An Intracellular Epitope of ASFV CD2v Protein Elicits Humoral and Cellular Immune Responses

Lu, Wenlong; Bai, Yilin; Zhang, Shuai; Zhao, Xuyang; Jin, Jiaxin; Zhu, Xiaojing; Wang, Rui; Wu, Yanan; Zhang, Angke; Zhang, Gaiping; Zhuang, Guoqing; Sun, Aijun

doi:10.3390/ani13121967

Cited by 4 publications

(3 citation statements)

References 50 publications

(72 reference statements)

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“…Although multiple linear B-cell epitopes of CD2v have recently been identified (34)(35)(36)(37)(38)(39)(40), their functions remain unknown (Table 1). To analyze the antigenic epitopes recognized by our anti-CD2v mAbs, we designed and constructed five CD2v extracellular domain truncation mutants (D1-D5) (Figures 6A-C).…”

Section: Discussionmentioning

confidence: 99%

Development of novel monoclonal antibodies for blocking NF-κB activation induced by CD2v protein in African swine fever virus

Fan,

Wei,

Zhang

et al. 2024

Front. Immunol.

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BackgroundCD2v, a critical outer envelope glycoprotein of the African swine fever virus (ASFV), plays a central role in the hemadsorption phenomenon during ASFV infection and is recognized as an essential immunoprotective protein. Monoclonal antibodies (mAbs) targeting CD2v have demonstrated promise in both diagnosing and combating African swine fever (ASF). The objective of this study was to develop specific monoclonal antibodies against CD2v.MethodsIn this investigation, Recombinant CD2v was expressed in eukaryotic cells, and murine mAbs were generated through meticulous screening and hybridoma cloning. Various techniques, including indirect enzyme-linked immunosorbent assay (ELISA), western blotting, immunofluorescence assay (IFA), and bio-layer interferometry (BLI), were employed to characterize the mAbs. Epitope mapping was conducted using truncation mutants and epitope peptide mapping.ResultsAn optimal antibody pair for a highly sensitive sandwich ELISA was identified, and the antigenic structures recognized by the mAbs were elucidated. Two linear epitopes highly conserved in ASFV genotype II strains, particularly in Chinese endemic strains, were identified, along with a unique glycosylated epitope. Three mAbs, 2B25, 3G25, and 8G1, effectively blocked CD2v-induced NF-κB activation.ConclusionsThis study provides valuable insights into the antigenic structure of ASFV CD2v. The mAbs obtained in this study hold great potential for use in the development of ASF diagnostic strategies, and the identified epitopes may contribute to vaccine development against ASFV.

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Section: Discussionmentioning

confidence: 99%

Development of novel monoclonal antibodies for blocking NF-κB activation induced by CD2v protein in African swine fever virus

Fan,

Wei,

Zhang

et al. 2024

Front. Immunol.

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“…Lymphocyte peptide libraries for anti-ASFV immunization were screened using surviving porcine peripheral blood mononuclear cells (PBMCs) and randomly divided into sub-peptide Libraries A and B for immunization, with both groups being found to stimulate the production of strong cellular immune responses [10]. The T cell epitope of the CD2V single antigen, which promotes CD8 + T cell metabolism, was found to induce significantly greater viral clearance than whole-protein immunization [45].…”

Section: Discussionmentioning

confidence: 99%

Analysis of the Immunogenicity of African Swine Fever F317L Protein and Screening of T Cell Epitopes

Huang,

Zhai,

Wang

et al. 2024

Animals

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The African swine fever virus (ASFV) encodes numerous proteins characterized by complex immune escape mechanisms. At present, the structure and function of these proteins, including the F317L protein, have yet to be fully elucidated. In this study, we examined the immunogenicity of the F317L protein. Mice were subcutaneously immunized with the F317L protein using initial and subsequent booster doses, and, at the 28th day post-treatment, we assessed the humoral and cellular immune responses of mice. The F317L protein stimulated production of specific antibodies and activated humoral immune responses. In addition, F317L stimulated the production of large amounts of IFN-γ by splenic lymphocytes, thereby activating cellular immune responses. Using informatics technology, we predicted and synthesized 29 F317L protein T cell epitopes, which were screened using IFN-γ ELISpot. Among these, the F25 (246SRRSLVNPWT255) peptide was identified as having a stronger stimulatory effect than the full-length protein. Collectively, our findings revealed that the ASFV F317L protein can stimulate both strong humoral and cellular immunity in mice, and that the F25 (246SRRSLVNPWT255) peptide may be a potential active T cell epitope. These findings will provide a reference for further in-depth studies of the F317L protein and screening of antigenic epitopes.

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“…The low surface density reduces the affinity of antibodies for virions and also reduces the immunogenicity of protective antigens ( Zhu, 2022 ). It has been noted that the low surface density of CD2v on extracellular virions may be a result of intracellular localization of majority of the expressed CD2v rather than on the cell surface ( Goatley and Dixon, 2011 ; Lu et al, 2023 ).…”

Section: The Major Structural Proteins In Asfv Vaccine Developmentmentioning

confidence: 99%

Advancement in the development of gene/protein-based vaccines against African swine fever virus

Wang,

Huang,

Zhang

et al. 2024

Current Research in Microbial Sciences

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An Intracellular Epitope of ASFV CD2v Protein Elicits Humoral and Cellular Immune Responses

Cited by 4 publications

References 50 publications

Development of novel monoclonal antibodies for blocking NF-κB activation induced by CD2v protein in African swine fever virus

Development of novel monoclonal antibodies for blocking NF-κB activation induced by CD2v protein in African swine fever virus

Analysis of the Immunogenicity of African Swine Fever F317L Protein and Screening of T Cell Epitopes

Advancement in the development of gene/protein-based vaccines against African swine fever virus

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