Self-assembling ferritin nanoparticles coupled with linear sequences from canine distemper virus haemagglutinin protein elicit robust immune responses

Abstract

Background Canine distemper virus (CDV), which is highly infectious, has caused outbreaks of varying scales in domestic and wild animals worldwide, so the development of a high-efficiency vaccine has broad application prospects. Currently, the commercial vaccine of CDV is an attenuated vaccine, which has the disadvantages of a complex preparation process, high cost and safety risk. It is necessary to develop a safe and effective CDV vaccine that is easy to produce on a large scale. In this study, sequences of CDV haemagglutinin (HA) from the Yanaka strain were aligned, and three potential linear sequences, termed YaH3, YaH4, and YaH5, were collected. To increase the immunogenicity of the epitopes, ferritin was employed as a self-assembling nanoparticle element. The ferritin-coupled forms were termed YaH3F, YaH4F, and YaH5F, respectively. A full-length HA sequence coupled with ferritin was also constructed as a DNA vaccine to compare the immunogenicity of nanoparticles in prokaryotic expression. Result The self-assembly morphology of the proteins from prokaryotic expression was verified by transmission electron microscopy. All the proteins self-assembled into nanoparticles. The expression of the DNA vaccine YaHF in HEK-293T cells was also confirmed in vitro. After subcutaneous injection of epitope nanoparticles or intramuscular injection of DNA YaHF, all vaccines induced strong serum titres, and long-term potency of antibodies in serum could be detected after 84 days. Strong anti-CDV neutralizing activities were observed in both the YaH4F group and YaHF group. According to antibody typing and cytokine detection, YaH4F can induce both Th1 and Th2 immune responses. The results of flow cytometry detection indicated that compared with the control group, all the immunogens elicited an increase in CD3. Simultaneously, the serum antibodies induced by YaH4F and YaHF could significantly enhance the ADCC effect compared with the control group, indicating that the antibodies in the serum effectively recognized the antigens on the cell surface and induced NK cells to kill infected cells directly. Conclusions YaH4F self-assembling nanoparticle obtained by prokaryotic expression has no less of an immune effect than YaHF, and H4 has great potential to become a key target for the easy and rapid preparation of epitope vaccines. Graphical Abstract

A biepitope, adjuvant-free, self-assembled influenza nanovaccine provides cross-protection against H3N2 and H1N1 viruses in mice

“…Ferritin is a SANP composed of 24 identical subunits with robust thermal and chemical stability [20]. Currently, our group has displayed various antigens on the surface of ferritin NPs related to many infectious diseases, such as influenza, acquired immunodeficiency syndrome, and canine distemper [21][22][23][24].…”

Self-assembling ferritin nanoplatform for the development of infectious hematopoietic necrosis virus vaccine

“…Helicobacter pylori self-assembling ferritin has been used for the development of safe subunit nano vaccines, against some human and other mammalian viruses such as Middle East respiratory syndrome-coronavirus (MERS-CoV) ( 19 ), Influenza A virus ( 21 ), HIV ( 22 ), rotavirus A ( 23 ), SARS-CoV-2 ( 24 ), Canine distemper virus ( 25 ), classical swine fever virus ( 39 ), foot-and-mouth disease virus ( 40 ), hepatitis C virus ( 41 ), and Zika Virus ( 42 ), which have been produced in bacteria, insects and mammalian cells systems. In this study, we have for the first time used self-assembling ferritin nanocages as a vaccine platform against a non-mammalian viral pathogen (IHNV).…”

“…Ferritin is a major intracellular iron storage protein present in most living organisms, with 24 identical subunits that spontaneously self-assemble and form NPs complexes (20). Ferritin nanocages can enhance the immunogenicity of antigens by displaying them on their outer surface in an orderly manner, similar to the whole organism vaccines, leading to long-lasting immunity, which recently has been proven to work well both in vitro and in vivo for some mammalian viruses (21)(22)(23)(24)(25). The vaccines have been developed using eukaryotic host cells such as human embryonic kidney cells, and prokaryotes (E. coli) expression systems, delivered successfully through different routes in animal models (21)(22)(23)(24)(25).…”

“…Ferritin nanocages can enhance the immunogenicity of antigens by displaying them on their outer surface in an orderly manner, similar to the whole organism vaccines, leading to long-lasting immunity, which recently has been proven to work well both in vitro and in vivo for some mammalian viruses (21)(22)(23)(24)(25). The vaccines have been developed using eukaryotic host cells such as human embryonic kidney cells, and prokaryotes (E. coli) expression systems, delivered successfully through different routes in animal models (21)(22)(23)(24)(25). The use of ferritin NPs as an antigen scaffold can therefore improve the immunogenicity of subunit vaccines, and can also be considered as a promising platform for oral vaccine development due to its unique structure and stability under harsh temperature and pH conditions (20,26).…”

Engineered biological nanoparticles as nanotherapeutics for tumor immunomodulation

“…214,246,247 Combining γδ-T-cell-derived exosomes (γδ-T-Exos) with radiotherapy overcomes radioresistance and preserves antitumor activities in immunosuppressive tumor microenvironments. 248 A photoactivatable silencing extracellular vesicle (PASEV) merges phototherapy with exosome-mediated small interfering RNA (siRNA) delivery against p21-activated kinase 4 (PAK4), an immune modulator involved in immune exclusion (Fig. 18A).…”