Werner I, Seitz-Merwald I, Kiessling A H, Kur F, Beiras-Fernandez A
Department of Thoracic and Cardiovascular Surgery, University Hospital Frankfurt, Goethe University, Frankfurt/Main, Germany.
Neovii Biotech GmbH, Gräfelfing, Germany.
Transplant Proc. 2014 Nov;46(9):3000-3. doi: 10.1016/j.transproceed.2014.07.016.
Antithymocyte globulin (ATG)-Fresenius (Neovii-Biotech, Graefelfing, Germany), a highly purified rabbit polyclonal antihuman T-lymphocyte immunoglobulin resulting from immunization of rabbits with the Jurkat T-lymphoblast cell line, is currently used for the prevention of acute rejection in patients receiving solid organ transplants. Our aim was to investigate the in vitro activity of ATG-Fresenius regarding the proliferation of peripheral blood mononuclear cells (PBMCs), an important mechanism of rejection after solid organ transplantation.
PBMCs were isolated from 6 healthy donors. Proliferation was assayed using [(3)H] thymidine incorporation. For analysis of mitogen-stimulated proliferation, the PBMCs were incubated at 37°C with various concentrations of ATG-Fresenius in the absence/presence of 40 μg/mL phytohemagglutinin. For analysis of the mixed lymphocyte reaction, PBMCs were incubated at 37°C with various concentrations of ATG-Fresenius for 3 days. On day 3, PBMCs (stimulator cells) from allogeneic donors were incubated with 25 μg/mL mitomycin C. The responder cells (preincubated with ATG-Fresenius) were then cultured at 37°C with the stimulator cells for 6 days. Groups were compared using ANOVA and the Tukey-Kramer multiple comparison test.
Preincubation of PBMCs with ATG results in concentration-dependent inhibition of phytohemagglutinin-stimulated proliferation. The effect was more pronounced after 2 and 3 days of treatment with ATG compared with 1 day. There was a concentration-dependent decrease in the mixed lymphocyte reaction-induced proliferation (up to 80%) at ATG-Fresenius concentrations as low as 0.05 to 0.5 μg/mL. No further effect on proliferation at ATG-Fresenius concentrations of 0.5 to 50 μg/mL was seen, and higher concentrations (>100 μg/mL) totally inhibited proliferation.
Our in vitro results provide more evidence of the beneficial effect of ATGs in the early phase of solid organ transplantation, by reducing effector cell proliferation.
抗胸腺细胞球蛋白（ATG）-费森尤斯（德国格雷费尔芬的Neovii生物技术公司）是一种高度纯化的兔多克隆抗人T淋巴细胞免疫球蛋白，通过用Jurkat T淋巴母细胞系免疫兔子制备而成，目前用于预防实体器官移植患者的急性排斥反应。我们的目的是研究ATG-费森尤斯对外周血单个核细胞（PBMC）增殖的体外活性，PBMC增殖是实体器官移植后排斥反应的一个重要机制。
从6名健康供体中分离PBMC。使用[³H]胸腺嘧啶核苷掺入法检测增殖情况。为分析有丝分裂原刺激的增殖，将PBMC在37°C下与不同浓度的ATG-费森尤斯在不存在/存在40μg/mL植物血凝素的情况下孵育。为分析混合淋巴细胞反应，将PBMC在37°C下与不同浓度的ATG-费森尤斯孵育3天。在第3天，将来自异体供体的PBMC（刺激细胞）与25μg/mL丝裂霉素C孵育。然后将反应细胞（预先与ATG-费森尤斯孵育）在37°C下与刺激细胞共培养6天。使用方差分析和Tukey-Kramer多重比较检验对各组进行比较。
PBMC与ATG预孵育会导致植物血凝素刺激的增殖受到浓度依赖性抑制。与处理1天相比，ATG处理2天和3天后这种作用更明显。在低至0.05至0.5μg/mL的ATG-费森尤斯浓度下，混合淋巴细胞反应诱导的增殖呈浓度依赖性降低（高达80%）。在ATG-费森尤斯浓度为0.5至50μg/mL时未观察到对增殖的进一步影响，而更高浓度（>100μg/mL）则完全抑制增殖。
我们的体外研究结果通过减少效应细胞增殖，为ATG在实体器官移植早期阶段的有益作用提供了更多证据。
Zotero 插件