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伴放线聚集杆菌引发的T淋巴细胞表型及功能具有血清型依赖性。

T-lymphocyte phenotype and function triggered by Aggregatibacter actinomycetemcomitans is serotype-dependent.

作者信息

Díaz-Zúñiga J, Melgar-Rodríguez S, Alvarez C, Monasterio G, Benítez A, Ciuchi P, Díaz C, Mardones J, Escobar A, Sanz M, Vernal R

机构信息

Periodontal Biology Laboratory, Dental School, Universidad de Chile, Santiago de Chile, Chile.

Department of Conservative Dentistry, Dental School, Universidad de Chile, Santiago de Chile, Chile.

出版信息

J Periodontal Res. 2015 Dec;50(6):824-35. doi: 10.1111/jre.12270. Epub 2015 Mar 30.

Abstract

BACKGROUND AND OBJECTIVE

Based on lipopolysaccharide (LPS) antigenicity, different Aggregatibacter actinomycetemcomitans serotypes have been described. Serotype b strains have demonstrated a stronger capacity to trigger cytokine production on dendritic cells (DCs). As DCs regulate the development of T-lymphocyte lineages, the objective of this investigation was to study the response of T lymphocytes after being stimulated with autologous DCs primed with different bacterial strains belonging to the most prevalent serotypes of A. actinomycetemcomitans in humans: a-c.

MATERIAL AND METHODS

Human DCs were primed with increasing multiplicity of infection (10(-1) -10(2) ) or the purified LPS (10-50 ng/mL) of A. actinomycetemcomitans serotypes a-c and then used to stimulate autologous naïve CD4(+) T lymphocytes. The T-helper (Th) type 1, Th2, Th17 and T-regulatory transcription factors T-bet, GATA-3, RORC2 and Foxp3, which are the master-switch genes implied in their specific differentiation, as well as T-cell phenotype-specific cytokine patterns were quantified by real-time reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. In addition, the intracellular expression of T-bet/interferon-γ, GATA-3/interleukin-4, RORC2/interleukin-17A and Foxp3/transforming growth factor-β1 was analysed by double staining and flow cytometry.

RESULTS

All the A. actinomycetemcomitans serotypes led to T-lymphocyte activation; however, when T lymphocytes were stimulated with DCs primed with the A. actinomycetemcomitans serotype b strain or their purified LPS, higher levels of Th1- and Th17-associated transcription factors and cytokines were detected compared with similar experiments with the other serotypes.

CONCLUSION

These results demonstrate that serotype b of A. actinomycetemcomitans has a higher capacity of trigger Th1 and Th17 phenotype and function and it was demonstrated that their LPS is a more potent immunogen compared with the other serotypes.

摘要

背景与目的

基于脂多糖(LPS)抗原性,已描述了不同的伴放线聚集杆菌血清型。b血清型菌株已显示出更强的触发树突状细胞(DCs)产生细胞因子的能力。由于DCs调节T淋巴细胞谱系的发育,本研究的目的是研究用人类中最常见的伴放线聚集杆菌血清型a - c的不同菌株致敏的自体DCs刺激后T淋巴细胞的反应。

材料与方法

用递增的感染复数(10⁻¹ - 10²)或伴放线聚集杆菌血清型a - c的纯化LPS(10 - 50 ng/mL)致敏人DCs,然后用于刺激自体初始CD4⁺ T淋巴细胞。通过实时逆转录 - 聚合酶链反应和酶联免疫吸附测定法定量T辅助(Th)1型、Th2型、Th17型和调节性T细胞转录因子T - bet、GATA - 3、RORC2和Foxp3,这些是其特定分化中所涉及的主开关基因,以及T细胞表型特异性细胞因子模式。此外,通过双重染色和流式细胞术分析T - bet/干扰素 - γ、GATA - 3/白细胞介素 - 4、RORC2/白细胞介素 - 17A和Foxp3/转化生长因子 - β1的细胞内表达。

结果

所有伴放线聚集杆菌血清型均导致T淋巴细胞活化;然而,当用伴放线聚集杆菌b血清型菌株或其纯化LPS致敏的DCs刺激T淋巴细胞时,与其他血清型的类似实验相比,检测到更高水平的Th1和Th17相关转录因子及细胞因子。

结论

这些结果表明,伴放线聚集杆菌b血清型具有更高的触发Th1和Th17表型及功能的能力,并且证明其LPS与其他血清型相比是一种更强效的免疫原。

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