Pre-treatment with anti-asialo GM1 depletes NK cells in spleen and airways before bleomycin injection.

Mice were injected with either anti-asialo GM1 or control rabbit sera at −24 h and −1 h before bleomycin administration (A) At day 0, before bleomycin injection, and (B) day 1 after bleomycin injection leukocytes were isolated from spleen and BAL fluid, stained and analyzed by flow cytometry. Leukocytes were identified by CD45 expression, and percentages and absolute numbers of T cells (TCRβ+), B cells (CD19+), NK cells (DX5+TCRβ-) and NKT cells (DX5+TCRβ+) were determined. In the BAL fluid analysis in (B), a single value in the quantification of absolute T cell numbers (data point: 2,453 T cells) in the sera-control group was determined by Dixon's Q test to be an outlier (>95% confidence), and was therefore excluded from subsequent analysis. Mean ± SEM of n = 4 mice per group. *p<0.05, ** p<0.01 by 2 way ANOVA followed by t-test.