Structural and functional characterisation of the ovine interferon gamma (IFNG) gene: Its role in nematode resistance in Rasa Aragonesa ewes

The ovine interferon gamma (IFNG) gene was structurally and functionally characterised, and the association between its variants and the response to Haemonchus contortus infection was examined in resistant and susceptible Rasa Aragonesa ewes in different physiological conditions. Sequential faecal egg counts (FECs) were performed on a naturally infected flock of 400 Rasa Aragonesa ewes and two groups of ewes selected for high (n = 38) or low (n = 33) FEC, which were subsequently challenged with H. contortus in the spring and autumn. The second challenge infection was performed to explore the extent to which the previously established resistance or susceptibility to H. contortus based on FEC reflected the worm burdens of ewes in different physiological states. The ovine IFNG gene was partially isolated and characterised, including the promoter and other regulatory regions. In addition, DNA sequencing of the resistant and susceptible Rasa Aragonesa ewes revealed 13 polymorphisms (12 SNPs and 1 microsatellite) in the IFNG gene. Association studies showed that a haplotype composed of C (SNP −641) and (GTTT)5 (microsatellite) was linked to the FEC and worm burden recorded in spring and autumn challenges, respectively, and thus to susceptibility to H. contortus in adult Rasa Aragonesa ewes. Our results also suggested that the IFNG gene was not responsible for the phenotypic variation in susceptibility to H. contortus but was linked to a QTL segregating near the IFNG gene. No significant differences in IFNG gene expression were found between either resistant and susceptible or pregnant and non-pregnant ewes. Finally, we studied the effects of the polymorphisms on IFNG gene expression, finding that the C/T SNP at −641 and the microsatellite showed significant effects in spleen. The C/T SNP at −641 was located within a possible trans-acting factor binding site. Further work is needed to elucidate the function of this SNP.