Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice
Objective: To evaluate the expression of DNA plasmid-harboring modified antibody gene that produces neutralizing human monoclonal antibodies against four serotypes of dengue virus (DENV) without enhancing activity in BALB/c mice. Methods: We constructed pFUSE-based vectors (pFUSE_1G7C2_ hVH and pFUS...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Wolters Kluwer Medknow Publications
2021-01-01
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| Series: | Asian Pacific Journal of Tropical Medicine |
| Subjects: | |
| Online Access: | http://www.apjtm.org/article.asp?issn=1995-7645;year=2021;volume=14;issue=7;spage=299;epage=308;aulast=Benjathummarak |
| _version_ | 1798037543448477696 |
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| author | Surachet Benjathummarak Atsushi Yamanaka Thanyaluk Krasae Chonlatip Pipattanaboon Subenya Injampa Pannamthip Pitaksajjakul Pongrama Ramasoota |
| author_facet | Surachet Benjathummarak Atsushi Yamanaka Thanyaluk Krasae Chonlatip Pipattanaboon Subenya Injampa Pannamthip Pitaksajjakul Pongrama Ramasoota |
| author_sort | Surachet Benjathummarak |
| collection | DOAJ |
| description | Objective: To evaluate the expression of DNA plasmid-harboring modified antibody gene that produces neutralizing human monoclonal antibodies against four serotypes of dengue virus (DENV) without enhancing activity in BALB/c mice.
Methods: We constructed pFUSE-based vectors (pFUSE_1G7C2_ hVH and pFUSE_1G7C2_hVL) containing genes encoding the variable domains of the heavy or light chain of the anti-dengue virus antibody 1G7C2, a human IgG1 that has been characterized for its neutralizing activity to DENV-1-4. Leucine (L) at positions 234 and 235 on the Fc CH2 domain in pFUSE_1G7C2_hVH was mutated to alanine (A) (LALA mutation) by site direct mutagenesis, and the new plasmid was termed pFUSE_1G7C2_hVH_LALA. An equal amount of pFUSE_1G7C2_hVL and 1G7C2_hG1-LALA plasmids were co-transfected into Chinese hamster ovary cells (CHO-K1) and a single dose of 100 μg 1G7C2_hG1-LALA plasmid was intramuscularly injected, followed by electroporation in BALB/c mice. The secreted 1G7C2_hG1-LALA antibodies in cell culture supernatant and mouse serum were examined for their biological functions, neutralization and enhancing activity.
Results: The co-transfection of heavy- and light-chain 1G7C2_ hG1-LALA plasmids in CHO-K1 cells produced approximately 3 900 ng/mL human IgG and neutralized 90%-100% all four DENV, with no enhancing activity. Furthermore, the modified human IgG was produced more than 1 000 ng/mL in mouse serum on day 7 post plasmid injection and showed cross-neutralization to four DENV serotypes. Subsequently, antibody production and neutralization decreased rapidly. Nevertheless, the secreted neutralizing 1G7C2_ hG1-LALA in mouse serum demonstrated complete absence of enhancing activities to all DENV serotypes.
Conclusions: These findings reveal that a new modified 1G7C2_ hG1-LALA expressing plasmid based on gene transfer is a possible therapeutic antibody candidate against DENV infection. |
| first_indexed | 2024-04-11T21:27:58Z |
| format | Article |
| id | doaj.art-c88d5482cd7a4ab59a374bd3bbf27e7f |
| institution | Directory Open Access Journal |
| issn | 2352-4146 |
| language | English |
| last_indexed | 2024-04-11T21:27:58Z |
| publishDate | 2021-01-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Asian Pacific Journal of Tropical Medicine |
| spelling | doaj.art-c88d5482cd7a4ab59a374bd3bbf27e7f2022-12-22T04:02:19ZengWolters Kluwer Medknow PublicationsAsian Pacific Journal of Tropical Medicine2352-41462021-01-0114729930810.4103/1995-7645.320520Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in miceSurachet BenjathummarakAtsushi YamanakaThanyaluk KrasaeChonlatip PipattanaboonSubenya InjampaPannamthip PitaksajjakulPongrama RamasootaObjective: To evaluate the expression of DNA plasmid-harboring modified antibody gene that produces neutralizing human monoclonal antibodies against four serotypes of dengue virus (DENV) without enhancing activity in BALB/c mice. Methods: We constructed pFUSE-based vectors (pFUSE_1G7C2_ hVH and pFUSE_1G7C2_hVL) containing genes encoding the variable domains of the heavy or light chain of the anti-dengue virus antibody 1G7C2, a human IgG1 that has been characterized for its neutralizing activity to DENV-1-4. Leucine (L) at positions 234 and 235 on the Fc CH2 domain in pFUSE_1G7C2_hVH was mutated to alanine (A) (LALA mutation) by site direct mutagenesis, and the new plasmid was termed pFUSE_1G7C2_hVH_LALA. An equal amount of pFUSE_1G7C2_hVL and 1G7C2_hG1-LALA plasmids were co-transfected into Chinese hamster ovary cells (CHO-K1) and a single dose of 100 μg 1G7C2_hG1-LALA plasmid was intramuscularly injected, followed by electroporation in BALB/c mice. The secreted 1G7C2_hG1-LALA antibodies in cell culture supernatant and mouse serum were examined for their biological functions, neutralization and enhancing activity. Results: The co-transfection of heavy- and light-chain 1G7C2_ hG1-LALA plasmids in CHO-K1 cells produced approximately 3 900 ng/mL human IgG and neutralized 90%-100% all four DENV, with no enhancing activity. Furthermore, the modified human IgG was produced more than 1 000 ng/mL in mouse serum on day 7 post plasmid injection and showed cross-neutralization to four DENV serotypes. Subsequently, antibody production and neutralization decreased rapidly. Nevertheless, the secreted neutralizing 1G7C2_ hG1-LALA in mouse serum demonstrated complete absence of enhancing activities to all DENV serotypes. Conclusions: These findings reveal that a new modified 1G7C2_ hG1-LALA expressing plasmid based on gene transfer is a possible therapeutic antibody candidate against DENV infection.http://www.apjtm.org/article.asp?issn=1995-7645;year=2021;volume=14;issue=7;spage=299;epage=308;aulast=Benjathummarakdengue virus; lala mutation; antibody-dependent enhancement (ade); antibody plasmid; electroporation |
| spellingShingle | Surachet Benjathummarak Atsushi Yamanaka Thanyaluk Krasae Chonlatip Pipattanaboon Subenya Injampa Pannamthip Pitaksajjakul Pongrama Ramasoota Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice Asian Pacific Journal of Tropical Medicine dengue virus; lala mutation; antibody-dependent enhancement (ade); antibody plasmid; electroporation |
| title | Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice |
| title_full | Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice |
| title_fullStr | Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice |
| title_full_unstemmed | Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice |
| title_short | Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice |
| title_sort | plasmid dna encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice |
| topic | dengue virus; lala mutation; antibody-dependent enhancement (ade); antibody plasmid; electroporation |
| url | http://www.apjtm.org/article.asp?issn=1995-7645;year=2021;volume=14;issue=7;spage=299;epage=308;aulast=Benjathummarak |
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