Tesis

Uji Tantang Seed Vaksin pcDNA3.1 -rpfB dan pcDNA3.1-rpfD Dalam Mencegah Infeksi Mycobacterium tuberculosis Pada Mencit Balb/c = Challenge Test of pcDNA3.1-rpfB and pcDNA3.1-rpfD Seed Vaccines in Preventing Mycobacterium tuberculosis Infection in Balb/c Mice.

Tuberkulosis (TBC) merupakan penyakit infeksi dengan angka mortalitas dan morbiditas yang tinggi secara global dan hingga saat ini hanya tersedia satu vaksin TBC yang berlisensi yaitu Bacille Calmette-Guerin (BCG). Vaksin BCG diketahui tidak dapat mencegah TBC pada individu dewasa dan tidak dapat digunakan pada individu imunokompromais sehingga membuka kesempatan pengembangan vaksin TBC baru. Seed vaksin pcDNA-rpfB dan pcDNA-rpfD yang telah dikembangkan oleh Departemen Mikrobiologi FKUI terbukti mampu menginduksi respon imun humoral dan spesifik terhadap TBC dan mampu menunda pertumbuhan M. tuberculosis secara in-vitro. Penelitian ini menganalisis kemampuan seed vaksin tersebut dalam mencegah infeksi M. tuberculosis secara in-vivo. Pengujian dilakukan dengan mengimunisasi mencit Balb/c dengan seed vaksin ataupun BCG sebagai kontrol, kemudian dilakukan uji tantang dengan introduksi M. tuberculosis. Respon humoral mencit dianalisis melalui deteksi IgG dengan pewarnaan sel pasca transfeksi seed vaksin. Beban basiler M. tuberculosis di organ paru dan limpa dianalisis secara seluler melalui kultur pada MGIT dan media LJ; serta secara molekuler melalui uji RT-PCR. Gambaran kerusakan paru juga dianalisis melalui pemeriksaan histopatologi. Seed vaksin pcDNA-rpfB dan pcDNA-rpfD mampu menginduksi respon humoral IgG dan memberikan gambaran minimnya infiltrasi sel-sel radang di organ paru dibandingkan dengan kontrol. Beban basiler M. tuberculosis tidak berhasil dianalisis secara seluler maupun molekuler. Dari hasil ini seed vaksin pcDNA-rpfB dan pcDNA-rpfD terbukti mampu mencegah inflamasi lokal di organ paru pada hewan coba.
Kata Kunci: Vaksin, Mycobacterium tuberculosis, BCG, Imunisasi, pcDNA.3.1 -rpfB, pcDNA3.1-rpfD


Tuberculosis (TBC) is an infectious disease with high mortality and morbidity rates globally and to date only one licensed TBC vaccine, Bacille Calmette-Guerin (BCG), is available. The BCG vaccine is not known to prevent TBC in adults and cannot be used in immunocompromised individuals, opening the opportunity to develop a new TBC vaccine. The pcDNA-rpfB and pcDNA-rpfD vaccine seeds that have been developed by the Department of Microbiology FKUI have been shown to be able to induce humoral and specific immune responses against TBC and to delay the growth of M. tuberculosis in-vitro. This study analyzed the ability of the seed vaccine to prevent M. tuberculosis infection in-vivo. The test was conducted by immunizing Balb/c mice with the seed vaccine or BCG as a control, then challenged with the introduction ofM. tuberculosis. Humoral response of mice was analyzed through IgG detection by cell staining after seed vaccine transfection. The bacillary burden of M. tuberculosis in the lung organs and spleen was analyzed cellularly through culture on MGIT and LJ media; and molecularly through RTPCR assay. The features of lung damage were also analyzed through histopathological examination. Seed vaccines pcDNA-rpfB and pcDNA-rpfD were able to induce IgG humoral response and showed minimal infiltration of inflammatory cells in the lung organs compared to controls. The bacillary burden of M. tuberculosis was not successfully analyzed cellularly or molecularly. From these results, the pcDNA-rpfB and pcDNA-rpfD vaccine seeds were shown to be able to prevent local inflammation in the lung organs in experimental animals.
Key words: Vaccine, Mycobacterium tuberculosis, BCG, Immunization, pcDNA3.1 -rpfB, pcDNA3.1-rpfD