Biodegradable polylactide microspheres enhance specific immune response induced by Hepatitis B surface antigen

Hepatitis B (HB) infection caused by Hepatitis B virus (HBV) is the most common liver disease in the world. HB vaccine, when administered in conjunction with alum adjuvants, induces Th2 immunity that confers protection against HBV. However, currently available vaccine formulations and adjuvants do not elicit adequate Th1 and CTL responses that are important for prevention of maternal transmission of the virus. Microspheres synthesized from poly (D, L-lactide-co-glycolide) (PLGA) or poly (D, L-lactide) (PLA) polymers have been considered as promising tools for in vivo delivery of antigens and drugs. Here we describe PLA microspheres synthesized by premix membrane emulsification method and their application in formulating a new microsphere based HB vaccine. To evaluate the immunogenicity of this microsphere vaccine, BALB/c mice were immunized with microsphere vaccine and a series of immunological assays were conducted. Results of Enzyme-linked ImmunoSpot (ELISPOT) assays revealed that the number of interferon-gamma (IFN-γ)-producing splenocytes and CD8+ T cells increased significantly in the microsphere vaccine group. Microsphere vaccine group showed enhanced specific cell lysis when compared with HB surface antigen (HBsAg) only group in 51Cr cytotoxicity assays. Moreover, microsphere vaccine elicited a comparable level of antibody production as that of HB vaccine administered with alum adjuvant. We show that phagocytosis of HBsAg by dendritic cells is more pronounced in microsphere vaccine group when compared with other control groups. These results clearly demonstrate the potential of using PLA microspheres as effective HB vaccine adjuvants for an enhanced Th1 immune response.

[1]  P. Cresswell,et al.  Enhanced and prolonged cross‐presentation following endosomal escape of exogenous antigens encapsulated in biodegradable nanoparticles , 2006, Immunology.

[2]  F. Chisari,et al.  Intracellular inactivation of the hepatitis B virus by cytotoxic T lymphocytes. , 1996, Immunity.

[3]  T. Umbreit,et al.  Significance of the type and the size of biomaterial particles on phagocytosis and tissue distribution. , 2001, Journal of biomedical materials research.

[4]  Chandan Thomas,et al.  Aerosolized PLA and PLGA nanoparticles enhance humoral, mucosal and cytokine responses to hepatitis B vaccine. , 2011, Molecular pharmaceutics.

[5]  F. Chisari,et al.  Polyclonality and multispecificity of the CTL response to a single viral epitope. , 1998, Journal of immunology.

[6]  M. Hilleman Overview of the pathogenesis, prophylaxis and therapeusis of viral hepatitis B, with focus on reduction to practical applications. , 2001, Vaccine.

[7]  J. Granados,et al.  Cellular and Humoral Mechanisms Involved in the Control of Tuberculosis , 2012, Clinical & developmental immunology.

[8]  M. S. Singh,et al.  Poly(lactide-co-glycolide) microparticles for the development of single-dose controlled-release vaccines. , 1998, Advanced drug delivery reviews.

[9]  Manmohan J. Singh,et al.  Microparticles as vaccine adjuvants and delivery systems , 2003, Expert review of vaccines.

[10]  W. Koff,et al.  Controlled release microparticles as a single dose hepatitis B vaccine: evaluation of immunogenicity in mice. , 1997, Vaccine.

[11]  Manmohan J. Singh,et al.  Cationic microparticles: A potent delivery system for DNA vaccines. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[12]  F. Chisari,et al.  Hepatitis B virus immunopathogenesis. , 1995, Annual review of immunology.

[13]  J. Reimann,et al.  Antibody and cytotoxic T-cell responses to soluble hepatitis B virus (HBV) S antigen in mice: implication for the pathogenesis of HBV-induced hepatitis , 1994, Journal of virology.

[14]  M. Shapiro,et al.  Viral clearance without destruction of infected cells during acute HBV infection. , 1999, Science.

[15]  R. Langer,et al.  A single-step immunization by sustained antigen release. , 1979, Journal of immunological methods.

[16]  P. Klenerman,et al.  Cellular immunity in children with successful immunoprophylactic treatment for mother-to-child transmission of hepatitis B virus , 2010, BMC infectious diseases.

[17]  B. Gander,et al.  Induction of a cytotoxic T lymphocyte response by immunization with a malaria specific CTL peptide entrapped in biodegradable polymer microspheres. , 1997, Vaccine.

[18]  W. Koff,et al.  Immunogenicity and protection in small-animal models with controlled-release tetanus toxoid microparticles as a single-dose vaccine , 1997, Infection and immunity.

[19]  Eldridge,et al.  Biodegradable and biocompatible poly(DL-lactide-co-glycolide) microspheres as an adjuvant for staphylococcal enterotoxin B toxoid which enhances the level of toxin-neutralizing antibodies , 1991, Infection and immunity.

[20]  Wei Wei,et al.  Uniform-sized PLA nanoparticles: preparation by premix membrane emulsification. , 2008, International journal of pharmaceutics.

[21]  J. Ulmer,et al.  Induction of Broad and Potent Anti-Human Immunodeficiency Virus Immune Responses in Rhesus Macaques by Priming with a DNA Vaccine and Boosting with Protein-Adsorbed Polylactide Coglycolide Microparticles , 2003, Journal of Virology.

[22]  Sven Frokjaer,et al.  Particle size and surface charge affect particle uptake by human dendritic cells in an in vitro model. , 2005, International journal of pharmaceutics.

[23]  S. Vyas,et al.  Strong systemic and mucosal immune responses to surface-modified PLGA microspheres containing recombinant hepatitis B antigen administered intranasally. , 2006, Vaccine.

[24]  Ding‐Shinn Chen,et al.  Hepatitis B Virus Infection , 2007 .