Sun Yao-Gui, Yin Wei, Fan Xin-Feng, Fan Kuo-Hai, Jiang Jun-Bing, Li Hong-Quan
Clinical Veterinary Medicine, College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi 030801, P R China.
Cell Commun Adhes. 2012 Oct;19(5-6):79-84. doi: 10.3109/15419061.2012.743999. Epub 2012 Nov 14.
The immune adherence (IA) between the porcine erythrocytes and the opsonized Escherichia coli carried green fluorescent protein gene (GFP-E.coli) were detected by the fluorescence microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) with an attempt to verify the existence of IA between the porcine erythrocytes and complemented-opsonized microbes. Under fluorescence microscopy, GFP-E.coli opsonized by fresh rabbit serum complement adhered to the erythrocytes and could not be detached by PBS washing, and no IA was observed between the erythrocytes and nonopsonized GFP-E.coli after co-incubation. SEM and TEM also revealed the existence of IA between the serum complement-opsonized GFP-E.coli membrane and the erythrocyte membrane. The partial complement receptor type 1 (CR1)-like gene from porcine was generated by RT-PCR and rapid amplification of cDNA 3' end (3' RACE) (157bp and 578bp), both of which have high similarity with published mammal's CR1 gene. The sequences were spliced based on homology comparison and submitted to GenBank (GenBank Accession No. JX033989). These results indicated that the porcine erythrocytes were able to bind to the opsonized microorganisms. Furthermore, the sequencing results confirmed that the CR1-like gene exists in porcine.
通过荧光显微镜、扫描电子显微镜（SEM）和透射电子显微镜（TEM）检测猪红细胞与携带绿色荧光蛋白基因的调理大肠杆菌（GFP-E.coli）之间的免疫黏附（IA），试图验证猪红细胞与补体调理微生物之间IA的存在。在荧光显微镜下，新鲜兔血清补体调理的GFP-E.coli黏附于红细胞，经PBS洗涤不能脱落，共孵育后红细胞与未调理的GFP-E.coli之间未观察到IA。SEM和TEM也揭示了血清补体调理的GFP-E.coli膜与红细胞膜之间存在IA。通过RT-PCR和cDNA 3'末端快速扩增（3' RACE）获得了猪的部分1型补体受体（CR1）样基因（157bp和578bp），二者与已发表的哺乳动物CR1基因具有高度相似性。基于同源性比较对序列进行拼接并提交至GenBank（GenBank登录号：JX033989）。这些结果表明猪红细胞能够与调理后的微生物结合。此外，测序结果证实猪中存在CR1样基因。
