The Effect of Nerve Growth Factor on Cell Proliferation and Expression of Its Receptors in Cultured Human Keratinocytes / 대한피부과학회지

BACKGROUND: Nerve growth factor(NGF) is a neurotrophic polypeptide necessary for the survival and growth of some central neurons, as well as sensory afferent and sympathetic neurons. In addition to its actions on the nervous system, it also has a significant biologic effects on cells of the immune-inflammatory compartment. Recent studies suggest that NGF is an important autocrine growth factor and survival factor for keratinocytes which express both high- and low-affinity receptors for NGF. OBJECTIVE: The purpose of this study is to detect NFG receptors on cultured human keratinocytes and to evaluate the effect of NGF on proliferation of cultured human keratinocytes. METHODS: Cultured human keratinocytes were examined for the expression of high affinity receptor TrkA and low affinity receptor p75 by Northern blot, Western blot and immunocytochemistry. The effects of NGF on proliferation of cultured human keratinocytes were also evaluated. To specify the NGF effect on proliferation of human keratinocytes, excess of anti-NGF neutralizing polyclonal antibody was added. RESULTS: 1) NGF significantly stimulated the proliferation of keratinocytes in both 1% of keratinocyte growth supplement(KGS)-added medium(100ng/ml) and 0.2% KGS-added media(50, 100, 500ng/ml), (p<0.05). The cell number was dose-dependently increased in 0.2% KGS-added media. 2) Whenever we added 500 ng/ml of anti-NGF polyclonal antibody to the growth media, the cell number was statistically higher in 100ng/ml NGF-added group of 1% KGS-added medium, but there was not any statistical significance in 0.2% KGS-added media group. 3) Immunocytochemical staining with specific antibodies to TrkA and p75 revealed positive findings for these receptors, but TrkB and TrkC were not detected. 4) We could not detect both the mRNA and protein of TrkA and p75 by Northern and Western blot methods. CONCLUSION: These results suggest that both high affinity- and low affinity receptors for NGF are expressed in cultured human keratinocytes and NGF can induce keratinocyte proliferation.

Effect of IGF-1 on the Proliferation of Cultured Normal Human Melanocytes / 대한피부과학회지

BACKGROUND: Human growth hormone(hGH) plays a central role in linear bone growth and body metabolism. Its mitogenic effect in human tissues is mediated via direct and indirect actions. As proposed by the "somatomedin hypothesis", many circulating GH-mediated effects are exerted indirectly and systemically via stimulation of hepatic synthesis of insulin-like growth factor 1(IGF-1). Given additional evidences for the expression of growth hormone receptor(GH-R) and IGF-1 receptor(IGF-1R) on many target tissues including keratinocytes, melanocytes, and fibroblasts, it is now evident that the GH can act via systemic IGF-1 secreted by the liver and locally produced IGF-1, as well as directly through the GH receptor. OBJECTIVE: The purpose of this study was to investigate not only the effect of IGF-1 on the morphologic changes, proliferation, and melanization of cultured human melanocytes but also on its signal transduction pathway through the IGF-1R. METHODS: Melanocytes were exposed to IGF-1 at 10, 25, 50, 75, 100ng/ml and we examined the changes of cell morphology, number of cells, [3H]-thymidine incorporation, MTS assay, and melanization according to the concentrations and exposure times of IGF-1. Also, the activity of p44/42 MAPK/ERK according to the various exposure times of IGF-1(25ng/ml) was examined using the Western blotting method to find out about the signal transdution pathway of IGF-1. RESULTS: The results were as follows: 1. There were no significant morphological changes of cells between the control and experimental groups according to the concentrations and exposure times of IGF-1. 2. The effects on melanocytes according to the concentrations of IGF-1 5 days after adding IGF-1 : 1) The number of cells, [3H]-thymidine incorporation, and MTS assay were significantly higher than those of control group in all experimental groups(p<0.05). 2) The melanin content showed an insignificant decrease in all experimental groups. 3) The melanocytes responded independent of the IGF-1 concentration in the assay of cell number, [3H]-thymidine incorporation and MTS. 3. The effects on melanocytes according to the exposure times(3 days, 5 days, 7 days) of IGF-1(25 ng/ml) : 1) The number of cells, [3H]-thymidine incorporation, and MTS assay increased as time went by, and was significantly higher than those of control group at all exposure times(p<0.05). 2) The melanin content decreased after exposure of IGF-1, especially that of 3 days exposure group showed a significant decrease(p<0.05). 4. The activities of p44/42 MAPK/ERK increased suddenly at 5 minutes with a peak at 60 minutes and then abruptly decreased at 120 minutes after adding IGF-1 CONCLUSION: In summary, this study demonstrates that IGF-1 has no effect on the morphology, but it does increase the proliferation and slightly decrease the melanization of cultured human melanocytes. In addition, it is suggested that IGF-1 plays a role in regulation of proliferation of melanocytes via the receptor PTK pathway with activation of p44/42 MAPK/ERK.

