Perivascular Lymphocyte Clusters Induced by Gastric Subserous Layer Vaccination Mediate Optimal Immunity against Helicobacter through Facilitating Immune Cell Infiltration and Local Antibody Response
Perivascular lymphocyte clusters contributed to reduction of colonization of H. felis in gastric mucosa of mice. The depletion of perivascular lymphocyte clusters and a parabiosis mice model was performed as follows: (a) all mice were performed with laparotomy to access the stomach and injected with alum-based vaccine into GSL. At day 28, these immunized mice were divided into two groups. For one group, half of the mice were administered with anti-CD90.2 antibody and cohoused with the other half. At day 42, the anti-CD90.2 antibody-treated mice and the other mice not injected with an antibody were surgically jointed. At day 56, the indicated partner was infected with H. felis. Another group was treated the same way but used IgG2b isotype. After the later challenge on day 3, those mice were harvested and analyzed. (b, c) CD90.2 cells of the blood from indicated partner mice were analyzed by flow cytometry. Representative FACS plots were shown. (d) Frozen sections of gastric tissue from IgG2b isotype and CD90.2 antibody-treated mice were stained with antibodies against CD31 (red) and CD90.2 (green) and DAPI (blue). Scale bars: 200 μm. (e) Mice were performed as indicated in the table; 3 days later, the mRNA level of H. felis-16S was detected by QRT-PCR. Values are the (). All indicated values were tested using the ANOVA analyses or t tests. ;; n.s.: not significant.
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