Thymus glands were collected on PND 21 and spleens on PND 21, 90, 6 months and 1 year. They were transferred from NCTR to the current lab coded so that the tests were conducted blind to treatment (++ for binding).

They were transferred in 1640 RPMI medium supplemented with 5% bovine calf serum and penicillin/streptomycin on ice overnight. Thymocytes and splenocytes were isolated by mechanical disruption and made into single cell suspensions in RPMI 1640 medium supplemented with 10% BCS and penicillin/streptomycin. Cells were counted on Beckman Coulter Counter. Cells were washed and prepared for flow cytometry. Flow cytometry is a well-established method, but samples could not be tested the day of collection. All tissues were processed the day after collection. (+ for methods).

Li, J. et al. 2018a

Risk of bias

HAWC