained brown distributed in the connective tissue were observed. This finding indicated that CD7 and CD14 analogs were explored in coelomocytes and somatic tissue cells, and had immunological functions of T cells and NK cells, which provides theoretical supports for immune evolution among organisms.
引文
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[4] 张峰,张莉,倪慧,等.仿刺参体腔细胞CD35的化学发光免疫检测[J].大连海洋大学学报,2011,26(2):176-179.
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[14] 沙爱龙.流式细胞仪在贝类研究中的应用[J].南方水产科学,2007,3(6):71-74.
[15] 张东华,李登举,冉丹,等.外周血干细胞移植前后淋巴细胞免疫表型变化[J].中国现代医学杂志,2003,13(24):44-46.
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[18] Kim D,Kim J Y.Anti-CD14 antibody reduces LPS responsiveness via TLR4 internalization in human monocytes[J].Molecular Immunology,2014,57(2):210-215.
[19] 童裳亮.海洋动物的细胞培养与应用[J].生物工程进展,1994,14(6):47-48.
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[22] Fyhn U E H,Costlow J D.Tissue cultures of cirripeds[J].The Biological Bulletin,1975,149(2):316-330.
[23] 常亚青,丁君,宋坚,等.海参、海胆生物学研究与养殖[M].北京:海洋出版社,2004.
[24] 聂竹兰,李霞,辛涛.仿刺参体壁的组织学和组织化学[J].大连海洋大学学报,2007,22(3):184-187.
[2] 武明欣,王雅平,李培玉,等.饲料中添加木聚糖酶对刺参幼参生长、消化和体腔液酶活力的影响[J].大连海洋大学学报,2018,33(3):329-335.
[3] 陈庭家,田景玉,丁月,等.刺参体腔细胞类型及其死亡率与温度的关系[J].大连工业大学学报,2016,35(6):407-410.
[4] 张峰,张莉,倪慧,等.仿刺参体腔细胞CD35的化学发光免疫检测[J].大连海洋大学学报,2011,26(2):176-179.
[5] Ashton-Alcox K A,Ford S E.Variability in molluscan hemocytes:a flow cytometric study[J].Tissue and Cell,1998,30(2):195-204.
[6] 尹洪滨,孙中武,孙大江,等.俄罗斯鲟的细胞遗传学分析[J].水产学报,2006,30(2):181-184.
[7] 张之晟,董成稳,赵俊,等.基于DNA含量的复合鲫倍性分析[J].华南师范大学学报:自然科学版,2006(1):99-103.
[8] 汪开毓,耿毅,黄小丽.用流式细胞术检测喹乙醇诱导的鲤肝细胞凋亡[J].中国水产科学,2005,12(5):648-651.
[9] Fisher W S.Flow cytometry:a tool for cell research in bivalve pathology[M]//Fisher W S.Disease Processes in Marine Bivalve Molluscs.Bethesda:American Fisheries Society,1988.
[10] Goedken M,De Guise S.Flow cytometry as a tool to quantify oyster defence mechanisms[J].Fish & Shellfish Immunology,2004,16(4):539-552.
[11] 石芳芳,李成华,宋林生,等.用流式细胞仪测定扇贝血细胞吞噬活性[J].生物技术通报,2006(S1):430-433.
[12] 崔巍,朱陵群,赵明镜,等.淋巴细胞免疫表型检测及其统计学分析[J].中国免疫学杂志,2001,17(10):538-540.
[13] Lin Wenyu,Zhang Haiyan,Beck G.Phylogeny of natural cytotoxicity:cytotoxic activity of coelomocytes of the purple sea urchin,Arbacia punctulata[J].Journal of Experimental Zoology,2001,290(7):741-750.
[14] 沙爱龙.流式细胞仪在贝类研究中的应用[J].南方水产科学,2007,3(6):71-74.
[15] 张东华,李登举,冉丹,等.外周血干细胞移植前后淋巴细胞免疫表型变化[J].中国现代医学杂志,2003,13(24):44-46.
[16] Milush J M,Long B R,Snyder-Cappione J E,et al.Functionally distinct subsets of human NK cells and monocyte/DC-like cells identified by coexpression of CD56,CD7,and CD4[J].Blood,2009,114(23):4823-4831.
[17] Zanoni I,Granucci F.Role of CD14 in host protection against infections and in metabolism regulation[J].Frontiers in Cellular and Infection Microbiology,2013,3:32.
[18] Kim D,Kim J Y.Anti-CD14 antibody reduces LPS responsiveness via TLR4 internalization in human monocytes[J].Molecular Immunology,2014,57(2):210-215.
[19] 童裳亮.海洋动物的细胞培养与应用[J].生物工程进展,1994,14(6):47-48.
[20] 顾立刚.医学免疫学与微生物学[M].北京:中国中医药出版社,2004.
[21] Huang Y S,Rousseau K,Le Belle N,et al.Opposite effects of insulin-like growth factors(IGFs) on gonadotropin(GtH-II) and growth hormone(GH) production by primary culture of European eel(Anguilla anguilla) pituitary cells[J].Aquaculture,1999,177(1-4):73-83.
[22] Fyhn U E H,Costlow J D.Tissue cultures of cirripeds[J].The Biological Bulletin,1975,149(2):316-330.
[23] 常亚青,丁君,宋坚,等.海参、海胆生物学研究与养殖[M].北京:海洋出版社,2004.
[24] 聂竹兰,李霞,辛涛.仿刺参体壁的组织学和组织化学[J].大连海洋大学学报,2007,22(3):184-187.