Figure 6. Early recruitment of T cells and the absence of neutrophils in the CNS of LGTV-infected Ccr5-deficient mice result in decreased neuropathology.

(A) A schematic diagram of the experimental layout for the T cell adoptive transfer experiment is displayed. Donor mice (WT or Ccr5^−/−) were infected with LGTV, and T cells were isolated from spleens harvested on day 8 post infection. 1×10⁶ T cells were transferred into LGTV-infected recipient mice (Ccr5^−/− mice) intravenously on day 5 post infection and evaluated for survival for 18 days (B). Kaplan-Meier analysis of recipient mice with 18–20 Ccr5^−/− mice per group. WT→Ccr5^−/− mice received T cells from WT mice and Ccr5^−/−→Ccr5^−/− received T cells from Ccr5^−/− mice. (C) The total number of CD3⁺ T cells (CD45^hiCD3⁺) was assessed by flow cytometry on day 8 post infection in the brains of Ccr5^−/− mice receiving either WT or Ccr5^−/− CD3⁺ T cells. Data are shown as mean ±SD for 4–6 mice per group. (D) Body weight was measured daily for isotype and anti-Ly6G (clone 1A8) treated LGTV-infected mice for 12 days post infection with 16-17 mice per group pooled from two independent experiments. (E) Ccr5^−/− mice were injected with either isotype or 1A8 antibody, and survival was assessed by a Kaplan-Meier analysis through day 18 post infection with 16-17 mice per group pooled from two independent experiments. The arrows in C and D indicate the days when the antibody was administered. (F) The viral load was quantified in the brains of isotype or 1A8 antibody-treated mice on day 12 post infection. Shown is the mean ±SD from 7–8 mice per group on day 12 post infection from two independent experiments. (G) Immunohistochemical analysis was performed on paraffin-embedded brain sections for myeloperoxidase (MPO) from mice treated with isotype or 1A8 antibody on day 12 post infection. Immunohistochemical analysis was performed on paraffin-embedded brain sections after staining for apoptosis (TUNEL stain, left panels) or LGTV (right panels) in the (H) meninges, (I) hippocampus, and (J) deep cortical layers for 3-5 mice per genotype and condition from two independent experiments. Representative images are shown at 20× magnification. * p<0.05.