Apoptosis of neutrophils was significantly downregulated in its early stages by H37Rv (P = 0.01) when compared with the control. Other strains did not influence the rate of early apoptosis (Table 1). Considering late apoptosis, H37Rv (P = 0.003)
and BCG (P = 0.01) induced significantly higher apoptosis when compared with Mw. When compared with control, there was an increasing trend in the rate of late apoptosis MK-2206 molecular weight of H37Rv-infected neutrophils, but the change was not significant (Table 1). Similarly, PMA (P = 0.001), BCG (P = 0.03) and H37Rv (P = 0.0005) significantly increased the necrotic cell population when compared to control. Also, H37Rv (P = 0.002) was able to significantly increase the necrosis of neutrophils selleck compound library compared with Mw (Table 1). A representative scatter plot of apoptosis is shown in Fig. 3. Figure 4 represents levels of pro-inflammatory cytokines in infected neutrophil supernatants. Significantly higher levels of TNF-α were observed in H37Rv-infected (P = 0.01) and PMA-stimulated (P = 0.03) neutrophils. Vaccine strains did not have profound effect on the release of TNF-α by neutrophils (a). None of the strains was able to modulate the secretion of the major pro-inflammatory cytokine IFN-γ by neutrophils (b).
Figure 5 depicts the expression of chemokine receptors CCR5 and CCR7 in representative histograms (a and b) and Box and Whisker plots (c and d). The expression of CCR5 was significantly upregulated in all conditions (PMA: P = 0.002, BCG: P = 0.003, Mw: P = 0.003, H37Rv: P = 0.01) (c). With PMA-stimulated Nu sups, significantly increased expression of CCR7 (P = 0.008) was observed on monocytes. Similarly, CCR7 showed significantly
higher expression on stimulation with Nu sups from H37Rv (P = 0.01) but not from BCG and Mw. Also, there was a significantly higher expression of CCR7 on monocytes stimulated with H37Rv-infected Nu sups (P = 0.03) when compared to Mw-infected sups (d). Figure 6 depicts the expression of CD 69 and CXCR3 in representative histograms (a and b) and Box and Whisker plots (c and d). The activation marker CD69 was found to be significantly upregulated when stimulated with H37Rv (P = 0.0008)-infected Nu sups. PMA-stimulated Nu sup was also found to significantly increase the expression of CD69 (P = 0.0003) when compared with control Isotretinoin (c). The expression of the chemokine receptor CXCR3 was not influenced on stimulation with any infected sup (d). The interaction of neutrophils with macrophages, as well as the downstream effects on T cell activity, could result in a range of outcomes from early clearance of infection to dissemination of viable bacteria together with an attenuated acquired immune response (Lowe et al., 2012). Neutrophils are rapidly recruited to sites of mycobacterial infection, where they phagocytose bacilli and induce chain of responses through various receptors to initiate the immune response against MTB.