Evaluation of the sensitivity of Ghanaian HIV-1 strains to protease inhibitors

• Project/Area Number: 15590426
• 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 Infectious Diseases
• Principal Investigator: TOKUNAGA Kenzo Natl.Inst.Infec.Dis., Dept.of Pathology, PI, 感染病理部, 主任研究官 (50342895)
• Co-Investigator(Kenkyū-buntansha): ISHIKAWA Koichi Natl.Inst.Infec.Dis., AIDS Research Center, PI, エイズ研究センター, 主任研究官 (50192480) ; SATA Tetsutaro Natl.Inst.Infec.Dis., Dept.of Pathology, Director, 感染病理部, 部長 (00162397)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: HIV-1 / Ghanaian strains / Subtypes / Protease inhibitors / Phylogenetic tree / Cassette vector / Drug sensitivity / Structural analysis / プロテアーゼ

Research Abstract

Now that highly active antiretroviral therapy(HAART) is being initiated on a large scale in West Africa, it remains controversial whether protease inhibitors(PIs), originally designed and tested against human immunodeficiency virus type 1(HTV-1) subtype B, are equally effective against the non-B subtypes that are prevalent in West African countries. In this study, we investigated whether Ghanaian HIV-1 isolates, as representatives of West African isolates, are susceptible to PIs. We first generated an HIV-1 protease cassette vector proviral DNA fragment carrying a luciferase gene, which allows patient-derived HIV-1 proteases to be inserted and to be subjected to both genotypic and phenotypic assays. HIV-1 protease genes derived from 39 treatment-naive Ghanaian patients were used in this experiment as representatives of West African strains. The cloned patient-derived HIV-1 protease genes were first sequenced and then genetically compared. Phenotypic analysis was performed with Ghanaian HTV-1 protease chimeric viruses in the presence of 6 different PIs. Structural models of an HIV-1 protease homodimers were constructed by the molecular modeling software. Genetic analysis of cloned patient-derived HIV-1 protease genes indicated that most of the Ghanaian HIV-1 proteases are placed as subtype CRF02_AG strains, which are phylogenetically distant from subtype B strains, and that Ghanaian HIV-1 proteases do not harbor known major mutations influencing drug resistance but commonly carry 2-3 minor mutations. Phenotypic analysis performed with HIV-1 protease-recombinant viruses in the presence of 6 different PIs revealed that Ghanaian HIV-1 proteases are differentially less susceptible to the PIs. In support of this finding of differential susceptibility, structural analysis showed a significant distortion of nelfinavir, but not of amprenavir, in the Ghanaian protease pocket, suggesting nelfinavir might be less insertable into the Ghanaian protease than into the protease of subtype B. These findings provide implications for the combination of PIs during the introduction of HAART into West Africa.

