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Clinical and Developmental Immunology
Volume 2013 (2013), Article ID 989673, 11 pages
PD-L1 Expression Induced by the 2009 Pandemic Influenza A(H1N1) Virus Impairs the Human T Cell Response
1Medical Research Unit on Immunochemistry (UIMIQ), Specialties Hospital, National Medical Centre “Siglo XXI,” Mexican Social Security Institute (IMSS), 06720 México, DF, Mexico
2Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, 11340 México, DF, Mexico
3Facultad de Química, Universidad Nacional Autónoma de México, 04510 México, DF, Mexico
4Unidad de Investigación en Epidemiología Clínica, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social (IMSS), 06720 México, DF, Mexico
5Instituto Nacional de Salud Pública, 62100 Cuernavaca, MOR, Mexico
Received 8 July 2013; Accepted 22 August 2013
Academic Editor: Oscar Bottasso
Copyright © 2013 Nuriban Valero-Pacheco et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Figure S1. Representative plots of sorted conventional dendritic cells (cDCs) and isolated memory CD4+ T cells (Tm). (a) Gating strategy reflecting the purity of isolated memory CD4+T cells (Tm) characterized by the CD3+CD4+CD45RO+CD45RA- phenotype (purity of 99.6%). (b) Gating strategy for sorting cDCs. (c) Gating strategy to phenotype cDCs, purity of 90.0% of lineage negative cDC population (CD3-, CD14-, CD19-, CD56-), HLA-DR+, and CD123dim.
Figure S2. The A(H1N1)pdm09 virus does not induce PD-1 expression;whereas PD-L1 expression on DCs is induced directly by the A(H1N1)pdm09 virus whilst PD-L1 expression on T cells is dependent on the presence of antigen-presenting cells. (a) PBMCs were stimulated with A(H1N1)pdm09 virus (pH1N1), seasonal influenza virus (H3N2), staphylococcal enterotoxin B or synthetic TLR7 agonist (CL264); PD-1 expression in DCs and T cells was analyzed by flow cytometry. Fold increase in PD-1 expression in cDCs and pDCs, CD4+ and CD8+ T cells after 18 h of stimulus. Enriched (HLA-DR+ cell-depleted)T cells and DCs (b) were stimulated with pH1N1, SEB or CL264; PD-L1 expression in DCs and T cells was analyzed by flow cytometry and representative histograms are shown. M: medium.
Figure S3. PD-L1 is expressed in cDCs and memory CD4+ T cells after 5 and 7 days of culture with A(H1N1)pdm09. (a) PD-L1 expression on isolated memory CD4+ T cells, 7 days after co-culture with sorted cDCs in the presence (blue) or absence (red) of pH1N1 virus. (b) PD-L1 expression on cDCs cultured for 5 days in the presence (blue) or absence (red) of pH1N1.
Figure S4. Gating strategy and representative plots of analyzed dendritic (DCs) and T cells from patients and healthy controls. Gating strategy and representative histograms of PD-L1 expression in cDCs (a, Lin-HLA-DR+CD123dim) and pDCs (b, Lin-HLA-DR+CD123+). Gating strategy and representative histograms of PD-L1 expression in CD4+ T cells (c, CD4+CD8-) and CD8+ T cells (d, CD4-CD8+). The shaded histogram represents PD-L1 expression in a healthy control, whereas the blue and red histograms are representative of two pH1N1+patients.