Locht H, Skogh T, Kihlström E
Department of Health and Environment, Division of Rheumatology, Faculty of Health Sciences, Linköping University, Sweden.
Clin Exp Immunol. 1999 Sep;117(3):568-73. doi: 10.1046/j.1365-2249.1999.01008.x.
Fifty-five serum samples from patients with reactive arthritis (ReA), 40 from patients with ankylosing spondylitis (AS) and three from patients with chronic sacroiliac joint arthritis were analysed for the presence of ANCA of IgG class by means of enzyme immunosorbent assay using lactoferrin (Lf), myeloperoxidase (MPO) and antigen extracted from azurophil granules ('alpha-antigen') containing proteinase 3 (PR3) as substrate. IgG-ANCA were found in 31 (56%) patients with ReA. Twenty-three (42%) had anti-Lf antibodies, nine (16%) had anti-MPO and eight (15%) had anti-alpha-antigen antibodies, none of which reacted with PR3. Only six (14%) AS or sacroiliac joint arthritis patients had ANCA (P < 0.001). Three (7%) had anti-Lf, two (5%) anti-MPO and two (5%) anti-alpha-antigen antibodies. Yersinia and Salmonella bacteria were separated by SDS-PAGE and blots were incubated with serum from rabbits immunized with human Lf. The hyperimmune serum recognized a band of 78 kD from both bacteria which was not seen when preimmune serum was used. The reaction to the 78-kD antigen could be completely inhibited when anti-Lf antibodies were absorbed on Lf coupled to cyanogen bromide-activated Sepharose, possibly indicating cross-reacting epitopes in Lf and enterobacterial antigen.
采用酶联免疫吸附测定法，以乳铁蛋白（Lf）、髓过氧化物酶（MPO）以及从含有蛋白酶3（PR3）的嗜天青颗粒中提取的抗原（“α抗原”）为底物，对55份反应性关节炎（ReA）患者的血清样本、40份强直性脊柱炎（AS）患者的血清样本以及3份慢性骶髂关节炎患者的血清样本进行IgG类抗中性粒细胞胞浆抗体（ANCA）检测。在31例（56%）ReA患者中检测到IgG-ANCA。23例（42%）有抗Lf抗体，9例（16%）有抗MPO抗体，8例（15%）有抗α抗原抗体，均不与PR3反应。仅有6例（14%）AS或骶髂关节炎患者有ANCA（P<0.001）。3例（7%）有抗Lf抗体，2例（5%）有抗MPO抗体，2例（5%）有抗α抗原抗体。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分离耶尔森菌和沙门菌，并将印迹与用人Lf免疫的兔血清孵育。免疫血清识别出两种细菌的一条78 kD的条带，而使用免疫前血清时未观察到该条带。当抗Lf抗体与偶联到溴化氰活化琼脂糖上的Lf结合后，对78-kD抗原的反应可被完全抑制，这可能表明Lf和肠道细菌抗原中存在交叉反应表位。
