Updated COVID-19 mRNA boosters protect against severe disease and death by SARS-CoV-2, especially among vulnerable populations. However, repeated mRNA vaccines/boosters have been linked to uni-directional IgG subclass switching of spike-specific antibodies towards IgG4 (IgG3→IgG1→IgG2→IgG4).
In contrast to IgG3 and IgG1, IgG4 binds poorly to Fc gamma receptors (FcγR) present on NK cells (FcγRIIIa) or phagocytes (FcγRIIa) limiting Fc-mediated non-neutralising anti-viral responses. However, its impacts across Omicron variants and sarbecoviruses remain underexplored. Furthermore, IgG4 undergoes Fab-arm exchange in vivo—forming non-symmetrical bispecific antibodies—which could weaken their binding to escape viral variants. Sequential subclass switching could also favor pre-existing biases from ancestral imprinting.
Here, we show that plasma and salivary IgG4 increased substantially following 4 exposures to mRNA vaccines/boosters, in either primary mRNA vaccinees (mRNA cohort; 2x mRNA vaccine + 2x mRNA boosters) or adenovirus-vector vaccinees (Vaxzevria cohort; 2x adenoviral-vector vaccine + 4x mRNA boosters).
Elevated class-switched IgG4 negatively correlated with plasma concentrations of IgG1 targeting Omicron spikes (BA.1, XBB.1.5, JN.1), peaking 4 weeks after their 4th mRNA dose in both cohorts (mRNA: r=-0.53 to -0.66, p<0.01; Vaxzevria: r=-0.65 to -0.71, p<0.01). While ancestral imprinting greatly influenced IgG4 spike responses—particularly towards the RBD—IgG4 responses trended similarly to total IgG, resulting in comparable proportions of IgG4 (%IgG4 / total IgG) across Omicron variants.
Omicron spike-specific plasma antibody engagement with FcγRIIa and FcγRIIIa improved overall following boosters, however these responses also negatively associated with rising IgG4 (mRNA: r=-0.74 to -0.86, p<0.0001; Vaxzevria: r=-0.73 to -0.88, p<0.001). Similar trends were observed in saliva, though IgG4 had a weaker impact. Depletion of IgG4 increased IgG1 binding against viral variants (+13-16%), highlighting how sequential subclass switching results in epitope competition. Similarly, IgG4 depletion also rescued Fc-receptor engagement across Omicron variants, particularly FcγRIIIa (+11-14% binding; p<0.0001).
Spike-specific IgG4 was also cross-reactive to sarbecoviruses found in bats (RaTG13, RsSHC014, HKU3) and pangolins (GX-P5L, GD-1). Increased proportions of cross-reactive IgG4 negatively associated with FcγRIIa and FcγRIIIa engagement across sarbecoviruses (mRNA: r=-0.54 to -0.93, p<0.01; Vaxzevria: r=-0.63 to -0.94, p<0.01), highlighting the wider implications of class-switched IgG4.
Through epitope blocking, we determined that a majority of ancestrally-imprinted RBD IgG4 (mRNA: 61-71%, p<0.0001; Vaxzevria: 43-52%, p<0.0001) targeted the receptor binding motif—the mutational hotspot binding ACE2 receptor. IgG4 depletion revealed that while IgG4 greatly neutralised ancestral SARS-CoV-2, its direct contribution towards BA.1 or XBB.1.5 neutralisation were more modest.
Our findings underscore the growing urgency to better understand and mitigate IgG4 class-switching following mRNA vaccines/boosters.