Injampa, Subenya

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Injampa, Subenya
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subenya.in@kmitl.ac.th

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    Item type:Publication,
    Human heavy chain antibody genes elicited in thai dengue patients during DENV2 secondary infection
    (2020-01-01)
    Thammasonthijarern, Nipa
    ;
    Puangmanee, Wilarat
    ;
    Sriburin, Pimolpachr
    ;
    Injampa, Subenya
    ;
    Chatchen, Supawat
    Dengue is one of the most serious mosquito-borne viral diseases occurring in humans. To combat the complexity of 4 antigenically distinct serotypes, the ideal vaccine for dengue should be able to stimulate cross-neutralizing antibodies. Recently, genetics-based immune responses have been studied to guide vaccine design against several viral pathogens. Despite a recent approval of dengue vaccine, information on genetics-based immune responses against dengue virus (DENV) is still limited. Consequently, we aimed to determine the profiles of immunoglobulin heavy chain genes from DENV2 infected patients. The immunoglobulin heavy chain variable region genes (IGHV) were amplified from peripheral blood mononuclear cells of DENV2 secondary infected patients in the acute, convalescence, and recovery phases. Antibody heavy chain genes were sequenced using next-generation sequencing, and analyzed to identify correlations with neutralizing and enhancing activities of the serum samples. IGHV1-69, 3-23, and 3-30 were frequently discovered in our Thai DENV2 infected patients. Our findings provide new data on the human B cell response during secondary DENV2 infections in Thai dengue patients that offer supportive information for dengue vaccine design and therapeutics development.
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    Item type:Publication,
    Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice
    (2021-07-01)
    Benjathummarak, Surachet
    ;
    Yamanaka, Atsushi
    ;
    Krasae, Thanyaluk
    ;
    Pipattanaboon, Chonlatip
    ;
    Injampa, Subenya
    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 1G7C2hG1-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.