Evaluation of cell death and proliferation in psoriatic epidermis.

Background: Previous studies of psoriatic epidermis using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL) method, a type of apoptotic detection method, showed that TUNEL-positive keratinocytes were abundantly distributed in all layers of the psoriatic epidermis, although psoriasis is a hyperproliferative disorder.

Objective: We sought to clarify the nature of cell kinetics in a psoriatic epidermis on the basis of differences in the reactivities in TUNEL and formamide-induced DNA denaturation assay combined with the detection of denatured DNA with a monoclonal antibody (MAb) against single-stranded DNA (formamide-MAb assay) between the normal and psoriatic epidermides.

Methods: The kinetics of keratinocytes was evaluated by the immunohistochemistry of Ki-67 for proliferation activity and by TUNEL, TUNEL combined with transmission electron microscopy (TUNEL/TEM), and formamide-MAb assay for apoptosis.

The activation of caspase-3 and DNA fragmentation in B cells phagocytosed by macrophages.

Apoptotic signaling of mammalian cells involves two pathways: the death receptor and mitochondrial pathways. In this in vivo study, we investigated apoptotic signaling of B cells in mouse germinal centers (GCs) of gut-associated lymphoid tissues (GALTs) using transmission electron microscopy (TEM), terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL), immunofluorescence of members of caspase family and cFLIP(L), and caspase activity assay. It was very difficult to ultrastructurally differentiate B cells undergoing apoptosis from B cells differentiating into memory cells or plasma cells among B cells constituting GCs.