ABSTRACT
Objective:To investigate the MUC1 specific immune response enhanced by thmosinα1(Tα1)using MUC1-MBP as the specific antigen,and to discuss the feasibility of MUC1-MBP as an adjuvant.Methods:The C57BL/6 mice were randomly divided into normal saline group,MUC1-MBP+BCG group and MUC1-MBP+Tα1 group.The T cellular immune activities,MUC1 special antibody and subclass,anti-tumor effect of Tα1 combined with MUC1-MBP were detected.4-7 d after the 3rd immunization,the spleen indexes of the mice in various groups were measured;the lymphocyte proliferation response was used to detect the stimulate index(SI)of the mice in various groups;the levels of specific cytokines IFN-γ,IL-2,IL-4 and IL-10 in the supernatant of spleen cells of the mice were detected by ELISA;the level of serum MUC1-specific antibody was detected by ELISA.The C57BL/6 mice were inoculated with B16-MUC1 7 d after the last immunization and the survival of the mice was observed.Results:Compared with normal saline group,the spleen index and SI of the mice in MUC1-MBP+Tα1 and MUC1-MBP+BCG groups were significantly increased(P< 0.05 or P<0.01);the specific SI of MUC1 were significantly increased(P< 0.05).Compared with normal saline group,the levels of IFN-γ and IL-2 in the supernatant of spleen cells of the mice in MUC1-MBP+BCG group were obviously increased(P<0.05);the levels of IL-4 and IL-10 were slightly increased,but there were no significant differences(P>0.05);compared with normal saline group,the level of IL-4 in MUC1-MBP+Tα1 group was obviously increased(P< 0.01);the levels of IFN-γ,IL-2 and IL-10 were slightly increased,but there were no significant differences(P>0.05).The titer of MUC1 specific antibody was increased with the increase of concentration of Tα1.The antibody subtype detection results showed that compared with normal saline group,the level of IgG1 in MUC1-MBP+BCG group was significantly increased(P< 0.05 or P< 0.01),and the level of IgG2a had no obvious change.The tumor prevention experiment results showed that compared with normal saline group,the survival rates of the mice in MUC1-MBP+BCG group and MUC1-MBP+Tα1 group had no significant differences.Conclusion:MUC1-MBP combined with Tα1 prefers to Th2 immune responses in the mice.It indicates that Tα1 can be used as an adjuvanted preventive vaccines,but not suitable for therapeutic vaccines.
ABSTRACT
Objective: To investigate the MUC1 specific immune response enhanced by thmosinal (Tα1) using MUC1-MBP as the specific antigen, and to discuss the feasibility of MUC1-MBP as an adjuvant. Methods: The C57BL/6 mice were randomly divided into normal saline group, MUC1-MBP + BCG group and MUC1-MBP + Tα1 group. The T cellular immune activities, MUC1 special antibody and subclass, anti-tumor effect of Tα1 combined with MUC1-MBP were detected. 4-7 d after the 3rd immunization, the spleen indexes of the mice in various groups were measured; the lymphocyte proliferation response was used to detect the stimulate index (SI) of the mice in various groups; the levels of specific cytokines IFN-γ, IL-2, IL-4 and IL-10 in the supernatant of spleen cells of the mice were detected by ELISA; the level of serum MUC1-specific antibody was detected by ELISA. The C57BL/6 mice were inoculated with B16-MUC1 7 d after the last immunization and the survival of the mice was observed. Results: Compared with normal saline group, the spleen index and SI of the mice in MUCl-MBP+Tα1 and MUC1-MBP+BCG groups were significantly increased (P0.05); compared with normal saline group, the level of IL-4 in MUCl-MBP+Tal group was obviously increased (P0.05). The titer of MUC1 specific antibody was increased with the increase of concentration of Tα1. The antibody subtype detection results showed that compared with normal saline group, the level of IgG1 in MUCl-MBP + BCG group was significantly increased (P< 0.05 or P< 0.01), and the level of IgG2a had no obvious change. The tumor prevention experiment results showed that compared with normal saline group, the survival rates of the mice in MUC1-MBP+BCG group and MUCl-MBP + Tal group had no significant differences. Conclusion: MUC1-MBP combined with Tα1 prefers to Th2 immune responses in the mice. It indicates that Tα1 can be used as an adjuvanted preventive vaccines, but not suitable for therapeutic vaccines.
ABSTRACT
Objective:To study the inductive effect of recombinant MUC1-MBP fusion protein combined with R848 on the immune activity of T cells in the mice,and to provide experimental basis for searching the suitable adjuvants of recombinant MUC1-MBP fusion protein.Methods:A total of 21 C57BL/6 mice were randomly divided into control group(treat with normal saline),MUC1-MBP+R848 group (treated with MUC1-MBP+R848),and MUC1-MBP+baillus calmette guerin(BCG) group(treated with MUC1-MBP+BCG)(n=7).After immunized for 4-7 d,the spleen tissue was taken and the spleen indexes of the mice in various groups were measured;the stimmulus index(SI) of the mice was detected by lymphocyte proliferation response;the levels of tumor necrosis factor-γ(TNF-γ) and interleukin-4(IL-4) were detected by ELISA method;the proprotions of T lymphocyte subsets in spleen cells were analyzed by flow cytometry.Results:Compared with control group,the spleen indexes,SI,and TNF-γ levels of the mice in MUC1-MBP+ R848 and MUC1-MBP+BCG groups were significantly increased (P0.05).Compared with control group,the proprotions of CD3+T,CD4+T,and CD8+ T cells of the mice in MUC1-MBP+ R848 group and MUC1-MBP+BCG group were significantly increased(P<0.05 or P<0.01).Conclusion:Both MUC1-MBP fusion protein combined with R848 and BCG can induce the Th1 type of immune activity in the mice,and R848 is the potential candidate adjuvant for MUC1-MBP.