核酸标记技术检测法

 

四、 核酸标记技术检测法

　　通过核酸标记技术可将细胞因子cDNA作为基因探钊检测细胞内细胞因子 基因组 DNA或mRNA。主要有以下几种方法：

1.应用同位素（或非同位素）标记的cDNA探针，检测经Northern blot后细胞因子mRNA水平或采用打点杂交法。

2.应用标记cDNA探钊与细胞或组织切片进行原位杂交，然后进行放射自显影。

3.细胞因子mRNA经反转录为cDNA，用特异性细胞因子引物经聚合酶链反应（PCR）扩增细胞因子cDNA,Southern blot后用标记探针检测特异细胞因子DNA水平。

（金伯泉）

参 考 文 献

1.Ared WP,et al.An IL-1 inhibitor from human monocytes production and characterization of biologic properties.J Immunol,1980;143:1851

2.Baggiolini M and Dahinden CA.CC chemokines in allergic inflammation.Immunol Today 1994;15(3):127

3.Balkwill FR and　Burke F.The cytokine network.Immunol Today,1989;10(9):299

4.Bazan JF.Haemopoietic receptors and helical cytokines .Immunol Today,1990;11(10):350

5.Cannon J G,et al.Circulating interleukin-1 and tumor necrosis of tumors.PNAS,1975;72:3666

7.Chan SH,et al.Induction of interferon γ production by natural killer cell stimulatory factor:characterization of the responder cells and synergy with other inducers.J Exp Med,1991;173:869

8.Chen WF,et al.IL-10:A novel cytotoxic T cell diffeentiation factor.J Immunol,1991;147(2):528

9.Cosman D,et al.A new cytokine receptor superfamily.TIBS,1990;15-July:265

10.Dinarello CA.Interleukin 1 and interleukin-1antagonism.Blood,1991;77(8):1627

11.Dower SK,et al.Molecular characterization of two types of interleukin-1 recep-tor coding peptide on murine and human cells.J Leukoc Biol(suppl.),1990;1:103

12.Durum SK and Mealy K.Hilton head revisited-cytokine explosion of the80s takes shape for the 90s.Immunol Today,1990;11(4):103

13.Durum SK,et al.New cytokines and receptors nake their debut in SanAntonio.Immunol Today,1991;12(2):54

14.Edward A,et al.How B and　Tcells talk to each other.Nature,1994;367:425

15.Eisenberg SP.et al.Primary structure and funtional expression from complementary DNA of a human inteleukin 1 receptor antagonist Nature,1990;3:341

16.Engelmann H,et al.A Tumor necrosis factor binding protein purified to homogeneity from human urine protects cells from tumor necrosis factor toxicity.J Biol Chem 1989;264:11974

17.Fiorentino DF,et al.IL-10 acts on the antigen-presenting cell to inhibit cytokine production by Th1 cells.J Immunol,1991;146(10):3444

18.Fong Y,et al. Antibodies to cachetin/TNF reduce IL-1β and IL-6 appearance during lethal bacteremia J Exp Med 1989;170:1627

19.Gasson JC.Molecular physiology of granulocyte-macrophage colony-stimulating factor.Blood,1991;77(6):1131

20.Gately MK,et al.Synergy between recombinant interleukin 2(r IL-2)and IL-2 depleted lymphokine containing supernatants in facilitating allogeneic human cytolytic T lymphocyte responses in vitro.j Immunol,1986;136:1274

21.Gately MK,et al.Regulation of human lymphocyte proliferation by a heterodimeric cytokine,IL-12(cyotoxic lymphocyte maturation factor).j Immunol,1991;147:874

22.Gubler V,et al.Coexpression of two distinct genesis requird to generate secreted bioactive cytotoxic lymphocyte maturation factor.PNAS,1991;88:4143

23.Hayashida K,et al.Molecular cloning of a second subunit of the receptor for human granulocyte-macrophage colony-stimulating factor(GM-CSF):reconstitution of a high-affinity GM-CSf receptor.PNAS,1990;87:9655

24.Himeno T,et al.Expression of endogenous tumor necrosis factor.Cancer Res,1990;50:4941

25.Hirano T,et al.Biological and clinical aspects of interleukin 6.Immunol Today,1990;11(12):443

26.Horuk R.The interleukin-8-receptor family:from chemokines to malaria.Immunol Today,1994;15(4):

27.Humnum CH et al.Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor.Nature,1990;343:336

28.James K.Interaction between cytokines and α2macroglobulin.Immunol Today,1990;11(50:163

29.Kishimoto T.The biology of interleukin-6.Blood,1989;74:1

tory factor(NKSF),a cytokine with multiple biologic effects on human lymphocytes.J Exp Med 1989;170:827

31.Krantz S.Erythropoietin.Blood,1991;77(3):419

32.Li