The Third Signal Cytokine IL-12 Rescues the Anti-Viral Function of Exhausted HBV-Specific CD8 T Cells

Abstract

Optimal immune activation of naïve CD8 T cells requires signal 1 mediated by the T cell receptor, signal 2 mediated by co-stimulation and signal 3 provided by pro-inflammatory cytokines. However, the potential for signal 3 cytokines to rescue anti-viral responses in functionally exhausted T cells has not been defined. We investigated the effect of using third signal cytokines IL-12 or IFN-α to rescue the exhausted CD8 T cell response characteristic of patients persistently infected with hepatitis B virus (HBV). We found that IL-12, but not IFN-α, potently augmented the capacity of HBV-specific CD8 T cells to produce effector cytokines upon stimulation by cognate antigen. Functional recovery mediated by IL-12 was accompanied by down-modulation of the hallmark inhibitory receptor PD-1 and an increase in the transcription factor T-bet. PD-1 down-regulation was observed in HBV but not CMV-specific T cells, in line with our finding that the highly functional CMV response was not further enhanced by IL-12. IL-12 enhanced a number of characteristics of HBV-specific T cells important for viral control: cytotoxicity, polyfunctionality and multispecificity. Furthermore, IL-12 significantly decreased the pro-apoptotic molecule Bim, which is capable of mediating premature attrition of HBV-specific CD8 T cells. Combining IL-12 with blockade of the PD-1 pathway further increased CD8 functionality in the majority of patients. These data provide new insights into the distinct signalling requirements of exhausted T cells and the potential to recover responses optimised to control persistent viral infections. Persistent viral infections continue to cause major morbidity and mortality; chronic hepatitis B virus infection alone accounts for more than a million deaths annually. Such infections are characterised by a failure of viral control perpetuated by exhaustion of the T cell response. Here we show that the cytokine IL-12 can act as a potent “third signal” to rescue antiviral function in exhausted T cells. IL-12 has previously been shown to enhance naïve T cell responses but this is the first demonstration of its capacity to boost the disabled antiviral response in a persistent viral infection. IL-12 was able to down-regulate PD-1, a key inhibitory receptor driving T cell exhaustion, resulting in the recovery of hepatitis B virus-specific responses able to mediate multiple antiviral functions. Control responses in the same patients directed against the well-controlled cytomegalovirus did not require IL-12 to function efficiently. Our findings therefore elucidate a role for IL-12 in re-programming functionally exhausted T cells in persistent viral infections.