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

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Nanoparticles of Conformation-stabilized Canine Distemper Virus Hemagglutinin are Highly Immunogenic and Induce Robust Immunity

10.21203/rs.3.rs-74893/v1 ◽

2020 ◽

Author(s):

Hao Feng ◽

Jingjian Dong ◽

Yan Chen ◽

Lili Shi ◽

Bing Shen ◽

...

Keyword(s):

Immune Responses ◽

Self Assembly ◽

Canine Distemper Virus ◽

Systemic Disease ◽

Neutralizing Antibody ◽

Canine Distemper ◽

Igg Antibody ◽

Organ Systems ◽

New Type ◽

H Protein

Abstract BackgroundCanine distemper virus (CDV) infection of ferrets, dogs, and giant pandas causes an acute systemic disease involving multiple organ systems, including the respiratory tract, lymphoid system, and central nervous system. In this study, we tested a new type candidate CDV vaccine–CDV nanoparticles–based on hemagglutinin protein. MethodsThe nanoparticles were generated from conformation-stabilized CDV hemagglutinin tetramers. Immune responses against CDV were evaluated in mice. Immunization was initiated 6 weeks after birth and boosted twice with 4-week intervals. The blood and mucosal samples were collected 2 weeks after each immunization. ResultsVaccination with CDV nanoparticles elicited high levels of IgG antibody titers in mice (approximately seven- to eight fold higher than that obtained with soluble CDV H protein), as well as mucosal immune responses, and developed increased CDV-specific neutralizing antibody. The mice that received nanoparticles showed significantly higher IFN-γ- and IL-4-secreting cell population in the spleen and lymph node compared with mice immunized with soluble H protein. The co-stimulatory molecular expression of CD80 and CD86 on the surface of DCs were also upregulated. ConclusionThe results demonstrate that self-assembly into nanoparticles can increase the immunogenicity of vaccine antigens, and nanoparticles assembled from conformation-stabilized CDV H protein has the potential to serve as a new type CDV vaccine.

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Severe canine distemper outbreak in unvaccinated dogs in Mozambique

Journal of the South African Veterinary Association ◽

10.4102/jsava.v87i1.1350 ◽

2016 ◽

Vol 87 (1) ◽

Cited By ~ 1

Author(s):

Julieta Zacarias ◽

Alberto Dimande ◽

Sara Achá ◽

Paula T. Dias ◽

Elisa M. Leonel ◽

...

Keyword(s):

Developing Countries ◽

Animal Welfare ◽

Low Income ◽

Immunohistochemical Staining ◽

Canine Distemper Virus ◽

Large Scale ◽

Clinical Signs ◽

Canine Distemper ◽

Animal Suffering ◽

Low Income Families

Although significant animal suffering caused by preventable diseases is frequently seen in developing countries, reports of this are scarce. This report describes avoidable animal suffering owing to a suspected canine distemper (CD) outbreak in unvaccinated dogs owned by low-income families in Mozambique that killed approximately 200 animals. Affected dogs exhibited clinical signs, and gross and microscopic lesions compatible with CD. Immunohistochemical staining confirmed the presence of canine distemper virus (CDV) in the kidney of one dog from the cohort. This brief communication again illustrates that large outbreaks of CDV in unvaccinated dogs occur and that large-scale avoidable suffering and threats to the health of dogs and wild canines continue. Mass vaccination supported by government and non-government organisations is recommended.Keywords: Canine distemper; dogs; outbreak; animal welfare; Mozambique

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A recombinant canine distemper virus expressing a modified rabies virus glycoprotein induces immune responses in mice

Virus Genes ◽

10.1007/s11262-015-1169-x ◽

2015 ◽

Vol 50 (3) ◽

pp. 434-441 ◽

Cited By ~ 10

Author(s):

Zhili Li ◽

Jigui Wang ◽

Daoli Yuan ◽

Shuang Wang ◽

Jiazeng Sun ◽

...

Keyword(s):

Immune Responses ◽

Rabies Virus ◽

Canine Distemper Virus ◽

Canine Distemper ◽

Rabies Virus Glycoprotein ◽

Virus Glycoprotein

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Nanoparticles of conformation-stabilized canine distemper virus hemagglutinin are highly immunogenic and induce robust immunity

Virology Journal ◽

10.1186/s12985-021-01702-0 ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Jingjian Dong ◽

Yan Chen ◽

Lili Shi ◽

Bing Shen ◽

Xianliang Sun ◽

...

