Specific Th2 immune response of MUC1 induced by thmosin α1 / 吉林大学学报(医学版)

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.

Specific Th2 immune response of MUC1 induced by thmosin α1 / 吉林大学学报(医学版)

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.

Murine basophils express highly in liver and regulate priming of na(i)ve CD4+ T cells to develop into Th2 cells / 中华微生物学和免疫学杂志

Objective To compare the levels of murine basophils distribution in different organs and analyze the characteristics of CD4+T cells activation/differentiation regulated by basophils.Methods Firstly,the mononuclear leukocytes were isolated separately from periphery blood,liver,spleen,lung and lymph nodes and the percentages of FcεR I+CD49b+ basophils in different organs were investigated.Secondly,liver basophils were purified by immuno-microbead strategy and co-clutured with different cytokines for 24 h.Apoptosis status of basophils was detected by Annexin V straining method.Thirdly,Mice were subcutaneously implanted with a miniosmotic pump containing 5 μg IL-3.At day 7 after implantation,liver mononuclear cells were harvested and the percentages of basophils were analyzed.Finally,Th1/Th2 polarization influenced by basophils orientated cytokines was investigated when naive OT-Ⅱ CD4 T cells were stimulated with spleen dentritic cells and OVA peptides.Results The percentage of murine basophils in mononuclear leukocytes of liver was significantly higher than that of periphery blood,spleen,lung and lymph nodes under physiological condition.Both of in vitro and ex vivo experiments showed that IL-3 played an inhibitory role on apoptosis of basophils.Cytokines secreted by basophils was investigated to induce Th2 polarization and inhibit Th1 response when naive OT-Ⅱ CD4 T cells were co-cultured with spleen dentritic cells and OVA peptides.Conclusion Th2 response was enhanced and Th1 response was down-regulated by murine basophils,which implied that basophils might participate in the construction of liver specific physiological immuno-tolerant microenvironment.

Analysis of Specific Th1/Th2 Helper Cell Responses and IgG Subtype Antibodies in Anti-CD4 Monoclonal Antibody Treated Mice with Autoimmune Cardiomyopathy / 华中科技大学学报（医学）（英德文版）

The cytokine repertoire of ADP/ATP carrier-specific humoral immune responses and the cytokine-dependent anti-ADP/ATP carrier antibody IgG subclasses were examined in a cohort of ADP/ATP carrier-immunized BALB/c mice treated with anti-CD4 monoclonal antibody. Eighteen male BALB/c mice (6-8 weeks old) were randomized into 3 groups: dilated cardiomyopathy (DCM)group, DCM-tolerance (Tol) group and control group. The mice in DCM group were immunized with the peptides derived from human ADP/ATP carrier protein for 6 months and mice in the control group were sham-immunized, while the mice in DCM-Tol group were immunized with ADP/ATP carrier protein and anti-CD4 McAb simultaneously. Serum autoantibody against ADP/ATP carrier and IgG subclasses were measured by ELISA, intracellular cytokines IFN-γ and IL-4 of Th cells were monitored with flow cytometry, and splenic T cell cytokines IFN-γ, IL-2, IL-4 and IL-6 were detected by using real-time fluorescent quantitative PCR. The results showed that the autoantibody against ADP/ATP carrier was found in all mice in DCM group, and the antibody level, serum IgG1 and IgG2a subclasses, cytokines in T cells and Th cells were all elevated in DCM group, as compared with those in control group (P＜0.01). On the other hand, in DCM-Tol group, the autoantibody level and contents of all the cytokines were significantly different from those in DCM group (P＜0.01), and were close to those in control group. And the levels of IgG1, IgG2a, IgG2b and IgG3 were influenced,to varying degrees, by anti-CD4 McAb as compared with those in DCM group. All these four types of IgG subclasses were substantially decreased in DCM-Tol group as compared with DCM group. It is concluded that the treatment with anti-CD4 McAb could prevent the activation of T cells, reverse the abnormal secretion of cytokines and the imbalance between Th1/Th2 cell subsets and abnormal production of autoantibody against ADP/ATP carrier, and eventually avoid myocardial injuries.

