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  • Title: Serum antibodies against native and denaturated hemagglutinin glycoproteins detected by ELISA as correlates of protection after influenza vaccination in healthy vaccinees and in kidney transplant recipients.
    Author: Grund S, Pietzonka S, Michel S, Adams O.
    Journal: J Virol Methods; 2013 Nov; 193(2):558-64. PubMed ID: 23896019.
    Abstract:
    The microneutralization assay is the standard method to investigate immune responses to influenza vaccination. However there remains some uncertainty as to whether ELISA results are a true measure of immunity in healthy or immuno-compromised vaccines. Furthermore it has been questioned if antibodies against native ("folded") and against denaturated ("unfolded") viral glycoproteins can equally be used as a marker of protection. In this study, two different quantitative IgG-ELISA assays detecting (i) antibodies against unfolded recombinant hemagglutinin (HA) (r-ELISA) and (ii) antibodies against the native HA on the influenza virus surface captured by fetuin-linkage (f-ELISA) were compared to microneutralization titers in sera from 29 healthy vaccinees (n=87 sera) and 39 kidney transplant recipients (n=117 sera) collected before, three weeks after and six months after vaccination against influenza A (H1N1) 2009. With both ELISAs a significant increase in antibody levels was detected after vaccination and linear regression analysis demonstrated that r-ELISA and f-ELISA correlated with microneutralization (R=0.622 for r-ELISA vs. R=0.56 for f-ELISA). For the healthy vaccinees both ELISAs were found to be adequate to distinguish protected from non-protected individuals (sensitivity and specificity: 87.5%/85.3% for r-ELISA and 87.5%/88.3% for f-ELISA). Results from the transplant recipients showed a slightly reduced sensitivity of 73.3% for r-ELISA while the f-ELISA demonstrated similar sensitivity and specificity as in the healthy vaccinees. However, in order to obtain these assay performances the cut-off-values for protection had to be adjusted for both assays and both investigation cohorts respectively limiting their application in routine laboratories.
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