Treatment for rheumatoid arthritis with immunogene therapy by angiostatin

本研究的目的是建立抗原特异性免疫基因治疗类风湿关节炎的血管抑素。首先，我们用磺基琥珀酰亚胺基4-(N-马来酰亚胺甲基)环己烷-1-羧酸酯(Sulo-SMCC)将IL-10表达载体与抗人ICAM-1的单抗结合。然后将结合物加入ICAM-1表达细胞(ICAM-1COS；稳定转导ICAM-1表达载体的COS细胞滑膜成纤维细胞系E11)。免疫基因作用72小时后，在表达ICAM-1的COS细胞上清液中检测到明显的IL-10，而在对照COS细胞中未检测到IL-10。将表达载体与抗体Fab片段偶联后，转染率提高了4倍。在培养液中加入天然抗体降低了转染率，支持这种方法的抗原特异性基因传递。我们计划将Angiostatin基因和ICAM-1单抗偶联并注入RA SCID小鼠(移植了人滑膜组织的SCID小鼠)

The aim of this research is establishment of antigen-specific immunogene therapy for rheumatoid arthritis with angiostatin. At first, we combined IL-10 expression vector and monoclonal antibodies against human ICAM-1 using sulfo-succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (sulfo-SMCC). Then we added the conjugates to ICAM-1 expressing cells (ICAM-1 COS ; COS cells stably trasfected with ICAM-1 expression vector, synovial fibroblast cell line E11). We detected significant IL-10 in the supernatant of ICAM-1 expressing cells but not in control COS cells after 72hour incubation with immunogene. The tranfection efficiency increased four times when the expression vector was conjugated to Fab fragment of antibody. The addition of native antibody to the medium reduced transfection efficiency, supporting antigen-specific gene delivery of this method. We are planning to conjugate angiostatin gene and ICAM-1 monoclonal antibody and infuse to RA SCID mice (SCID mice transplanted with human synovial tissue)