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Production of interleukin-8 by human trichilemmoma and squamous cell carcinoma cell lines
394
PR0D"CTION
OF A SPEClFIC
INHIBITOR
OF INTERLEUKIN
1 BY HUMAAN
THE EFFECTS OF PEMPHIGUS ANTIBODY ON THE MOUSE EPIDERHAL
EPIDERMAL KERATINOCYTES
KERATINOCYTE
HIDEAKI TAKEMATSU AND HACHIRO TAGAMI Department of Dermatology, Tohoku University School of Medicine, Sendai
SHOZO FUTAMURA, YUKI TAKAGI AND YASUO ASADA. Department of Dermatology, kansai Medical Unviersity, Osaka.
Interleukin 1 (IL-l) is produced by a variety of cells, including keratinocytes.Profound tissue damage can result from unrestricted production of IL-l; however, nws'cinflammatory processes are self-limited.A heterogeneous group of IL-1 inhibitors have been reported. In the present study, we found that cultured human epidermal keratinocytes constitutivelv released an IL-1 inhibitor. It was eluted from qel chromatographyat a M of 100 kDa at pH 7.4, and 25 kD: at pH 2.9, and showed noreffect on IL-*-dependentproliferation of mouse thymocytes and a T lymphocyte cell line, or spontaneous proliferation of other cell lines. The biochemical characterizationof the IL-1 inhibitor, i.e., the dissociation into components with a Mr of 25 kDa at acidic pH, strongly suggests that TGF_B 1s the IL-1 inhibitor released from normal human epidermal keratinocytes.(CO-inVestigat0rs: DZS. N. Isono and K. Kumagai)
MODULATION OF GROWTH AND DIFFERENTIATION IN NORMAL HUMAN KERATINOCYTES BY TRANSFORMING GROWTH FACTOR-S 1 MAKOTO HASHIRO, HIDENOBU OKUMURA, KUNIO MATSUMOTO, KOJI HASHIMOTO AND KUNIHIKO YOSHIKAWA Department of Dermatology, Osaka University School of Medicine, Osaka. The effect of transforming growth factor-6 1 (TGF-6 1) on the growth and differentiationof normal human keratinocytes cultured in serum-free medium was investigated. Addition of TGF-6 1 at 2 "g/ml inhibited the proliferation of keratinocytes both under low Ca2+ condition and high Ca2+ condition almost completely. The inhibition of keratinocyte proliferation by TGF-6 1 was also accompanied by decrease of the transient c-myc mRNA expression. The evaluation of keratinocyte differentiationwas expressed by the involucrin staining and cell sorting. The involucrin expression was increased by TGF-6 1 at 20 ng/ml under high Ca2+ condition but decreased at the came concentrationunder low Ca2+ condition. These data indicate that growth inhibition of normal human keratinocytes by TGF-8 1 was associated with decreased expression of c-myc gene and inductive potential of differentiationwas not associated with growth inhibition.
PRODUCTION OF INTERLEUKIN-8 BY HUMAN TRICHILEMMOMA AND SQUAMOUS CELL CARCINOMA CELL LINES. HIKARU ETO, KOHZOH YONEMOTO, KENSEI KATSUOKA.SHIGEO NISHIYAMA.TOHRU AKAHOSHI*, AND TAMOTSU KANZAKI**. Depts. of Dermatol. Med.*,Kitasato Univ. and Int. Sagamihara. and Dept. of Dermatol. Sch. of Med., Nagoya City Univ. Medical Sch.**. Nagoya, Japan. Interleukin-8(IL-8) is a cytokine mainly responsible for neutrophil chemotaxis. Previously we reported that trichilemmoma cell line. K-TL-1. produce neutrophil chemotactic factor in viva and In the present study. we surveyed in vitro. whether K-TL-1 and other epidermal tumor cell lines can generate IL-8 by means of RIA. Among various IK-TL-2 and LV-SCC cell lines tested, K-TL-1. secreted IL-8 into culture supernatants at the range of 21-38ng/ml. Other cell lines including did not melanomas, produce detectable level of IL-8. When recombinant IL-8(0.1-lOOng/ml) and/or anti-IL-8 antibody were added to K-TL-1 culture, the cell growth was not altered. These data suggest that certain transformed epidermal cell lines can produce IL-8. however, IL-8 does not affect proliferation of these cells.
TREATED
WITH
CYTOCHALASIN
D.
The present study is made a plan to investigate the role of actin filaments (AF) on pemphigus IgG (P-G) induced acantholysis in mouse keratinocyte (MK). AF, keratin filament (KFI and desmosomal component were studied by immunofluorescence (IFI. MK were treated with cytochalasin D (CD) prior to culture in low Ca++ media containing P-G. CD treatment caused the aggregation of AF but did not affect distribution of keratin filaments (KF). On the untreated cells the reorganizationof KF or AF and cell-cell detachment occured rapidly after transfer to the low Ca++ media with P-G. On the other hand, in MK treated with CD, the cell-cell detachment could not be observed after incubation with after incubation in low Ca++ media with P-G. These results indicate that AF may have a important role on the disassembly of desmosomes induced by P-G.
