猪肺炎支原体感染诱导的固有免疫应答研究进展

猪肺炎支原体是引起猪支原体肺炎的病原。由于缺乏成熟的猪肺炎支原体感染动物模型,使得猪肺炎支原体相关的抗感染免疫研究进展较为缓慢。本文从猪肺炎支原体感染后的炎症反应、固有免疫系统对猪肺炎支原体的识别、固有免疫细胞的作用、补体系统、抗菌肽、自噬以及细胞凋亡7个方面进行综述,旨在阐明固有免疫系统各组分在猪肺炎支原体感染中发挥的作用的研究进展,并对今后猪肺炎支原体感染的固有免疫应答研究的重点方向进行展望。     

The effect of Mycoplasma arthritidis infection on the kinetics of colloidal carbon clearance in mice

Abstract The activity of phagocytes from A/J mice was estimated by the carbon clearance test following injection of Mycoplasma arthritidis . Phagocytic activity was significantly depressed 12 h post-infection ( P =0.001) and returned to normal values at 24 h. For animals examined 2 and 7 days post-infection, the overall phagocytic activity increased significantly ( P <10⁻⁴). Phagocytic activity gradually decreased and returned to that of the control group by the end of the fourth week. The relative weights of liver and spleen were significantly increased from the 2nd day post infection ( P =0.0028 and P =0.0014 respectively) and remained increased until the end of the experiment. The early depressive effect on phagocytic activity may be related to superantigen activity with the production of mediators such as macrophage deactivating factor. The later expansion of the macrophage population might bring about the stimulation of autoreactive clones of T and B cells and be responsible for the chronic arthritis that developed in the mycoplasma treated mice.     

Interactions of cultured rat synovial and ocular ciliary body cells with two strains of mycoplasma arthritidis

Summary Strains ofMycoplasma arthritidis differ in their ability to cause joint and ocular inflammations. Although the reasons for this difference are not fully understood, pathogenic myćoplasmas commonly require close associations with the cells they damage. Using³H-uridine labeled mycoplasma, we compared cellular interactions of in vitro cultivated rat synovial and ocular ciliary body epithelial cells with two American Type Culture Collection strains ofM. arthritidis shown to differ in their virulence. Radiolabeling assays gave evidence of á stronger retention capability on cultured cells by the more pathogenic strain, 14152. Scanning electron microscopy demonstrated cellular associations with the two strains of mycoplasma, with more of the 14152 adhering to both cell types. Examination by transmission electron microscopy showed evidence of contact between the more virulent 14152 strain and both cell types, but no similar evidence with the comparatively less virulent strain, 19611. The pathogenicity of different strains ofM. arthritidis may vary according to their ability to closely associate with specific target cells involved in the disease process. This work was supported by funding from the National Society for the Prevention of Blindness, Fight for Sight, Retinitis Pigmentosa International, grant EY05415 from the National Institutes of Health, Bethesda, MD, to Thirkill, and, in part, by an unrestricted grant from Research to Prevent Blindness.     

Identification of an isoschizomer of the HhaI DNA methyltransferase in Mycoplasma arthritidis

The genome of Mycoplasma arthritidis strain 158 has modified cytosine residues at AGCT sequences that render the DNA resistant to digestion with the AluI restriction endonuclease. The DNA methyltransferase responsible for the base modification has previously been designated MarI. From the complete genome sequence of M. arthritidis , we identify Marth_orf138 as a candidate marI gene. Marth_orf138 was cloned in Escherichia coli and its TGA codons converted to TGG. DNA isolated from E. coli cells expressing the modified Marth_orf138 gene was degraded by the AluI nuclease, indicating that Marth_orf138 does not code for MarI. However, the DNA from E. coli was found to have acquired resistance to the restriction endonuclease HhaI. Genomic DNA from M. arthritidis was also found to be resistant to HhaI (recognizes GCGC). The M. arthritidis isoschizomer of the HhaI DNA methyltransferase, coded by Marth_orf138, is designated MarII. Transformation of M. arthritidis was not significantly affected by modification of plasmid at HhaI sites, indicating that the mycoplasma lacks a restriction endonuclease that recognizes GCGC sites.     

