van der Geld Ymke M, Hellmark Thomas, Selga Daina, Heeringa Peter, Huitema Minke G, Limburg Pieter C, Kallenberg Cees G M
Department of Clinical immunology, University Medical Center Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands.
Ann Rheum Dis. 2007 Dec;66(12):1679-82. doi: 10.1136/ard.2006.064626. Epub 2007 Jul 20.
In this study, we employed chimeric human/mouse Proteinase 3 (PR3) proteins as tools to induce an autoantibody response to PR3 in rats and mice.
Rats and mice were immunised with recombinant human PR3 (HPR3), recombinant murine PR3 (mPR3), single chimeric human/mouse PR3 (HHm, HmH, mHH, mmH, mHm, Hmm) or pools of chimeric proteins. Antibodies to mPR3 and HPR3 were measured by ELISA. Antibodies to rat PR3 were determined by indirect immunofluorescence (IIF) on rat white blood cells. Urinalysis was performed by dipstick analysis. Kidney and lung tissue was obtained for pathological examination.
In mice, immunisation with the chimeric human/mouse PR3 Hmm led to an autoantibody response to mPR3. Rats immunised with the chimeric human/mouse PR3 Hmm, HmH and mmH, or a pool of the chimeric human/mouse PR3 proteins, produced antibodies selectively binding to rat granulocytes as detected by IIF. No gross pathological abnormalities could be detected in kidney or lungs of mice or rats immunised with chimeric human/mouse PR3.
Immunisation with chimeric human/mouse proteins induces autoantibodies to PR3 in rats and mice. Chimeric proteins can be instrumental in developing experimental models for autoimmune diseases.
在本研究中,我们使用人/鼠嵌合蛋白酶3(PR3)蛋白作为工具,在大鼠和小鼠中诱导针对PR3的自身抗体反应。
用重组人PR3(HPR3)、重组鼠PR3(mPR3)、单一的人/鼠嵌合PR3(HHm、HmH、mHH、mmH、mHm、Hmm)或嵌合蛋白池免疫大鼠和小鼠。通过酶联免疫吸附测定(ELISA)检测针对mPR3和HPR3的抗体。通过对大鼠白细胞进行间接免疫荧光法(IIF)测定针对大鼠PR3的抗体。通过试纸条分析进行尿液分析。获取肾脏和肺组织进行病理检查。
在小鼠中,用人/鼠嵌合PR3 Hmm免疫导致对mPR3产生自身抗体反应。用人/鼠嵌合PR3 Hmm、HmH和mmH或人/鼠嵌合PR3蛋白池免疫的大鼠,通过IIF检测产生了选择性结合大鼠粒细胞的抗体。在用嵌合人/鼠PR3免疫的小鼠或大鼠的肾脏或肺中未检测到明显的病理异常。
用人/鼠嵌合蛋白免疫可在大鼠和小鼠中诱导针对PR3的自身抗体。嵌合蛋白有助于开发自身免疫性疾病的实验模型。