THE SERUM AND BLISTER ECP (EOSINOPHILE CATIONIC PROTEIN) LEVELS IN PATIENTS WITH PEMPHIGUS AND BULLOUS PEMPHIGOID. KUNIKO KAWAMURA, TAKASHI HORIKOSHI, NIRO HANADA, HIROAKI EGUCHI. Department of Dermatology, Sapporo Medical College, Sapporo, Japan ECP levels in serum and blister fluid were measured in patients with pemphigus (vulgaris; 5 csses, folisceus; 3 cases, erythematosus; 1 case) and bullous pemphigoid (2 CSSSS). Normal value of serum ECP was less than 10 u g/L. Serum ECP in patients were 10 = 30 u g/L. ECP values in blister, however, were more than 2000 LL g/L. When normal skin was incubated for 36 hrs with blister fluid of bullous pemphigoid, the hydropic degeneration of bssal cells was observed. These changes were reduced when anti-ECP antibody was added. The acantholysis of epidermsl keratinocytes ss seen in pemphigus was not observed. These data indicated that ECP secreted by eosinophils may play roles in the formation of blister in bullous pemphigoid and in the enlargement of blister in pemphigusvulgaris.
THE &PRESSION OF 170 KD AND 230 KD BULLOUS PPIPHIGOID (BP) ANTIGENS IN CULTURED EPIDERMAL CELLS WITHOUT HPiIDESMOSOME FORMATION. SHIGERU TANIGUCHI, TAKAE HIRONE, TAKASHI HASHIMOTO#, TAKFJI NISHIKAWA#. Department of Dermatology, Kanazawa University, nazewa and 9 . Department of Dermatology, Keio University, Tokyo Recently, the heterogeneity of bullous pemphigoid (BP) antigens has been reported and 170 kd and 230 kd bands have been most frequently observed by irmnunoblotting. It was also shown that epidermal cells cultured on collagen gel produce hemidesmosomes (HDe) but cells cultured on cellulose membrane do not. In this study, the expression of 170 kd and 230 kd BP antigens in the epidermal cells cultured on these substrata was investigated to clarify a role of HD in the BP antigens expression. Immunofluorescentstaining revealed a linear fluorescent band at the interface of the cellulose membrane and cultured epidermal cells with 170 kd and 230 kd BP antibodies. The extracts of epidermis, cells on collagen gel and cells on cellulose membrane showed the same pattern by immunoblottine with these antibodies. The evidence indicates that HD is not-necessary for the expressions of 170 kd and 230 kd BP antigens, and that the BP antigens expressed in cells without HD have the same molecular weight as those with HD.
PR0D"CTION
OF A SPEClFIC
INHIBITOR
OF INTERLEUKIN
1 BY HUMAAN
THE EFFECTS OF PEMPHIGUS ANTIBODY ON THE MOUSE EPIDERHAL
EPIDERMAL KERATINOCYTES
KERATINOCYTE
HIDEAKI TAKEMATSU AND HACHIRO TAGAMI Department of Dermatology, Tohoku University School of Medicine, Sendai
SHOZO FUTAMURA, YUKI TAKAGI AND YASUO ASADA. Department of Dermatology, kansai Medical Unviersity, Osaka.
Interleukin 1 (IL-l) is produced by a variety of cells, including keratinocytes.Profound tissue damage can result from unrestricted production of IL-l; however, nws'cinflammatory processes are self-limited.A heterogeneous group of IL-1 inhibitors have been reported. In the present study, we found that cultured human epidermal keratinocytes constitutivelv released an IL-1 inhibitor. It was eluted from qel chromatographyat a M of 100 kDa at pH 7.4, and 25 kD: at pH 2.9, and showed noreffect on IL-*-dependentproliferation of mouse thymocytes and a T lymphocyte cell line, or spontaneous proliferation of other cell lines. The biochemical characterizationof the IL-1 inhibitor, i.e., the dissociation into components with a Mr of 25 kDa at acidic pH, strongly suggests that TGF_B 1s the IL-1 inhibitor released from normal human epidermal keratinocytes.(CO-inVestigat0rs: DZS. N. Isono and K. Kumagai)
MODULATION OF GROWTH AND DIFFERENTIATION IN NORMAL HUMAN KERATINOCYTES BY TRANSFORMING GROWTH FACTOR-S 1 MAKOTO HASHIRO, HIDENOBU OKUMURA, KUNIO MATSUMOTO, KOJI HASHIMOTO AND KUNIHIKO YOSHIKAWA Department of Dermatology, Osaka University School of Medicine, Osaka. The effect of transforming growth factor-6 1 (TGF-6 1) on the growth and differentiationof normal human keratinocytes cultured in serum-free medium was investigated. Addition of TGF-6 1 at 2 "g/ml inhibited the proliferation of keratinocytes both under low Ca2+ condition and high Ca2+ condition almost completely. The inhibition of keratinocyte proliferation by TGF-6 1 was also accompanied by decrease of the transient c-myc mRNA expression. The evaluation of keratinocyte differentiationwas expressed by the involucrin staining and cell sorting. The involucrin expression was increased by TGF-6 1 at 20 ng/ml under high Ca2+ condition but decreased at the came concentrationunder low Ca2+ condition. These data indicate that growth inhibition of normal human keratinocytes by TGF-8 1 was associated with decreased expression of c-myc gene and inductive potential of differentiationwas not associated with growth inhibition.
