Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 3154-3163 |
| Number of pages | 10 |
| Journal | Vaccine |
| Volume | 19 |
| Issue number | 23-24 |
| DOIs | |
| Publication status | Published - Apr 30 2001 |
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Keywords
- Atomic force microscopy
- HBsAg L particles
- Immune responses
- Pre-S1 region
- Yeast-derived hepatitis B vaccine
ASJC Scopus subject areas
- Immunology
- Microbiology
- Virology
- Infectious Diseases
- Public Health, Environmental and Occupational Health
- veterinary(all)
Cite this
Physicochemical and immunological characterization of hepatitis B virus envelope particles exclusively consisting of the entire L (pre-S1 + pre-S2 + S) protein. / Yamada, Tadanori; Iwabuki, Hidehiko; Kanno, Takashi; Tanaka, Hiroyuki; Kawai, Tomoji; Fukuda, Hideki; Kondo, Akihiko; Seno, Masaharu; Tanizawa, Katsuyuki; Kuroda, Shun'ichi.
In: Vaccine, Vol. 19, No. 23-24, 30.04.2001, p. 3154-3163.Research output: Contribution to journal › Article
}
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
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 -