Role of AIM in Corynebacterium-induced granuloma formation in mice
© Kuwata et al; licensee BioMed Central Ltd 2004
Published: 14 January 2004
Apoptosis inhibitor expressed by macrophages (AIM) is a murine macrophage-specific protein and belongs to the macrophage scavenger receptor cysteine-rich domain superfamily. AIM has been introduced as the inducer of resistance to thymocyte apoptosis . Because apoptosis of inflammatory cells plays a pivotal role in inflammation , we have applied a mouse model to address potential involvement of AIM in the process of granulomatous inflammation in vivo.
Mice deficient in AIM (AIM-/-) were generated by disruption exon 3 of the AIM gene (1). AIM-/- and wild-type (AIM+/+) mice were used. Heat-killed Corynebacterium parvum (C. parvum), 0.5 mg, was injected into the tail vein. All mice were killed under diethyl ether anesthesia at various time intervals after injection.
Formaldehyde-fixed and paraffin embedded livers were sectioned and stained with hematoxylin and eosin for light microscopy.
Flow Cytometric Analysis and Detection of Apoptosis
The surface phenotype of leukocytes obtained from livers was analyzed using fluorescein isothiocyanate- (FITC), phycoerythrin-, or biotin-conjugated anti-CD3 and anti-NK1.1 monoclonal antibodies in conjunction with a two- or three-color immunofluorescence test. To determine the percentage of cells undergoing apoptosis, FITC-labeled Annexin-V was used.
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