Research Products

• [Journal Article] Interleukin-4 up-regulates T-tropic human immunodeficiency virus type 1 transcription in primary CD4(+) CD38(+) T-lymphocyte subset (2005)
  • Author(s): Li, Y.G., Iwabu, Y., Warachit, J., Kinomoto, M., Ibrahim, M.S., Tsuji, S., Mukai, T., Kameoka, M., Tokunaga, K., Sata, T., Ikuta, K.
  • Journal Title: Microbiol.Immunol. 49 Pages: 155-165
  • NAID: 10014430004
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Amino acid 36 in HIV-1 gp41 ectodomain controls fusogenic activity : Implications for the molecular mechanism of viral escape from a fusion inhibitor. (2005)
  • Author(s): Kinomoto, M., Yokoyama, M., Sato, H., Kojima, A., Kurata, T., Ikuta, K., Sata, T., Tokunaga, K.
  • Journal Title: J.Virol. 79 Pages: 5996-6004
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] HIV-1 proteases from drug-naive West African patients are differentially less susceptible to protease inhibitors. (2005)
  • Author(s): Kinomoto, M., Appiah-Opong, R., Brandful, J.A.M., Yokoyama, M., Nii-Trebi, N., Ugly-Kwame, E., Sato, H., Ofori-Adjei, D., Kurata, T., Barre-Sinoussi, F., Sata, T., Tokunaga, K.
  • Journal Title: Clin.Infect.Dis. 41 Pages: 243-251
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Interleukin-4 upregulates T-tropic human immunodeficiency virus type 1 transcription in primary CD4^+CD38^+ T-lymphocyte subset. (2005)
  • Author(s): Yong-Gang, L., Iwabu, Y., Warachit, J., Kinomoto, M., Ibrahim, M.S., Tsuji, Y, Mukai, T., Kameoka, M., Tokunaga, K., Sata, T., Ikuta, K.
  • Journal Title: Microbiol.Immunol. 49 Pages: 155-165
  • NAID: 10014430004
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] HIV-1 proteases from drug-naive West African patients are differentially less susceptible to protease inhibitors. (2005)
  • Author(s): Kinomoto, M., Appiah-Opong, R., Brandful, J.A.M., Yokoyama, M., Nii-Trebi, N., Ugly-Kwame, E., Sato, H., Ofori-Adjei, D., Kurata, T., Barre-Sinoussi, F., Sata, T., Tokunaga.K.
  • Journal Title: Clin.Infect.Dis. 41 Pages: 243-251
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Nucleolin and the packaging signal, psi, promote the budding of human immunodeficiency virus type-1 (HIV-1). (2005)
  • Author(s): Li, Y.G., Iwabu, Y., Warachit, J., Kinomoto M., Ibrahim, M.S., Tsuji, S., Mukai, T., Kameoka, M., Tokunaga, K., Sata, T., Ikuta, K.
  • Journal Title: Microbiol.Immunol. 48 Pages: 111-118
• [Journal Article] HIV-1 proteases from drug-naive West African patients are differentially less susceptible to protease inhibitors. (2005)
  • Author(s): Kinomoto, M., Appiah-Opong, R., Brandful, J.A.M., Yokoyama, M., Nii-Trebi, N., Ugly-Kwame, E., Sato, H., Ofori-Adjei, D., Kurata, T., Barre-Sinoussi, F., Sata, T., Tokunaga, K.
  • Journal Title: Clin.Infect.Dis. In press
• [Journal Article] Amino acid 36 in HIV-1 gp41 ectodomain controls fusogenic activity : Implications for the molecular mechanism of viral escape from a fusion inhibitor. (2005)
  • Author(s): Kinomoto, M., Yokoyama, M., Sato, H., Kojima, A., Kurata, T., Ikuta, K., Sata, T., Tokunaga, K.
  • Journal Title: J.Virol. In press
• [Journal Article] Nucleolin, and the packaging signal, Ψ, promote the budding of human immunodeficiency virus type 1 (HIV-1) (2004)
  • Author(s): Ueno, T., Tokunaga, K., Sawa, H., Maeda, M., Chiba, J., Kojima, A., Hasegawa, H., Shoya, Y., Sata, T., Kurata, T., Takahashi, H.
  • Journal Title: Microbiol.Immunol. 48 Pages: 111-118
  • NAID: 50000003890
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Human APOBEC3F is another host factor that blocks HIV-1 replication. (2004)
  • Author(s): Zheng, Y.-H., Irwin, D., Kurosu, T., Tokunaga, K., Sata, T., Peterlin, B.M.
