Journal: Frontiers in Cellular and Infection Microbiology
Article Title: Persistent Herpes Simplex Virus Type 1 Infection of Enteric Neurons Triggers CD8 + T Cell Response and Gastrointestinal Neuromuscular Dysfunction
doi: 10.3389/fcimb.2021.615350
Figure Lengend Snippet: CD3 + cells infiltrate the LMMP following HSV-1 infection. (A) LMMP preparations were obtained from the ileum of sham- and HSV-1-infected mice as described in Methods. The samples were enzymatically digested, and the resulting cell suspensions were labeled with anti-CD11c and anti-F4/80 antibodies and analyzed using flow cytometry. Cells were first selected on a forward scatter and side scatter dot plot and then double positive cells were recorded. The data are reported as the number of CD11c + F4/80 + cells detected in 10 5 events. The experiments were repeated 2 times; n=4 mice per group. (B) Cell suspension obtained as described in (A) were labeled with anti-CD3 antibody and analyzed by flow cytometry. Data are reported as the percentage of CD3 + cells in 10 5 events on side scatter dot plot. The experiments were repeated 2 times; n=4 mice per group. * denotes p < 0.05 vs sham infected mice. (C) Sections of ileum obtained from the sham or HSV-1 infected mice were subjected to immunohistochemistry using anti-CD3 antibody. Scale bars: 50 μm. Representative images of four separate experiments; n=3 mice per group. (D) Cell suspensions obtained from the LMMP, as described in (A) , were labeled with anti-CD3 and anti-CD4 antibodies (E) or with anti-CD3 and anti-CD8 antibodies (F) or with anti-CD3 and anti-CD69 antibodies. The samples were analyzed using flow cytometry. Cells were first selected on a forward scatter and side scatter dot plot and then double positive cells were recorded in 10,000 events. Data are reported as the percentage of double positive cells. Experiments were repeated 3 times n=4 mice per group. * denotes p < 0.05 vs sham infected mice.
Article Snippet: CD4 (rabbit) Cy7 conjugate , 50134-R001 , Sino Biological Inc , FC.
Techniques: Infection, Labeling, Flow Cytometry, Immunohistochemistry
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