Anti cd11b

For ICC assays, BV2 microglial cells were exposed to 200 ng/mL LPS or PBS for 30 min before treatment with 50 μM donepezil or vehicle (1% DMSO) for 5.5 h. The cells were then fixed for 10 min in 4% PFA, washed with PBS, and incubated overnight at 4 °C with anti-CD11b (Abcam, Cambridge, UK) and anti-p-NFκBser536 (Cell Signaling Technology, Danvers, MA, USA) or anti-CD11b and anti-p-STAT3ser727 (Abcam, Cambridge, UK). After washing 3× for 10 min with PBS, the cells were incubated at room temperature with Alexa Fluor 488-conjugated anti-rat and Alexa Fluor 555-conjugated anti-rabbit antibodies (1:200, Molecular Probes, Eugene, OR, USA) for 2 h. The immunostained cells were washed with PBS for 10 min and mounted on glass slides using fluorescence mounting medium (Agilent Technologies, Santa Clara, CA USA). Images of the cells were acquired by fluorescence microscopy (DMi8, Leica Microsystems, Wetzlar, Germany), and fluorescence intensity was analyzed using Image J (Version 1.53a, National Institutes of Health, Bethesda, MD, USA).

Longitudinal stomach tissue strips from H. felis-infected or uninfected mice were fixed with 10% formaldehyde, embedded in paraffin, and cut into 4-μm-thick sections. Hematoxylin and eosin staining was performed as per standard protocols, and the presence of lymphoid follicles and lymphoepithelial lesions (LELs) were recorded. Sections of murine stomach or human MALT lymphoma tissues were subjected to heat-induced epitope retrieval and then incubated overnight with anti-TCRγδ (Abcam, Cat. no. ab118864), anti-IL-17 (Abcam, Cat. no. ab79056), anti-IL-1β (Abcam, Cat. no. ab9722), anti-IL-23 (Santa Cruz, Cat. no. sc-50303), anti-Arg-1 (Proteintech, Cat. no. 16001-1-AP), anti-iNOS (Proteintech, Cat. no. 18985-1-AP), anti-Gr-1 (R&D systems, Cat. no. MAB1037-100), anti-CD11b (Abcam, Cat. no. ab133357), anti-CD33 (Abcam, Cat. no. ab11032), and anti-CD11b (Abcam, Cat. no. ab133357) primary antibodies at 4°C. The following day, the sections were probed with biotin–streptavidin horseradish peroxidase-conjugated secondary antibody, and stained using 3,3′-diaminobenzidine reagent. For immunofluorescence, fluorochrome-conjugated secondary antibodies were used. Positively stained cells were counted in five random non-overlapping fields (400× magnification; Nikon, Ni-U) using ImageJ software. The histoscore of each biomarker was evaluated according to Table 2.

Brain sections were blocked with 10% goat serum in PBS containing 0.2% Triton X-100 and then immunostained with microglial marker mouse anti-CD11b (1:200, Abcam, Cambridge, UK), rabbit anti-Iba1 (1:400, Wako Chemical), rat anti-CD68 (1:200, AbD Serotec) and astrocyte marker mouse anti-GFAP (1:200, Merck Millipore) in blocking solution at 4 °C overnight. Omission of the primary antibody incubation step and/or isotype-matched control antibodies was used to ascertain staining specificity. Brain sections were then incubated at room temperature for 1 hour with AlexaFluor goat anti-mouse IgG-488, goat anti-rabbit IgG-488 and goat anti-rat IgG-555 (Molecular Probes, Life Technologies, USA) at a dilution of 1:200. Brain sections were counterstained with DAPI (4′,6-diamidino-2-phenylindole, 1:2000, Sigma) and mounted using Prolong Gold (Molecular Probes).