Compartmentalized ocular lymphatic system mediates eye–brain immunity

The eye, an anatomical extension of the central nervous system (CNS), exhibits many molecular and cellular parallels to the brain. Emerging research demonstrates that changes in the brain are often reflected in the eye, particularly in the retina1. Still, the possibility of an immunological nexus between the posterior eye and the rest of the CNS tissues remains unexplored. Here, studying immune responses to herpes simplex virus in the brain, we observed that intravitreal immunization protects mice against intracranial viral challenge. This protection extended to bacteria and even tumours, allowing therapeutic immune responses against glioblastoma through intravitreal immunization. We further show that the anterior and posterior compartments of the eye have distinct lymphatic drainage systems, with the latter draining to the deep cervical lymph nodes through lymphatic vasculature in the optic nerve sheath. This posterior lymphatic drainage, like that of meningeal lymphatics, could be modulated by the lymphatic stimulator VEGFC. Conversely, we show that inhibition of lymphatic signalling on the optic nerve could overcome a major limitation in gene therapy by diminishing the immune response to adeno-associated virus and ensuring continued efficacy after multiple doses. These results reveal a shared lymphatic circuit able to mount a unified immune response between the posterior eye and the brain, highlighting an understudied immunological feature of the eye and opening up the potential for new therapeutic strategies in ocular and CNS diseases.


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For brain tissues, tissues were collected and incubated in a digestion cocktail containing 1 mg ml−1 collagenase D (Roche) and 30 μg ml−1 DNase I (Sigma-Aldrich) in RPMI at 37 °C for 45 min.Tissues were pipetted to break tissue down and filtered through a 70-μm filter.Then, cells were mixed in 3 ml of 25% Percoll (Sigma-Aldrich) solution and centrifuged at 580g for 15 min without brake.The Percoll layer was removed, and cell pellets were treated with 0.5ml ACK buffer and spun for 5 minutes at 500g.Then the cell pellets were resuspended in FACS buffer (PBS +2% FBS+ 1mM EDTA) for staining.
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