After the incubation period, the caspase-8 activity was measured using Caspase Glo Assay kit

perature for 2 hours and then studied in the EIA for H1N1 binding IgG-Celgosivir antibodies described above. The inhibition percent51-/OD value multiplied 100. Hemagglutination inhibition test for antibodies to H1N1 Hemagglutination inhibition test for antibodies against H1N1 influenza A/ California/07/2009 vaccine strain and epidemic A/Finland/814/01 and A/ Finland/715/00 virus strains was performed as previously described. The neutralizing antibodies to Polio Sabin 1 virus were determined with a standard microneutralization assay as previously described. Sandwich EIA for antibodies to Pandemrix derived H1N1 viral proteins Polystyrene plates were coated with rabbit antibodies to HA, NA, anti-NA MAb and anti-NP PAb all from Sino Biological, China), 1 mg/mL in PBS, and kept overnight at +4 C 0.5% BSA-PBS was used for post-coating. After washing with PBS, Pandemrix antigen suspension was added and incubated in the antibody coated wells. After washing, mouse monoclonal antibodies to HA, NA or NP, anti-NA Mab or anti- NP MAb were added. In the EIA for IgGantibodies to Pandemrix derived NP, plasma samples were diluted 1:300 in 0.5% bovine serum albumin -PBS. The binding to antibodies was detected with alkaline phosphatase conjugated anti-human IgG-antibody or anti-mouse IgGantibody. After adding the substrate the color reaction was read. EIA for antibodies to recombinant H1N1 viral proteins Polystyrene plates were coated with recombinant HA from H1N1 influenza PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19684114 A/ California/07/2009 vaccine strain or NP from A/Puerto Rico/8/1934 in PBS, 0.1 mg/well. 0.5% BSA-PBS was used for post-coating, washing buffer was either 0.5% BSA-PBS or 0.5% BSA-0.05% PBSpolysorbate 80, and plasma samples were diluted 1:100 in 0.5% BSA-PBS or 0.5% BSA-0.05%PBS-polysorbate 80. Alkaline phosphatase conjugated anti-human IgG-antibody or anti-mouse IgG antibody was used as a conjugate. 6 / 23 Influenza A Vaccine Antigen and Narcolepsy Risk Genotyping of HLA DQB106:02 risk allele of narcolepsy HLA genotyping for the selected DQB1 and DQA1 alleles was performed using sequence-specific oligonucleotide hybridization as earlier described. MES-PAGE analysis All PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19683642 polyacrylamide gels were run by the NOVEX Highresolution Bolt Mini Gel Tank System with ready-made Bolt 412% Bis-Tris Plus gradient gels under MES buffer conditions according to the instructions of the manufacturer. Gels were stained by PageSilver Silver Staining Kit from Fermentas Life Sciences according to the instructions of the manufacturer. Pre-stained standard SeeBlue Plus 2 was used for molecular weight calculations under reduced conditions. Nonreducing conditions allowed no molecular weight estimations. Recombinant Influenza A virus H1N1 nucleoprotein and HA were used as control proteins. Western blot analysis All polyacrylamide gels were run by the NOVEX Highresolution Bolt Mini Gel Tank System with ready-made Bolt 412% Bis-Tris Plus gradient gels under MES buffer conditions according to the instructions of the manufacturer. After the gel electrophoresis each gel was blotted by the iBlot Dry Blotting System with the pre-installed P0 program with the 10 minutes transfer selection according to the manufacturer instructions. Ready-made iBlot Gel Transfer Regular Nitrocellulose Stacks were used. Milk powder diluted in TBS-0.05% Tween20 was used for blocking, and mouse monoclonal antibodies against HA or NP or anti- NP MAb, Sino Biologicals) diluted at the concentration of 1 mg/mL in Tween- TBS was used for the identification