The pathomechanism of postmenopausal osteoporosis-the involvement of MIF and therapeutic approach using MIF-DNA vaccine

1)为了建立以巨噬细胞移动抑制因子为靶点的主动免疫疗法，我们将巨噬细胞移动抑制因子（MIF）cDNA序列的第二环出区替换为破伤风毒素p30的cDNA序列，构建了MIF DNA疫苗（MIF-DV）。作为对照疫苗（CV），我们使用没有插入物的质粒。当将这些疫苗给予4周龄雌性BALB/c小鼠（每组n = 5）时，MIF-DV组在给药后4周显示血清抗MIF抗体滴度显著增加，而CV组没有显示这种增加。当给予这些小鼠抗II型胶原抗体以诱导关节炎时，MIF-DV组的关节炎评分显著低于自然病程组或CV组。类似地，在使用IL-1Ra敲除小鼠的实验中，这些小鼠发生自发性自身免疫性关节炎，关节炎的发病率、关节炎评分、关节炎的发生率、关节炎的发生率、关节炎的 ...更多信息 MIF-DV组的足肿胀度和后足肿胀度明显低于自然病程组和CV组。这些表明，MIF-DV的施用可以作为针对关节炎的预防方法是有希望的。（Arthritis Rheum 2007）2）使用上述方法，我们对3周龄雌性BALB/c小鼠施用MIF-DV和CV。在确认DV组中血清抗MIF自身抗体水平的增强后，我们对MIF-DV或CV施用小鼠进行卵巢切除术（OVX）或假手术。即，将小鼠分为4组，即，MIF-DV-假手术、MIF-DV-OVX、CV-假手术和CV-OVX。术后4周处死动物，行显微CT和骨组织形态计量学分析。在CV给药的小鼠中，OVX诱导骨小梁体积的损失和破骨细胞面积和数量的增加，即，高骨转换状态，如在正常小鼠中所见。另一方面，在MIF-DV给药的小鼠中，OVX未能诱导骨小梁体积的损失或破骨细胞面积或数量的增加。MIF-DV的管理也可能是一个有前途的预防方法，对骨质疏松症引起的雌激素缺乏。（疫苗，提交中）减

1) Aiming at establishing an active immunization therapy targeting macrophage migration inhibitory factor, we have developed a MIF-DNA vaccine (MIF-DV) by inserting into plasmid a construct that consists of MIF cDNA sequence, in which the 2nd loopout region of MIF cDNA sequence was replaced by the cDNA sequence of tetanus toxin p30 as helper T epitope. As a control vaccine (CV), we used plasmid without insert. When these vaccines were administered to 4-weeks old female BALB/c mice (n=5, each group), the MIF-DV group showed significant increase in serum anti-MIF antibody titer at 4 weeks after administration, while the CV group showed no such increase. When these mice were given anti-type II collage antibody in order to induce arthritis, the arthritis score was significantly lower in MIF-DV group than in natural course group or in CV group. Similarly, in experiments using IL-1Ra knockout mice, which develop spontaneous autoimmune arthritis, the incidence of arthritis, arthritis score, a … More nd hindfoot swelling were significantly lower in MIF-DV group than in natural course group or in CV group. These suggested that the administration of MIF-DV could be promising as a prophylactic approach against arthritis. (Arthritis Rheum 2007)2) Using the methods described above, we administered MIF-DV and CV against 3-weeks old female BALB/c mice. After confirming the enhancement of serum anti-MIF autoantibody levels in DV group, we performed ovariectomy (OVX) or sham surgery to MIF-DV or CV-administered mice. Namely, the mice were divided into 4 groups, i.e., MIF-DV-sham, MIF-DV-OVX, CV-sham, and CV-OVX. They were sacrificed at 4 weeks after surgery, and the femur were subjected to micro CT analysis and bone histomorphometric analysis. In CV-administered mice, OVX induced loss of trabecular bone volume and the increase in osteoclast area and number, i.e., high bone turnover state, as seen in normal mice. On the other hand, in MIF-DV-administered mice, OVX failed to induce loss of trabecular bone volume or increase in osteoclast area or number. The administration of MIF-DV might also be a promising prophylactic approach against osteoporosis induced by estrogen deficiency. (Vaccine, under submission) Less