Human Transbodies to Reverse Transcriptase Connection Subdomain of HIV-1 Gag-Pol Polyprotein Reduce Infectiousness of the Virus Progeny
HIV-1 progeny are released from infected cells as immature particles that are unable to infect new cells. Gag-Pol polyprotein dimerization via the reverse transcriptase connection domain (RTCDs) is pivotal for proper activation of the virus protease (PR protein) in an early event of the progeny viru...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-08-01
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| Series: | Vaccines |
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| Online Access: | https://www.mdpi.com/2076-393X/9/8/893 |
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| author | Watee Seesuay Siratcha Phanthong Jaslan Densumite Kodchakorn Mahasongkram Nitat Sookrung Wanpen Chaicumpa |
| author_facet | Watee Seesuay Siratcha Phanthong Jaslan Densumite Kodchakorn Mahasongkram Nitat Sookrung Wanpen Chaicumpa |
| author_sort | Watee Seesuay |
| collection | DOAJ |
| description | HIV-1 progeny are released from infected cells as immature particles that are unable to infect new cells. Gag-Pol polyprotein dimerization via the reverse transcriptase connection domain (RTCDs) is pivotal for proper activation of the virus protease (PR protein) in an early event of the progeny virus maturation process. Thus, the RTCD is a potential therapeutic target for a broadly effective anti-HIV agent through impediment of virus maturation. In this study, human single-chain antibodies (HuscFvs) that bound to HIV-1 RTCD were generated using phage display technology. Computerized simulation guided the selection of the transformed <i>Escherichia coli</i>-derived HuscFvs that bound to the RTCD dimer interface. The selected HuscFvs were linked molecularly to human-derived-cell-penetrating peptide (CPP) to make them cell-penetrable (i.e., become transbodies). The CPP-HuscFvs/transbodies produced by a selected transformed <i>E. coli</i> clone were tested for anti-HIV-1 activity. CPP-HuscFvs of transformed <i>E. coli</i> clone 11 (CPP-HuscFv11) that presumptively bound at the RTCD dimer interface effectively reduced reverse transcriptase activity in the newly released virus progeny. Infectiousness of the progeny viruses obtained from CPP-HuscFv11-treated cells were reduced by a similar magnitude to those obtained from protease/reverse transcriptase inhibitor-treated cells, indicating anti-HIV-1 activity of the transbodies. The CPP-HuscFv11/transbodies to HIV-1 RTCD could be an alternative, anti-retroviral agent for long-term HIV-1 treatment. |
| first_indexed | 2024-03-10T08:18:24Z |
| format | Article |
| id | doaj.art-3906b8d8c27244999560c36917b7900c |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-10T08:18:24Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-3906b8d8c27244999560c36917b7900c2023-11-22T10:07:36ZengMDPI AGVaccines2076-393X2021-08-019889310.3390/vaccines9080893Human Transbodies to Reverse Transcriptase Connection Subdomain of HIV-1 Gag-Pol Polyprotein Reduce Infectiousness of the Virus ProgenyWatee Seesuay0Siratcha Phanthong1Jaslan Densumite2Kodchakorn Mahasongkram3Nitat Sookrung4Wanpen Chaicumpa5Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandCenter of Excellent Research of Allergy and Immunology, Department of Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandGraduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandCenter of Research Excellence on Therapeutic Proteins and Antibody Engineering, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandCenter of Research Excellence on Therapeutic Proteins and Antibody Engineering, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandCenter of Research Excellence on Therapeutic Proteins and Antibody Engineering, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandHIV-1 progeny are released from infected cells as immature particles that are unable to infect new cells. Gag-Pol polyprotein dimerization via the reverse transcriptase connection domain (RTCDs) is pivotal for proper activation of the virus protease (PR protein) in an early event of the progeny virus maturation process. Thus, the RTCD is a potential therapeutic target for a broadly effective anti-HIV agent through impediment of virus maturation. In this study, human single-chain antibodies (HuscFvs) that bound to HIV-1 RTCD were generated using phage display technology. Computerized simulation guided the selection of the transformed <i>Escherichia coli</i>-derived HuscFvs that bound to the RTCD dimer interface. The selected HuscFvs were linked molecularly to human-derived-cell-penetrating peptide (CPP) to make them cell-penetrable (i.e., become transbodies). The CPP-HuscFvs/transbodies produced by a selected transformed <i>E. coli</i> clone were tested for anti-HIV-1 activity. CPP-HuscFvs of transformed <i>E. coli</i> clone 11 (CPP-HuscFv11) that presumptively bound at the RTCD dimer interface effectively reduced reverse transcriptase activity in the newly released virus progeny. Infectiousness of the progeny viruses obtained from CPP-HuscFv11-treated cells were reduced by a similar magnitude to those obtained from protease/reverse transcriptase inhibitor-treated cells, indicating anti-HIV-1 activity of the transbodies. The CPP-HuscFv11/transbodies to HIV-1 RTCD could be an alternative, anti-retroviral agent for long-term HIV-1 treatment.https://www.mdpi.com/2076-393X/9/8/893human single-chain antibodies (HuscFvs)cell-penetrating antibodies (transbodies)human immunodeficiency virus 1 (HIV-1)reverse transcriptase connection subdomain (RTCD)Gag-Pol polyprotein |
| spellingShingle | Watee Seesuay Siratcha Phanthong Jaslan Densumite Kodchakorn Mahasongkram Nitat Sookrung Wanpen Chaicumpa Human Transbodies to Reverse Transcriptase Connection Subdomain of HIV-1 Gag-Pol Polyprotein Reduce Infectiousness of the Virus Progeny Vaccines human single-chain antibodies (HuscFvs) cell-penetrating antibodies (transbodies) human immunodeficiency virus 1 (HIV-1) reverse transcriptase connection subdomain (RTCD) Gag-Pol polyprotein |
| title | Human Transbodies to Reverse Transcriptase Connection Subdomain of HIV-1 Gag-Pol Polyprotein Reduce Infectiousness of the Virus Progeny |
| title_full | Human Transbodies to Reverse Transcriptase Connection Subdomain of HIV-1 Gag-Pol Polyprotein Reduce Infectiousness of the Virus Progeny |
| title_fullStr | Human Transbodies to Reverse Transcriptase Connection Subdomain of HIV-1 Gag-Pol Polyprotein Reduce Infectiousness of the Virus Progeny |
| title_full_unstemmed | Human Transbodies to Reverse Transcriptase Connection Subdomain of HIV-1 Gag-Pol Polyprotein Reduce Infectiousness of the Virus Progeny |
| title_short | Human Transbodies to Reverse Transcriptase Connection Subdomain of HIV-1 Gag-Pol Polyprotein Reduce Infectiousness of the Virus Progeny |
| title_sort | human transbodies to reverse transcriptase connection subdomain of hiv 1 gag pol polyprotein reduce infectiousness of the virus progeny |
| topic | human single-chain antibodies (HuscFvs) cell-penetrating antibodies (transbodies) human immunodeficiency virus 1 (HIV-1) reverse transcriptase connection subdomain (RTCD) Gag-Pol polyprotein |
| url | https://www.mdpi.com/2076-393X/9/8/893 |
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