Keyword(s):

Immune Responses ◽

Self Assembly ◽

Canine Distemper Virus ◽

Systemic Disease ◽

Neutralizing Antibody ◽

Multiple Organ ◽

Canine Distemper ◽

Igg Antibody ◽

Organ Systems ◽

H Protein

Abstract Background Canine distemper virus (CDV) infection of ferrets, dogs, and giant pandas causes an acute systemic disease involving multiple organ systems, including the respiratory tract, lymphoid system, and central nervous system. In this study, we tested a new candidate CDV vaccine-CDV nanoparticles-based on hemagglutinin protein. Methods The nanoparticles were generated from conformation-stabilized CDV hemagglutinin tetramers. Immune responses against CDV were evaluated in mice. Immunization was initiated 6 weeks after birth and boosted two times with 4-week intervals. The blood and mucosal samples were collected 2 weeks after each immunization. Results Vaccination with CDV nanoparticles elicited high levels of IgG antibody titers in mice (approximately sevenfold to eightfold higher than that obtained with soluble CDV H protein) and mucosal immune responses and developed increased CDV-specific neutralizing antibody. The mice that received nanoparticles showed significantly higher IFN-γ- and IL-4-secreting cell population in the spleen and lymph node compared with mice immunized with soluble H protein. The co-stimulatory molecular expression of CD80 and CD86 on the surface of DCs was also upregulated. Conclusion The results demonstrate that self-assembly into nanoparticles can increase the immunogenicity of vaccine antigens, and nanoparticles assembled from conformation-stabilized CDV H protein can serve as a new CDV vaccine.

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Correlation between humoral immune responses and presence of virus in the CNS in dogs experimentally infected with canine distemper virus

Archives of Virology ◽

10.1007/bf01316739 ◽

1991 ◽

Vol 121 (1-4) ◽

pp. 1-8 ◽

Cited By ~ 24

Author(s):

B. K. Rima ◽

N. Duffy ◽

W. J. Mitchell ◽

B. A. Summers ◽

M. J. G. Appel

Keyword(s):

Immune Responses ◽

Canine Distemper Virus ◽

Canine Distemper ◽

Humoral Immune ◽

Humoral Immune Responses

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IMMUNE RESPONSES TO CANINE DISTEMPER VIRUS IN JOINT DISEASES OF DOGS

Rheumatology ◽

10.1093/rheumatology/33.1.27 ◽

1994 ◽

Vol 33 (1) ◽

pp. 27-31 ◽

Cited By ~ 12

Author(s):

C. MAY ◽

S. D. CARTER ◽

S. C. BELL ◽

D. BENNETT

Keyword(s):

Immune Responses ◽

Canine Distemper Virus ◽

Joint Diseases ◽

Canine Distemper

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DNA vaccine encoding nucleocapsid and surface proteins of wild type canine distemper virus protects its natural host against distemper

Vaccine ◽

10.1016/s0264-410x(00)00119-5 ◽

2000 ◽

Vol 18 (26) ◽

pp. 2927-2936 ◽

Cited By ~ 30

Author(s):

Pascal Cherpillod ◽

Andrea Tipold ◽

Monika Griot-Wenk ◽

Carmen Cardozo ◽

Ines Schmid ◽

...

Keyword(s):

Dna Vaccine ◽

Canine Distemper Virus ◽

Natural Host ◽

Surface Proteins ◽

Canine Distemper ◽

Wild Type

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Inactivated Recombinant Rabies Viruses Displaying Canine Distemper Virus Glycoproteins Induce Protective Immunity against Both Pathogens

Journal of Virology ◽

10.1128/jvi.02077-16 ◽

2017 ◽

Vol 91 (8) ◽

Cited By ~ 12

Author(s):

Renata da Fontoura Budaszewski ◽

Andrew Hudacek ◽

Bevan Sawatsky ◽

Beate Krämer ◽

Xiangping Yin ◽

...

