We, for the first time, analyzed T cell and natural killer (NK) cell subsets in the cerebrospinal fluid (CSF) in nine patients with human T-lymphotropic virus type 1 (HTLV-1)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) treated with interferon-alpha (IFN-alpha). The CD4/CD8 ratio in CSF was significantly lower after the therapy, which is mainly attributable to the significant reduction in CD4+ cells, especially CD25+CD4 and CD45RO+CD4 cell subsets. Meanwhile, NK, natural T and NKT cell subsets in the CSF remained unchanged. There was no CSF lymphocyte subset significantly associated with the clinical efficacy of IFN-alpha in this small-scale study, and more patients should be analyzed to ascertain the link.