This is expected since granular cells act as surveillance cells a

This is expected since granular cells act as surveillance cells altering the activity of plasmatocytes during an immune reaction in lepidopteran larvae [69] and [50]. check details Attachment levels of hemocytes

from CTB-injected insects ( Fig. 5C) showed similar patterns to the total number of adhering hemocytes and differential counts as insects injected with corresponding CTX levels. The changes in the actual magnitude of total hemocyte counts were not as pronounced implying hemocytes were either less sensitive to CTB than CTX, or the involvement of the holotoxin’s A-subunit. CTA had no effect on the adhesive abilities of hemocytes from injected insects ( Fig. 5D), which was anticipated because lacking CTB it could not enter the hemocytes. No microaggregates were observed on the slides from any of the experiments, suggesting either no aggregates attached to the glass or the protonodules were not removed from the hemocoel during exsanguination. To determine the proportions of hemocytes capable of adhesion the ratios of absolute total circulating hemocytes from insects injected with

CTX, CTB and CTA (Fig. 5A) to absolute total attached hemocytes from the corresponding treatments and concentrations (Fig. 5B–D) were determined. As expected, hemocytes from larvae injected with 6 nM CTX showed significantly lower levels of adhesion (p<0.05) compared with the PBS injection ( Fig. 5E), implying, as did the in vitro results, Doxacurium chloride that loss of adhesion occurs at lower concentrations of Protein Tyrosine Kinase inhibitor CTX. However, hemocytes from insects injected with 12 and 60 nM CTX showed similar (p>0.05) adhesion capabilities to glass compared with PBS-injected larvae ( Fig. 5E), implying that the increases in levels of circulating hemocytes in the hemocoel (see Fig. 5A) are not necessarily due to loss of hemocyte–tissue adhesion. The discharge of hemocyte stores during infection increases the number of hemocytes available in the hemolymph for adhesion-based immune reactions [32]. Increased circulation of adhesive hemocytes in the hemocoel of CTX-injected larvae might reflect discharge and/or a hemocyte

mobilization from reservoirs. The absence of increasing individually attached hemocytes in vitro (see Fig. 2A), unlike the in vivo response, is likely due to the absence of hemocyte stores, as existent hemocytes microaggregate lowering individual hemocyte counts. That granular cell attachment was more pronounced at the higher CTX concentrations ( Fig. 5B) suggests granular cells were primarily released into the plasma and/or mobilized from tissues. The adhesive hemocyte levels from CTB-injected insects were the same at all CTB concentrations ( Fig. 5E). The adhesive properties of hemocytes from insects injected with CTA were the same at all concentrations ( Fig. 5E), and since CTA had no effect on the concentration of circulating hemocytes ( Fig.

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