Effect of Interleukin-4 and Histamine on the Fibroblast in the Nasal Mucosa of Allergic Rhinitis and Non-Allergic Rhinitis / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Fibrosis of the subepithelial layer is a characteristic finding in chronic allergic rhinitis. Heterogeneity of the fibroblasts in the nasal mucosa was already clarified and a different response of the fibroblasts can be expected from various stimulations occurring within the mucous membrane of the nose. The aim of this study was to investigate the characteristics of fibroblasts in differentiation and function of fibroblasts in allergic mucosa of the nose. MATERIALS AND METHODS: Using the 3rd passage of fibroblasts taken from the inferior turbinates of allergic and non-allergic patients, we measured the proliferative potential by comparing cell growth in the culture system of fibroblasts, and calculated the doubling time and the percent change. We also compared the production of stem cell factor (SCF) and granulocyte macrophage colony stimulating factor (GM-CSF) after stimulation with interleukin (IL)-4 and/or histamine. Morphologic differences were examined by transmission electron microscopy. RESULTS: In the case of non-stimulated fibroblasts, proliferation was prominent in the non-allergic group (NAG). However, the proliferation was remarkably increased in the allergic group (AG) on day 6 when the fibroblasts were stimulated with IL-4 and/or histamine. On the production of GM-CSF, both non-stimulated and histamine stimulated fibroblasts were more prominent in the AG than in the NAG, and the production of SCF in the AG was similar to that in the NAG in the non-stimulated fibroblasts. Also, the production of GM-CSF and SCF were remarkably increased in the AG on day 4 after histamine stimulation. Morphologic differences were demonstrated between the AG and the NAG. CONCLUSION: These results suggest that IL-4 could be involved mainly in promoting the proliferation of allergic fibroblasts in the exponential period, and the production of GM-CSF and SCF could be remarkable in the early stage of the culture period by histamine stimulation.

Role of mononuclear cells of IgA nephropathy on ICAM-1 expression in mesangial cells

OBJECTIVES: To investigate the possible role of mononuclear cells and their products in the pathogenesis of IgA nephropathy, in vitro expression of ICAM-1 on cultured mouse mesangial cell (MC) was examined after stimulation with mononuclear cell culture supernatant from patients with IgA nephropathy. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated and cultured from 18 patients with primary IgA nephropathy, 8 normal controls and 5 patients with non-IgA nephropathy (FSGS 1, MGN 3, MPGN 1). ICAM-1 expression on cultured mouse MC by TNF-alpha, IL-1 beta and culture supernants of PBMC were analyzed using a cell ELISA method. The concentration of IL-1 beta and TNF-alpha in culture supernatants was measured by using a commercially available radioimmunoassay kit. RESULTS: Addition of human recombinant TNF-alpha induced an increased ICAM-1 expression in a dose-dependent manner. The expression of ICAM-1 was further increased after co-stimulation with TNF-alpha and IL-1 beta. Addition of PBMC culture supernatants into mouse MC induced significantly higher expression of ICAM-1 by supernatants from the patients with IgA nephropathy compared with that from normal controls. The concentration of TNF-alpha and IL-1 beta in supernatants from the patients with IgA nephropathy was significantly higher than that from those with non-IgA nephropathy. CONCLUSION: TNF-alpha and IL-1 released from mononuclear cells induced the up-regulation of ICAM-1 expression and this may be related to the immune pathogenesis of IgA nephropathy.

