Di Genaro M S, Cargnelutti D E, Castro D O, Eliçabe R J, Gutiérrez J V, Correa S G, de Guzmán A M S
Laboratory of Microbiology, Faculty of Chemistry, Biochemistry and Pharmacy, National University of San Luis, 5700 San Luis, Argentina.
Scand J Rheumatol. 2007 Jan-Feb;36(1):28-35. doi: 10.1080/03009740600906651.
To study the role of IL-12p40 at the onset of reactive arthritis (ReA) after Yersinia enterocolitica O:3 infection, and analyse relevant microbial antigens and articular expression of Toll-like receptor (TLR) mRNA.
Wild-type C57BL/6 and IL-12p40-deficient (IL-12p40-/-) mice were orogastrically infected with Y. enterocolitica O:3. Early (day 3) and late (day 21) after infection, the number of bacteria were determined in Peyer's patches (PP), mesenteric lymph nodes (MLN), the spleen, and joints. Histological studies of joints were performed. Collagen-specific and anti-Yersinia antibodies were measured by enzyme-linked immunosorbent assay (ELISA). The presence of Yersinia antigens was studied by dot blot. Induction of articular mRNA of TLR2, TLR4, and tumour necrosis factor (TNF)-alpha was analysed by reverse transcription-polymerase chain reaction (RT-PCR). TNFalpha protein levels were measured by ELISA.
At day 3, bacterial recovery in PP, MLN, and spleen was significantly increased in IL-12p40-/- mice. Histopathological changes were observed in IL-12p40-/- mice at day 21 after infection, and correlated with higher antibody response against type II collagen. Although live bacteria could not be isolated at day 21 after infection, articular microbial components, especially from the outer membrane (OM), were detected. Moreover, intra-articular immunoglobulins to Yersinia antigens were significantly higher in IL-12p40-/- mice. Furthermore, mRNA levels for TLR2, TLR4 and TNFalpha, and TNFalpha protein were increased in joints from IL-12p40-/- mice.
We concluded that IL-12p40 influences the resistance against Yersinia-triggered ReA. Bacterial products such as Yersinia OM could contribute to the ReA by induction of articular TLR expression, which results in an inflammatory response in the joint.
研究白细胞介素-12p40在小肠结肠炎耶尔森菌O:3感染后反应性关节炎（ReA）发病中的作用，并分析相关微生物抗原及Toll样受体（TLR）mRNA的关节表达情况。
将野生型C57BL/6小鼠和白细胞介素-12p40缺陷型（IL-12p40-/-）小鼠经口胃感染小肠结肠炎耶尔森菌O:3。在感染后的早期（第3天）和晚期（第21天），测定派伊尔结（PP）、肠系膜淋巴结（MLN）、脾脏和关节中的细菌数量。对关节进行组织学研究。通过酶联免疫吸附测定（ELISA）检测胶原特异性抗体和抗耶尔森菌抗体。通过斑点印迹研究耶尔森菌抗原的存在情况。通过逆转录-聚合酶链反应（RT-PCR）分析关节中TLR2、TLR4和肿瘤坏死因子（TNF）-α mRNA的诱导情况。通过ELISA测定TNFα蛋白水平。
在第3天，IL-12p40-/-小鼠的PP、MLN和脾脏中的细菌回收量显著增加。在感染后第21天，IL-12p40-/-小鼠出现组织病理学变化，且与针对II型胶原的更高抗体反应相关。尽管在感染后第21天无法分离出活菌，但检测到关节中的微生物成分，尤其是来自外膜（OM）的成分。此外，IL-12p40-/-小鼠关节内针对耶尔森菌抗原的免疫球蛋白显著更高。此外，IL-12p40-/-小鼠关节中TLR2、TLR4和TNFα的mRNA水平以及TNFα蛋白均升高。
我们得出结论，IL-12p40影响对耶尔森菌引发的ReA的抵抗力。耶尔森菌OM等细菌产物可通过诱导关节TLR表达导致ReA，进而在关节中引发炎症反应。
