Citrate cpt tubes

Umbilical cord blood was obtained from the placenta of uncomplicated term pregnancies after delivery into citrate CPT tubes (BD). Blood was processed per manufacturer's instructions within 24 h of collection, and resulting CBMC cryopreserved. Infants aged 1–24 months undergoing elective surgery at OHSU Doernbecher Children's Hospital were enrolled. Infants had not received BCG vaccination, which is not part of the childhood immunization series in the United States. Blood was drawn into citrate CPT tubes (BD) and processed per manufacturer's instructions to isolate PBMC within 24 h of collection, and PBMC subsequently cryopreserved. Adult PBMC were obtained by leukapheresis from healthy adult donors and cryopreserved. Adult participants lacked any detectable responses to Mycobacterium tuberculosis (Mtb) immunodominant antigens CFP-10 and ESAT-6 and therefore were assumed to be Mtb uninfected.

Children from age 2–60 months hospitalized with lower respiratory infection (LRTI) from etiologies other than TB were recruited from Mulago Hospital Pediatric Wards. All children were HIV-negative and had no known recent TB contacts. Children were reassessed 2 months following enrollment to establish vital status and ensure clinical improvement. Following completion of the study, data from all participating children was reviewed by study investigators (DAL, CLL, SK) to confirm diagnosis of non-TB LRTI. Blood was drawn into citrate CPT tubes (BD) and processed per manufacturer's instructions to isolate PBMC within 24 h of collection, and PBMC subsequently cryopreserved. Adult participants were recruited from the National TB Treatment Center at Mulago Hospital, hospital staff, and the community surrounding Kampala, Uganda, between 2001 and 2014. Participants were HIV uninfected and included individuals with (tuberculin skin test, TST ≥ 10) or without (TST <10 mm) evidence of Mtb infection. All had the absence of signs or symptoms of TB (prolonged cough, hemoptysis, fever, weight loss, and night sweats). Heparinized blood was collected for PBMC isolation by means of density gradient centrifugation.