CAJ | 학술논문

目的观察鼠疫耶尔森菌F1抗原(F1Ag)单克隆抗体制备过程中,F1Ag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、25μg组(第1次接种为皮下多点注射,F1Ag分别为150、100、50、25μg,第2、3次接种分别为皮下多点注射和腹腔注射,F1Ag量均为100 μg)、皮下接种组(3次F1Ag接种均采用皮下多点注射,每次100 μg)、腹腔接种组(3次F1Ag接种均采用腹腔注射,每次100 μg),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、25μg)组小鼠血清F1抗体效价(DAgS--ELISA法:G=12 173.87、13 440.37、15 024.19、4466.72;IHA微量法:G=19 972.32、18 089.40、23 170.47、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、50 μg组小鼠血清F1抗体效价明显高于25μg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、100μg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、13 440.37:IHA微量法:G=13 777.25、16 384.00、18 089.40),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50μg,加强免疫(腹腔注射)100μg,可以缩短整个免疫周期,免疫效果良好,抗体水平较高.
목적 관찰서역야이삼균F1항원(F1Ag)단극륭항체제비과정중,F1Ag면역Balb/c소서적면역제량화방법,탐색조작성강、용시단화면역효과호적방법.방법 7～9주령Balb/c소서48지,안체질량수궤분성6조:150、100、50、25μg조(제1차접충위피하다점주사,F1Ag분별위150、100、50、25μg,제2、3차접충분별위피하다점주사화복강주사,F1Ag량균위100 μg)、피하접충조(3차F1Ag접충균채용피하다점주사,매차100 μg)、복강접충조(3차F1Ag접충균채용복강주사,매차100 μg),매조8지.수차면역접충F1Ag+등량불씨완전좌제(CFA)적유화제;3주후제2차접충F1Ag+등량불씨불완전좌제(IFA)적유화제;1주후제3차접충F1Ag(불가좌제);1주후취소서미혈,용쌍항원협심매련면역흡부시험(DAgS-ELISA)화간접혈구응집시험(IHA)미량법검측F1항체.결과 불동제량(150、100、50、25μg)조소서혈청F1항체효개(DAgS--ELISA법:G=12 173.87、13 440.37、15 024.19、4466.72;IHA미량법:G=19 972.32、18 089.40、23 170.47、4871.08)비교,차이유통계학의의(DAgS-ELISA법:F=3.11,P<0.05;IHA미량법:F=4.11,P<0.05).150、100、50 μg조소서혈청F1항체효개명현고우25μg조(DAgS-ELISA법:t치분별위2.18、2.39、2.73,P<0.05;IHA법:t치분별위2.54、2.73、3.13,P<0.05).불동접충도경조건하,피하주사조、복강접충조、100μg조소서혈청F1항체효개정점차증고추세(DAgS-ELISA법:G=8933.44、9986.16、13 440.37:IHA미량법:G=13 777.25、16 384.00、18 089.40),단차이무통계학의의(F치분별위0.66、0.25,P>0.05).결론 F1Ag면역소서수차피하다점주사50μg,가강면역(복강주사)100μg,가이축단정개면역주기,면역효과량호,항체수평교고.
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 25 μg F1Ag inoculated group, having multipoint hypodermic inoculation of F1Ag of 150, 100, 50 and 25 μg followed by multipoint hypodermic inoculation of F1Ag of 100 μg for a second time and then intraperitoneal injection of 100 μg. Next, hypodermically inoculated group received F1Ag of 100 μg for three times in multiple points. Finally, the intraperitoneal injection group was intraperitoneally inoculated with F1Ag of 100 μg 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 μg 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 μg was almost the same (P> 0.05), and excelled as compared with that in 25 μg 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 μg 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, all P > 0.05). Conclusions Hyodermical inoculation of F1Ag with the first dose of 50 μg in multiple points for mouse is appropriate, and a strengthening dose of 100 μg in an intraperitoneal injection may shorten the immune period.