Study on immunization of Balb/c mice with F1 antigen of Yersinia pestis / 中国地方病学杂志

ABSTRACT

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.