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The primary antibodies, mouse monoclonal anti-human CD3 (Leica Biosystems, dilution 1:100), CD19 (Dako, dilution 1:50), CD68 (Dako, dilution 1:100), occludin (Santa Cruz Biotechnology, dilution 1:100) and claudin-1 (Santa Cruz Biotechnology, dilution 1:100), were added and incubated for 1 h at room temperature