TRANSPLANT IMMUNOSUPPRESSION WITH ANTISENSE TECHNOLOGY

描述：（改编自申请人的摘要）虽然生存的 移植肾1年移植率达80-90%，5年移植率达70- 80 存活大鼠中，大多数同种异体移植物（50-80%）受到影响， 移植后的前三个月，急性排斥反应发作。 环孢霉素（CsA）为基础的免疫抑制剂治疗窗窄， 从而导致毒性或过度免疫抑制。 本研究 专注于应用一种新的基因靶向和无毒 免疫抑制治疗有望诱导移植耐受。 细胞基因表达被设计成 通过沃森-克里克配对杂交特异性信使RNA。 反义 硫代磷酸寡核苷酸（PS-oligo）被设计成靶向 参与细胞间粘附的分子以及所需的分子 用于激活T和B细胞。 先前的研究表明， 细胞间粘附分子-1（ICAM-1），血管粘附分子-1 （VCAM-1）或c-raf反义PS-oligo可延长心脏存活时间 以剂量依赖性和序列特异性的方式进行同种异体移植。 本 实验将检测内皮细胞-白细胞粘附分子 （E-选择素），白细胞功能相关抗原分子-1（LFA-1）， 肾脏、心脏和小肠同种异体移植物的存活。 T细胞识别 通过由α和β组成的T细胞受体（TCR）的同种抗原 链;每条链由可变区（V）和恒定区组成。 因此，Vbeta 8 反义PS-oligo可阻断移植器官的排斥反应， 同种异体抗原特异性方式。 B7反义PS-寡核苷酸（第二次活化 信号）或/和白细胞介素（IL）-2/干扰素（IFN）-μ反义 PS-寡核苷酸（阻断T辅助细胞1）可能诱导移植耐受。 的 将测试c-raf、蛋白C（PKC）、Ras和Lck PS寡聚物来阻断T 细胞功能和心脏移植排斥反应。 B细胞识别同种抗原 通过免疫球蛋白（IG）B细胞受体（BCR），即IgM（μ）、IgG 1 γ 1）、IgG 2a（γ 2a）、IgG 2b（γ 2b）或IgG 3（γ 3）BCR;反义 靶向这些不同表位（μ，γ 1，γ 2a，γ 2b， γ 3）可以选择性地阻断针对同种异体抗原的抗体产生。 在 此外，γ 1和IL-4反义PS-寡核苷酸组合可抑制 IG类转换为IgG 1; γ 2a和T细胞生长因子-β（TGF-β） IgGa的反义PS-寡核苷酸;和 IgG 2b。 PS-oligo技术提供了有效和无毒的基因靶向 免疫抑制，这可能会彻底改变器官移植治疗方案 移植 与单克隆抗体相比，PS-寡核苷酸不 诱导产生抗寡核苷酸特异性抗体。

DESCRIPTION: (Adapted from the applicant's abstract) Although survival of kidney allografts reached a remarkable 80-90% one year and 70-80% five year survival rats, the majority of allografts (50-80%) are affected, during first three months postgrafting, by an acute rejection episode. Cyclosporine (CsA)-based immunosuppression has narrow therapeutic window, thereby resulting in toxicity or over immunosuppression. Present study focuses on application of a new gene-targeted and non-toxic immunosuppressive treatments poised to induce transplantation tolerance. Cellular gene expression is inhibited by oligonucleotide designed to hybridize a specific messenger RNA by Watson-Crick pairing. The antisense phosphorothioated oligonuleotides (PS-oligo) are designed to target molecules involved in cell-to-cell adhesion as well as molecules required for activation of T and B cells. Previous study showed that treatment with intercellular adhesion molecule-1 (lCAM-1), vascular adhesion molecule-1 (VCAM-1), or c-raf antisense PS-oligo prolonged the survival of heart allografts in a dose-dependent and sequence specific fashion. Present experiments will examine endothelial-leukocyte adhesion molecule (E-selectin), leukocyte function-associated antigen molecule-1 (LFA-1) on the survival of kidney, heart and small bowel allografts. T cells recognize alloantigens through T cell receptor (TCR) composed of alpha and beta chains; each chain consists variable (V) and constant regions. Thus, Vbeta8 antisense PS-oligo may block the rejection of organ allografts in alloantigen-specific fashion. B7 antisense PS-oligos (second activation signal) or/and with interleukin (IL)-2/interferon (IFN)-mu antisense PS-oligos (blocking T helper 1) may induce transplantation tolerance. The c-raf, protein C (PKC), Ras, and Lck PS-oligos will be tested to block T cell functions and heart allograft rejection. B cells recognize alloantigen through immunoglobulin (Ig) B cell receptor (BCR), namely the IgM(mu), IgG1 gamma1), IgG2a(gamma2a), IgG2b (gamma2b), or IgG3 (gamma3) BCR; antisense PS-oligo targeting these different epitopes (mu, gamma1, gamma2a, gamma2b, gamma3) may selectively block antibody production to alloantigens. In addition, in combination of gamma1 and IL-4 antisense PS-oligos may inhibit Ig class switching to IgG1; gamma2a and T cell growth factor-beta (TGF-beta) antisense PS-oligos to IgGa; and gamma2b and IFN-y antisense PS-oligos to IgG2b. PS-oligo technology offer potent and nontoxic gene-targeted immunosuppression, which may revolutionize therapeutic protocols for organ transplantation. In contrast to monoclonal antibodies PS-oligos do not induce production of anti-oligo specific antibodies.