Effect of L - tyrosine on Proliferation and Melanization in Cultured Normal Human Melanocytes / 대한피부과학회지

BACKGROUND: The biosynthesis of melanin is initiated by the enzymatic oxidation of L-tyrosine to L-dopa by tyrosinase. Some precursors of melanin are cytotoxic, and melanoma cells are killed as a risk of exposare to excess tyrosine or dopa in the culture medium. However, there have been few observations of the effects of L-tyrosine on cultured normal human melanocyte. OBJECTIVE: In order to investigate whether exogenous tyrosine induces cytotoxicity in cultured normal human melanocytes as in melanoma cells, we examined the effects of L-tyrosine on proliferation and melanization in normal human melanocytes. METHODS: A melanocyte culture was produced with a modified TIC medium. L-tyrosine was added to the culture medium, 100, 200, 400, and 800uM. After 2 days of incubation, the proliferation was measured by methylthiazol tetrazolium(MTT) assay and sulforhodamine B(SRB) assay. The melanin contenis were also measured by the modified Whittaker's method. RESULTS: On MTT assay, the proliferation of melanocytes had been stirnulated significantly (p< 0.05) in all L-tyrosine added groups. On SRB assay, the proliferation of melanocytes had heen stimulated significantly (p<005) in 200, 400, 800uM of L-tyrosine added groups. The melanin contents had increased in all L-tyrosine added groups, and had increased significantly (p<0.05) in 400uM of L-tyrosine added group. CONCLUSION: L-tyrosine is not toxic to normal melanocytes, It stimulates the proliferation and melnization of cultured normal human melanocytes.

Effect of L - tyrosine on Proliferation and Melanization in Cultured Normal Human Melanocytes / 대한피부과학회지

BACKGROUND: The biosynthesis of melanin is initiated by the enzymatic oxidation of L-tyrosine to L-dopa by tyrosinase. Some precursors of melanin are cytotoxic, and melanoma cells are killed as a risk of exposare to excess tyrosine or dopa in the culture medium. However, there have been few observations of the effects of L-tyrosine on cultured normal human melanocyte. OBJECTIVE: In order to investigate whether exogenous tyrosine induces cytotoxicity in cultured normal human melanocytes as in melanoma cells, we examined the effects of L-tyrosine on proliferation and melanization in normal human melanocytes. METHODS: A melanocyte culture was produced with a modified TIC medium. L-tyrosine was added to the culture medium, 100, 200, 400, and 800uM. After 2 days of incubation, the proliferation was measured by methylthiazol tetrazolium(MTT) assay and sulforhodamine B(SRB) assay. The melanin contenis were also measured by the modified Whittaker's method. RESULTS: On MTT assay, the proliferation of melanocytes had been stirnulated significantly (p< 0.05) in all L-tyrosine added groups. On SRB assay, the proliferation of melanocytes had heen stimulated significantly (p<005) in 200, 400, 800uM of L-tyrosine added groups. The melanin contents had increased in all L-tyrosine added groups, and had increased significantly (p<0.05) in 400uM of L-tyrosine added group. CONCLUSION: L-tyrosine is not toxic to normal melanocytes, It stimulates the proliferation and melnization of cultured normal human melanocytes.

Effects of REtinoids on Keratinocytes HLA - DR and ICAM - 1 Expression Induced by Interferon - gamma / 대한피부과학회지

BACKGROUND: Retinoids exert a wide range of effects on cell growth and development and have important effects on keratinocytes differentiation in vvc and in vitro. Besides the effects on epithelial differeotiation, modulation of cellular and amoral responses of lymphocytes, changes in natural killer and T-killer cell activities and riodulation of antigen presenting properties were shown by retionds. OBJECTIVE: We studied to investigate the immunologic role of etinoids. METHODS: With foreskin. the effects of 13-cis retinoic acid and etretinate on interferony induced HLA-DR and ICAM-1 expression of kratinocytes were!ev luated. RESULTS: 1. When keratinocytes were grown in low calcium media, priliferation was inhibited only with 8 x 10 M of 13-cis retinoic acid. 8 x 10 , 8 x 10 M of 13-cis retinoic acid and 8 x 10 , 8 x 10, 8 x 10 M of etretinate had no effect on keratinocytes roiferation. When cultured in 0.15 mM calcium media or 1.0 mM calcium media, 13-cis retino acid and etretinate had no effection keratinocytes proliferation. 2. Keratinocytes HLA-DR expression was decreased with 8 x 10 , 8 x 10 M of 13-cis retinoic acid in 0.15 mM carcium media and 8 x 10 , 8 x 10 M of 13-cis retinoic acid in 1.0 mM calcium media. Etretinate had no effect on keratinocytes HLA-DR expression. 3. Keratinocytes ICAM- 1 expression was increased with 8 x 10 M of 13-cis retinoic acid in low calcium. media. When cultured on 0.15 mM carcium media, ICAM-1 expression was increased with 8 x 10 M of 13-cis retinoic acid and tetinate. When cultured in 1.0 mM calcium media, ICAM-1 expression was increased with 8 x 10, 8 x 10 , 8 x 10 M of 13-cis retinoic acid and 8 x 10, 8 x 10 , 8 x 10 M of etretinate. CONCLUSION: These results suggest that retiniods have an immunomodulating effect as well as effects on epithelial differentiation. Clarification of the mechanism of increased expression of ICAM-1 and decreased expression of HLA-DR remains to the proved.

