Differential expression of Toll-like receptors on human alveolar macrophages and autologous peripheral monocytes.
Differential expression of Toll-like receptors on human alveolar macrophages and autologous peripheral monocytes.
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DOI：
10.1186/1465-9921-11-2
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发表时间：
2010-01-05
影响因子：
5.8
通讯作者：
Torres M
中科院分区：
文献类型：
--
作者：
Juarez E;Nuñez C;Sada E;Ellner JJ;Schwander SK;Torres M
Toll样受体（TLR）是调节肺免疫应答和识别呼吸道病原体如结核分枝杆菌（M.tb）的关键组分。通过对人肺泡巨噬细胞的检测，本研究试图更好地确定TLR 2、TLR 4和TLR 9在人肺室中的表达谱，这些表达谱至今仍不清楚。16名健康受试者接受静脉穿刺，11名受试者接受额外的支气管肺泡灌洗，分别获得外周血单核细胞和支气管肺泡细胞。通过荧光激活细胞分选和qRT-PCR评估TLR的表面和细胞内表达。用TLR特异性配体刺激细胞，并通过ELISA和细胞因子珠阵列评估细胞因子产生。肺泡巨噬细胞上TLR 2的表面表达显著低于血液单核细胞（1.2 ± 0.4%对57 ± 11.1%，相对平均荧光强度[rMFI]：0.9 ± 0.1对3.2 ± 0.1，p < 0.05）。肺泡巨噬细胞和单核细胞中TLR 4和TLR 9表达细胞的比例和TLR 4的rMFI相当。然而，TLR 9在肺泡巨噬细胞上的表面表达高于单核细胞（rMFI，218.4 ± 187.3对4.4 ± 1.4，p < 0.05），而受体的细胞内表达和TLR 9阳性细胞的比例在两种细胞类型中相似。TLR 2、TLR 4和TLR 9 mRNA在支气管肺泡细胞中的表达低于单核细胞。Pam 3Cys、LPS和M.tb DNA上调支气管肺泡细胞和单核细胞中TLR 2、TLR 4和TLR 9 mRNA。与TLR 2表面和mRNA表达的降低相对应，Pam 3Cys诱导支气管肺泡细胞产生的TNF-α、IL-1β和IL-6低于单核细胞。尽管TLR 4在两种细胞类型上的表达相当，但LPS在单核细胞中比在肺泡巨噬细胞中诱导更高水平的IL-10。结核分枝杆菌DNA，TLR 9的配体，在两种细胞类型中诱导相似水平的细胞因子。自体人肺泡巨噬细胞和单核细胞的TLR表达谱是不相同的，因此可能有助于肺和全身的区室化免疫应答。这些差异可能对靶向呼吸道的TLR刺激佐剂疫苗的设计和功效评估具有重要意义。
Toll-like receptors (TLRs) are critical components in the regulation of pulmonary immune responses and the recognition of respiratory pathogens such as Mycobacterium Tuberculosis (M.tb). Through examination of human alveolar macrophages this study attempts to better define the expression profiles of TLR2, TLR4 and TLR9 in the human lung compartment which are as yet still poorly defined. Sixteen healthy subjects underwent venipuncture, and eleven subjects underwent additional bronchoalveolar lavage to obtain peripheral blood mononuclear and bronchoalveolar cells, respectively. Surface and intracellular expression of TLRs was assessed by fluorescence-activated cell sorting and qRT-PCR. Cells were stimulated with TLR-specific ligands and cytokine production assessed by ELISA and cytokine bead array. Surface expression of TLR2 was significantly lower on alveolar macrophages than on blood monocytes (1.2 ± 0.4% vs. 57 ± 11.1%, relative mean fluorescence intensity [rMFI]: 0.9 ± 0.1 vs. 3.2 ± 0.1, p < 0.05). The proportion of TLR4 and TLR9-expressing cells and the rMFIs of TLR4 were comparable between alveolar macrophages and monocytes. The surface expression of TLR9 however, was higher on alveolar macrophages than on monocytes (rMFI, 218.4 ± 187.3 vs. 4.4 ± 1.4, p < 0.05) while the intracellular expression of the receptor and the proportion of TLR9 positive cells were similar in both cell types. TLR2, TLR4 and TLR9 mRNA expression was lower in bronchoalveolar cells than in monocytes. Pam3Cys, LPS, and M.tb DNA upregulated TLR2, TLR4 and TLR9 mRNA in both, bronchoalveolar cells and monocytes. Corresponding with the reduced surface and mRNA expression of TLR2, Pam3Cys induced lower production of TNF-α, IL-1β and IL-6 in bronchoalveolar cells than in monocytes. Despite comparable expression of TLR4 on both cell types, LPS induced higher levels of IL-10 in monocytes than in alveolar macrophages. M.tb DNA, the ligand for TLR9, induced similar levels of cytokines in both cell types. The TLR expression profile of autologous human alveolar macrophages and monocytes is not identical, therefore perhaps contributing to compartmentalized immune responses in the lungs and systemically. These dissimilarities may have important implications for the design and efficacy evaluation of vaccines with TLR-stimulating adjuvants that target the respiratory tract.
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影响因子：
5.5
作者：
Brekke, Ole-Lars;Christiansen, Dorte;Mollnes, Tom E.
通讯作者：
Mollnes, Tom E.
影响因子：
3.1
作者：
Cario, E;Podolsky, DK
通讯作者：
Podolsky, DK
影响因子：
4.4
作者：
Henning, Lisa N.;Azad, Abul K.;Schlesinger, Larry S.
通讯作者：
Schlesinger, Larry S.
DOI：
10.1165/rcmb.2002-0163oc
发表时间：
2003-07-01
影响因子：
6.4
作者：
Fenhalls, G;Squires, GR;Lukey, PT
通讯作者：
Lukey, PT
DOI：
10.1164/rccm.200503-411oc
发表时间：
2006-01-15
影响因子：
24.7
作者：
Carranza, C;Juárez, E;Schwander, SK
通讯作者：
Schwander, SK