阻断CD40-CD40配体途径可增强供体来源的树突状细胞祖细胞诱导心脏同种异体移植长期存活的能力。

Blockade of the CD40-CD40 ligand pathway potentiates the capacity of donor-derived dendritic cell progenitors to induce long-term cardiac allograft survival.

作者信息

Lu L, Li W, Fu F, Chambers F G, Qian S, Fung J J, Thomson A W

机构信息

Thomas E. Starzl Transplantation Institute and Department of Surgery, University of Pittsburgh, Pennsylvania 15213, USA.

出版信息

Transplantation. 1997 Dec 27;64(12):1808-15. doi: 10.1097/00007890-199712270-00031.

DOI:10.1097/00007890-199712270-00031

PMID:9422424

Abstract

BACKGROUND

Failure of costimulatory molecule-deficient donor dendritic cells (DCs) to induce indefinite allograft acceptance may be a result of the 'late" up-regulation of these molecules on the DCs after interaction with host T cells. Ligation of CD40 on antigen-presenting cells by its cognate ligand CD40L is thought to induce expression of CD80 (B7-1) and CD86 (B7-2). We examined the influence of anti-CD40L monoclonal antibody (mAb) on the capacity of donor-derived DC progenitors to induce long-term allograft survival.

METHODS

High purity DC progenitors were grown from B10 (H2b) mouse bone marrow in granulocyte-macrophage colony-stimulating factor and transforming growth factor beta1 (TGFbeta1). Mature DC were propagated in granulocyte-macrophage colony-stimulating factor and interleukin-4. Their phenotype was characterized by flow cytometric analysis and their function by mixed leukocyte reactivity. Anti-donor cytotoxic T lymphocyte activity in grafts and spleens of vascularized heart allograft recipients was also assessed.

RESULTS

The TGFbeta3-cultured cells were (1) DEC 205-positive, MHC class II-positive, CD80dim, CD86dim, and CD40dim, (2) poor stimulators of naive allogeneic T-cell proliferation, and (3) able to prolong significantly B10 cardiac allograft survival in C3H (H2k) recipients when given (2 x 10[6] i.v.) 7 days before organ transplantation (median survival time [MST] 26 days vs. 12 days in controls, and 5 days in interleukin-4 DC-treated animals). Their allostimulatory activity was further diminished by addition of anti-CD40L mAb at the start of the mixed leukocyte cultures. Anti-CD40L mAb alone (250 microg/mouse, i.p.; day -7) did not prolong cardiac graft survival (MST 12 days). In contrast, TGFbeta-cultured DCs + anti-CD40L mAb extended graft survival to a MST of 77 days, and inhibited substantially the anti-donor cytotoxic T lymphocyte activity of graft-infiltrating cells and host spleen cells assessed 8 days after transplant.

CONCLUSIONS

The CD40-CD40L pathway appears important in regulation of allogeneic DC-T-cell functional interaction in vivo; its blockade increases markedly the potential of costimulatory molecule-deficient DCs of donor origin to induce long-lasting allograft survival.

摘要

背景

共刺激分子缺陷的供体树突状细胞（DC）不能诱导同种异体移植物的长期存活，这可能是由于这些分子在与宿主T细胞相互作用后在DC上“晚期”上调所致。抗原呈递细胞上的CD40与其同源配体CD40L结合被认为可诱导CD80（B7-1）和CD86（B7-2）的表达。我们研究了抗CD40L单克隆抗体（mAb）对供体来源的DC祖细胞诱导长期同种异体移植物存活能力的影响。

方法

从B10（H2b）小鼠骨髓中在粒细胞-巨噬细胞集落刺激因子和转化生长因子β1（TGFβ1）存在的情况下培养出高纯度的DC祖细胞。成熟的DC在粒细胞-巨噬细胞集落刺激因子和白细胞介素-4中增殖。通过流式细胞术分析其表型，通过混合淋巴细胞反应分析其功能。还评估了血管化心脏同种异体移植受体的移植物和脾脏中抗供体细胞毒性T淋巴细胞活性。

结果

TGFβ3培养的细胞（1）DEC 205阳性、MHC II类阳性、CD80弱阳性、CD86弱阳性和CD40弱阳性，（2）对幼稚同种异体T细胞增殖的刺激能力较差，（3）在器官移植前7天静脉注射（2×10[6]）时，能够显著延长C3H（H2k）受体中B10心脏同种异体移植物的存活时间（中位存活时间[MST]为26天，而对照组为12天，白细胞介素-4处理的DC组为5天）。在混合淋巴细胞培养开始时加入抗CD40L mAb可进一步降低其同种异体刺激活性。单独使用抗CD40L mAb（250μg/小鼠，腹腔注射；第-7天）不能延长心脏移植物的存活时间（MST为12天）。相反，TGFβ培养的DC+抗CD40L mAb可将移植物存活时间延长至MST为77天，并在移植后8天显著抑制移植物浸润细胞和宿主脾脏细胞的抗供体细胞毒性T淋巴细胞活性。