Effect of Radiation on Cultured Human Normal Keratinocytes and Melanocytes / 대한피부과학회지

BACKGROUND: Radiation has been used in t,he medical field of dragnosis and treatment. There is widely used ionizing radiat:ion such as naturally occuring r-rays or machine-made X-ray. This radiation is able to induce the structural and functional alterations of the mammalian cells. But we have few detailed reports on the effects of radiation on epidermal cells and their immune functions. OBJECTIVE: This study was performed to investigate the effect of radiation on cultured human keratinocytes and melanocytes. MATERIALS AND METHODS: Cultured human keratinocytes and melanocytes were irradiated with 2,6, l0Gy from a Co saurce and stimulated by 100 U/ml of ekratinocyte immediately after irradiation. We investigated cell numbers and morphological changes, DNA synthesis and HLA-DR antigen expression. RESULTS: After exposure to r-ray, the proliferation of keratinocytes and melanocytes decreased in a time and dose dependent fashion to each control group. Tliey showed decreased density, a larger size and a round appearance after radiation exposure and an especially shortened and decreased number of dendrites in the melanocytes. In DNA synthesis counted using [H]-thymidine incorporation, the keratinocvtes decreased values in a dose depen(lent manner at 24 and 72 hours after irradiation but no differense was observed at 168 hours. In melanocytes, there was a greater decrease than that of keratinocytes. The melanin content/cell in all radiation exposed groups increased in a time and dose dependent fashion t,o each contr ol group. HLA-DR antigen expression on keratinocytes after radiat,ion exposure decreased to the control group, but there were no significant differences acccirding to the dose of radiation, And there were no significant diifferences of HLA-DR antigen expression on the melanocytes betweer. controls and the radiation exposed groups. CONCLUSION: Antiproliferative activity was dependent on the exposure time and dose of r-ray exposure. According to the time after radiation exposure, melanogenic activity was stimulated. The expression of HLA-DR, antigen decreased in keratinocyte after radiation exposure but there was no decrease in melanocytes.

Effect of Radiation on Cultured Human Normal Keratinocytes and Melanocytes / 대한피부과학회지

BACKGROUND: Radiation has been used in t,he medical field of dragnosis and treatment. There is widely used ionizing radiat:ion such as naturally occuring r-rays or machine-made X-ray. This radiation is able to induce the structural and functional alterations of the mammalian cells. But we have few detailed reports on the effects of radiation on epidermal cells and their immune functions. OBJECTIVE: This study was performed to investigate the effect of radiation on cultured human keratinocytes and melanocytes. MATERIALS AND METHODS: Cultured human keratinocytes and melanocytes were irradiated with 2,6, l0Gy from a Co saurce and stimulated by 100 U/ml of ekratinocyte immediately after irradiation. We investigated cell numbers and morphological changes, DNA synthesis and HLA-DR antigen expression. RESULTS: After exposure to r-ray, the proliferation of keratinocytes and melanocytes decreased in a time and dose dependent fashion to each control group. Tliey showed decreased density, a larger size and a round appearance after radiation exposure and an especially shortened and decreased number of dendrites in the melanocytes. In DNA synthesis counted using [H]-thymidine incorporation, the keratinocvtes decreased values in a dose depen(lent manner at 24 and 72 hours after irradiation but no differense was observed at 168 hours. In melanocytes, there was a greater decrease than that of keratinocytes. The melanin content/cell in all radiation exposed groups increased in a time and dose dependent fashion t,o each contr ol group. HLA-DR antigen expression on keratinocytes after radiat,ion exposure decreased to the control group, but there were no significant differences acccirding to the dose of radiation, And there were no significant diifferences of HLA-DR antigen expression on the melanocytes betweer. controls and the radiation exposed groups. CONCLUSION: Antiproliferative activity was dependent on the exposure time and dose of r-ray exposure. According to the time after radiation exposure, melanogenic activity was stimulated. The expression of HLA-DR, antigen decreased in keratinocyte after radiation exposure but there was no decrease in melanocytes.

