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Antigens in the posterior eye elicit immune responses in the brain. a, Schematic of the schedule of procedures for the experiments described below. b, Wild-type C57BL/6J mice were immunized using heat-inactivated HSV-2 injection through i.p., i.c., AC and IVT administration. Survival was monitored after i.c. challenge with a lethal dose of HSV-2 30 days later (naive, n = 18; i.p., n = 12; i.c., n = 6; AC, n = 6; IVT, n = 18). c, dCLNs of mice were ligated using a cauterizer. Seven days later, mice were injected through the IVT route with heat-inactivated HSV-2. Their survival was monitored after i.c. challenge with a lethal dose of HSV-2 30 days later (naive, n = 5; IVT immunized, n = 5; LN ligation, n = 6). d, Schematic of the parabiosis mouse model and treatment plans. e, Mice were injected through the IVT route with heat-inactivated HSV-2. Four weeks later, the immunized mice were joined to naive mice. The immunized mice or naive mice were challenged through the i.c. route with a lethal dose of HSV-2 after 3 weeks, and their survival was monitored (naive, n = 4; IVT, n = 4; IVT–naive (IVT challenge), n = 6; IVT–naive (naive challenge), n = 2; naive–naive (naive challenge), n = 2). f, Anti-HSV-specific antibody was measured by enzyme-linked immunosorbent assay after different routes of HSV-2 immunization (i.p., n = 12; i.c., n = 6; AC, n = 10; IVT, n = 10). Data are shown as mean ± s.e.m. g, Wild-type C57BL/6J mice were injected with heat-inactivated HSV-1 through i.p., i.c., AC or IVT administration. Their survival was monitored after i.c. challenge with a lethal dose of HSV-1 30 days later (naive, n = 15; i.p., n = 6; i.c., n = 6; AC, n = 6; IVT, n = 18). h, As in a, but S. pneumoniae strain TIGR4 was used (naive, n = 8; i.p., n = 5; i.c., n = 5; AC, n = 8; IVT, n = 8). i, Mice were inoculated through the i.c. route with 50,000 GL261 luciferase-expressing (GL261–Luc) brain tumour cells, treated with irradiated GL261–Luc cells through s.c., i.c., AC or IVT administration (day 7) along with anti-PD1 (RMP1-14) antibodies (days 7, 9 and 11) and monitored for survival (naive, n = 6; s.c., n = 6; i.c., n = 6; AC, n = 6; IVT, n = 12). Data are representative of two independent experiments. The graphics in a,d were created with BioRender.com. Source Data
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