Tetrazolium reduction methods for assessment of substrate oxidation and strain differentiation among mycoplasmas, with particular reference to Mycoplasma bovigenitalium and some members of the Mycoplasma mycoides cluster

Aims:  To apply a rapid nitroblue tetrazolium (NBT) reduction assay of substrate metabolism by mycoplasmas that would help to differentiate Mycoplasmas .
Methods and Results:  Growth, substrate preferences and tetrazolium reduction were assessed for 18 strains of Mycoplasma bovigenitalium and Mycoplasma ovine serogroup 11. NBT reduction was detectable in 1 h with 10⁸ CFU ml⁻¹. Use of α-ketobutyrate, lactate and pyruvate to support growth and NBT reduction were correlated: pyruvate was preferred and lactate was used by only four of the 18 strains. Selected members of the Mycoplasma mycoides cluster were also assessed and monotetrazoles tested as alternatives to NBT. The NBT method was applied to a further 19 species.
Conclusions:  This simple and reproducible method requires only small amounts of cells, enabling routine assessment of substrate use within 1 h, and the rapid assignment of numerous mycoplasmas to one of six physiological groups. The four physiological groups of M. bovigenitalium and Mycoplasma serogroup 11 strains were indistinguishable from each other, which supports the view that these belong to the same species.
Significance and Impact of the Study:  Strain-specific substrate-utilization patterns by mycoplasmas can be obtained rapidly and reliably. The method has potential as a large-scale semi-automated procedure to monitor numerous strains and substrates simultaneously.     

大学生尿沉渣中解脲脲原体和人型支原体感染情况分析

目的了解解脲脲原体(Ureaplasma urealytium,Uu)和人型支原体(Mycoplasma homanis,Mh)在大学生泌尿道中的感染情况.方法采用培养方法对37例正常的在校大学生的尿沉渣进行Uu和Mh的培养.结果采用培养方法检测21.6%(8/37)的大学生Uu感染为阳性,其中分离阳性者全部是女生(8/20,40%),13.50%(5/37)大学生Mh培养为阳性,其中阳性者亦全部为女生,占女生人数的25%(5/20).结论Uu和Mh可能是女性泌尿道的正常寄生的微生物之一.     

3类抗菌药物体外抗解脲脲原体和人型支原体的作用

目的比较四环素类、大环内酯类和喹诺酮类3类抗菌药物,对临床分离株解脲脲原体(Uu)和人型支原体(Mh)的抗菌作用,为临床用药提供参考依据.方法采用直接肉汤药盘法测定了临床标本中,195株Uu和1218株Mh对3类抗菌药物中的8种抗生素的敏感性.结果 195株Uu对四环素、多西环素、米诺环素、罗红霉素、阿齐霉素、交沙霉素、氧氟沙星和司帕沙星的敏感率,分别为21.0%、48.2%、39.5%、79.0%、88.7%、74.9%、28.2%和64.6%.118株Mh对四环素、多西环素、米诺环素、罗红霉素、阿齐霉素、交沙霉素、氧氟沙星、司帕沙星的敏感率分别为5.1%、33.9%、26.3%、0.0%、0.0%、89.8%、70.3%和64.4%.960例混合感染的Uu Mh,对交沙霉素最敏感(79.2%).结论泌尿生殖道支原体的药敏监测对指导临床治疗具有重要意义.     

The relaxing ori-ter balance of Mycoplasma genomes

Mycoplasma are wall-less bacteria with small genomes, which are thought to have resulted from massive genome reductive processes, during which the ori-ter balance may be disrupted. For technical difficulties, ori and ter have been located only in a few Mycoplasma strains. Using the Z curve method, we were able to locate turning points on the Mycoplasma genomes, with the minimum and maximum points co-locating with ori or ter in the reference genomes. Assuming Z curve correctly located ori and ter, we calculated the distances from ori to ter in both directions on the circular genome and calculated the ori-ter balance status. The Mycoplasma genomes were not balanced, possibly as a result of close association of Mycoplasma with hosts, where there would be no other microbes for Mycoplasma to compete with for nutrients, so fastest possible growth related to balanced genomes might not be needed by Mycoplasma, leading to a relaxing ori-ter balance.     

肺炎支原体P1蛋白羧基端基因片段在大肠杆菌中克隆的研究

在大肠杆菌中克隆肺炎支原体P1蛋白羧基端基因片段,为P1蛋白基因片段的扩增、表达及探讨羧基端基因片段功能打基础.采用PCR扩增方法获取P1结构基因.扩增产物用SalI和EcoRI酶切消化,回收1kb大小的DNA片段并与pUC19DNA连接,转入大肠杆菌JM109菌株.用X-gal平板及质粒图谱分析方法筛选重组克隆株,再用限制性核酸内切酶酶切图谱分析鉴定.经PCR扩增MPDNA获得1条5.0kbDNA片段.重组质粒限制性内切酶指纹图谱显示出2条带,1条为pUC19载体DNA带,另1条是1kb的插入片段.实验获得肺炎支原体P1蛋白结构基因及含P1蛋白羧基端DNA片段的重组克隆株.