Central nervous system recruitment of effector memory CD8⁺ T lymphocytes during neuroinflammation is dependent on α4 integrin

Clonally expanded CD8⁺ T lymphocytes are present in multiple sclerosis lesions, as well as in the cerebrospinal fluid of patients with multiple sclerosis. In experimental autoimmune encephalomyelitis, CD8 ⁺ T lymphocytes are found in spinal cord and brainstem lesions. However, the exact phenotype of central nervous system-infiltrating CD8 ⁺ T lymphocytes and the mechanism by which these cells cross the blood-brain barrier remain largely unknown. Using cerebrospinal fluid from patients with multiple sclerosis, spinal cord from experimental autoimmune encephalomyelitis and coronavirus-induced encephalitis, we demonstrate that central nervous system-infiltrating CD8⁺ T lymphocytes are mostly of the effector memory phenotype (CD62L^- CCR7^- granzymeB ^hi). We further show that purified human effector memory CD8 ⁺ T lymphocytes transmigrate more readily across blood-brain barrier-endothelial cells than non-effector memory CD8⁺ T lymphocytes, and that blood-brain barrier endothelium promotes the selective recruitment of effector memory CD8⁺ T lymphocytes. Furthermore, we provide evidence for the recruitment of interferon-γ-and interleukin-17-secreting CD8⁺ T lymphocytes by human and mouse blood-brain barrier endothelium. Finally, we show that in vitro migration of CD8⁺ T lymphocytes across blood-brain barrier-endothelial cells is dependent on α4 integrin, but independent of intercellular adhesion molecule-1/leucocyte function-associated antigen-1, activated leucocyte cell adhesion molecule/CD6 and the chemokine monocyte chemotactic protein-1/CCL2. We also demonstrate that in vivo neutralization of very late antigen-4 restricts central nervous system infiltration of CD8⁺ T lymphocytes in active immunization and adoptive transfer experimental autoimmune encephalomyelitis, and in coronavirus-induced encephalitis. Our study thus demonstrates an active role of the blood-brain barrier in the recruitment of effector memory CD8 ⁺ T lymphocytes to the CNS compartment and defines α4 integrin as a major contributor of CD8⁺ T lymphocyte entry into the brain.