TLR4

We hypothesized that blood transfusion could induce platelet-mediated innate immune reactions caused by the interaction between TLR4 and damage-associated molecular patterns (DAMPs). To test this hypothesis, we assessed the levels of TLR4, lipopolysaccharide (LPS)-binding protein, and certain DAMPs including high mobility group box 1 (HMGB1), S100 calcium-binding protein A8 (S100A8, MRP8, and calgranulin A), S100 calcium-binding protein A9 (S100A9, MRP14, and calgranulin B), and serum amyloid A (SAA) in a simulated cross-reaction ex vivo. Furthermore, pro-inflammatory cytokines downstream of TLR4 signaling, including TNF-α, IL-1β, and IL-6, were evaluated. The excel sheet include demographic variables of the four groups (control, M, S, and I). The sheet named Fig 3 was the raw data of Effect of Toll-like receptor 4 (TLR4) expression after 0.9% saline was mixed with washed platelets . The sheet named Fig 4 was the raw data of effect of Toll-like receptor 4 (TLR4) expression on matched blood type mixing (Group M), uncross-matched ABO type-specific mixing (Group S), and ABO incompatible blood mixing (Group I) after mixing donor red blood cells and washed platelets.The sheet named Fig 5 was the raw data of Effect of DAMPs (HMGB1, S100A8, S100A9, and SAA) in the plasma prepared from mixing donor red blood cells and recipient whole blood on total blood mixing, matched blood type mixing (Group M), uncross-matched ABO type-specific mixing (Group S), and ABO incompatible blood mixing (Group I). The sheet named Fig 6 was the raw data of Effect of LPS-binding protein and pro-inflammatory cytokines in the plasma, prepared from mixing donor red blood cells and recipient whole blood, on matched blood type mixing (Group M), uncross-matched ABO type-specific mixing (Group S), and ABO incompatible blood mixing (Group I).

本研究提出假说：输血可诱发血小板介导的固有免疫反应，该反应由Toll样受体4（Toll-like receptor 4, TLR4）与损伤相关分子模式（damage-associated molecular patterns, DAMPs）的相互作用所介导。为验证该假说，本研究通过体外交叉反应模拟实验，检测了TLR4、脂多糖（lipopolysaccharide, LPS）结合蛋白，以及高迁移率族蛋白B1（high mobility group box 1, HMGB1）、S100钙结合蛋白A8（S100 calcium-binding protein A8, S100A8、MRP8、钙粒蛋白A）、S100钙结合蛋白A9（S100 calcium-binding protein A9, S100A9、MRP14、钙粒蛋白B）、血清淀粉样蛋白A（serum amyloid A, SAA）等多种DAMPs的水平。此外，本研究还对TLR4信号通路下游的促炎细胞因子——肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）、白细胞介素-1β（interleukin-1β, IL-1β）及白细胞介素-6（interleukin-6, IL-6）进行了检测。该Excel文件包含4组实验对象（对照组、M组、S组及I组）的人口统计学变量数据。其中，名为"Fig 3"的工作表为0.9%生理盐水与洗涤血小板混合后，Toll样受体4（TLR4）表达变化影响的原始检测数据；名为"Fig 4"的工作表为供者红细胞与洗涤血小板混合后，TLR4表达在同型血型混合（M组）、ABO血型非交叉配型特异性混合（S组）以及ABO血型不相容性混合（I组）场景下的影响原始检测数据；名为"Fig 5"的工作表为供者红细胞与受者全血混合制备血浆后，DAMPs（HMGB1、S100A8、S100A9及SAA）在全血混合、同型血型混合（M组）、ABO血型非交叉配型特异性混合（S组）以及ABO血型不相容性混合（I组）场景下的影响原始检测数据；名为"Fig 6"的工作表为供者红细胞与受者全血混合制备血浆后，LPS结合蛋白及促炎细胞因子在同型血型混合（M组）、ABO血型非交叉配型特异性混合（S组）以及ABO血型不相容性混合（I组）场景下的影响原始检测数据。