Kemampuan Infeksi Virus Kimera Simian Human Immunodeficiency Virus (SHIVst) pada Kultur Sel HT-29

Abstract

ABSTRAK
Simian Human Immunodeficiency Virus (SHIV) merupakan virus kimera yang berasal dari
rekombinasi antara HIV yang dapat menginfeksi manusia dan SIV yang dapat menginfeksi primata
non manuisa. Pada penelitian ini, dilakukan substitusi sekuens gen virus kimera pada sekuens gen
gag, yaitu Cyclophilin A binding domain, gen vif, dan gen nef. Hasil rekombinasi pada virus kimera
SHIV ini dilakukan dan diinfeksikan pada sel manusia yaitu sel HT-29. Uji infektivitas virus kimera
SHIVst ini bertujuan untuk melihat kemampuan infeksi virus kimera pada sel manusia melalui
kinetika replikasi virus yaitu melalui keberadaan materi genetic yaitu RNA virus dan protein kapsid,
yaitu p24. Hasil yang diperoleh virus kimera dapat dihasilkan melalui transfeksi pada sel CHO.
Analisis keberadaan materi genetic melalui tahapan isolasi RNA yang divisualisasi melalui proses
amplifikasi dengan PCR konvensional dan reverse transcription PCR (RT-PCR) terdapat pita RNA
virus yang terbentuk. Namun, pada hasil infeksi virus kimera yang dilakukan pada sel HT-29,
keberadaan RNA virus melalui tahapan amplifikasi baik PCR konvensional maupun RT PCR tidak
menunjukkan terbentuknya pita spesifik RNA virus.
Kata kunci: Infeksi, HIV, SIV, SHIVst, Kultur Sel HT-29

ABSTRACT
Simian Human Immunodeficiency Virus (SHIV) is a chimeric virus originating from recombination
between HIV which can infect humans and SIV which can infect non-human primates. In this study,
the chimera virus gene sequence was substituted by gag, specifically Cyclophilin A binding domain
gene, vif, and nef gene. The result of recombination in the SHIV chimera virus was carried out and
infected in human cells, namely HT-29 cells which have an alternative molecule that can recognize
the gp120 receptor, galactocylceremide. The infectivity test of the SHIVst chimera virus aims to see
the ability of chimera virus infection through viral replication kinetics which is observed through the
presence of viral RNA as genetic material and capsid protein, p24. The results obtained by the
chimera virus can be produced by transfection in CHO cells. Analysis of the presence of genetic
material through the stages of RNA isolation was visualized through the amplification process by
conventional PCR and reverse transcription PCR (RT-PCR) showed that viral RNA bands were
formed. However, in the results of chimera virus infection carried out on HT-29 cells, the presence of
viral RNA through the amplification stages of both conventional PCR and RT PCR did not show the
formation of viral RNA-specific bands.
Keywords: Infectivity, HIV, SIV, SHIVst, HT-29 Cell Culture.