Journal: Scientific Reports

Article Title: CEACAM1 mediates B cell aggregation in central nervous system autoimmunity

doi: 10.1038/srep29847

Figure Lengend Snippet: ( A,B ) CEACAM1 expression on B cells in the CNS in acute ( A ) and chronic ( B ) MP4-induced EAE. ( C ) CEACAM1 expression on B cells in lymph nodes of MP4-immunized mice. ( D ) Negative control staining in the absence of primary antibody. Scale bars: 50 μm. ( E ) Inhibition of murine B cell aggregation induced by 25 μg/ml LPS + 5 U/ml IL-4 after treatment with 200 μg/ml anti-CEACAM1 antibody (mCC1). For each experiment n = 3 B6 mice were pooled. B cell aggregation was induced over a period of 72 h (compare images of untreated and LPS + IL-4-stimulated B cells). Data are representative of four independent experiments with similar results. Statistical significance was determined by Wilcoxon rank sum test. Data are expressed as mean ± s.d. * P < 0.05; **P < 0.01; *** P < 0.001. ( F ) Clinical course of EAE after treatment of MP4-immunized B6 mice with mCC1 ( n = 8) or mIgG1 isotype control antibody ( n = 7). Treatment was started with disease onset and given every three days. Results are from three independent experiments. Statistical significance was determined by one-way ANOVA for day-to-day comparisons (* P < 0.05) or by Wilcoxon rank sum test for the entire time course ( ### P < 0.001). Data are expressed as mean ± s.d. ( G ) Representative image of perivascular infiltrates in the cerebellum of MP4-immunized mice (left panel, circles) and their number after treatment with mCC1 vs . mIgG (right panel). ( H–K ) Percentage of non-B cell infiltrates ( H ), diffuse B cell infiltrates ( I ) and B cell aggregates ( J ) and size of B cell aggregates ( K ) in the cerebellum of MP4-immunized mice after treatment with mCC1 vs . mIgG1. Statistical significance was determined by Wilcoxon rank sum test. * P < 0.05.

Article Snippet: MP4-immunized mice were treated by intraperitoneal injections of 150 μg anti-CEACAM1 mAb CC1 (mouse IgG1; produced and purified by inVivo BioTech Services) or isotype control mIgG1 (inVivo BioTech Services) every third day for 27.6 ± 5.4 days after disease onset.

Techniques: Expressing, Negative Control, Staining, Inhibition

Journal: Scientific Reports

Article Title: CEACAM1 mediates B cell aggregation in central nervous system autoimmunity

doi: 10.1038/srep29847

Figure Lengend Snippet: ( A ) B cell ELISPOT assay of draining inguinal lymph nodes for the detection of MP4-specific antibodies and total IgG in MP4-immunized mice treated with either mCC1 ( n = 8) or mIgG1 ( n = 7). ( B ) Serum ELISA for the detection of MP4-specific antibodies in MP4-immunized mice treated with either mCC1 ( n = 8) or mIgG1 ( n = 7). Statistical significance was determined by Wilcoxon rank sum test. ( C ) Representative images of IHC staining of inguinal draining lymph nodes from MP4-immunized mice treated either with mCC1 ( n = 8) or mIgG1 ( n = 7). Scale bars: 50 μm.

Article Snippet: MP4-immunized mice were treated by intraperitoneal injections of 150 μg anti-CEACAM1 mAb CC1 (mouse IgG1; produced and purified by inVivo BioTech Services) or isotype control mIgG1 (inVivo BioTech Services) every third day for 27.6 ± 5.4 days after disease onset.

Techniques: Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay, Immunohistochemistry