Effects of 8 - Methoxypsoralen ( 8 - MOP ) on the Proliferation and Melanization of Cultured Normal Human Melanocytes without UVA / 대한피부과학회지

BACKGROUND: Psoralen has been used in the treatment of certain hypojigmentary disorders with UVA or solar irradiation. However trecent report proposed the actions of psiralens are direct and do not require the presence of ultraviolet light. The report also suggested that tze specific receptors other than DNA would be present. OBJECTIVE: This study was done ta identify the effects of 8-methoryporalen(8-MOP) on the proliferation and melanization of cultured normal human melanocytes without UVA. METHODS: Melanocytes were cultured in melanocyte culture medium neluding 16% or 5% FBS. We added 8-MOP by their concentrations from 10 M to 10 M. After 8 hours treatment, we investigated the melanocytes proliferation and Lhe melanin contents. RESULTS: We could not detecet any significant differences of melanoytes proliferation and melanin contents between the control end experimental groups. CONCLUSION: There were no effect on the proliferation and the milanization of cultured normal human melanocytes with 8-MOP only.

Effects of Panax ginseng Saponins and Water Extract on the Growth of Cultured Human Keratinocytes and Melanocytes / 대한피부과학회지

BACKGROUND: Diagnosis of paucibacillary leprosy is difficult owing to lack of sensitive diagnostic tools. Recently, several investigators have studied the use of polymerase chain reaction(PCR) to detect Mycabacterium leprae. Using nested-PCR the sensitivity and specif city of DNA amplification is considerably improved. OBJECTIVE: The purpose on investigation is to assess the efficacy if nested-PCR which is applied to formalin-fixed, paraffin-embedded biopsies material of patients with 1 prosy. METHODS: Biopsy samples were taken from patients with lepremc tous(11 patients) and tuberculoid (10 patients) leprosy, fixed in formalin, and embedded in parafin. The DNA from samples was extracted and amplified through 25 cycles by using the outside pairs of primer(L and L). The second amplification was allowed thproceed through 15 cycles using insice gairs of primer(L and L4). RESULTS: All twenty one samples showed 347-base-pair products. To confirm that the 347-bp product did correspond to the expected portion of the M. leprae groE gene, the amplified product was digested with Pst I. Pst I dipestion yielded 254-and 93-bp fragmerts, as predicted from the sequence of the M. leprae gene. The senilivity was that a single organism was idntified by nested-PCR. CONCLUSION: The nested-PCR is sensitive, specific, and simple diagiostic tool for leprosy.

The Effects of Multiple UVB Exposures on Proliferation and Melanization in Cultured Human Melanocytes / 대한피부과학회지

BACKGROUND: In the skin, the major stimulus for cutaneous pigmentation is ultraviolet radiation. The most important physiologic role of melanin is protection against harmful UV radiation to skin. It is known there are some differences in melanization between a single and multiple exposures of UVB, in vivo. Little if known about the functions of the melanocyte alone in cutaneous pigmentation after ultraviolet exposure, because of the complexity of interactions in the whole epidermis. OBJECTIVE: To investigate the effects of multiple exposures at various dosages of UVB, and to compare the effect of UVB in multiple divided exposures with a single exposure at the same total dosage of UVB on proliferation and melanization in cultured human melanocyte. METHODS: Melanocytes were cultured by modified TIC medium. The melanoctes were exposed daily for three consecutive days to UVB at 2, 4, 8 and 16 mJ/cm2and a single exposure at 24 mJ/cm2. The morphologic changes were examined by phase contrast microscopy. The melanocytes were counted by hemocytometer and melanin contents were assayed by spectrophotometer. RESULTS: 1. The effects of multiple UVB exposures: 1) The morphologic changes were as follows: With three time exposures at a dosage of 8 mJ/cm2, themelanocytes enlarged in size, and elongated their dendrites slightly; with three time exposures at a dosage of 16 mJ/cm2, enlargement in sized and elongation of dendrited were more significant. 2) With three time exposures at dosages of 2 nd 4 mJ/cm2, the proliferation of melanocytes was stiumlated significantly(p<0.05). However, with three time exposures at dosages of 8 and 16 mJ/cm2the proliferation was inhibited(p<0.05). 3) With three time exposures at dosages of 2 and 4 mJ/cm2, the melanin contents were decreased. However, with three tiem exposures at a dosage of 16 mJ/cm2, the melanin contents were highly increased(p<0.01). 2. The comparison between multiple divided exposures and a single exposure at the same toal dosage of UVB: 1) There were no morphologic differences of dendrities between with three time exposures at a dosage of 8 mJ/cm2 and with a single exposure at a dosage of 24 mJ/cm2. However enlarged melanocytes were more numerous with a single exposure. 2) The proliferation of melanocytes was more inhibited with a single exposure than with multiple divided exposures(p<0.05). 3) The melanin contents were more increased with a single exposure than with multiple divided exposures(p<0.05). CONCLUSION: With multiple exposures at lower dosages of UVB, the proliferation of melanocytes was stimulated, and melanization was decreased. However, with multiple exposures at higher dosages of UVB, the proliferation was inhibited, and melanization was increased. At the same total dosage of UVB, the proliferation was more inhibited, and the melanization was more increased with a single exposure than with multiple divided exposures.

