αcd8 clone 53 6

Antibodies and reagents used for histological, biochemical experiments and flow cytometry are as follows: F4/80 (clone BM8, eBioscience, CALTAG/Invitrogen), αCD4 (clone RM4-5, eBioscience), αCD8 (clone 53-6.7, eBioscience), αCD25 (clone PC61.5, eBioscience), αFoxP3 (clone NRRF-30, eBioscience), αCD3 (clone 145-2C11, eBioscience), αTCRβ (clone H57-597, eBioscience), αCD69 (clone H1.2F3, Pharmingen), αVβ8.2 (clone MR5-2, eBioscience) and αCD16/32 (clone 93, eBioscience). Secondary antibodies and related reagents are: Alexa fluor 488 conjugated anti-rat IgG (Molecular Probes), DAPI (Roche) or Hoechst33342 (Invitrogen) for nuclear staining, Cytofix/Cytoperm solution (BD Bioscience) to permeabilize cells and G-Block (Genostaff Inc., Tokyo, Japan) as a blocking reagent. Specimens were analyzed using a confocal microscope, FV10i-DOC (Olympus, Tokyo). Stained cells were analyzed using an SH800 flow cytometer (Sony Inc.), LSRII (BD) or FlowJo.

α-CD4-clone GK1.5 (catalog no. BE0003-1), α-CD8-clone 2.43 (catalog no. BE0061), and isotype control-clone LTF2 (catalog no. BE0090) depletion antibodies were purchased from Bio X Cell Inc. C57BL6/J WT mice were treated every other day, starting at −2 dpi, with 400 μg each of α-CD4 and α-CD8 antibodies, or 800 μg of isotype control, until 6 dpi. On day 7 after infection, mice were humanely euthanized, and T cells were analyzed by flow cytometry using α-CD4-clone RM4-5 (BD Pharmingen) and α-CD8-clone 53-6.7 (eBioscience) antibodies.