"[
ABSTRAKDemam berdarah dengue (DBD) merupakan penyakit yang disebabkan karena
infeksi virus dengue (DENV), yang banyak ditemukan di Indonesia. Belum ada
terapi yang spesifik dalam pengobatan DBD. Upaya pengembangan vaksin
dengue yang efektif sangat diperlukan. Pada penelitian ini dilakukan analisa
imunogenisitas kandidat vaksin DNA prM-E dengue serotipe 2 (pUMD2.kl.20)
dengan menganalisis sel T CD4, sel T CD8, IFN-γ dan TNF-α. pada sel U937 dan
PBMC secara in vitro, bertujuan untuk mengetahui respon imun ketika vaksin
diinjeksikan ke dalam tubuh manusia. Dasar penelitian ini adalah sel U937
ditransfeksi dengan pUMD2.kl.20 menggunakan lipofectamin. Sel U937 akan
berperan sebagai APCs yang akan mengekspresikan protein prM-E DENV-2 dan
mempresentasikan protein tersebut melalui molekul MHC class I dan MHC class
II kepada Peripheral Blood Mononuclear Cell (PBMC) manusia. Tahapan kerja
yang dilakukan dalam penelitian ini terdiri dari: (a) kultur galur sel U937, (b)
transfeksi sel U937 dengan pUMD2.kl.20, (c) transfeksi sel U937 dengan plasmid
s(pUMVC), (d) infeksi sel U937 dengan DENV-2 strain DS.18/09, (e) pewarnaan
dan pengamatan hasil pewarnaan menggunakan alat semi flowcytometri (TALI).
pUMD2.kl.20 mengaktivasi sel T CD4 dan sel T CD8 untuk berploriferasi. Hasil
penelitian ini menunjukkan bahwa konsentrasi sel T CD8 lebih tinggi dari
konsentrasi sel T CD4 dan konsentrasi sel positif yang mensekresikan IFNγ lebih
tinggi dari konsentrasi sel positif yang mensekresikan TNFα. Kondisi optimal dari
aktivasi sel T CD4 oleh pUMD2.kl.20 adalah 24 jam setelah penambahan PBMC
setelah transfeksi (8,53x10⁴sel/ml), untuk sel T CD8 adalah 24 jam setelah
penambahan PBMC setelah transfeksi (49,4x10⁴ sel/ml). Kondisi optimal dari
aktivasi sel+ yang mensekresikan IFNγ oleh pUMD2.kl.20 adalah 24 jam setelah
penambahan PBMC 48 jam setelah transfeksi (9,86x10⁴ sel/ml), dan untuk sel+
yang mensekresikan TNFα adalah 2 jam setelah penambahan PBMC 48 jam
setelah transfeksi (2,1x10⁴ sel/ml). Dari penelitian ini dapat disimpulkan bahwa
pUMD2.kl.20 bersifat imunogenik.
ABSTRACTDengue hemorrhagic fever (DHF) is a disease caused by infection with dengue
virus (DENV), which is found in Indonesia. There is no specific therapy in the
treatment of DHF. An effort to develop an effective dengue vaccine is needed. In
this research, analysis of the immunogenicity of the DNA vaccine candidate prME
dengue serotype 2 (pUMD2.kl.20) by analyzing the CD4 T cells, CD8 T cells,
IFN-γ and TNF-α in U937 cells and PBMC in vitro, aims to determine the
immune response when the vaccine is injected into the human body. This research
approach is based on transfection of U937 cells with pUMD2.kl.20 using
lipofectamin. U937 cells acts as APCs which will express the protein prM-E
DENV-2 and presenting these proteins through the MHC class I and MHC class II
molecule to the Peripheral Blood Mononuclear Cell (PBMC) of human body.
Stages of the work in this study consisted of: (a) culture cell line U937, (b)
transfection of U937 cells with pUMD2.kl.20, (c) transfection of U937 cells with
plasmid (pUMVC), (d) infection of U937 cells with DENV-2 strains DS.18/09,
(e) staining and observation results of staining using a semi-flow cytometri
(TALI). pUMD2.kl.20 activate CD4 T cells and CD8 T cells to proliferate. CD8 T
cells concentration higher than the concentration of CD4 T cells and the secretion
of INFγ-positive cells concentration higher than the concentration of the secretion
of TNFα-positive cells. Optimal condition of CD4 T cells activation by
pUMD2.kl.20 is 24 hours after the addition of PBMC after transfection (8,53x10⁴
cells/ml), for CD8 T cells was 24 hours after the addition of PBMC after
transfection (49,4x10⁴ cells/ml). Optimal conditions secretion of IFNγ-positive
cells were activated by pUMD2.kl.20 is 24 hours after the addition of PBMC 48
hours after transfection (9,86x10⁴ cells/ml), and for the secretion of TNFα-
positive cells were activated by pUMD2.kl.20 is 2 hours after the addition of
PBMC 48 hours after transfection (2,1x10⁴ cells/ml). From this study it can be
concluded that the pUMD2.kl.20 immunogenic, Dengue hemorrhagic fever (DHF) is a disease caused by infection with dengue
virus (DENV), which is found in Indonesia. There is no specific therapy in the
treatment of DHF. An effort to develop an effective dengue vaccine is needed. In
this research, analysis of the immunogenicity of the DNA vaccine candidate prME
dengue serotype 2 (pUMD2.kl.20) by analyzing the CD4 T cells, CD8 T cells,
IFN-γ and TNF-α in U937 cells and PBMC in vitro, aims to determine the
immune response when the vaccine is injected into the human body. This research
approach is based on transfection of U937 cells with pUMD2.kl.20 using
lipofectamin. U937 cells acts as APCs which will express the protein prM-E
DENV-2 and presenting these proteins through the MHC class I and MHC class II
molecule to the Peripheral Blood Mononuclear Cell (PBMC) of human body.
Stages of the work in this study consisted of: (a) culture cell line U937, (b)
transfection of U937 cells with pUMD2.kl.20, (c) transfection of U937 cells with
plasmid (pUMVC), (d) infection of U937 cells with DENV-2 strains DS.18/09,
(e) staining and observation results of staining using a semi-flow cytometri
(TALI). pUMD2.kl.20 activate CD4 T cells and CD8 T cells to proliferate. CD8 T
cells concentration higher than the concentration of CD4 T cells and the secretion
of INFγ-positive cells concentration higher than the concentration of the secretion
of TNFα-positive cells. Optimal condition of CD4 T cells activation by
pUMD2.kl.20 is 24 hours after the addition of PBMC after transfection (8,53x10⁴
cells/ml), for CD8 T cells was 24 hours after the addition of PBMC after
transfection (49,4x10⁴ cells/ml). Optimal conditions secretion of IFNγ-positive
cells were activated by pUMD2.kl.20 is 24 hours after the addition of PBMC 48
hours after transfection (9,86x10⁴ cells/ml), and for the secretion of TNFα-
positive cells were activated by pUMD2.kl.20 is 2 hours after the addition of
PBMC 48 hours after transfection (2,1x10⁴ cells/ml). From this study it can be
concluded that the pUMD2.kl.20 immunogenic]"
