Bernatoniene Jolanta, Zhang Qibo, Dogan Semih, Mitchell Tim J, Paton James C, Finn Adam
Institute of Child Life and Health, University of Bristol, Bristol, United Kingdom.
J Infect Dis. 2008 Dec 15;198(12):1823-33. doi: 10.1086/593177.
Choline-binding protein A (CbpA) and pneumolysin (Ply) can induce the expression and release of chemokines by human cells, which might modulate specific immune responses. In dendritic cells (DCs), such effects could be important for the size and character of the immunity induced if administered as vaccines. We studied the induction of CCL and CXCL chemokines by CbpA and Ply in DCs and related signaling pathways.
Proteins derived from bacterial cultures and cloning were used as stimulants. DCs were generated from CD14+ human monocytes by negative selection, followed by coculture with recombinant human granulocyte-macrophage colony-stimulating factor and recombinant interleukin-4. The role played by Toll-like receptors (TLRs) was assessed using anti-TLR antibodies. Likewise, specific inhibitors (given in parentheses) of signaling molecules were used: NF-kappaB (SN50), extracellular signal-regulated kinase (PD98059), p38 (SB203580), and Jun N-terminal kinase (SP600125).
Both CbpA and Ply significantly up-regulated DC mRNA of several CCL (2, 4, 5, and 8) and CXCL (8 and 10) chemokines studied as well as the expression of 3 proteins studied: CCL2, CCL5, and CXCL8. Ply stimulation was blocked by anti-TLR4. Inhibition of NF-kappaB and several mitogen-activated protein kinase signaling pathways also reduced chemokine release.
Chemokine induction in DCs by CbpA and Ply may be important for their potential use in future pneumococcal vaccines.
胆碱结合蛋白A（CbpA）和肺炎球菌溶血素（Ply）可诱导人细胞表达和释放趋化因子，这可能调节特异性免疫反应。在树突状细胞（DCs）中，如果作为疫苗给药，此类效应对于诱导的免疫反应的规模和特性可能很重要。我们研究了CbpA和Ply在DCs中对CCL和CXCL趋化因子的诱导作用及相关信号通路。
将细菌培养物和克隆获得的蛋白质用作刺激物。通过阴性选择从CD14 +人单核细胞生成DCs，随后与重组人粒细胞-巨噬细胞集落刺激因子和重组白细胞介素-4共培养。使用抗Toll样受体（TLR）抗体评估TLR的作用。同样，使用信号分子的特异性抑制剂（括号内给出）：核因子κB（SN50）、细胞外信号调节激酶（PD98059）、p38（SB203580）和Jun氨基末端激酶（SP600125）。
CbpA和Ply均显著上调了所研究的几种CCL（2、4、5和8）和CXCL（8和10）趋化因子的DC mRNA以及所研究的3种蛋白质的表达：CCL2、CCL5和CXCL8。抗TLR4可阻断Ply刺激。抑制核因子κB和几种丝裂原活化蛋白激酶信号通路也可减少趋化因子释放。
CbpA和Ply在DCs中诱导趋化因子可能对其未来在肺炎球菌疫苗中的潜在应用很重要。
