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Immunohistochemical characterization of the cellular infiltrate in alopecia areata with or without atopic diathesis
DEVELOPMENT OF
HUMAN
NAZUKO
FORMATION
HAIR
OF
A MONOCLONAL
ANTIBODY
TO
THE
CELLS
FOLLICLE
YAMAGAMI,
MASAE
AND T‘WASHI
TAKAHASHI,
TEZUKA
Department of Dermatology, Kinki University School of Medicine, Osaka, Japan Using human hair follicles as an antigen, we developed a monoclonal antibody (TYHF-1)to the cells of human harr follicle. W&B/c mice were immunized with homoqinized human hair follicular cells. The spleen cells were hybridized with SpZ/O mouse myeloma cells. Antibody-producing hybridomas were cloned by limiting dilution method. By immunofluorescencestaining techniques,TYHF-1 stained the cytoplasm of the cells in hair follicle in frozen sections of normal human. The class of this antibody was 1qM. On immunoblot analysis, TYHF-1 reacted with 46 and 74KD proteins in extract of normal human hair fallicular The staining pattern of immunofluorescent study Cells. after absorbing test using keratin was not changed. these results suggest that TYHF-1 recognues Therefore, non-keratln protein of the follicular cell.
the
Effects of androgens on human outer root sheath cells cultured in a defined, serum-free medium. SOTARO
KURATA,
SATOSHI
ITAMI,
AND
SUSUMU
TAKAYASU.
Department of Dermatology, Medical College of Oita, Oita, Japan. Outer root sheath cells(ORSC) obtained from plucked hair follicles could be serially cultured in a defined medium without serum. ORSCs grew well on type I as well as type IV collagen coated dishes, Optimal as in the case of epidermal keratinocytes. concentrations of calcium and EGF for the arowth of ORSC were similar to those reported for epidermal keratinocytes. ORSCs could be passaged for five subcultures. Dihydrotestosterone, R1881, aqd testosterone stimulated the incorporation of “Hthvmidine into beard ORSCs and this stimulation wa; inhibited by cyproterone acetate. On the osher hand, androgens showed little or no effect on Hthymidine uptake into occipital scalp ORSCs. These results sucvxst that the srowth of beard ORSCs is __ regulated by androgens.
THE
EFFECT
OF
MINOXIDIL
ON
HAIR
GROWTH
IN
PARTIAL PURIFICATION AND PROPERTIES OF NEUTROPHIL AND LYM PBOCYTKCHEMOTACTJCFLCTORS PROM MURJNHFIAIR FOLLICLES YURU SOW’ “, YOR:KO MIYi.~i+W’” , KEN.11T?.KA!lORI“, HIDEOKI OGAWA", Juntendo University School of Hedicine,China-Japan Friendship Hospital,Departmentof Dermatology Our cheaotactic studies, using marine anagen hair bulbs of C3H strain as a chernoattractant demonstrated the presence of potent PIN and lymphocyte chemoattractants in hair bulb extr acts (HRB) 1” vitro and in viva assay systems. The PMN chemoattractant was a high molecular weight protein (54Kd). and its tert.iary structure may he essential for activity. On the other hand, the lymphocyte chemoattractant consisted of two kinds of high molecular weight, substances (54Kd and 25Kd). 54Kd substances was heat resistant and 25Kd sllbstancewas heat labile. These findings suggest that HRE contains two kind of lymphocyte chemotactic factors with different properties. Intradermal injection of 54Kd and 25Kd substances induced an accumulation of PNNs and lymphocytes, and lyw phocytes, respectively.
IMMUNOHISTOCHEMICALCHARACTERIZATIONOF THE CELLULAR INFILTRATE IN ALOPECIA AREATA WITH OR WITHOUT ATOPIC DIATHESIS AKIRA ARAI, TOSHIHIKO NOGUCHI, KENSEI KATSUOKA Department of Dermatology, Kitasato Univ. School of Medicine, Kanagawa. Recent therapeutic studies and laboratory findings suggest that alopecia areata(AA) is caused by autoimmunity. Direct evidence for an autoimnune etiology in AA is the presence of mononuclear cells infiltrate around the lower portions of hair follicles in the early stages of the disease. On the other hand, some people think that the atopic diathesis is important factor for AA. In the results of our investigation,54% of AA had atopic diathesis. The immunohistochemicalanalysis of T-cell subsets in the peribulbar and intrabulbar infiltrates revealed that the proportion of OKT 4' cells was higher than that of OKT f3+cells in the cases with atopic diathesis,and OKT 8' cells were predominant in the group without atopic diathesis. In addition, the difference in the expression of HLA-DR antigen between atopic and non-atopic groups was examined and we found that there are some variations of HLA-DR expression and T-cell subsets among the stage of leasions with or without atopic diathesis.