An experimental study of epidermal separation by dispase under negative pressure

No abstract available.

The effects of recombinant interferon-?on Proliferation and HLA-DR antigen expression in cultured human melanocyts / 대한피부과학회지

BACKGROUND: The main function of melanocyte is known to protect the skin from hazardous sunlight. But, some investigators have claimed lately that melanocytes are also related to the immunologic role in the epidermis because these cells produce IL-1 activity and IL-1beta convertase activity, in vitro. OBJECTIVE: Our purposes were to investigate the effects of rIFN-gammaon the proliferation of melanocytes, melanin content, and the expression of HLA-DR antigen on melanocytes after a rIFN-gammaexposure. MEHTODS: The number of melanocytes, the melanin content, and the expression of HLA-DR antigen were evaluated on cultured human melanocytes according to a time sequence and various concentrations of rIFN-gamma. RESULTS: Antiproliferative activity on melanocytes was dependent on the exposure time and the concentration of rIFN-gamma. According to the exposure time and the concentration of rIFN-gamma, melanogenic acivity was inhibitd or stimulated. Normal melanocytes didn't express HLA-DR antigen, but when normal melanocytes were exposed to rIFN-gamma, the expression of HLA-DR antigen increased in a time-and concentration-dependent fashion. After the removal of rIFN - gammafrom the culture media, the expression of HLA-DR antigen on melanocytes also disappeared. CONCLUSION: In our study, melanocytes seem to be related to the immunologic role in the epidermis because these cells expressed HLA-DR antigen after rIFN-gammaexposure and we think that study could help to investigate between melanocytes and immunologic mechanisms in various inflammatory skin diseases.

Effect of r-interferon-gamma on inerleukin-1 production by keratinocytes / 대한피부과학회지

It now appears that in addition to being a target for immunologic injury, the epidermis actively participate in various immunologic processes and thus may serve as the peripheral limb of the immune system. For example, keratinocytes produce the cytokines including IL-1 and in certain inflammatory diseases characterized by accumulation of T lymphocytes, kerationchtes aberrantly express HLA-DR antigen. In this study, the effect of interferon-gamma on keratinocytes IL-1alpha production were invesigated by radioimmunoassay, using as a model cultured keratinocytes stimulated with 30U/ml of recombinant-Interferon-gamma(IFN), IFN and 1microgram/ml of PMA(IFN-PMA), IFN-gamma and 25microgram/ml of LPS(IFN-LPS), PMA, and LPS. The expression of HLA-DR antigen and the effect of IFN-gammaon the proliferation of kerationcytes were also evaluated. The results were as follows: 1. The proliferation of keratinocytes was significantly inhibited in the IFN, IFN-PMA, and IFN-LPS groups compared with the control. The proliferation of kerationcytes in PMA and LPS groups was not inhibited. 2. The percent of HLA-DR positive kerationcytes was 62.5+/-3.1%, and 65.8+/-2.6% in IFN, IFN-PMA, and IFN-LPS group, respectively. There were no HLA-DR expression in the control, PMA, and LPS groups. 3. The amounts of IL-lalpha were 22.58+/-8.41, 49.32+/-13.01, 57.02+/-14.99, 96.98+/-43.17, 22.30+/-4.26, and 44.60+/-20.51 fmol in supernatant of control, IFN, IFN-PMA, IFN-LPS, PMA, and LPS group, respectively. The differences between IFN and control, IFN-PMA and PMA, and IFN-LPS and LPS were significant (P<0.05). The produciton of IL-1alphawas enhanced by IFN, but not PMA and LPS. The IL-1alpha induced by IFN and PMA was higher than that induced by IFN or PMA alone. The IL-1alpha induced by IFN and LPS was higher tthan that induced by IFN or LPS alone. 4. The amounts of IL-1alpha were 59.82+/-11.57, 70.15+/-25.22, 73.50+/-17.15, 63.67+/-32.38, 48.62+/-4.81, and 50.92+/-15.01 fmol in cell lysate of control, IFN, IFN-PMA, IFN-LPS, PMA, PMA, and LPS groups, respectively. The amount of IL-1alpha in IFN-PMA was significantly increased as compared to PMA(P<0.05). 5. The total amounts of IL-1alpha were 82.40+/-8.98, 119.47+/-21.88, 130.52+/-8.12, 160.66+/-34.51, 70.92+/-1.15, and 95.53+/-27.89 fmol in the control, IFN, IFN-PMA, IFN-LPS, Pma, and LPS groups, respectively, The differences between IFN and control, control, IFN-PMA and PMA and IFN-LPS and LPS were signigicant (P<0.05) the production of IL-1alpha was enhanced by IFN, but not PMA and LPS. The IL-1alpha induced by IFN and PMA was higher than that induced by PMA. The IL-1alpha induced by IFN and LPS was higher than that induced by IFN or LPS alone. In summary, the results indicate that when kerationcytes are activated by IFN-gamma, they are actively participating in the immunologic reaction by the increasing production of IL-1alpha.

