CAJ | 학술논문

应用噬菌体展示技术筛选人巨细胞病毒糖蛋白M新的中和抗原表位
응용서균체전시기술사선인거세포병독당단백M신적중화항원표위
A Novel Neutralizing epitope of Human cytomegalovirus glycoprotein M Screened by Phage Display

万方数据

生物化学与生物物理进展生物化學與生物物理進展 생물화학여생물물리진전
PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
2009年 2期 220-227 ,共8页

王本旭%刘玉%刘展%高亚萍%王芳%潘和平%杨光%徐华%沈倍奋%柳川%邵宁生

왕본욱%류옥%류전%고아평%왕방%반화평%양광%서화%침배강%류천%소저생

人巨细胞病毒%糖蛋白M%中和表位人巨細胞病毒%糖蛋白M%中和錶位
인거세포병독%당단백M%중화표위
human cytomegalovirus%glycoprotein M%neutralizing epitope

人巨细胞病毒(HCMV)糖蛋白复合物Ⅱ包括两种蛋白,即糖蛋白M(gM)和糖蛋白N(gN).尽管来自于HCMV阳性病人血清中的糖蛋白复合物Ⅱ的IgG抗体能够中和HCMV粒子,但迄今为止,还没有gM中和性抗原表位的相关研究.应用消减杂交技术,通过噬菌体肽库筛选获得gM抗原的一个表位,即MAD.MAD氨基酸序列与gM第32～38位序列高度同源.将MAD与钥孔血蓝蛋白偶联免疫小鼠可产牛抗MAD多抗,该多抗小仅结合天然HCMV病毒粒子,而且特异结合重组表达的gM30～78多肽.ELISA结果表明MAD能够特异结合HCMV阳性的病人血清.病毒巾和实验结果进一步证明抗MAD多抗能够抑制HCMV AD169株病毒感染人胚肺细胞.总之,MAD表位有可能成为HCMV病毒疫苗潜在的保护性抗原.
인거세포병독(HCMV)당단백복합물Ⅱ포괄량충단백,즉당단백M(gM)화당단백N(gN).진관래자우HCMV양성병인혈청중적당단백복합물Ⅱ적IgG항체능구중화HCMV입자,단흘금위지,환몰유gM중화성항원표위적상관연구.응용소감잡교기술,통과서균체태고사선획득gM항원적일개표위,즉MAD.MAD안기산서렬여gM제32～38위서렬고도동원.장MAD여약공혈람단백우련면역소서가산우항MAD다항,해다항소부결합천연HCMV병독입자,이차특이결합중조표체적gM30～78다태.ELISA결과표명MAD능구특이결합HCMV양성적병인혈청.병독건화실험결과진일보증명항MAD다항능구억제HCMV AD169주병독감염인배폐세포.총지,MAD표위유가능성위HCMV병독역묘잠재적보호성항원.
Human cytomegalovirus glycoprotein complex Ⅱ (gC Ⅱ ) consists of two glycoproteins, gM and gN. Although gC Ⅱ specific IgG purified from HCMV positive patient sera can neutralize HCMV, there has been no report on the generation of virus-neutralizing antibodies by immunizing with one epitope of gM. The epitope, termed MAD, was screened from random phage peptide library by subtractive strategy. The peptide sequence of MAD was highly homologous with 32~38 amino acids of HCMV gM. Mice immunized with MAD coupled with keyhole limpet hemocyanin (KLH) could produce specific antibodies against MAD, and the antibodies obtained could bind not only native HCMV particles, but also the recombinant gM30～78 peptide. ELISA analysis results showed that MAD could specifically bind HCMV-positive human serum samples. Virus-neutralizing assay results demonstrated that the antibodies against MAD could inhibit HCMV strain AD169 entering the human embryonic lung cells. The results suggested that MAD could be used as a new potential protective antigen in the development of HCMV vaccine.