Male balb c mice

BALB/c male mice, 4–6 week old (breeding pairs originally obtained from Jackson Laboratory, Bar Harbor, ME) were bred and housed under pathogen-free conditions and were used at 7–8 week of age. The mice were immunized by intramuscular (IM) injection with a vaccine consisting of 50 μg of conjugate vaccine in 100 μl PBS mixed with 750 μg of alum. Three weeks later they received a booster injection with the same vaccine with alum as adjuvant. The same tissue site was used for both the prime and boost vaccine administrations. Blood was collected at weeks 2, 4, 6, 8, 12, 16 after initial immunization and allowed to clot at room temperature, and then centrifuged at 10,000 g for 20 minutes. The resulting serum was stored at −80°C. All animal studies were conducted under protocols reviewed and approved by Baylor’s Animal Care and Use Committee.

BALB/c male mice (9–10 weeks old, Jackson Laboratory) were housed in groups of five with food and water available ad libitum in a temperature-controlled room maintained on 12 h light/dark cycle (lights on 07:00 to 19:00h, 22±1°C, 50–60% humidity) and used after at least one week of acclimatization. All animal procedures were approved by the Boston Children’s Hospital Institutional Animal Care and Use Committee (IACUC).

C57BL/6 and Balb/c male mice at 6–8 weeks of age were purchased from Jackson Laboratory (Bar Harbor, ME). All mice were housed in UCLA animal facility under specific pathogen-free conditions, received humane care according to the criteria outlined in the “Guide for the Care and Use of Laboratory Animals” (NIH publication 86–23 revised 1985).