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. RESULTS: The number of Ki-67-positive cells in the psoriatic epidermis was significantly higher than that in the normal epidermis. In the normal epidermis, both TUNEL and formamide-MAb assay showed a similar distribution pattern, that is, both TUNEL and formamide-MAb assay-positive keratinocytes were present only in the upper granular layer. In the psoriatic epidermis, most keratinocytes were negative for the formamide-MAb assay, while TUNEL-positive cells were abundantly distributed in all layers of the psoriatic epidermis. TUNEL/TEM method clearly demonstrated that many immunogold particles that stain the sites of 3'-OH DNA ends were evenly distributed on the euchromatin in psoriatic keratinocyte nuclei, in contrast to their presence on the peripheral condensed chromatin in normal keratinocyte nuclei. CONCLUSION: The increased TUNEL reactivity in psoriatic lesions is due to the increase in the number of DNA nicks resulting from active DNA replication but not due to DNA double-strand breaks produced during the apoptotic process, and the formamide-MAb assay is a reliable method for the detection of apoptosis, particularly in the epidermis. Copyright 2004 Japanese Society for Investigative Dermatology
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. RESULTS: The number of Ki-67-positive cells in the psoriatic epidermis was significantly higher than that in the normal epidermis. In the normal epidermis, both TUNEL and formamide-MAb assay showed a similar distribution pattern, that is, both TUNEL and formamide-MAb assay-positive keratinocytes were present only in the upper granular layer. In the psoriatic epidermis, most keratinocytes were negative for the formamide-MAb assay, while TUNEL-positive cells were abundantly distributed in all layers of the psoriatic epidermis. TUNEL/TEM method clearly demonstrated that many immunogold particles that stain the sites of 3'-OH DNA ends were evenly distributed on the euchromatin in psoriatic keratinocyte nuclei, in contrast to their presence on the peripheral condensed chromatin in normal keratinocyte nuclei. CONCLUSION: The increased TUNEL reactivity in psoriatic lesions is due to the increase in the number of DNA nicks resulting from active DNA replication but not due to DNA double-strand breaks produced during the apoptotic process, and the formamide-MAb assay is a reliable method for the detection of apoptosis, particularly in the epidermis. Copyright 2004 Japanese Society for Investigative Dermatology