PRODUCTION OF INTERLEUKIN-8 BY HUMAN TRICHILEMMOMA AND SQUAMOUS CELL CARCINOMA CELL LINES. HIKARU ETO, KOHZOH YONEMOTO, KENSEI KATSUOKA.SHIGEO NISHIYAMA.TOHRU AKAHOSHI*, AND TAMOTSU KANZAKI**. Depts. of Dermatol. Med.*,Kitasato Univ. and Int. Sagamihara. and Dept. of Dermatol. Sch. of Med., Nagoya City Univ. Medical Sch.**. Nagoya, Japan. Interleukin-8(IL-8) is a cytokine mainly responsible for neutrophil chemotaxis. Previously we reported that trichilemmoma cell line. K-TL-1. produce neutrophil chemotactic factor in viva and In the present study. we surveyed in vitro. whether K-TL-1 and other epidermal tumor cell lines can generate IL-8 by means of RIA. Among various IK-TL-2 and LV-SCC cell lines tested, K-TL-1. secreted IL-8 into culture supernatants at the range of 21-38ng/ml. Other cell lines including did not melanomas, produce detectable level of IL-8. When recombinant IL-8(0.1-lOOng/ml) and/or anti-IL-8 antibody were added to K-TL-1 culture, the cell growth was not altered. These data suggest that certain transformed epidermal cell lines can produce IL-8. however, IL-8 does not affect proliferation of these cells.
TREATED
WITH
CYTOCHALASIN
D.
The present study is made a plan to investigate the role of actin filaments (AF) on pemphigus IgG (P-G) induced acantholysis in mouse keratinocyte (MK). AF, keratin filament (KFI and desmosomal component were studied by immunofluorescence (IFI. MK were treated with cytochalasin D (CD) prior to culture in low Ca++ media containing P-G. CD treatment caused the aggregation of AF but did not affect distribution of keratin filaments (KF). On the untreated cells the reorganizationof KF or AF and cell-cell detachment occured rapidly after transfer to the low Ca++ media with P-G. On the other hand, in MK treated with CD, the cell-cell detachment could not be observed after incubation with after incubation in low Ca++ media with P-G. These results indicate that AF may have a important role on the disassembly of desmosomes induced by P-G.
THE SERUM AND BLISTER ECP (EOSINOPHILE CATIONIC PROTEIN) LEVELS IN PATIENTS WITH PEMPHIGUS AND BULLOUS PEMPHIGOID. KUNIKO KAWAMURA, TAKASHI HORIKOSHI, NIRO HANADA, HIROAKI EGUCHI. Department of Dermatology, Sapporo Medical College, Sapporo, Japan ECP levels in serum and blister fluid were measured in patients with pemphigus (vulgaris; 5 csses, folisceus; 3 cases, erythematosus; 1 case) and bullous pemphigoid (2 CSSSS). Normal value of serum ECP was less than 10 u g/L. Serum ECP in patients were 10 = 30 u g/L. ECP values in blister, however, were more than 2000 LL g/L. When normal skin was incubated for 36 hrs with blister fluid of bullous pemphigoid, the hydropic degeneration of bssal cells was observed. These changes were reduced when anti-ECP antibody was added. The acantholysis of epidermsl keratinocytes ss seen in pemphigus was not observed. These data indicated that ECP secreted by eosinophils may play roles in the formation of blister in bullous pemphigoid and in the enlargement of blister in pemphigusvulgaris.
THE &PRESSION OF 170 KD AND 230 KD BULLOUS PPIPHIGOID (BP) ANTIGENS IN CULTURED EPIDERMAL CELLS WITHOUT HPiIDESMOSOME FORMATION. SHIGERU TANIGUCHI, TAKAE HIRONE, TAKASHI HASHIMOTO#, TAKFJI NISHIKAWA#. Department of Dermatology, Kanazawa University, nazewa and 9 . Department of Dermatology, Keio University, Tokyo Recently, the heterogeneity of bullous pemphigoid (BP) antigens has been reported and 170 kd and 230 kd bands have been most frequently observed by irmnunoblotting. It was also shown that epidermal cells cultured on collagen gel produce hemidesmosomes (HDe) but cells cultured on cellulose membrane do not. In this study, the expression of 170 kd and 230 kd BP antigens in the epidermal cells cultured on these substrata was investigated to clarify a role of HD in the BP antigens expression. Immunofluorescentstaining revealed a linear fluorescent band at the interface of the cellulose membrane and cultured epidermal cells with 170 kd and 230 kd BP antibodies. The extracts of epidermis, cells on collagen gel and cells on cellulose membrane showed the same pattern by immunoblottine with these antibodies. The evidence indicates that HD is not-necessary for the expressions of 170 kd and 230 kd BP antigens, and that the BP antigens expressed in cells without HD have the same molecular weight as those with HD.