Further, since surface PF-01367338 concentration attachment of M. genitalium to culture flasks often correlates with adherence to eukaryotic cells, we tested the TIM207 strain for hemadsorption with sheep erythrocytes, a technique that we routinely use to assess the adherence of this species, and compared its phenotype with G37. TIM207 showed a hemadsorption positive phenotype similar to that of wild type G37 (data not shown), suggesting that there is no difference between these two strains with regard to adherence to eukaryotic cells. To evaluate this further, the ability of TIM207 strain to adhere/invade epithelial cells was also assessed by infecting
HeLa cells. Confocal microscopic analysis of the infected cells revealed that both G37 and TIM207 strains exhibit similar Selleckchem MK1775 levels of adherence/invasion, although the control strain TIM262 was little different from these by showing relatively higher levels of adherence/invasion (Figure 4). The reason for this difference is unknown at present. However, the fact that both G37 and TIM207 show more or less similar phenotype reiterates that the partial non-adherence to culture flasks by TIM207 strain has no correlation with its adherence to or invasion of eukaryotic cells. It has been shown [35] that invasion of M. genitalium into cultured Selleckchem QNZ HeLa and EM42 cervical epithelial cell lines occurs within
30 minutes after postinfection and the invaded bacteria are localized within nuclei. Interestingly, this study has also reported that only a subset of the bacteria (M. genitalium) invades the cells. This fact was confirmed by another group that used electron microscopy and they estimated that the invading
bacteria would be around 50% of the total bacteria showing adherence [50]. In this context, it will be of interest to know whether there exists any difference between the wild type and TIM207 in the quantity of invading bacteria and this question will be addressed in our future studies. Figure 4 Microscopic observation of adherence/invasion of M. genitalium strains to HeLa cells: FITC labeled M. genitalium strains were used to infect HeLa cells with MOI enough of 1:25 for 1 h as described in material and methods and observed with confocal microscopy. G37, TIM207 and TIM262 indicate infection of cells with M. genitalium wild type G37 strain, MG_207 mutant strain and control strain TIM262, respectively. PBS indicates uninfected control. Nevertheless, the partial culture flask non-adherence phenotype that we observed with the TIM207 strain is different from that of the completely culture flask non-adhering phenotype of M. genitalium strain reported earlier [51]. Feldner et al. [52] reported that adherence of mycoplasma to culture flasks are based on electrostatic forces rather than adhesion mediated.
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