The effect of ovine peripheral blood mononuclear cells on Haemonchus contortus larval morbidity in vitro.

The objective of this study was to determine the effects of peripheral blood mononuclear cells (PBMC), derived from parasite-resistant St. Croix (STC) hair sheep and parasite-susceptible Suffolk (SUF) sheep, on Haemonchus contortus L3 stage larval death in vitro, with or without autologous serum. Larval morbidity was quantified by measuring larval ATP concentration following incubation with PBMC. Larvae exposed to either STC- or SUF-derived PBMC had lower ATP than live larvae (0.12 µmol/L ATP and 0.16 µmol/L ATP vs 0.27 µmol/L ATP, respectively) (P<.001) and greater ATP of dead larvae (0.03 µmol/L ATP) (P<.001). Breed differences were observed with addition of autologous serum. Larvae exposed to SUF-derived PBMC with autologous serum were not significantly different from live larval ATP. STC-derived serum did not significantly reduce larval ATP compared to PBMC alone (0.11 µmol/L ATP), but was significantly reduced compared to live larvae (0.22 µmol/L ATP) and SUF-derived PBMC with autologous serum (0.23 µmol/L ATP) (P<.001). These data indicate that a cellular response alone is capable of significantly reducing larval ATP in a breed-independent manner. However, addition of serum to SUF-PBMC failed to reduce larval ATP, indicating breed-dependent humoral response to H. contortus.

Effects of peripheral blood mononuclear cells on Haemonchus contortus larval motility in vitro.

The objective of this experiment was to determine effects of peripheral blood mononuclear cells (PBMC), derived from parasite-resistant St. Croix (STC) and parasite-susceptible Suffolk (SF) sheep, on motility of Haemonchus contortus L3 stage larvae in vitro. Peripheral blood mononuclear cells were collected from 10 lambs of each breed, 5 naïve and 5 which had received a priming infection with H. contortus. Larval motility was quantified using a MIF Nikon Sweptfield microscope and NIS Elements AR software, and measurements included path length (PL) (µm), velocity (VEL) (µm/s) and acceleration (ACC) (µm/s2 ). After 18 h of incubation, PL and VEL were greatest in larvae cultured with SF-derived PMBC and were significantly different from all other groups (P < 0·01). No difference was observed in PL or VEL between larvae exposed to naïve or primed STC-derived PBMC and primed-SF PBMC. Differences in ACC were detected between larvae cultured with primed STC-derived PBMC (10·91 µm/s2 ) and naïve SF-derived PBMC (45·7 µm/s2 ) (P = 0·035). These data indicate an innate ability of STC-derived PBMC to severely inhibit larval motility.