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.Liu Y and Mullbacher A.Hypothesis:immunological help is reciprocally delivered between different subpopulations of lymphocytes.Scand j Immunol,1989;30:277

　　33.Lowenthal JM,et al.Binding and internalization of interleukin 1 by Tcells direct evidence for high and low affinity classes of interleukin-1receptor.J Exp Med 1986;164:1060

　　34.Moore KW,et al.Homology of cytokine synthesis inhibtory factor(IL-10)to the Epstein-Barr virus gene BCRFI.Science,1990;248:1230

　　35.Morgan RA,et al.Selective in vitro growth of t lymphocytes.Science,1976;193:1007

　　36.Murphy PM.The molecular biology of leukocyte chemoattractant receptors.Annu Rev Immunol,1994;12:593

　　37.Musash M,et al.Direct and synergistic effects of interleukin 11 on murine hemopoiesis in culture.PNAS,1991;88:765

　　38.NacNeil IA,et al.IL-10,A novel growth factor for mature and immatureT cell.J Immunol,1990;145:4167

　　39.Namen AE,et al.Stimulation of B-cell progenitors by cloned murine interleukin 7.Nature,1988;33:571

　　40.Paul SR,et al.Molecular cloning of a cDNA encoding interleukin 11,a stromal cell-derived lymphopoietic and hematopoietic cytokine.PNAS,1990;87:7512

　　41.Pizzolo G,et al.Soluble interleukin 2 receptor in the serum of patients with Hodgkin's disease.Br J Cancer 1987;55:427

　　42.Ralop P.A summary of cytokine aspects of the intermational symposium of molecular hematology.1990;Lymphokine and Cytokine research,1991;10(1):77

　　43.Rot A.Endothelial cell binding of NAP-1/IL-8 role in neutrophil emigration.Immunol Today 1992;13(8):291

　　44.Rubin LA,et al.The soluble interlenkin-2 receptor biology,function and clinical application.Annals of Internal.Medicine 1990;113(8):619

　　45.Rubin LA,et al.Soluble interleukin 2 receptors are released from activated human　lymphoid cells in vitro.J Immunol.1985;135:3172

　　46.Sims JE,et al.cDNA expression cloning of the IL-1 receptor,a member of the immunoglobulin superfamily.Science 1988;241:585

　　47.Smith KA.Interleukin-2:inception,impact and implications.Science,

　　1988;240:1169

　　48.Snouwaert JN,et al.Effects of site-specific mutations on biologic activity of recombinant human IL-6.J Immunol,1991;146(2):585

　　49.Stern AS,et al.Purification to homogencity and　partial characterization of cytotoxic lymphocyte maturation factor from human B-lypmhoblastoid cell.PNAS,1990;87:6808

　　50.Stotter H,et al.IL-7induces human lymphokine-activated killer cell activity and is regulated by IL-4.J Immunol 1991;146(1):150

　　51.Takaki S,et al.Molecular cloning and expression of the murine interleukin-5 receptor.EMBO,1990;(13):4367

　　52.Taub DD and Oppenheim JJ.Review of the chemokine meeting the third international symposium of chemotactic cytokines.Cytokine,1993;5(3):175

　　53.Uckun FM,et al.Interleukin 7 receptor ligation stimulates tyrosine phosphorylation,inositol phospholipid turnover,and clonal proliferation ofhuman B-cell precursors.PNAS,1991;88:3589

　　54.Uyttenhove C,et al.Functional and structural characterization of P40,a mouse glycoprotein with T cell-growth factor activity.PNAS,1988;85:6934

　　55.William E,et al.Structure and functional expression of a human interleukin-8 receptor.Science,1991;253:1278

　　56.Wong HL,et al.Characterization of a factor(s) which synergizes with recombinant interleukin 2 promoting allogenic human cytolytic T-lymphocyte responses in vitro.Cell Immunol,1988;11:39

　　57.Xing X and Yong SC:Human recombinant IL-3 is a growth factor for normal B cells.J Immunol,1992;148(2):491

　　58.Yin T,et al.Involvement of IL-6 signal transducer gp130 in IL-11-mediated signal transduction.J Immunol,1993;151:2555

　　59.侯云德.干扰素的不同亚型与临床应用。中国生物物品学杂志1991；6（4）：145

　　60.曹广文、杜平.生药活性多糖对淋巴细胞/细胞素的调节与抗肿瘤作用。免疫学杂志，1992；8（4）：273

　　61.孙红、施渭康.白血病抑制因子.细胞生物学，1994；16（2）49

　　62.段海清、黄翠芳.抑瘤素M研究进展.因外医学免疫学分册，1994；17（4）：185

　　63.朱学军、曹雪涛.肿瘤坏死因子拮抗剂的种类性质及其治疗作用.国外医学免疫分册，1994；17（1）：21

　　64.舒翠玲、金伯泉.白细胞介素12（IL-12）的研究概况.免疫学杂志，1992；8（2）：133

　　65.孙忱、金伯泉.可溶性细胞因子受体研究进展.国外医学免疫学分册，1994；17（2）：70

　　66.孙凯、金伯泉.细胞因子受体中的共用链现象及信号转导

　　67.黄传书、金伯泉.IL-3、IL-5和GM-CSF受体研究的进展。国外医学免疫学分册，1995；18（3）：121

　　68.张新海、金伯泉. 趋化因子研究进展-趋化因子家族（一）。国外医学免疫学分册，1995；18（3）：130