  • Journal Title: J.Virol. 78 Pages: 6073-6076
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Nucleolin and the packaging signal, Ψ, promote the budding of human immuno- deficiency virus type-1(HIV-1). (2004)
  • Author(s): Ueno, T., Tokunaga, K., Sawa, H., Maeda, M., Chiba, J, Kojima, A., Hasegawa, H., Shoya, Y., Sata, T., Kurata, T.,, Takahashi, H.
  • Journal Title: Microbiol.Immunol. 48 Pages: 111-118
  • NAID: 50000003890
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Human APOBEC3F is another host factor that blocks human immunodeficiency virus type 1 replication. (2004)
  • Author(s): Zheng, Y.H., Irwin, D., Kurosu, T., Tokunaga, K., Sata, T., Peterlin, B.M.
  • Journal Title: J.Virol. 78 Pages: 6073-6076
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Human APOBEC3F is another host factor that blocks HIV-1 replication. (2004)
  • Author(s): Zheng, Y.-H., Irwin, D., Kurosu, T., Tokunaga, K., Sata.T., Peterlin, B.M.
  • Journal Title: J.Virol. 78 Pages: 6073-6076
• [Journal Article] Human topoisomerase I promotes HIV-1 proviral DNA synthesis : implications for the species specificity of HIV-1 infection. (2003)
  • Author(s): Shoya, Y., Tokunaga, K., Sawa, H., Maeda, M., Ueno, T., Yoshikawa, T., Sata, T., Kurata, T., Cullen, B.R., Takahashi, H.
  • Journal Title: Proc.Natl.Acad.Sci.U.S.A. 100 Pages: 8442-8447
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Interleukin-8 cross-desensitizes cellular responses to both CCR5 and CXCR4 but inhibitsHIV-1 infection only to CCR5 : role of receptor cross-internalization and signal strength. (2003)
  • Author(s): Richardson, R.M., Tokunaga, K., Marjoram, R., Sata, T., Snyderman, R
  • Journal Title: J.Biol.Chem. 278 Pages: 15867-15873
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Interleukin-8 cross-desensitizes cellular responses to both CCR5 and CXCR4 but inhibits HIV-1 infection only to CCR5 : role of receptor cross-internalization and signal strength. (2003)
  • Author(s): Richardson, R.M., Tokunaga, K., Marjoram, R., Sata, T., Snyderman, R.
  • Journal Title: J.Biol.Chem. 278 Pages: 15867-15873
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Human topoisomerase I promotes HIV-1 proviral DNA synthesis : implications for the species specificity of HIV-1 infection. (2003)
  • Author(s): Shoya, Y., Tokunaga, K.(co-corresponding author), Sawa, H., Maeda, M., Ueno, T., Yoshikawa, T., Sata, T., Kurata, T., Cullen, B.R., Takahashi, H.
  • Journal Title: Proc.Natl.Acad.Sci.U.S.A. 100 Pages: 8442-8447
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Richardson, R.M., Tokunaga K., Marjoram, R., Sata, T., Snyderman, R: "Interleukin-8 cross-desensitizes cellular responses to both CCR5 and CXCR4 but inhibits HIV-1 infection only to CCR5 : role of receptor cross-internalization and signal strength."J.Biol.Chem.. 278. 15867-15873 (2003)
• [Publications] Shoya, Y., Tokunaga, K., Sawa, H., Maeda, M., Ueno, T., Yoshikawa, T., Sata, T., Kurata, T., Cullen, B.R., Takahashi, H.: "Human topoisomerase I promotes HIV-1 proviral DNA synthesis : implications for the species specificity of HIV-1 infection."Proc.Natl.Acad.Sci.U.S.A.. 100. 8442-8447 (2003)
• [Publications] Ueno, T., Tokunaga, K., Sawa, H., Maeda, M., Chiba, J., Kojima, A., Hasegawa, H., Shoya, Y., Sata, T., Kurata, T., Takahashi, H.: "Nucleolin, and the packaging signal, Ψ, promote the budding of human immunodeficiency virus type 1(HIV-1)"Microbiol.Immunol.. 48. 111-118 (2004)
• [Publications] Zheng, Y.-H., Irwin, D., Kurosu, T., Tokunaga, K., Sata, T., Peterlin, B.M.: "Human APOBEC3F is another host factor that blocks HIV-1 replication."J.Virol.. In press. (2004)