Human sp a

Human SP-A (BioVendor, Brno, Czech Republic), mouse eotaxin-1 (CCL11, eBioscience, Vienna, Austria) and eotaxin-2 (CCL24, R&D Systems, Minneapolis MN) assays were performed on BALF according to manufacturers’ protocols. Samples for CCL11 and CCL24 were diluted 1:2 and samples for Human SP-A were assayed undiluted. Briefly, BALF samples were incubated on a 96-well plate coated with the capture antibody and detected with a Streptavidin-HRP:biotin-conjugated secondary antibody complex on a plate reader (BioTek Instruments, Winooski VT).

The ELISA measurements were performed as per manufacturer’s protocol. Mouse ELISAs. Mouse SP-D (Cat # 028248) and SP-A (Cat # 028239) from USBiological Life Sciences and mouse CC16 (Cat# EKU03200) ELISA was purchased from Biomatik. Mouse albumin ELISA (Cat# E90-134) was from Bethyl Laboratories, Inc. Human SP-A, SP-D and CC16 were also measured by ELISAs (BioVendor). Human ELISAs. The plasma sample collected closest to study enrollment was measured using the following assays. Human SP-D (Cat # RD194059101) SP-A (Cat# RD191139200R) and CC16 (Cat# RD191022200) kits were obtained from BioVendor and ELISA measurements were performed as per manufacturer’s protocol. Plasma was diluted 11-fold for SP-D measurement and diluted 25-fold for CC16 measurement, and the final OD was measured at 450 nm using Synergy 2 plate reader from BioTek. Proteins are reported in absolute concentrations (pg/mL or ng/mL plasma).