Virus like particle of hepatitis B virus pre-S protein
View Patent ↗As disclosed herein, the preS antigen on infectious hepatitis B virus (HBV) particles can provide B and T ceil epitopes that promote the humoral and cellular responses and enhance the seroprotection rate by overcoming non-responsiveness to the S antigen-only vaccines. Therefore, compositions and methods are disclosed using the preS antigen to develop vaccines and immune therapies for treating or preventing hepatitis B infection, in particular, virus-like particles (VLPs) are disclosed that contain the preS antigen on its surface. These VLPs can be used alone or in combination with vaccines containing the hepatitis B surface antigen (HBsAg) to vaccinate subjects against HBV as well as to activate T ceils for adoptive T cell therapy to eradicate HBV infected hapatocytes.
1. A virus like particle (VLP) comprising
a) a fusion protein comprising
i) a signal peptide,
ii) a hepatitis B preS antigen,
iii) a heterologous hemagglutinin (HA) transmembrane domain; and
b) an influenza virus matrix protein 1 (M1),
wherein the signal peptide is fused at the N-terminus of the hepatitis B preS antigen, wherein the hepatitis B preS antigen is fused at the N-terminus of the heterologous HA transmembrane domain, and
wherein the preS antigen comprises the amino acid sequence SEQ ID NO:1, or an antigenic variant thereof having at least 90% identity to SEQ ID NO:1.
2. The VLP of claim 1 , wherein the preS antigen comprises the amino acid sequence SEQ ID NO: 1.
3. The VLP of claim 1 , wherein the fusion protein comprises the amino acid sequence SEQ ID NO:2.
4. The VLP of claim 1 , wherein the signal peptide comprises the signal peptide from HA.
5. The VLP of claim 1 , produced by coinfecting insect cells with one or more recombinant baculoviruses expressing the matrix protein and the fusion protein, culturing the insect cells under physiological conditions, and purifying the VLPs from insect cell culture supernatants.
6. The VLP of claim 5 , produced by introducing into mammalian cells one or more expression vectors expressing the matrix protein and the fusion protein, culturing the mammalian cells under appropriate conditions, and purifying the VLPs from cell culture supernatants.
7. A vaccine comprising an effective amount of the VLP of claim 1 in a pharmaceutically acceptable excipient.
8. The vaccine of claim 7 , further comprising an adjuvant.
9. A method of vaccinating a subject for hepatitis B comprising administering the vaccine of claim 7 to a subject in need thereof by intranasal, intramuscular, subcutaneous, transdermal, or sublingual administration.
10. The method of claim 9 , further comprising administering to the subject a vaccine comprising a hepatitis B surface antigen (HBsAg).
11. A method for activating CD8+ T cells for adoptive cell therapy (ACT), comprising co-culturing CD8+ T cells and dendritic cells with the VLPs of claim 1 .
12. The VLP of claim 1 , wherein the heterologous HA further comprises a cytoplasmic tail.
13. The VLP of claim 12 , wherein the heterologous HA comprises the amino acid sequence SEQ ID NO:15, or a variant thereof having at least 90% identity to SEQ ID NO:15.