1.Analysis of immunogenicity of African swine fever virus p37 recombinant protein in mice
Ying HUANG ; Wenzhu ZHAI ; Chunhao TAO ; Yuheng HE ; Zhen WANG ; Yuanyuan CHU ; Zhongbao PANG ; Hongfei ZHU ; Hong JIA
Chinese Journal of Veterinary Science 2025;45(5):889-895
The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P<0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P<0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.
2.Analysis of immunogenicity of African swine fever virus p37 recombinant protein in mice
Ying HUANG ; Wenzhu ZHAI ; Chunhao TAO ; Yuheng HE ; Zhen WANG ; Yuanyuan CHU ; Zhongbao PANG ; Hongfei ZHU ; Hong JIA
Chinese Journal of Veterinary Science 2025;45(5):889-895
The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P<0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P<0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.
3.Expression of African swine fever virus pp62 protein in HEK 293F cells and analy-sis ofinduced immune response
Wenzhu ZHAI ; Ying HUANG ; Chunhao TAO ; Yuheng HE ; Yuanyuan CHU ; Zhen WANG ; Zhongbao PANG ; Hongfei ZHU ; Hong JIA
Chinese Journal of Veterinary Science 2024;44(12):2514-2520,2555
African swine fever(ASF)is a highly contagious disease caused by the African swine fe-ver virus(ASFV).To evaluate the immunogenicity of the pp62(CP530R)protein of ASFV,the recombinant CP530R protein was expressed in HEK 293F cells transfected with the plasmid pMAL-Fc-CP530R.Six-week-old female C57BL/6J mice were immunized with 10 μg of purified pp62 protein via subcutaneous injection,followed by a booster immunization with the same dosage at 21 days(enhanced).The humoral and cellular immune responses in the mice were then assessed using ELISA,flow cytometry,and ELISpot assays.Western blot analysis confirmed that the pp62 protein was successfully expressed,with a molecular weight of approximately 118.5 kDa.ELISA results indicated that a high level of specific antibodies was detected in the immunized mice,with antibody titers reaching up to 1∶1 638 400 at 7 days after the secondary immunization.The pro-portion of CD8+T lymphocytes in the immunized mice increased compared to the control group(P<0.05).Results from the Q-PlexTM Mouse Cytokine Screen demonstrated that the secretion levels of IFN-γ,IL-2,IL-4,and IL-10 in serum were significantly upregulated in the immunized mice following secondary immunization(P<0.001).In summary,these findings indicate that the pp62 protein can significantly stimulate both humoral and cellular immunity in mice,laying the groundwork for further studies on the function of the ASFV pp62 protein and the identification of novel vaccine antigens for ASF.
4.Expression of African swine fever virus pp62 protein in HEK 293F cells and analy-sis ofinduced immune response
Wenzhu ZHAI ; Ying HUANG ; Chunhao TAO ; Yuheng HE ; Yuanyuan CHU ; Zhen WANG ; Zhongbao PANG ; Hongfei ZHU ; Hong JIA
Chinese Journal of Veterinary Science 2024;44(12):2514-2520,2555
African swine fever(ASF)is a highly contagious disease caused by the African swine fe-ver virus(ASFV).To evaluate the immunogenicity of the pp62(CP530R)protein of ASFV,the recombinant CP530R protein was expressed in HEK 293F cells transfected with the plasmid pMAL-Fc-CP530R.Six-week-old female C57BL/6J mice were immunized with 10 μg of purified pp62 protein via subcutaneous injection,followed by a booster immunization with the same dosage at 21 days(enhanced).The humoral and cellular immune responses in the mice were then assessed using ELISA,flow cytometry,and ELISpot assays.Western blot analysis confirmed that the pp62 protein was successfully expressed,with a molecular weight of approximately 118.5 kDa.ELISA results indicated that a high level of specific antibodies was detected in the immunized mice,with antibody titers reaching up to 1∶1 638 400 at 7 days after the secondary immunization.The pro-portion of CD8+T lymphocytes in the immunized mice increased compared to the control group(P<0.05).Results from the Q-PlexTM Mouse Cytokine Screen demonstrated that the secretion levels of IFN-γ,IL-2,IL-4,and IL-10 in serum were significantly upregulated in the immunized mice following secondary immunization(P<0.001).In summary,these findings indicate that the pp62 protein can significantly stimulate both humoral and cellular immunity in mice,laying the groundwork for further studies on the function of the ASFV pp62 protein and the identification of novel vaccine antigens for ASF.
5.Measurement of dose field distribution around mobile cone beam computed tomography (CBCT)
Rui ZONG ; Nan WANG ; Wenzhu BO ; Lei LIU ; Hezheng ZHAI ; Jie GAO ; Wentian CAO ; Xinguang YU ; Zipo ZHAI
Chinese Journal of Radiological Health 2021;30(5):638-642
Objective To measure the peripheral dose distributions of the mobile head cone beam computed tomography (CBCT) and evaluate the impact of CBCT on the surrounding personnel and environment, and provide data support for clinical radiation protection management. Methods Combined with the structural characteristics of CBCT, AT1123 was used in the direction of 0° (counterclockwise), 45°, 90°, 135°, 180°, 225°, 270° and 315° in front of CBCT to measure the ambient dose equivalent rate of 30 cm, 80 cm and 130 cm away from the ground when the equipment was normally out of the beam, and the boundary of the temporary control area was drawn. At the same time, the dose level behind the lead screen 1 m away from the external surface of the equipment was measured and analyzed. Results The dose field around CBCT was symmetrically distributed with the dividing line of 0° and 180°, and the radiation dose level of 5.5 m in the direction of 0°, 3.5 m in the direction of 45°, 0.5 m in the direction of 90° and within 1.0 m in the direction of 180° (inside the "spoon" type) was higher than 2.5 μSv/h. The radiation dose levels of CT aperture 0° (straight forward), 45° and 315° behind the lead screen 1 m away from the equipment surface were 0.37 μSv/h, 0.22 μSv/h and 0.54 μSv/h, respectively. Conclusion The results show that the radiation dose around the mobile head cone beam CT is in a low dose level, the distribution of the dose field can provide necessary reference for the administrative and medical personnel to strengthen the radiation safety management. At the same time, it is suggested that lead screens should be set up in the clinical use of mobile CT to ensure the health and safety of the surrounding people and the environment.

Result Analysis
Print
Save
E-mail