CD8⁺ T cells retain protective functions despite sustained inhibitory receptor expression during Epstein-Barr virus infection in vivo

by Bithi Chatterjee, Yun Deng, Angelika Holler, Nicolas Nunez, Tarik Azzi, Liliana Danusia Vanoaica, Anne Müller, Hana Zdimerova, Olga Antsiferova, Andrea Zbinden, Riccarda Capaul, Johannes H. Dreyer, David Nadal, Burkhard Becher, Mark D. Robinson, Hans Stauss, Christian Münz

Epstein Barr virus (EBV) is one of the most ubiquitous human pathogens in the world, persistently infecting more than 90% of the adult human population. It drives some of the strongest human CD8⁺ T cell responses, which can be observed during symptomatic primary infection known as infectious mononucleosis (IM). Despite high viral loads and prolonged CD8⁺ T cell stimulation during IM, EBV enters latency and is under lifelong immune control in most individuals that experience this disease. We investigated whether changes in T cell function, as frequently characterized by PD-1 up-regulation, occur during IM due to the prolonged exposure to high antigen levels. We readily detected the expansion of PD-1 positive CD8⁺ T cells together with high frequencies of Tim-3, 2B4, and KLRG1 expression during IM and in mice with reconstituted human immune system components (huNSG mice) that had been infected with a high dose of EBV. These PD-1 positive CD8⁺ T cells, however, retained proliferation, cytokine production, and cytotoxic abilities. Multiple subsets of CD8⁺ T cells expanded during EBV infection, including PD-1⁺Tim-3⁺KLRG1⁺ cells that express CXCR5 and TCF-1 germinal center homing and memory markers, and may also contain BATF3. Moreover, blocking the PD-1 axis compromised EBV specific immune control and resulted in virus-associated lymphomagenesis. Finally, PD-1⁺, Tim-3⁺, and KLRG1⁺ CD8⁺ T cell expansion coincided with declining viral loads during low dose EBV infection. These findings suggest that EBV infection primes PD-1 positive CD8⁺ T cell populations that rely on this receptor axis for the efficient immune control of this ubiquitous human tumor virus.