Abstract

 Background and Objectives

 The specific activation of the immune system especially T lymphocytes has been a long-term goal of cancer immunotherapy. In an approach to induce the patient’s own immune cells, chimeric antigen receptors (CAR) were used to activate cytotoxic T lymphocytes. The CAR consists of an antigen binding domain of antibody molecule linked through an extracellular linker to transmembrane and cytoplasmic domains of lymphocyte-triggering moieties.

 Materials and Methods

 In our research group, variable domains of heavy chain antibodies (VHH or nanobody) derived from camelids were used instead of the antigen-binding domain in chimeric receptors constructs. To obtain a stable and increased expression of VHH harboring CAR, the PhiC31 integrase system was used for CAR gene integration into the T-cell genome. Constructs expressing CAR and PhiC31 integrase were co-transfected into the Jurkat cell line and CAR expression was quantified after one day and 30 days after transfection by a semi-quantitative RT-PCR method.

 Results

 Our results on day 30 confirmed that co-transfection of PhiC31 integrase constructs and CAR expressing constructs resulted in stable and high expression of the receptors.

 Conclusions

 Based on our results, we conclude that PhiC31 integrase enzyme can be used for immunotherapy of cancer mediated by T cells expressing chimeric recetors.