Kusters J G, Gerrits M M, Van Strijp J A, Vandenbroucke-Grauls C M
Department of Medical Microbiology, School of Medicine, Vrije Universiteit, Amsterdam, The Netherlands.
Infect Immun. 1997 Sep;65(9):3672-9. doi: 10.1128/iai.65.9.3672-3679.1997.
Helicobacter pylori can transform from its normal helical bacillary morphology to a coccoid morphology. Since this coccoid form cannot be cultured in vitro, it has been speculated that it is a dormant form potentially involved in the transmission of H. pylori and in a patient's relapse after antibiotic therapy. In this study we determined the effects of aging, temperature, aerobiosis, starvation, and antibiotics on the morphologic conversion rate and culturability of H. pylori. Aerobiosis and the addition of a bactericidal antibiotic to the culture medium resulted in the highest conversion rate. During the conversion to coccoid forms, the cultures always lost culturability at the stage where 50% of the organisms were still in bacillary form; this result indicated that culturability and coccoid morphology are two separate but related entities. Independent of the conditions used to induce the conversion into coccoids, the morphological conversion was accompanied by several marked antigenic and ultrastructural changes. Also, both the total amounts and the integrity of RNA and DNA were significantly reduced in coccoid forms. With the potential-sensitive probe diOC(5)-3, a clear loss of membrane potential in coccoid forms was observed. Inhibition of protein or RNA synthesis by the addition of bacteriostatic antibiotics did not prevent the conversion to coccoid forms but resulted in an increased conversion rate. Hence, we conclude that conversion of H. pylori from the bacillary to the coccoid form is a passive process that does not require protein synthesis. Our data suggest that the coccoid form of H. pylori is the morphologic manifestation of bacterial cell death.
幽门螺杆菌可从其正常的螺旋杆菌形态转变为球形形态。由于这种球形形态无法在体外培养,因此据推测它是一种潜在的休眠形式,可能参与幽门螺杆菌的传播以及患者在抗生素治疗后的复发。在本研究中,我们确定了老化、温度、需氧性、饥饿和抗生素对幽门螺杆菌形态转化率和可培养性的影响。需氧性以及向培养基中添加杀菌抗生素导致最高的转化率。在向球形形态转变的过程中,培养物总是在50%的生物体仍处于杆菌形态的阶段失去可培养性;这一结果表明可培养性和球形形态是两个独立但相关的实体。无论用于诱导转化为球形的条件如何,形态转化都伴随着一些明显的抗原和超微结构变化。此外,球形形态的RNA和DNA总量及完整性均显著降低。使用电位敏感探针diOC(5)-3,观察到球形形态的膜电位明显丧失。添加抑菌抗生素抑制蛋白质或RNA合成并不能阻止向球形形态的转化,但会导致转化率增加。因此,我们得出结论,幽门螺杆菌从杆菌形态向球形形态的转化是一个被动过程,不需要蛋白质合成。我们的数据表明,幽门螺杆菌的球形形态是细菌细胞死亡的形态学表现。