Puskás L G, Tiszlavicz L, Rázga Zs, Torday L L, Krenács T, Papp J Gy
Laboratory of Functional Genomics, Biological Research Center, Hungarian Academy of Sciences, P.O. Box 521, H-6701 Szeged, Hungary.
Acta Biol Hung. 2005;56(3-4):233-45. doi: 10.1556/ABiol.56.2005.3-4.7.
Recent and historical evidence is consistent with the view that atherosclerosis is an infectious disease or microbial toxicosis impacted by genetics and behavior. Because small bacterial-like particles, also known as nanobacteria have been detected in kidney stones, kidney and liver cyst fluids, and can form a calcium apatite coat we posited that this agent is present in calcified human atherosclerotic plaques. Carotid and aortic atherosclerotic plaques and blood samples collected at autopsy were examined for nanobacteria-like structures by light microscopy (hematoxylin-eosin and a calcium-specific von Kossa staining), immuno-gold labeling for transmission electron microscopy (TEM) for specific nanobacterial antigens, and propagation from homogenized, filtered specimens in culture medium. Nanobacterial antigens were identified in situ by immuno-TEM in 9 of 14 plaque specimens, but none of the normal carotid or aortic tissue (5 specimens). Nanobacteria-like particles were propagated from 26 of 42 sclerotic aorta and carotid samples and were confirmed by dot immunoblot, light microscopy and TEM. [3H]L-aspartic acid was incorporated into high molecular weight compounds of demineralized particles. PCR amplification of 16S rDNA sequences from the particles was unsuccessful by traditional protocols. Identification of nanobacteria-like particles at the lesion supports, but does not by itself prove the hypothesis that these agents contribute to the pathogenesis of atherosclerosis, especially vascular calcifications.
动脉粥样硬化是一种受遗传和行为影响的感染性疾病或微生物中毒症。由于在肾结石、肾囊肿和肝囊肿液中检测到了类似细菌的小颗粒，即纳米细菌，且它们能形成磷酸钙外壳，因此我们推测这种病原体存在于人类动脉粥样硬化钙化斑块中。通过光学显微镜（苏木精-伊红染色和钙特异性的冯·科萨染色）对尸检时采集的颈动脉和主动脉粥样硬化斑块及血液样本进行检查，以寻找类似纳米细菌的结构；利用针对特定纳米细菌抗原的免疫金标记进行透射电子显微镜（TEM）检查；并将匀浆、过滤后的标本接种于培养基中进行培养。在14个斑块标本中有9个通过免疫TEM在原位鉴定出纳米细菌抗原，但在5个正常颈动脉或主动脉组织标本中均未发现。从42个硬化主动脉和颈动脉样本中的26个样本中培养出了类似纳米细菌的颗粒，并通过斑点免疫印迹、光学显微镜和TEM进行了确认。[3H]L-天冬氨酸掺入了脱矿颗粒的高分子量化合物中。按照传统方法，从这些颗粒中进行16S rDNA序列的PCR扩增未成功。在病变处鉴定出类似纳米细菌的颗粒支持了这些病原体参与动脉粥样硬化发病机制（尤其是血管钙化）的假说，但仅凭这一点并不能证明该假说。