Kinetics of HLA-DR and ICAM-1 on cultured human keratinocytes and their influence on in vitro alloimmune responses / 대한피부과학회지

BACKGROUND: Kerationcytes make up the vast majority of cells within the epidermis. Recent attention has focused on the role keratinocytes may play in the induction of T cell mediated inflammatory responses in skin, particularily because keratinocytes, when activated by immunologic stimuli, express MHC class llAg and secrete cytokines. But in experimentally induced lichenoid tissue reaction by interferon-nu, MHC class ll Ag was not essential for the enhanced T cell trafficking. There is growing evidence that keratinocyte intercellular adhesion molecule-1 (ICAM-1) expression is involved in the epidermal trafficking of T lymphocytes. OBJECT: To investigate the kinetics of expression of the HLA-DR and ICAM-1 on cultured human keratinocytes by recombinant-interferon-nu(IFN) and phorbol myristate acetate(PMA), and the influence of the HLA-DR and ICAM-1 and the stimulation of peripheral blood mononuclear leukocytes(PBML). RESULTS: 1. 1 U/ml of IFN can induce HLA-DR and ICAM-1 on keratinocytes. Expression of both antigens were increased in a dose and exporsure time dependent fashion. But expression of HLA-DR was less sensitive to IFN than ICAM-1 2. ICAM-1 induction was more rapid than HLA-DR. Keratinocytes expressed HLA-DR 6hours after IFN treatment amd increased rapidly after 12 hours. 3. HLA-DR positive keratinocytes were decreased more rapidly that ICAM-1 positive kerationcytes. 4. Proliferations of PBML were slightly inhibited when cultured with keratinocytes which were treated or not treated with IFN. But IFN treated keratinocytes stimulated the PBML more than untreated keratinocytes. Proliferaton of PBML by IFN treated keratinocytes were inhibited by anti-ICAM monoclonal antibody 5. PMA treated keratinocytes stimulated the PBML more than untreated keratinocytes. Proliferation of PBML by PMA treated keratinocytes was inhibited by anti-ICAM monoclonal antibodies and anti-LFA-1 monoclonal antibodies. CONCLUSION: These results suggest that keratinocytes can express not only HLA-DR but also ICAMP1 may play a important role in initiating immunologic response. Complete clarification of the function of HLA-DR and ICAM-1 positive keratinocytes requires furthe5r study

The effects of a single exposure of UVB on morphologic changes and proliforation in cultured normal melanocytes / 대한피부과학회지

In humans, the major stimulus for cutaneous pigmentation is ultraviolet radiation. Little is known about the mechanism underlying this response, in part, because of the complexity of the interactions involving the whole epidermis. The present stucy was undertake to evaluate the effects of a single exposure of UVB on cultured normal melanocytes. Melanocytes were exposed to UVB from 5.1 mJ/cm to 203 mJ/cm. The results were as follows : 1. The main morphologic changes in UVB-exposed groups w re larger sized cells, more blunted dendrites, and shorter dendrites than in the control group. These cells increased sized according to the increased doses of VVB, but above 101.5 mJ/cm, the melanocytes shrunk and were destroyed. 2. From 20.3 mJ/cm of UVB, the proliferation of melanocyte was decreased, Especially, there was statistical!y significant difference above 50.8 mJ/cm (p<0.05, p<0.01). 3. The antiproliferativo effect increased with the passage of tirie after UVB exposure. So, cell count could not be done in 101.5 mJ/cm and 203 mJ/cm on the third day, and in 50.8 mJ/cm, 101.5 m J/cm and 203 mJ/cm on the seventh day. 4. Statistically the melanin content per well was significantl dicreased to 11-28% of each control group with dose above 50.8 mJ/cm (p<0.05, p<0.01). The melanin content per cell was increased to 107-128% of each control group when doses were below 20.3 mJ/cm and decreased to 49-79% of each control group when above 0.8 mJ/cm on the third day, but there was no statistically significant difference. In summary, when melarocytes were exposed to UVB, morphclogic changes progressed to cell differentiation. The results also suggested that a low or dose of UVB has an antiproliferative arid mild melanogenic effect, and a higher dose of UVB has a direct cytotoxic effect.

