TY - JOUR
T1 - Physicochemical and immunological characterization of hepatitis B virus envelope particles exclusively consisting of the entire L (pre-S1 + pre-S2 + S) protein
AU - Yamada, Tadanori
AU - Iwabuki, Hidehiko
AU - Kanno, Takashi
AU - Tanaka, Hiroyuki
AU - Kawai, Tomoji
AU - Fukuda, Hideki
AU - Kondo, Akihiko
AU - Seno, Masaharu
AU - Tanizawa, Katsuyuki
AU - Kuroda, Shun'ichi
N1 - Funding Information:
We thank Professor Yuji Goto and Ms Miyo Sakai (Osaka University) for performing the analytical ultracentrifugation, Professor Hiroko Tada (Okayama University) for helpful advices, and Ms Yayoi Wada and Mr. Kazumasa Fukao for their technical assistance. This study was supported in part by research grants from TERUMO Life Science Foundation, Kowa Life Science Foundation, and Naito Foundation, and a Grant-in-Aid for Scientific Research on the Osaka University Center of Excellence (COE) program “Creation of Highly Harmonized Functional Materials” from the Ministry of Education, Science, Sports and Culture of Japan.
PY - 2001/4/30
Y1 - 2001/4/30
N2 - The hepatitis B virus (HBV) envelope (env) protein is composed of three regions; the 108- or 119-residue pre-S1 region involved in the direct interaction with hepatocytes, the 55-residue pre-S2 region associated with the polymerized albumin-mediated interaction, and the major 226-residue S protein region. Thus, to improve the immunogenic potency of conventional HB vaccines, development of a new vaccine containing the entire pre-S1 region in addition to pre-S2 and S is desired. We previously reported the efficient production of the HBV env L (pre-S1 + pre-S2 + S) protein in the recombinant yeast cells [J Biol Chem 267 (1992) 1953]. In this study, the HBV env L protein produced as nano-particles in yeast has been purified and characterized. By equilibrium sedimentation, an average molecular weight of L particle was estimated to be approximately 6.4 × 106, indicating that about 110 molecules of L proteins are assembled into an L particle. By atomic force microscopy in a moist atmosphere, the L particles were observed as large spherical particles with a diameter of 50-500 nm. The L particles were stable on short-time heating at a high temperature and long-time storage at a low temperature but rather unstable on repeated freezing and thawing and treatment with dithiothreitol. When immunized in mice, L particles elicited efficiently and simultaneously the anti-S, anti-pre-S2, and anti-pre-S1 antibodies. The ED50 values in mice for the anti-S and anti-pre-S2 antibodies were similar to those elicited by the M (pre-S2 + S) particles. Furthermore, the anti-pre-S1 rabbit antibodies were found to recognize various segments of the pre-S1 region, including the pre-S1 (21-47) segment. These results show the high ability of L particles to induce all antibodies against HBV env proteins, hence promising the future application of L particles for the next generation HB vaccine.
AB - The hepatitis B virus (HBV) envelope (env) protein is composed of three regions; the 108- or 119-residue pre-S1 region involved in the direct interaction with hepatocytes, the 55-residue pre-S2 region associated with the polymerized albumin-mediated interaction, and the major 226-residue S protein region. Thus, to improve the immunogenic potency of conventional HB vaccines, development of a new vaccine containing the entire pre-S1 region in addition to pre-S2 and S is desired. We previously reported the efficient production of the HBV env L (pre-S1 + pre-S2 + S) protein in the recombinant yeast cells [J Biol Chem 267 (1992) 1953]. In this study, the HBV env L protein produced as nano-particles in yeast has been purified and characterized. By equilibrium sedimentation, an average molecular weight of L particle was estimated to be approximately 6.4 × 106, indicating that about 110 molecules of L proteins are assembled into an L particle. By atomic force microscopy in a moist atmosphere, the L particles were observed as large spherical particles with a diameter of 50-500 nm. The L particles were stable on short-time heating at a high temperature and long-time storage at a low temperature but rather unstable on repeated freezing and thawing and treatment with dithiothreitol. When immunized in mice, L particles elicited efficiently and simultaneously the anti-S, anti-pre-S2, and anti-pre-S1 antibodies. The ED50 values in mice for the anti-S and anti-pre-S2 antibodies were similar to those elicited by the M (pre-S2 + S) particles. Furthermore, the anti-pre-S1 rabbit antibodies were found to recognize various segments of the pre-S1 region, including the pre-S1 (21-47) segment. These results show the high ability of L particles to induce all antibodies against HBV env proteins, hence promising the future application of L particles for the next generation HB vaccine.
KW - Atomic force microscopy
KW - HBsAg L particles
KW - Immune responses
KW - Pre-S1 region
KW - Yeast-derived hepatitis B vaccine
UR - http://www.scopus.com/inward/record.url?scp=0035972033&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0035972033&partnerID=8YFLogxK
U2 - 10.1016/S0264-410X(01)00017-2
DO - 10.1016/S0264-410X(01)00017-2
M3 - Article
C2 - 11312011
AN - SCOPUS:0035972033
VL - 19
SP - 3154
EP - 3163
JO - Vaccine
JF - Vaccine
SN - 0264-410X
IS - 23-24
ER -
