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Updated: Aug 7, 2026
Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions
Published on: April 20, 2016
Normal mouse skin lymphocyte, Langerhans cell, and keratinocyte responses to intradermal injections of
1Department of Dermatology, University of Texas Medical School at Houston 77030, USA.
To investigate the skin immune responses induced by intradermal injections of interferon-alpha (IFN-alpha) and interferon-gamma (IFN-gamma), normal C3H mice were injected intradermally with murine recombinant interferon-alpha (rIFN-alpha), interferon-gamma (rIFN-gamma), or the combination of rIFN-alpha and rIFN-gamma, three times per week for 2 or 4 weeks. Contralateral sites were similarly injected with placebo (saline). The results showed that after 6 or 12 injections of rIFN-alpha (10,000 IU/per injection), epidermal ATPase+ and Ia+ Langerhans cells (LGs) decreased significantly (p < 0.01 and p < 0.05) but dermal lymphocytic infiltrates, including CD3+ (pan-T cells), L3T4+ (T helper cells), Lyt-2+ (T suppressor/cytotoxic cells), MOMA+ (macrophages and monocytes), and dermal Ia+ cells, increased significantly (p < 0.01) compared with both saline injection sites and untreated skin. In contrast to rIFN-alpha, 6 injections of 10,000 IU rIFN-gamma failed to induce significant changes in either epidermal LGs or dermal lymphocytic infiltrates, except for an increase in dermal MOMA+ cells. High-dose rIFN-gamma injections (50,000 IU) strongly enhanced the expression of Ia antigen in epidermal keratinocytes (KCs), increased dermal lymphocytic infiltrates, and decreased the LGs in a similar pattern to that of rIFN-alpha. Furthermore, rIFN-gamma (50,000 IU) injections induced Ia antigen expression on the KCs in the contralateral saline injection areas, suggesting a systemic effect. Injections of a combination of rIFN-alpha and rIFN-gamma failed to show synergism for induction of skin immune responses.
To investigate the skin immune responses induced by intradermal injections of interferon-alpha (IFN-alpha) and interferon-gamma (IFN-gamma), normal C3H mice were injected intradermally with murine recombinant interferon-alpha (rIFN-alpha), interferon-gamma (rIFN-gamma), or the combination of rIFN-alpha and rIFN-gamma, three times per week for 2 or 4 weeks. Contralateral sites were similarly injected with placebo (saline). The results showed that after 6 or 12 injections of rIFN-alpha (10,000 IU/per injection), epidermal ATPase+ and Ia+ Langerhans cells (LGs) decreased significantly (p < 0.01 and p < 0.05) but dermal lymphocytic infiltrates, including CD3+ (pan-T cells), L3T4+ (T helper cells), Lyt-2+ (T suppressor/cytotoxic cells), MOMA+ (macrophages and monocytes), and dermal Ia+ cells, increased significantly (p < 0.01) compared with both saline injection sites and untreated skin. In contrast to rIFN-alpha, 6 injections of 10,000 IU rIFN-gamma failed to induce significant changes in either epidermal LGs or dermal lymphocytic infiltrates, except for an increase in dermal MOMA+ cells. High-dose rIFN-gamma injections (50,000 IU) strongly enhanced the expression of Ia antigen in epidermal keratinocytes (KCs), increased dermal lymphocytic infiltrates, and decreased the LGs in a similar pattern to that of rIFN-alpha. Furthermore, rIFN-gamma (50,000 IU) injections induced Ia antigen expression on the KCs in the contralateral saline injection areas, suggesting a systemic effect. Injections of a combination of rIFN-alpha and rIFN-gamma failed to show synergism for induction of skin immune responses.
