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

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.