Human Subjects

PBMCs for the reference samples were obtained from buffy coats of healthy blood donors at Massachusetts General Hospital and were cryopreserved according to IRB approved protocols. HIV-infected subjects were recruited from outpatient clinics at local Boston hospitals and referred from providers throughout the United States, following institutional review board approval and written informed consent (IRB protocol #2003P001894). For the second part of the study, BAL samples with donor-matched peripheral blood samples from HIV-infected patients were used. Participants were recruited from existing cohorts at Massachusetts General Hospital following institutional review board approval and written informed consent (IRB protocol #2013P002436). Bronchoscopy was performed under conscious sedation via standard technique. Two BALs were performed with 120 mL of normal saline each in a segment of the lingula and a segment of the right middle lobe. Peripheral blood from the study subjects was obtained on the same day as the bronchos-copy and all samples were processed immediately and stained for CyTOF analysis, without cryopreservation.