TRANSPLANT IMMUNOSUPPRESSION WITH ANTISENSE TECHNOLOGY

利用反义技术进行移植免疫抑制

• 批准号: 2672641
• 负责人: Stanislaw M Stepkowski
• 金额: $ 20.15万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2672641
• 关键词: B cell receptor; T cell receptor; antisense nucleic acid; biological signal transduction; cell cell interaction; cyclosporines; gastrointestinal transplantation; gene expression; heart transplantation; helper T lymphocyte; immunosuppression; interferon gamma; interleukin 2; isoantigen; kidney transplantation; laboratory mouse; laboratory rat; leukocyte activation /transformation; leukocyte adhesion molecules; messenger RNA; monoclonal antibody; oligonucleotides; protein kinase C; selectins; transplant rejection

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.

描述：（改编自申请人的摘要）尽管 同种异体肾移植一年达到 80-90%，五年达到 70-80% 存活大鼠中，大多数同种异体移植物（50-80%）受到影响，在 移植后前三个月，出现急性排斥反应。 基于环孢素（CsA）的免疫抑制治疗窗窄， 从而导致毒性或过度免疫抑制。 目前的研究 专注于新型基因靶向、无毒的应用 免疫抑制治疗有望诱导移植耐受。 细胞基因表达被设计为的寡核苷酸抑制 通过 Watson-Crick 配对杂交特定的信使 RNA。 反义 硫代磷酸寡核苷酸（PS-oligo）旨在靶向 参与细胞间粘附的分子以及所需的分子 用于激活 T 细胞和 B 细胞。 先前的研究表明，治疗 细胞间粘附分子-1 (lCAM-1)、血管粘附分子-1 (VCAM-1) 或 c-raf 反义 PS-oligo 延长心脏存活 以剂量依赖性和序列特异性方式进行同种异体移植。 展示 实验将检查内皮白细胞粘附分子 (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) 产生同种异体抗原 链条；每条链由可变区 (V) 和恒定区组成。 因此，Vbeta8 反义 PS-oligo 可能会阻止同种异体器官移植的排斥反应 同种异体抗原特异性时尚。 B7 反义 PS-oligos（二次激活 信号）或/和白细胞介素（IL）-2/干扰素（IFN）-mu反义 PS-oligos（阻断 T 辅助蛋白 1）可能会诱导移植耐受。 这 将测试 c-raf、蛋白 C (PKC)、Ras 和 Lck PS-oligos 来阻断 T 细胞功能和心脏同种异体移植排斥。 B 细胞识别同种异体抗原 通过免疫球蛋白（Ig）B细胞受体（BCR），即IgM（mu）、IgG1 γ1)、IgG2a(γ2a)、IgG2b(γ2b)或IgG3(γ3)BCR；反义 PS-oligo 靶向这些不同的表位（mu、gamma1、gamma2a、gamma2b、 gamma3)可以选择性地阻断同种异体抗原的抗体产生。 在 此外，结合 gamma1 和 IL-4 反义 PS-oligos 可能会抑制 Ig 类别切换为 IgG1； gamma2a 和 T 细胞生长因子-β (TGF-β) IgGa 反义 PS-oligos；和 gamma2b 和 IFN-y 反义 PS-oligos IgG2b。 PS-oligo 技术提供有效且无毒的基因靶向 免疫抑制，这可能会彻底改变器官的治疗方案 移植。 与单克隆抗体相比，PS-oligos 不 诱导抗寡特异性抗体的产生。

项目成果

Nucleotide sequences of three distinct complementary DNA clones encoding rat class II major histocompatibility complex RT1.D beta-chain proteins.

编码大鼠 II 类主要组织相容性复合体 RT1.D β 链蛋白的三个不同互补 DNA 克隆的核苷酸序列。

• 发表时间: 1999-07
• 影响因子: 3.2
• 作者: Tian, L;Wang, M;Yu, J;Kahan, B D;Stepkowski, S M
• 通讯作者: Stepkowski, S M

Development of antisense oligodeoxynucleotides for transplantation.

用于移植的反义寡脱氧核苷酸的开发。

• 发表时间: 2000-06
• 作者: Stepkowski; S M
• 通讯作者: S M

Inhibition of C-raf expression by antisense oligonucleotides extends heart allograft survival in rats.

反义寡核苷酸抑制 C-raf 表达可延长大鼠同种异体心脏移植物的存活率。

• 发表时间: 2000-08-27
• 影响因子: 6.2
• 作者: Stepkowski, S M;Qu, X;Wang, M E;Tian, L;Chen, W;Wancewicz, E V;Johnston, J F;Bennett, C F;Monia, B P
• 通讯作者: Monia, B P

ICAM-1 antisense oligodeoxynucleotide improves islet allograft survival and function.

ICAM-1 反义寡脱氧核苷酸可改善胰岛同种异体移植物的存活和功能。

• 发表时间: 2000-11
• 影响因子: 3.3
• 作者: Katz, S M;Bennett, F;Stecker, K;Clark, J H;Pham, T;Wang, M E;Kahan, B D;Stepkowski, S M
• 通讯作者: Stepkowski, S M

Perfusion of kidneys with unformulated "naked" intercellular adhesion molecule-1 antisense oligodeoxynucleotides prevents ischemic/reperfusion injury.

用未配制的“裸”细胞间粘附分子-1反义寡脱氧核苷酸灌注肾脏可预防缺血/再灌注损伤。

• 发表时间: 1999-09-27
• 影响因子: 6.2
• 作者: Chen, W;Bennett, C F;Wang, M E;Dragun, D;Tian, L;Stecker, K;Clark, J H;Kahan, B D;Stepkowski, S M
• 通讯作者: Stepkowski, S M