中国血吸虫病防治杂志
中國血吸蟲病防治雜誌
중국혈흡충병방치잡지
CHINESE JOURNAL OF SCHISTOSOMIASIS CONTROL
2014年
4期
394-398
,共5页
王馨茁%洪炀%韩宏晓%李长健%曹晓丹%陆珂%李浩%朱传刚%傅志强%林矫矫
王馨茁%洪煬%韓宏曉%李長健%曹曉丹%陸珂%李浩%硃傳剛%傅誌彊%林矯矯
왕형촬%홍양%한굉효%리장건%조효단%륙가%리호%주전강%부지강%림교교
日本血吸虫%肌钙蛋白T%克隆%表达%免疫保护%效果评估
日本血吸蟲%肌鈣蛋白T%剋隆%錶達%免疫保護%效果評估
일본혈흡충%기개단백T%극륭%표체%면역보호%효과평고
Schistosoma japonicum%Troponin T%Clone%Expression%Immuno-protection%Efficacy evaluation
目的：克隆、表达日本血吸虫肌钙蛋白T（SjTnT）编码cDNA，评估重组抗原抗血吸虫感染的免疫保护效果。方法利用PCR技术扩增SjTnT基因，构建重组表达质粒pET28a（+）-SjTnT并诱导表达，用重组蛋白rSjTnT免疫BALB/c小鼠制备免疫血清。利用免疫印迹试验（Western blotting）和酶联免疫吸附试验（ELISA）检测rSjTnT诱导的免疫反应。采用rSjTnT免疫小鼠，评估其诱导的免疫保护效果。结果成功克隆、表达了日本血吸虫SjTnT基因。Western blotting显示rSjTnT能被该重组蛋白免疫小鼠血清识别，具有良好的免疫原性。rSjTnT能诱导小鼠产生高水平的特异性IgG抗体。动物免疫保护试验表明，rSjTnT能诱导小鼠产生33.89%的减虫率以及43.94%的肝脏减卵率。结论制备的rSjTnT在小鼠体内能诱导产生部分抗血吸虫感染的免疫保护效果，为评估SjTnT的生物学功能奠定了基础。
目的：剋隆、錶達日本血吸蟲肌鈣蛋白T（SjTnT）編碼cDNA，評估重組抗原抗血吸蟲感染的免疫保護效果。方法利用PCR技術擴增SjTnT基因，構建重組錶達質粒pET28a（+）-SjTnT併誘導錶達，用重組蛋白rSjTnT免疫BALB/c小鼠製備免疫血清。利用免疫印跡試驗（Western blotting）和酶聯免疫吸附試驗（ELISA）檢測rSjTnT誘導的免疫反應。採用rSjTnT免疫小鼠，評估其誘導的免疫保護效果。結果成功剋隆、錶達瞭日本血吸蟲SjTnT基因。Western blotting顯示rSjTnT能被該重組蛋白免疫小鼠血清識彆，具有良好的免疫原性。rSjTnT能誘導小鼠產生高水平的特異性IgG抗體。動物免疫保護試驗錶明，rSjTnT能誘導小鼠產生33.89%的減蟲率以及43.94%的肝髒減卵率。結論製備的rSjTnT在小鼠體內能誘導產生部分抗血吸蟲感染的免疫保護效果，為評估SjTnT的生物學功能奠定瞭基礎。
목적：극륭、표체일본혈흡충기개단백T（SjTnT）편마cDNA，평고중조항원항혈흡충감염적면역보호효과。방법이용PCR기술확증SjTnT기인，구건중조표체질립pET28a（+）-SjTnT병유도표체，용중조단백rSjTnT면역BALB/c소서제비면역혈청。이용면역인적시험（Western blotting）화매련면역흡부시험（ELISA）검측rSjTnT유도적면역반응。채용rSjTnT면역소서，평고기유도적면역보호효과。결과성공극륭、표체료일본혈흡충SjTnT기인。Western blotting현시rSjTnT능피해중조단백면역소서혈청식별，구유량호적면역원성。rSjTnT능유도소서산생고수평적특이성IgG항체。동물면역보호시험표명，rSjTnT능유도소서산생33.89%적감충솔이급43.94%적간장감란솔。결론제비적rSjTnT재소서체내능유도산생부분항혈흡충감염적면역보호효과，위평고SjTnT적생물학공능전정료기출。
Objective To clone cDNA encoding troponin T of Schistosoma japonicum(SjTnT),and evaluate the protective efficacy induced by recombinant SjTnT in BALB/c mice against S. japonicum challenge infection. Methods The SjTnT gene was amplified from 28-day-schistosome cDNAs by PCR and then subcloned into pET28a(+). The recombinant SjTnT protein (rSjTnT)was expressed in Escherichia coli BL21(DE3)cells. The serum specific to rSjTnT was prepared by immunized BALB/c mice with the recombinant antigen,and the immunogenicity of rSjTnT was detected by Western blotting and ELISA. The immuno-protective efficacy induced by rSjTnT in BALB/c mice was evaluated according to the reduction in worm and egg counts. Results The cDNA encoding SjTnT was successfully cloned and expressed in E. coli. Western blotting showed that rSjTnT had a good immunogenicity. The high level of specific IgG antibodies was detected,and 33.89% worm reduction and 43.94% liver egg reduction were obtained in mice vaccinated with rSjTnT combined with Seppic 206 adjuvant compared with those in the adjuvant control group. Conclusions rSjTnT could induce partial immuno-protection against S. japonicum infec-tion in BALB/c mice. This study provided a basic for understanding the biological function of SjTnT.
