Cpg odn 1826

LRV1c + SFI: 168 μL of 1.2 mg/mL LRV1c protein solution in PBS (Sigma) (202 μg of protein) were added to 832 μL of SFI (0.2% DMSO (Applichem) in sterile 0.9% NaCl (SFI, Braun).
LRV1c + SWE: 168 μL of 1.2 mg/mL LRV1c protein solution in PBS were diluted into 232 μL of WFI. The solution was added to 500 μL of SWE. The mixture was vortexed 5s and left standing at least 30 minutes at room temperature before administration.
LRV1c + SM: 168 μL of 1.2 mg/mL LRV1c protein solution in PBS were diluted into 216 μL of WFI and 300 μL of the solution were added to 700 μL of SM. The mixture was vortexed 5s and left standing at least 30 minutes at room temperature before administration.
LRV1c + LQ: 168 μL of 1.2 mg/mL LRV1c protein solution in PBS were diluted into 432 μL of sterile PBS. 600 μL of the solution were added to 400 μL of LQ and left standing at least 30 minutes at room temperature before administration.
LRV1c + Montanide ISA 720: 700 μL of Montanide ISA-720 were added to 168 μL of 1.2 mg/mL of LRV1c protein solution in PBS by using two syringes connected by a female luer lock adapter until a homogenous emulsion was obtained (50 times).
LRV1c + CpG: 168 μL of 1.2 mg/mL LRV1c protein solution in PBS were diluted into 782 μL in PBS (Sigma) and added 50 μL (20mg/mL) of CpG ODN-1826 (InvivoGen), for a final amount of 10 μg of LRV1c and 50 μg of CpG ODN-1826 for each mouse.

C. abortus strains CP16 (Chinese Institute of Veterinary Drug Control, Beijing, China) and B577 (ATCC^® VR-656™ (obtained from Dr. Bernhard Kaltenboeck, Auburn University, Alabama) were propagated and purified in Buffalo Green Monkey Kidney (BGMK) cells (ATCC cat# PTA-4594) as previously described (Li et al., 2005 (link)). Organisms were purified as elementary bodies (EBs) and stored in aliquots at −80°C until use. C. abortus antigen was prepared by UV-inactivation of EBs for 3 h followed by sonication using a Sonic Dismembrator Model 505 (Thermo Fisher Scientific, Rockford, IL). VCG were generated by genetic inactivation following protein E-mediated lysis as described previously (Eko et al., 2000 (link)). VCG preparations were stored at room temperature until use. Purified mouse Fms-like tyrosine kinase 3 (Flt3) ligand (FL) was obtained from R&D Systems (Minneapolis, MN), CpG 1826 ODN was obtained from InvivoGen (San Diego, CA) and lipopolysaccharide (LPS) was purchased from Sigma (St. Louis, MO).