Predominant Th2 type immune response in bronchoalveolar lavage fluid of patients with Mycoplasma pneumoniae pneumonia / 천식및알레르기

OBJECTIVE: Mycoplasma pneumoniae infection is known as one of the frequent causes of exacerbation of bronchial asthma and it can also be a trigger for the initiation of asthma. However, little is known about the pathogenesis of respiratory M. pneumoniae infection. Furthermore, there is little data on human cytokine production and its involvement in the pathogenesis of M. pneumoniae infection. In order to investigate the immunopathogenesis of M. pneumoniae infection, we investigated the cytokine production in the bronchoalveolar lavage ( BAL ) fluid of patients with M. pneumoniae pneumonia and viral pneumonia, and compared the results with those of control subjects. SUBJECT AND METHOD: BAL was performed with fiberoptic bronchoscopy in patients with M. pneumoniae pneumonia( n=9 ), viral pneumonia( n=9 ), and control subjects( n=6 ) aged 3 years to 9 years. M. pneumoniae pneumonia was documented by polymerase chain reaction and serologic analysis. Four respiratory viruses ( adenovirus, influenza A, influenza B, parainfluenza ) were detected by culture method. Cell pellets and supernatants were separated by centrifugation and Interleukin( IL ) - 2, Interferon( IFN )-r, IL-4, and IL-5 levels were measured in concentrated BAL supernatants by ELISA. RESULTS: Analysis of cytokines revealed significantly increased production of IL-4 ( p< 0.0001 ) and IL-2 ( p< 0.0001 ), in patients with M. pneumoniae pneumonia and significantly increased production of IL-2 (p <0.0001) in patients with viral pneumonia compared with those of the control subjects. Ratio of IL-4/IFN-r was significantly increased in patients with M. pneumoniae pneumonia ( p< 0.005 ) but not in patients with viral pneumonia compared with that of the control subjects. CONCLUSION: IL-4 production and IL-4/IFN-r ratio were increased in the BAL fluid of patients with M. pneumoniae infection. These findings suggest that predominant Th2 immune response could play an important role in the pathogenesis of M. pneumoniae infection.

The study on polarization of Th1 and Th2 immune responses in BALB/c mice infected with toxoplasma gondii / 中国免疫学杂志

Objective:To explore the polarization of Th1 and Th2 immune response and the changes of transcription factor(T-bet and GATA-3) in BALB/c mice infected with toxoplasma gondii.Methods:Sixty BALB/c mice were infected with toxoplasma tachyzoite(1?104/m,0.5 ml) by intraperitoneal injection and thirty BALB/c mice only received an injection of same amount of normal saline as control.After injection,two mice of infection group and one mouse of control group were killed in odd number days.Then,the serum IFN-? and IL-4 were measured by ELISA,and the mRNA expression of T-bet and GATA-3 in mice spleens were detected by Real-Time RT-PCR.Results:In the infection group of mice,the IFN-? was remarkable increased at the fourth day and kept the peak from the 5th day to 7th day.Then,it decreased to normal level at 9th day postinjection;but IL-4 was remarkable increased at the 8th day and reached to the peak at 9th day.Then it decreased to normal level at 15th day postinjection.The mRNA of T-bet expression was increased at the third day and kept the peak from the 5th day to 7th day.Then,it decreased to normal level at 9th day postinjection.The mRNA of GATA-3 expression was increased at the 7th day and rised to the peak at 11th day.Then,it decreased to normal level at 13th day postinjection.There were not obvious changes of IFN-? and IL-4 or T-bet and GATA-3 in normal mice.Conclusion:The Th1 immune response is dominant in acute infection when mice are infected by toxoplasma.Then it biases toward Th2 response at 9th day to 13th day postinfection.The shift of Th1/Th2 polarization influences the finale of toxoplasma infection.