B6.129p2 il10tm1cgn j b6 il 10

For embryo transfer (ET) recipients, 8–10 week old female C57BL/6J (The Jackson Laboratory, Bar Harbor, ME), C57BL/6NTac (Taconic Biosciences, Inc., Cambridge, IN facilities), and Crl:CD1 (Charles River Laboratories, Wilmington, MA) mice were purchased and allowed to acclimate for 1 week prior to use. Embryos from 8 week-old female B6.129P2-Il10^tm1Cgn/J (B6 IL-10^−/−) and C3Bir.129P2(B6)-Il10^tm1Cgn/J (C3H IL-10^−/−; The Jackson Laboratory) mice were harvested from colonies maintained on site. Vasectomized, 8–10 week old Crl:CD1 male mice (Charles River Laboratories) were co-housed to induce pseudopregnancy and intrauterine embryo transfer was performed (see section Embryo Collection and Transfer). All mice were housed in microisolator cages on ventilated racks (Thoren, Hazelton, PA) on a 14:10 light dark cycle, and provided ad libitum access to 5,058 irradiated breeder chow (LabDiet, St. Louis, MO) and acidified autoclaved water.

The NSV, TE12 and TE strains of SINV [12 (link)] were grown and assayed by plaque formation in BHK-21 cells. C57BL/6J (B6 WT) and B6.129P2-Il10^tm1Cgn/J (B6 IL10-/-) mice were purchased from Jackson Laboratories and bred in-house. Next, 4- to 6-week-old sex and age-matched mice were infected intranasally with 10⁵ plaque-forming units (pfu) NSV diluted in PBS. Disease severity was assessed using a previously described scoring system [22 (link)]. Briefly, 0 = no signs of disease; 1 = abnormal hind limb and tail posture, ruffled fur, and/or hunched back; 2 = one hind-limb paralyzed; 3 = both hind-limbs or full-body paralysis; 4 = death.
For the collection of cervical lymph node (CLN), brain and spinal cord tissues, mice were anesthetized with isoflurane and perfused with ice-cold PBS. Tissues were used fresh or frozen at −80 °C. Subsequently, 10% weight per volume tissue homogenates were made in PBS and clarified by centrifugation. All experiments were performed according to protocols approved by the Johns Hopkins University Animal Care and Use Committee.