Mycobacteria Exploit Host Hyaluronan for Efficient Extracellular Replication
Figure 5
The effect of 3 hyaluronan synthases on the growth of BCG and M. tuberculosis.
(A), Established transfectant cells (Rat 3Y1 fibroblasts) with control vector (Mock) or vector to express hyaluronan synthase 1 (HAS1), HAS2 (HAS2), or HAS3 (HAS3) were cultured in the presence of BCG-Luc or media alone. The growth of bacteria was monitored by luciferase activity. RLU, relative luciferase unit. The results are expressed as mean±the standard deviation (n = 3). For statistical analysis, a two-way ANOVA with Bonferroni Post tests were used to obtain P-values for each time point, comparing the various growth conditions to the control. *P<0.01. (B), Hyaluronidase (HAase) treatment enhances the growth of BCG after infection to HAS1-tranfected cells. After 16 hours exposure of BCG-Luc to transfected cells with control vector (Mock) or vector expressing HAS1 (HAS1), unbound bacteria were washed and cultured in the presence or absence of 2 units/ml of hyaluronidase (HAase). Bacterial growth was monitored by the luciferase activity (RLU). Cntl, HAS1-transfectant cells without infection of BCG-luc. The results are expressed as mean±the standard deviation (n = 3). For statistical analysis, a two-way ANOVA with Bonferroni Post tests were used to obtain P-values for each time point, comparing the various growth conditions to the control. *P<0.01. (C), The growth of M. tuberculosis H37Rv after infection to transfectant 3Y1 fibroblasts with control vector (Mock) or vector to express hyaluronan synthase 1 (HAS1), HAS2 (HAS2), or HAS3 (HAS3) was monitored by CFU. The results are expressed as mean±the standard deviation (n = 3). For statistical analysis, a two-way ANOVA with Bonferroni Post tests were used to obtain P-values for each time point, comparing the various growth conditions to the control. *P<0.01.