Fig. 1
From: HLA-associated protection of lymphocytes during influenza virus infection
Protein Synthesis by Influenza Virus (IAV)-exposed Human PBMC. These representative images show results using cells from HLA-A2-homozygous donor number 1 in part b and HLA-A2-homozygous donor number 2 in part c. a Autoradiograms (monocytes/macrophages, lanes 1, 2; purified lymphocytes, lanes 3, 4) show representative protein synthesis results using PBMC from an HLA-A2-.heterozygous donor after sham-exposure or exposure of PBMC to IAV at an MOI = 10. Odd-numbered lanes show lysates of sham-exposed cells, and even-numbered lanes show lysates of IAV-exposed cells. HA = hemagglutinin, NA/NP = neuraminidase and nucleoprotein, which co-migrate, and M = the matrix protein. Numbers show positions of standard proteins having the indicated Mr. × 10− 3. b Autoradiograms show representative protein synthesis results using PBMC from an HLA-A2-homozygous donor. After sham-exposure (lanes 1, 2 and 5, 6) or exposure to IAV as PBMC at an MOI = 10 (lanes 3, 4 and 7, 8), purified monocytes/macrophages (lanes 1–4) and lymphocytes (lanes 5–8) were obtained and pulse-labeled and analyzed. Odd-numbered lanes show total cell lysates, and even-numbered lanes show lysates immunoprecipitated with mouse monoclonal anti-NP antibody. c Autoradiograms show cell lysates from an HLA-A2-homozygous donor (lanes 1–14) and an HLA-A1,2 donor (lanes 15–28) that were immunoprecipitated using mouse monoclonal anti-NP antibody and polyclonal anti-HA and anti-NA antibodies. After sham-exposure (odd-numbered lanes) or exposure to IAV (even-numbered lanes) as unseparated PBMC, purified lymphocytes and monocytes/macrophages were obtained, pulse-labeled and collected. Lymphocytes were pulse-labeled 0–2 h (lanes 1, 2 and 15, 16), 2–4 h (lanes 3, 4 and 17, 18), 4–6 h (lanes 5, 6 and 19, 20), 6–8 h (lanes 7, 8 and 21, 22), 8–10 h (lanes 9, 10 and 23, 24), and 22–24 h (lanes 11, 12 and 25, 26) after exposure. Monocytes/macrophages (lanes 13, 14 and 27, 28) were pulse-labeled 4–6 h after exposure. Lane 25 is blank (lysate not available). Lane 29 shows positions of standard Mr. proteins