Optimized polypeptide for a subunit vaccine against avian reovirus

An avian reovirus and vaccine technology, which is used in viral peptides, antiviral agents, veterinary vaccines, etc.

Pending Publication Date: 2018-08-21
GAVISH GALILEE BIO APPL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these vaccines are ineffective against multiple ARVs found in the field (Jones 2000; Goldenberg et al. 2010; Lublin et al. 2011)

Method used

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  • Optimized polypeptide for a subunit vaccine against avian reovirus
  • Optimized polypeptide for a subunit vaccine against avian reovirus
  • Optimized polypeptide for a subunit vaccine against avian reovirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example 1. SC protein expression

[0097] After analyzing the SC protein structure, a cDNA fragment encoding three polypeptide residues: 1-326, 122-326, 192-326 was generated, cloned and expressed in E. coli. A polyhistidine tag was added at the 5' end to allow detection and purification of the expressed protein. The resulting protein was partially purified. As determined on a 12% SDS-polyacrylamide gel, the expressed SC fragments were 36kD, 23kD and 15kD with the expected sizes, respectively.

[0098] Figure 1 shows the efficient expression of the 122-326 fragment from four different reovirus isolates: SC1133, SC528, SC5223 and SC5215.

Embodiment 2

[0099] Example 2. Antibody Response to Vaccination

[0100] After vaccination, the immunogenicity of the SC protein and the ability of the antibodies generated to detect the virus were tested by ELISA. Using SC122-326 as the antigen, antibodies obtained after vaccination with the proteins SC122-326 and SC192-326 detected the antigen at a high titer of 1:800,000. The average value of bird titers in this group was significantly higher than that of the negative control group (P figure 2 ).

[0101] Antibody levels against the whole virus reached 0.2 or less after inoculation with SC192-326 or the negative control, whereas antibodies obtained after inoculation with SC1-326 or SC122-326 were positive (0.25 and 0.5, respectively) ( image 3 ). The S / P of SC122-326 group was significantly higher than other groups.

[0102] Similarly, following vaccination, SC122-326 fragments of four different RI groups (SEQ ID NO: 2, 3, 4 and 5) were tested for immunogenicity and the ability of t...

Embodiment 3

[0103] Example 3. Cell Proliferation

[0104] Splenocytes from vaccinated chickens were examined for antigen-specific cell proliferation. Cells were treated with ConA as a control for T cell proliferation, LPS as a control for T cell proliferation (positive control), PBS (negative control) and ARV (specific antigen). In the CTB assay, the measured absorbance is proportional to the number of viable cells. Results are expressed as SI (mitogen or antigen stimulated / non-stimulated cells).

[0105] Positive control treatments exhibited proliferation after stimulation. Cells stimulated with ARV showed significant differences among treatment groups. Compared with the negative control, the cells in the group immunized with virus or SC122-326 showed significant proliferation (P Figure 4 ).

[0106] Similarly, antigen-specific cell proliferation was examined in splenocytes from chickens vaccinated with combinations comprising polypeptides of SEQ ID NO: 2, 3, 4 and 5.

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Abstract

An isolated polypeptide comprising an amino acid sequence corresponding to the amino acid residues forming a full or partial alpha-helical domain, the hinge domain, the beta-triple spiral domain and afull or partial globular head domain of an avian reovirus sigma C protein, and lacking the amino acid sequence that is N-terminal to said alpha-helical domain is provided. Furthermore, a vaccine comprising, or a viral vector expressing, at least one of the isolated polypeptides of the present invention is provided.

Description

technical field [0001] The present invention generally relates to vaccines against avian reoviruses. Background technique [0002] Avian reovirus (ARV) is a member of the genus Orthoreovirus in the family Reoviridae. It is associated with a number of diseases, most notably the viral arthritic syndrome (tenosynovitis), characterized by swelling of the hock joints and damage to the gastrocnemius tendon, and causes considerable economic loss to the poultry industry. Susceptibility to ARV occurs mainly in young chickens (1-2 weeks old). Control of viral tenosynovitis in broiler chickens is conferred by antibodies that are transferred to offspring following vaccination of the maternal flock. Available live attenuated and inactivated vaccines against ARV are based on the s1133 strain (van der Heide et al. 1983), as well as isolates belonging to a single serotype (Goldenberg et al. 2010). However, these vaccines are ineffective against multiple ARVs found in the field (Jones 200...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/14A61K39/15C12N15/63
CPCC12N15/63A61K39/12A61K2039/552C12N2720/12034C07K14/005C12N2720/12022A61P31/12A61K39/15
Inventor J·皮特柯务斯基D·戈尔登贝格
Owner GAVISH GALILEE BIO APPL
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