THE FUNCTION OF THE
MICE.
AND SHOZO TAKAHASHI, MASAAKI MOROHASHI Department of Dermatology, Toyama University, Toyama
AND
MASAYORI
Medical
KAGOURA.
and Pharmaceutical
Minoxiail is now examined the effectiveness to alopecia patients in Japan. In order to clarify the effect of minaxidil on hair growth experimentally, we studied the histologlcal changes of hair follicles in mice. Two% muvaxidil were smeared once a day on the back of 6 week-old BDF male mouse. And the biopsy specimens were processed for light and electron microscopy. Early stage of anagen hairs gradually increased in number at 1 and 2 weeks. At 3 weeks many middle stage of anagen hairs with well developed hair bulb were observed. Late stage of anaqen hairs could be seen only minoxidil group at 3 and 4 weeks. Electron microscopic findings showed the slight activated hair follicle cells at 1 week. We confirmed that minoxidil showed the higher ratio of anagen hairs especially well developed hair follicles in comparison to that of control group and also effected the mild morphological changes electron microscopically.
CHEMICAL
KUMIKO
STRATUM TREATMENTS.
KUMASAKA
AND
HACHIRO
Department of Dermatology, cine,
CORNEUM
UNDER
TAGAMI. Tohoku University
VARIOUS
PHYSICAL
School
of Medi-
Sendai
We assessed the effects of various drastic physical and chemical treatments on the stratum corneum functions using a simulation model for in viva stratum cornea. Physical treatments including heating up to 90°C. freezing, ultraviolet irradiation and repeated exposure to high and low humidities did not induce any parmanent functional derangements in the stratum corneum. In contrast, chemical treatments such as application of aceton/ether, chloroform/methanol or 10% sodium lauryl sulfate produced various degrees of functional damages in the stratum corneum depending upon the duration of the application time. The present studies demonstrated that the stratum corneum is more resistant to various drastic physical treatments than the chemical treatments.
HUMAN
NAZUKO
FORMATION
HAIR
OF
A MONOCLONAL
ANTIBODY
TO
THE
CELLS
FOLLICLE
YAMAGAMI,
MASAE
AND T‘WASHI
TAKAHASHI,
TEZUKA
Department of Dermatology, Kinki University School of Medicine, Osaka, Japan Using human hair follicles as an antigen, we developed a monoclonal antibody (TYHF-1)to the cells of human harr follicle. W&B/c mice were immunized with homoqinized human hair follicular cells. The spleen cells were hybridized with SpZ/O mouse myeloma cells. Antibody-producing hybridomas were cloned by limiting dilution method. By immunofluorescencestaining techniques,TYHF-1 stained the cytoplasm of the cells in hair follicle in frozen sections of normal human. The class of this antibody was 1qM. On immunoblot analysis, TYHF-1 reacted with 46 and 74KD proteins in extract of normal human hair fallicular The staining pattern of immunofluorescent study Cells. after absorbing test using keratin was not changed. these results suggest that TYHF-1 recognues Therefore, non-keratln protein of the follicular cell.
the
Effects of androgens on human outer root sheath cells cultured in a defined, serum-free medium. SOTARO
KURATA,
SATOSHI
ITAMI,
AND
SUSUMU
TAKAYASU.
Department of Dermatology, Medical College of Oita, Oita, Japan. Outer root sheath cells(ORSC) obtained from plucked hair follicles could be serially cultured in a defined medium without serum. ORSCs grew well on type I as well as type IV collagen coated dishes, Optimal as in the case of epidermal keratinocytes. concentrations of calcium and EGF for the arowth of ORSC were similar to those reported for epidermal keratinocytes. ORSCs could be passaged for five subcultures. Dihydrotestosterone, R1881, aqd testosterone stimulated the incorporation of “Hthvmidine into beard ORSCs and this stimulation wa; inhibited by cyproterone acetate. On the osher hand, androgens showed little or no effect on Hthymidine uptake into occipital scalp ORSCs. These results sucvxst that the srowth of beard ORSCs is __ regulated by androgens.