Keyword(s):

Immune Responses ◽

Fusion Protein ◽

Rabies Virus ◽

Canine Distemper Virus ◽

Canine Distemper ◽

Wild Type ◽

Attachment Protein ◽

Recombinant Viruses ◽

Virus Particles ◽

Residual Virulence

ABSTRACT The development of multivalent vaccines is an attractive methodology for the simultaneous prevention of several infectious diseases in vulnerable populations. Both canine distemper virus (CDV) and rabies virus (RABV) cause lethal disease in wild and domestic carnivores. While RABV vaccines are inactivated, the live-attenuated CDV vaccines retain residual virulence for highly susceptible wildlife species. In this study, we developed recombinant bivalent vaccine candidates based on recombinant vaccine strain rabies virus particles, which concurrently display the protective CDV and RABV glycoprotein antigens. The recombinant viruses replicated to near-wild-type titers, and the heterologous glycoproteins were efficiently expressed and incorporated in the viral particles. Immunization of ferrets with beta-propiolactone-inactivated recombinant virus particles elicited protective RABV antibody titers, and animals immunized with a combination of CDV attachment protein- and fusion protein-expressing recombinant viruses were protected from lethal CDV challenge. However, animals that were immunized with only a RABV expressing the attachment protein of CDV vaccine strain Onderstepoort succumbed to infection with a more recent wild-type strain, indicating that immune responses to the more conserved fusion protein contribute to protection against heterologous CDV strains. IMPORTANCE Rabies virus and canine distemper virus (CDV) cause high mortality rates and death in many carnivores. While rabies vaccines are inactivated and thus have an excellent safety profile and high stability, live-attenuated CDV vaccines can retain residual virulence in highly susceptible species. Here we generated recombinant inactivated rabies viruses that carry one of the CDV glycoproteins on their surface. Ferrets immunized twice with a mix of recombinant rabies viruses carrying the CDV fusion and attachment glycoproteins were protected from lethal CDV challenge, whereas all animals that received recombinant rabies viruses carrying only the CDV attachment protein according to the same immunization scheme died. Irrespective of the CDV antigens used, all animals developed protective titers against rabies virus, illustrating that a bivalent rabies virus-based vaccine against CDV induces protective immune responses against both pathogens.

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Immune responses in mice induced by multi-epitope DNA vaccine and protein vaccine of Crimean-Congo Hemorrhagic Fever Virus

10.1101/719724 ◽

2019 ◽

Author(s):

Meilipaiti Yusufu ◽

Alai Shalitanati ◽

Huan Yu ◽

Abulimiti Moming ◽

Yijie Li ◽

...

Keyword(s):

Immune Responses ◽

Dna Vaccine ◽

Hemorrhagic Fever ◽

Control Group ◽

Protein Vaccine ◽

Crimean Congo Hemorrhagic Fever ◽

Humoral Immune ◽

Humoral Immune Responses ◽

Significant Difference ◽

Double Copy

AbstractCrimean-Congo Hemorrhagic Fever (CCHF), caused by the CCHF virus (CCHFV), is a severe tick borne zoonosis widely distributed in over 30 countries and regions. Currently, there is no licensed vaccine available for CCHF in China. To evaluate the cellular and humoral immune responses induced by multi-epitope DNA and protein vaccine of CCHF in BALB/c mice, a multi-epitope gene (MEPX) segment with tandem including six highly conservative and immunedominant B cell epitopes was designed based on the analysis of hydrophilicity and antigenic determinant sites in amino acid sequences of nucleoprotein and glycoprotein from CCHFV strain YL04057. The single and double-copy multi-epitope gene (MEPX and MEPX2) were respectively cloned into the eukaryotic expression vector pVAX I to construct the recombinant (r) plasmid pVAX-MEPX and pVAX-MEPX2 as DNA vaccines. The results of immunofluorescence in vitro showed that the pVAX-MEPX and pVAX-MEPX2 could be expressed in 293T cells. The recombinant prokaryotic plasmid pET-32a-MEPX and pET-32a-MEPX2 constructed previously were transformed them into E. coli BL21 (DE3), and recombinant multi-epitope proteins (rMEPX and rMEPX2) were obtained and purificated by Nickel affinity chromatography. Western blot results showed that rMEPX and rMEPX2 had good antigenicity. BALB/c mice were immunized with DNA vaccine alone, protein vaccine alone, and DNA prime followed by recombinant protein boost immunization strategy, respectively. After three immunizations, MTT assay, cytokine content assay, and ELISA assay for antibody titers were used to evaluate the immune response. The proliferation of mouse specific T lymphocytes in the enhanced by pVAX-MEPX2 combined with rMEPX2 boosting group was significant, and the expression levels of serum IFN-γ and IL-4 in mice were as high as 118.67 pg/mL and 135.33 pg/mL with significant difference compared to the control group (p<0.01), and serum antibody titer could reach up to 4.1×105. Double-copy multi-epitope vaccines groups (pVAX-MEPX2+ rMEPX2) generated better cellular and humoral immune responses by DNA prime-protein vaccine boost combinatorial immunization. This result could lay the foundation for the development of CCHFV multi-epitope vaccine candidates.

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