鼠疫耶尔森菌F1抗原免疫Balb／c小鼠方法的研究  

Study on immunization of Balb/c mice with F1 antigen of Yersinia pestis

作　　者：刘合智[1] 杨晓燕[1] 胡乐乐[1] 史献明[1] 王海峰[1] 董国润[1] 李玉贵[1] 

机构地区：[1]河北省鼠疫防治所检验科,张家口075000

出　　处：《中国地方病学杂志》2009年第4期369-372,共4页Chinese Jouranl of Endemiology

基　　金：河北省卫生厅医学科学研究重点项目（2005080）;河北省医学适用技术跟踪项目（GL200635）

摘　　要：目的观察鼠疫耶尔森菌F1抗原（F1Ag）单克隆抗体制备过程中，FIAg免疫Balb／c小鼠的免疫剂量和方法，探索操作性强、用时短和免疫效果好的方法。方法7～9周龄Balb／c小鼠48只．按体质量随机分成6组：150、100、50、25ug组（第1次接种为皮下多点注射，F1Ag分别为150、100、50、25ug，第2、3次接种分别为皮下多点注射和腹腔注射，F1Ag量均为100ug）、皮下接种组（3次F1Ag接种均采用皮下多点注射，每次100txg）、腹腔接种组（3次F1Ag接种均采用腹腔注射，每次100ug），每组8只。首次免疫接种F1Ag＋等量弗氏完全佐剂（CFA）的乳化剂；3周后第2次接种F1Ag＋等量弗氏不完全佐剂（IFA）的乳化剂；1周后第3次接种F1Ag（不加佐剂）；1周后取小鼠尾血，用双抗原夹心酶联免疫吸附试验（DAgS—ELISA）和间接血球凝集试验（IHA）微量法检测F1抗体。结果不同剂量（150、100、50、25Ixg）组小鼠血清F1抗体效价（DAgS-ELISA法：G=12173．87、13440．37、15024．19、4466．72；IHA微量法：G=19972．32、18089．40、23170．47、4871．08）比较，差异有统计学意义（DAgS—ELISA法：F=3．11，P〈0．05；IHA微量法：F=4．11，P〈0．05）。150、100、50ug组小鼠血清F1抗体效价明显高于25ug组（DAgS—ELISA法：t值分别为2．18、2．39、2．73，P〈0．05；IHA法：t值分别为2．54、2．73、3．13，P〈0．05）。不同接种途径条件下，皮下注射组、腹腔接种组、100ug组小鼠血清F1抗体效价呈渐次增高趋势（DAgS—ELISA法：G：8933．44、9986．16、13440．37；IHA微量法：G=13777．25、16384．00、18089．40），但差异无统计学意义（F值分别为0．66、0．25，P〉0．05）。结论F1Ag免疫小鼠首次皮下多点注射50ug，加强免疫（腹腔注射）100ug，可以缩短整个免疫周期，免疫效果良好，抗体水平较高。Objective To study the doses and methods of F1 antigen（F1Ag） to immune Balb/c mice during the establishment of hybridoma cell strains. Secreting McAbs against F1Ag of Yersinia pestis. Methods Balb/c mice of seven to nine weeks old were randomly divided into six groups. The first four groups were 150, 100, 50 and 25ug F1Ag inoculated group, having muhipoint hypodermic inoculation of F1Ag of 150, 100, 50 and 25ug followed by muhipoint hypodermic inoculation of F1Ag of 100 ug for a second time and then intraperitoneal injection of 100 ug. Next, hypodermically inoculated group received F1Ag of 100 ug for three times in multiple points. Finally, the intraperitoneal injection group was intraperitoneally inoculated with F1Ag of 100ug for three times. Emulsification liquid of F1Ag ＋ Complete Frednd＇s adjuvant （ CFA ） of equivalence was used in the first inoculation, emulsification liquid of F1Ag ＋ Incomplete Frednd＇s adjuvant（IFA） balanced mix in the second, F1Ag liquid in the third. One week afterwards, tail blood of the mice was collected to test antibody titers of anti-F1Ag by double antigens sandwich enzyme linked immunosorbent assay （DAgS-ELISA） and trace indirect hemagglutination assay（IHA）. Results The levels of antibody of anti-F1Ag in 150,100,50 and 25 ug groups had statistics difference （DAgS-ELISA method： G = 12 173.87,13 440.37,15 024.19 and 4466.72, F = 3.11, P 〈 0.05; IHA： G = 19 972.32,18 089.40,23 170.47 and 4871.08, F= 4.11, P 〈 0.05）. Immune effect of the 3 groups of 150, 100 and 50 ug was almost the same （P 〉 0.05）, and excelled as compared with that in 25 ug group with statistics difference（DAgS-ELISA method ： t = 2.18,2.39,2.73, P 〈 0.05; IHA： t = 2.54,2.73,3.13, P〈 0.05）. The titer of F1 antibody had an increasing trend from the 100 ug group to hypodermic group and intraperitoneal injection groups, but without statistics difference （DAgS-ELISA method： G = 8933.44, 9986.16,13 440.37; IHA： G = 13 777.25,16 384.00,18 089.40, F = 0.66,0.25,

关 键 词：耶尔森菌 鼠疫 F1抗原 免疫 

分 类 号：R686[医药卫生—骨科学]