Effects of calcipotriol(MC 903), a novel synthetic derivative of vitamin D3 on the growth of cultured human keratinocytes and melanocytes / 대한피부과학회지

The cutaneous synthesis of vitamin D in response to ultraviolet radiation exposure is the most important factor in maintaining vitamin D balance in Man. The skin is not only the site of vitamin D synthesis, but also a target organ for calcitriol(1, 25-dihydroxy-vitamin D) which is naturally occuriag, hormonally active form of vitamin E. It is now known that calcitriol inhibits the proliferation of epidermal cells and induces her differentiation. In this study, epidermal keratinocytes and melanocytes were isilated from the neonatal foreskin, and were culturod using a MCDB 153 and modified TIC media, respectively. And then various concentratioris of calcipotriol(MC 903), a synthetic aralogue of calcitriol, were added to each culture. The effects of calcipotriol on the growth of human keratinocytes and melanocytes were investigated. The results were as follows : 1. The addition of calopotriol to human keratinocyte and melalocyte cultures inhibited their proliferation in a dosdependent manner. 2. Calcipotriol had no effects on the melanization process of the melanocyte. 3. Calcipotriol was found to inhibit the proliferation, however it induced the terminal differentiation of cultured keratinocytes, as judged by morphologicai changes. The decreased density of kerationcytes, The formation of cornified cells, and the cellular destruction in a concentration of 10 M of calcipotriol were observed. 4. By using the light atid the electron microscope, we observed that the epidermal thickness was decreased and terminal differentiation was facilitateir. Living Skin Equivalent (LSE) according to the increasing concentration of calcipotriol. A]i)parent cytotoxic effects were observed in 10 M, 10 M of calcipotriol. In summary, the above results indicate that the addition of calcipotriol to the in vitro culture system of human keratinocyte and melanocyte induces the biologic process of terminal differentiation of keratinocytes and inhibits proliferation of keratinoytes and melanocytes in a dose-dependent manner.

Effects of azelaic acid on melanocytes / 대한피부과학회지

The authors investigsted the effects of azelaic acid on human melanoma cells (G 361 melanoma cell line) and cultured normal melanocytes obtained from the prepuce of newborn. The results were as follows : 1. The proliferation of melanoma cells was decreased, but not in a dose-and time- ependent fashion. The cell population of melanoma cells after 2, 4, and 6 days of culture was decreased to 3.80 x 10(5)cells/well, 4.55 x 10(5)cells/well, and 4.30 X 10(5)cells/ well, respectively, in the presence of 10(-2)M azelaic acid. The proliferation of mormal elanocytes of normal melanocytes was decreased in a dose-dependent, but not in a time-dependent fashion. The cell population of normal melanocytes after 2, 4, and 6 days of culture was significantly decreased to 6.38 x 10(5)+/-1.37 x 10(5) cells/well, 5.33 x 10(5)+/-0.73x10(5) cells/well, and 7.20x10(5)+/-1,11 x10(5)cells/well, respectively, in the presence of 10(-2) M azelaic acid(p0.05) and 6953+/-1217 CPM (p<0.05) after 4 and 6 days of culture in the prensence of 10(-2) Mazelaic acid, 3. There was no difference in melanin content per well at various concentrations in either group, but the melanin content per individual melanocyte of the melanoma cells was increased to 0.0303 ng/cell, 0.0253 ng/cell, and 0.0377 ng/cell, respectively, and that of normal melanocytes was signifieantly increased to 0.0754+/-0.0215 ng/cell, 0.0719+/-0.0144 ng/cell(p<0.05), and 0.1089+/-0.0185 ng/cell(p<0.05), respectively, after 2, 4, and 6 days of culture in the presence of 10(-2) M azelaic acid.

Effect of HLA-DR expression on IL_1 production by keratinocytes / 대한피부과학회지

No abstract available.