Murine Models of Helminth Infection

Female BALB/c and IL-4gfp 4get reporter mice on the BALB/c background were bred in-house and maintained under specific pathogen-free conditions at the University of Edinburgh. Mice were used at 6–12 weeks of age, and randomly assigned to experimental groups. The L. sigmodontis life cycle was maintained in gerbils (Meriones unguiculatus) using the mite vector Ornithonyssus bacoti [22 (link)]. Mice were infected s.c. on the upper back with 30 L. sigmodontis L3 larvae. Adult or larval parasites were recovered by lavage of the thoracic cavity. L. sigmodontis antigen (LsAg) was prepared by collecting the PBS-soluble fraction of homogenized adult male and female worms. To quantify blood microfilariae, 30 μL of tail blood was collected in FACS lysing solution (Becton-Dickinson), and microfilaria counted using a dark field optical microscopy (Axiovert 25, Zeiss). N. brasiliensis was maintained in Sprague-Dawley rats as previously described [23 (link)]. Mice were infected by s.c. injection with 200 N. brasiliensis L3 larvae. The parathymic, posterior, mediastinal and paravertebral LN, were taken as a source of thoracic LN (tLN) draining the pleural cavity (PleC). PleC cells were recovered by lavage. TLN cells were dissociated and washed in RPMI-1640 (invitrogen) supplemented with 0.5% mouse sera (Caltag-Medsystems), 100 U/ml penicillin, 100 μg/ml streptomycin and 2 mM L-glutamine.