Overcoming the Challenge of Transduction of Human T-cells with Chimeric Antigen Receptor (CAR) Specific for ERBB2 Antigen

Abstract

were isolated from the peripheral blood mononuclear cells (PBMCs). The constructed CAR gene was inserted into a lentiviral plasmid containing the green fluorescent protein (GFP) tag and lentiviral particles were produced. These lentiviral particles were used to transduce activated T-cells by spinoculation. T-cells were activated using Dynabeadconjugated CD3/CD28 human T-cell activator and interleukin-2 (IL-2) before transduction. CD3+ T-cells were selected and GFP expression, which indicated transduction, was observed. Future studies will focus on in vitro and in vivo models to determine the efficiency of CAR-T cells in specifically targeting ERBB2-expressing cells. Keywords: Breast cancer; CD3+ T-cells; chimeric antigen receptor (CAR); immunotherapy ABSTRAK Kanser payudara adalah salah satu kanser yang kerap melanda kaum wanita. Kajian saintifik telah mengaitkan lebihan ekspresi antigen ERBB2 pada barah kanser yang lebih agresif. Untuk menangani masalah ini, teknologi reseptor kimera antigen (CAR) boleh digunakan. Untuk ini, sel T manusia ditransduksi dengan urutan gen pengekodan CAR yang khusus untuk antigen berkaitan-barah (TAA). Sel T yang ditransduksi dengan CAR (CAR-T) secara genetik dapat mensasar kepada antigen kanser tanpa memerlukan pengenalan kompleks kehistoserasian utama (MHC), menjadikan ia pilihan terapi imun berpotensi umum. Walau bagaimanapun, transduksi gen terapeutik ke dalam sel T manusia dan pengembangan