Brodeur B R, Johnson W M, Johnson K G, Diena B B
Infect Immun. 1977 Feb;15(2):560-7. doi: 10.1128/iai.15.2.560-567.1977.
As a basis for studies of gonococcal pathogenicity, tissue culture cells were infected with type 1 or type 4 Neisseria gonorrhoeae to determine intracellular viability. A simple and objective means of measurement was devised, based on the uptake of tritiated protein and deoxyribonucleic acid precursors by cycloheximide-inhibited cells infected with gonococci. Cycloheximide was found to inhibit protein synthesis by over 97% tissue culture cells at a concentration of 100 microng/ml. In contrast, N, gonorrhoeae was found to be highly resistant to this antibiotic, and protein synthesis was unaffected by concentrations up to 1,000 microng/ml. Extracellular gonococci were eliminated by treatment with high concentrations of penicillin during cycloheximide inhibition and prior to the addition of radioisotope. Levels of protein and deoxyribonucleic acid synthesis by N. gonorrhoeae in the cycloheximide-treated cells were significantly higher in T1-infected cells (RE2, HeLa, or HEp-2) than in the corresponding T4-infected cells. No differences were observed in tissue cell susceptibility to gonococcal infection. Intracytoplasmic localization of N. gonorrhoeae was confirmed by electron microscopy.
作为淋病奈瑟菌致病性研究的基础，用1型或4型淋病奈瑟菌感染组织培养细胞以确定细胞内存活能力。基于被淋病奈瑟菌感染的环己酰亚胺抑制细胞对氚标记蛋白质和脱氧核糖核酸前体的摄取，设计了一种简单而客观的测量方法。发现环己酰亚胺在浓度为100微克/毫升时可抑制组织培养细胞中超过97%的蛋白质合成。相比之下，发现淋病奈瑟菌对这种抗生素具有高度抗性，浓度高达1000微克/毫升时蛋白质合成不受影响。在环己酰亚胺抑制期间且在添加放射性同位素之前，用高浓度青霉素处理可消除细胞外淋病奈瑟菌。在经环己酰亚胺处理的细胞中，1型感染细胞（RE2、HeLa或HEp-2）中淋病奈瑟菌的蛋白质和脱氧核糖核酸合成水平显著高于相应的4型感染细胞。未观察到组织细胞对淋病奈瑟菌感染的易感性存在差异。通过电子显微镜证实了淋病奈瑟菌的胞质内定位。
