DEVELOPMENT OF NOVEL HIV-1 SUBTYPING METHOD USING RECOMBINANT GP120S EXPRESSED BY SENDAI VIRUS VECTOR

• Project/Area Number: 12670288
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Virology
• Research Institution: NATIONAL INSTITUTE OF INFECTIONS DISEASES
• Principal Investigator: TAKEBE YUTAKA NATIONAL INSTITUTE OF INFECTIONS DISEASES, AIDS RESEARCH CENTER, CHIEF, エイズ研究センター第1室, 室長 (50126116)
• Project Period (FY): 2000 – 2001
• Project Status: Completed (Fiscal Year 2001)
• Budget Amount: ¥3,400,000 (Direct Cost: ¥3,400,000); Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
• Keywords: HIV-1 subtype / HIV-1 envelope / Sendai virus vector / EIA / serotyping / HIV-1エンベロープタンパク質 / EIA / ELISA

Research Abstract

We developed Sendai virus (SeV) vectors that express HIV-1 env gp120s of subtypes A through E. In order to assess the expression level ofgp120, we developed the sandwich enzyme immunoassay (ElA) using purified polyclonal sera against C-terminus of gp120 (first antibody) and pooled HIV(+) sera (second antibody). Employing this assay system, we evaluated the effect of codon optimization and of fusion with CD5 leader sequence on expression level of gp120.We found that codon-optimization enhanced the expression of gp120 by 3-fold and the fusion to CD5 leader sequence showed additional enhancement of 1.5-fold.
EIA using the recombinant gp120 expressed by SeV vectors could differentiate the subtypes between B and E. Since high-level of cross-reactivity were observed between other subtypes, we found that the universal serological subtyping of HIV-1 subtypes is not feasible to achieve.

Research Products

• [Publications] Toriyoshi, H.Shioda, T., Sato, H., Sakaguchi, M., Eda, Y., Tokiyoshi, S., Kato, K., Nohtomi, K., Kusagawa, S., Taniguchi, K., Shiino, T., Kato, A., Foongladda, S., Linkanonsakul, S., Oka, S., Iwamoto, I., Wasi, C., Nagai, Y., Takebe, Y.: "Sendai virus based production of HIV-1 subtype B and subtype E gp120 antigens and their use for highly sensitive detection of subtype-specific serum antibodies"AIDS Res and Human Retroviruses. 15. 1109-1120 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Toriyoshi, H. Shioda, T., Sato, H., Sakaguchi, M., Eda, Y., Tokiyoshi, S., Kato, K., Nohtomi, K., Kusagawa, S., Taniguchi, K., Shiino, T., Kato, A., Foongladda, S., Linkanonsakul, S., Oka, S., Iwamoto, L, Wasi, C., Nagai, Y., and Takebe, Y.: "Sendai virus based production of HIV-1 subtype B and subtype E gp120 antigens and their use for highly sensitive detection of subtype-specific serum antibodies"AIDS Res and Human Retroviruses. 15. 1109-1120 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Toriyoshi, H., Shioda, T., Sato, H., Takebe, Y., et al.: "Sendal virus based production of HIV-1 subtype B and subtype E gp12O antigens and their use for highly sensitive detection of subtype-specific serum antibodies"AIDS Research and Human Retroviruses. 15. 1109-1120 (1999)
• [Publications] Toriyoshi,H.,Shioda,T.,Sato,H.,Takebe,Y., et al.: "Sendai virus based production of HIV-1 subtype B and subtype E gp120 antigens and their use for highly sensitive detection of subtype-specific serum antibodies."AIDS Research and Human Retroviruses.. 15. 1109-1120 (1999)
• [Publications] Motomura,K.,Kusagawa,Kato,K.,Takebe,Y., et al.: "Emergence of new forms of human immunodeficiency virus type 1 intersubtype recombinants in Central Myanmar."AIDS Research and Human Retroviruses.. 16. 1831-1843 (2000)
• [Publications] Kato,K.,Kusagawa,S.,Motomura,K.,Takebe.Y., et al.: "Closely related HIV-1 CRF01_AE variants among injecting drug users in northem Vietnam : Evidence of HIV spread across the Vietnam-China border"AIDS Research and Human Retroviruses. 17. 113-123 (2001)