THE
EFFECT
OF
MINOXIDIL
ON
HAIR
GROWTH
IN
PARTIAL PURIFICATION AND PROPERTIES OF NEUTROPHIL AND LYM PBOCYTKCHEMOTACTJCFLCTORS PROM MURJNHFIAIR FOLLICLES YURU SOW’ “, YOR:KO MIYi.~i+W’” , KEN.11T?.KA!lORI“, HIDEOKI OGAWA", Juntendo University School of Hedicine,China-Japan Friendship Hospital,Departmentof Dermatology Our cheaotactic studies, using marine anagen hair bulbs of C3H strain as a chernoattractant demonstrated the presence of potent PIN and lymphocyte chemoattractants in hair bulb extr acts (HRB) 1” vitro and in viva assay systems. The PMN chemoattractant was a high molecular weight protein (54Kd). and its tert.iary structure may he essential for activity. On the other hand, the lymphocyte chemoattractant consisted of two kinds of high molecular weight, substances (54Kd and 25Kd). 54Kd substances was heat resistant and 25Kd sllbstancewas heat labile. These findings suggest that HRE contains two kind of lymphocyte chemotactic factors with different properties. Intradermal injection of 54Kd and 25Kd substances induced an accumulation of PNNs and lymphocytes, and lyw phocytes, respectively.
IMMUNOHISTOCHEMICALCHARACTERIZATIONOF THE CELLULAR INFILTRATE IN ALOPECIA AREATA WITH OR WITHOUT ATOPIC DIATHESIS AKIRA ARAI, TOSHIHIKO NOGUCHI, KENSEI KATSUOKA Department of Dermatology, Kitasato Univ. School of Medicine, Kanagawa. Recent therapeutic studies and laboratory findings suggest that alopecia areata(AA) is caused by autoimmunity. Direct evidence for an autoimnune etiology in AA is the presence of mononuclear cells infiltrate around the lower portions of hair follicles in the early stages of the disease. On the other hand, some people think that the atopic diathesis is important factor for AA. In the results of our investigation,54% of AA had atopic diathesis. The immunohistochemicalanalysis of T-cell subsets in the peribulbar and intrabulbar infiltrates revealed that the proportion of OKT 4' cells was higher than that of OKT f3+cells in the cases with atopic diathesis,and OKT 8' cells were predominant in the group without atopic diathesis. In addition, the difference in the expression of HLA-DR antigen between atopic and non-atopic groups was examined and we found that there are some variations of HLA-DR expression and T-cell subsets among the stage of leasions with or without atopic diathesis.
THE FUNCTION OF THE
MICE.
AND SHOZO TAKAHASHI, MASAAKI MOROHASHI Department of Dermatology, Toyama University, Toyama
AND
MASAYORI
Medical
KAGOURA.
and Pharmaceutical
Minoxiail is now examined the effectiveness to alopecia patients in Japan. In order to clarify the effect of minaxidil on hair growth experimentally, we studied the histologlcal changes of hair follicles in mice. Two% muvaxidil were smeared once a day on the back of 6 week-old BDF male mouse. And the biopsy specimens were processed for light and electron microscopy. Early stage of anagen hairs gradually increased in number at 1 and 2 weeks. At 3 weeks many middle stage of anagen hairs with well developed hair bulb were observed. Late stage of anaqen hairs could be seen only minoxidil group at 3 and 4 weeks. Electron microscopic findings showed the slight activated hair follicle cells at 1 week. We confirmed that minoxidil showed the higher ratio of anagen hairs especially well developed hair follicles in comparison to that of control group and also effected the mild morphological changes electron microscopically.
CHEMICAL
KUMIKO
STRATUM TREATMENTS.
KUMASAKA
AND
HACHIRO
Department of Dermatology, cine,
CORNEUM
UNDER
TAGAMI. Tohoku University
VARIOUS
PHYSICAL
School
of Medi-
Sendai
We assessed the effects of various drastic physical and chemical treatments on the stratum corneum functions using a simulation model for in viva stratum cornea. Physical treatments including heating up to 90°C. freezing, ultraviolet irradiation and repeated exposure to high and low humidities did not induce any parmanent functional derangements in the stratum corneum. In contrast, chemical treatments such as application of aceton/ether, chloroform/methanol or 10% sodium lauryl sulfate produced various degrees of functional damages in the stratum corneum depending upon the duration of the application time. The present studies demonstrated that the stratum corneum is more resistant to various drastic physical treatments than the chemical treatments.