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目的通过分子佐剂C3d3增强人绒毛膜促性腺激素(hCG)β基因疫苗在不同品系小鼠的体液免疫效应。方法分别用pCMV4-hCG-βC3d3、pCMV4-hCGβ和pCMV4-hCGβ+pCMV4-C3d3联合免疫以辅助性T细胞(Th)2型优势的BALB/c小鼠和Th1型优势的C57BL/6小鼠,免疫剂量分别为5、10、50、100和500 pmol,同一剂量免疫两次、间隔3周。初次及加强免疫后每周采血,间接酶联免疫吸附试验(ELISA)测定血清抗hCGβ抗体水平。硫氰酸钠洗脱法ELISA分析抗hCGβ抗血清亲和力。结果两种品系小鼠pCMV4-hCG-βC3d3免疫产生抗hCGβ抗体的时间和效价均显著早(高)于pC-MV4-hCGβ和联合免疫组。在50 pmol免疫剂量时,C3d3在两种品系小鼠呈现出最强的佐剂活性,分别使hCGβ的免疫原性增强了400倍(BALB/c)和251倍(C57BL/6)。pCMV4-hCG-βC3d3组两个品系小鼠亲和力成熟较快。结论在Th2型及Th1型免疫优势的不同品系小鼠,分子佐剂C3d3均能增强hCGβ基因疫苗体液免疫效应。
Abstract：Objective: To enhance humoral immunity of hCGβ DNA immunizaiton in different strains of mice by gene fusion of the molecular adjuvant C3d3 to hCGβ. Methods: BALB/c and C57BL/6 mice were inoculated with eukaryotic expression plasmids pCMV4-hCGβ-C3d3,pCMV4-hCGβ and pCMV4-hCGβ plus pCMV4-C3d3 respectively,followed by a boost in a 3-weeks interval.The immunizing doses were 5,10,50,100 and 500 pmol respectively in each group.The antibody titers of the antisera were determined by indirect enzyme-linked immunosorbent assay(ELISA).Avidity of anti-hCGβ antibody was analyzed using sodium thiocyanate(NaSCN)-displacement ELISA.Results: The pCMV4-hCGβ-C3d3 immunization achieved an earlier and stronger antibody response than pCMV4-hCGβ alone or pCMV4-hCGβ plus pCMV4-C3d3 both in BALB/c and C57BL/6 mice.C3d3 showed the strongest adjuvant efficiency at immunizing dose of 50 pmol and the 400-and 251-fold increment of hCGβ immunogenicity respectively in BALB/c and C57BL/6 mice.The avidity maturation of the antibody was accelerated both in BALB/c and C57BL/6 mice inoculated with pCMV4-hCGβ-C3d3 than that with pCMV4-hCGβ alone or pCMV4-hCGβ plus pCMV4-C3d3. Conclusions: Molecular adjuvant C3d3 enhances the antibody responses to hCGβ in DNA immunization in mice of different strains,and this approach may be one way to overcome the poor immunogenicity and immunogenicity difference of hCGβ contraceptive vaccine from person to person.
[1]Talwar GP,Singh O,Pal R,et al.A vaccine that preventspregnancy in women[J].Proc Natl Acad Sci USA,1994,91(18):8532-8536.
[2]Singh M,Das SK,Suri S,et al.Regain of fertility andnormality of progeny born during below protective thresholdantibody titers in women immunized with the HSD-hCGvaccine[J].Am J Reprod Immunol,1998,39(6):395-398.
[3]Dempsey PW,Allison MED,Akkaraju S,et al.C3d of com-plement as a molecular adjuvant:bridging innate and acquiredimmunity[J].Science,1996,271(5247):348-350.
[4]Ross TM,Xu Y,Bright RA,et al.C3d enhancement ofantibodies to hemagglutinin accelerates protection againstinfluenza virus challenge[J].Nat Immunol,2000,1(2):127-131.
[5]Ross TM,Xu Y,Green TD,et al.Enhanced avidity matura-tion of antibody to human immunodeficiency virus envelope:DNA vaccination with gp120-C3d fusion proteins[J].AIDSRes Hum Retroviruses,2001,17(9):829-835.
[6]赵欣荣,李大金,蔡立荣,等.真核细胞表达质粒pCMV4增强hCG-βC3d3DNA疫苗的表达效率和免疫效力[J].中国免疫学杂志,2004,20(4):228-230.
[7]赵欣荣,李大金,蔡立荣,等.分子佐剂C3d增强hCGβ基因免疫体液免疫效应[J].中国免疫学杂志,2003,19(9):619-624.
[8]Green TD,Montefiori DC,Ross TM.Enhancement of anti-bodies to the human immunodeficiency virus type 1 envelopeby using the molecular adjuvant C3d[J].J Virol,2003,77(3):2046-2055.
[9]Hess MW,Schwendinger MG,Eskelinen EL,et al.Tracinguptake of C3dg-conjugated antigen in B cells via complementreceptor type 2(CR2,CD21)[J].Blood,2000,95(8):2617-2623.
[10]Kozono Y,Abe R,Kozono H,et al.Cross-linking CD21/CD35 or CD19 increases both B7-1 and B7-2 expression onmurine splenic B cells[J].J Immunol,1998,160(4):1565-1572.
[11]Kozono Y,Duke RC,Schleicher MS.Co-ligation of mousecomplement receptors 1 and 2 with surface IgM rescues splen-ic B cells and WEHI-231 cells from anti-surface IgM-inducedapoptosis[J].Eur J Immunol,1995,25(4):1013-1017.
[12]Roberts T,Snow EC.Cutting edge:recruitment of theCD19/CD21 coreceptor to B cell antigen receptor is requiredfor antigen-mediated expression of Bcl-2 by resting and cyc-ling hen egg lysozyme transgenic B cells[J].J Immunol,1999,62(8):4377-4380.
基本信息:
中图分类号:R392
引用信息:
[1]余敏,李大金,袁敏敏,等.C3d3与hCGβ基因融合质粒对不同品系小鼠DNA免疫增强抗hCGβ体液免疫效应[J].生殖医学杂志,2007(01):40-45.
基金信息:
国家自然科学基金(30271235);; 上海市自然科学基金(00ZB14058);; 教育部博士学科点专项科研基金(9941);; 卫生部优秀青年人才专项科研基金(97031)资助
2007-02-15
2007-02-15