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Retinoic acid stimulated mouse transformed epidermal cell line(Pam212) induced mast cell like colonies from bone marrow cells
567
568
THE EFFECTS OF INTERLEUKIN 1 (IL-l) ALPHA AND BETA ON CULTURED NORMAL HUMAN EPIDERMAL KERATINCCYTES(NHEK)
ADDITIVE EFFECT OF ANTI ICAM- AND ANTI NAC 1 IN ThE INDUCTIONOF IdEUTROPHIL-IIEDIATED SCC CELL DISSOCIATION
NORIKO TAKAHASHI, YORO KAWA, SHINICHI TAKANASHI, TERUYOSHI TAKENAKA, EIKO ICHIKAWA, MASATO KASHIMA, SHINICHI WATANABE AND MASAKO MIZOGUCHI
EIROSHI KATAYAMA AND HIDE0 YAOITA Department of Dermatology,Jichi Medical School, Tochigi
Although IL-l alpha and beta are distinct gene products, they recognize the same receptor and share almost the same biologicalproperties.To find out the effects of both forms of IL-l on proliferation and differentiation of keratinocytes,NHEK were cultured with IL-l alpha, IL-1 beta or Ca2t. To estimate their proliferation, the number of cultured NHEK was counted and the 3H-thymidine uptake by NHEK was measured. To estimate their differentiation, epithelial keratins were studied by immunoblottingusing two monoclonal anticytokeratinantibodies (PKK-1 and MA-904), a monoclonal antihuman filaggrin antibody, and rabbit antiserum against involucrin. The results of these experiments show that only IL-l alpha, but not beta, suppresses the proliferation of NHEK and produces differentiationof them. These findings suggest that some biological properties may be different in IL-l alpha and beta.
In the iestructionof cancer cells by effector cells, direct contact of them is supposed to be necessary.l:owever, precise mechanism is unkonwn. We previouslyreported that direct contact of PMN (neutrophils)with SCC cells (Dj?l-1) evoked Dn4-1 dissociationin 48h in vitro. After that we found that addition of nreactivatedneutrwhils to DJ!4-1 culture in the presence'ofanti ICAX-1 ind;ced DJH-1 dissociationmuch earlier than nonactivatedones. This time we found that anti Mac 1 (CDllb)significantlyenhanced the dissociationin this system. Namely, addition af both anti Hat I and anti ICAEI-1to the reaction system of GMCSFpretreatedneutrophilswith upregulatedMac 1 on IFN -ipretreatedDJM-1 sheet with upregulatedICA?-1 enhanced DJM -1 dissociationthan the addition of anti ICAM- alone. Furthermoresuch dissociationwas partially inhibitedby anti laninin,which DJM-1 hai an its surface.
569
570
GENERATION MICE AFTER
OF H-2 CLASS II-REACTIVE CD8+ CELLS IN CLASS II-DISPARATE SKIN GRAFT REJECTION
A histopathological,immunohistochemical, and ultrastructuralstudy.
KAZUHIRO KAWAI, MASAAKI IT0 AND YOSHIO SAT0 Department of Dermatology, Niigata University School of Medicine, Niigata
MASA IWASAKI1' *, SHIN-ICHIRO
NISHIYAMA
1 Department
of
TAKEZRKIl AND SHIGEO of LJermatology, Kltaaato Univ. School
Medicine, Sagamihara 2 Departmentof Dermatology,Sakurai Hospital,Mlzusawa
Primary rejection of H-2 class II-dispar+atE,;:,in grafts is known to be mediated by CD4 predominant T cell subset involved in HOWeVer, second-set rejection of class II-disparate skin In the grafts has not been well documented. of present study, we have shown that depletion CD4+ cells by in viva monoclonal antibody treatment results in prolongation of isolated class II-disparate skin graft survival in naive mice but CD8+ cells from these not in sensitized mice. sensitized mice depleted of CD4+ cells expressed These results class II reactivity in vitro. indicate the possibility that class II-reactive CD8+ cells generated in primary rejection of class II-disparate skin grafts could mediate second-set rejection in sensitized mice without CD4+ cells.
Lymphomatoidpapulosis (LyP) was studied by immunohistochemistry,and light and electron microscopy in four cases. According to histologic types of LyP (Willemzeet al. 1982), 2 cases were classifiedas type A. 1 as type B, and 1 as atypical large transitionalfarm. ~mmunoh~stochemically, cells expressedT cell antigens in all cases. However, in two cases these cells lacked expressionof some T cell antigens. In all casesthese cells had several activated antigen (CD25,~~30, CD71, HLA DR), hut in one case did not have EMA (epithelialmembrane antigen). Electron microscopy showed that atypical lymphoid cells were admixed with numerous macrophaqes.
571
J72
IMNUNOHISTOCHEMICALLOCALIZATIONOF BASIC FIBROBLASTGROWTH FACTOR IN SEVERAL SKIN DISEASES
RETINOIC ACID STI!.%X,ATED MXSE TRANSFORNEDEPIDERMAL CELL LINE(PAM212)INWCED MAST CELL LIKE COLONIES FROM BONE MARROWCELLS
HITOSHI YAGUCHI, RYOJI TSUBOI, RIE "EKI AND HIDEOKI OGAWA Departmentof Dermatology,Juntendo Univ. School of Medicine, Tokyo
ICHIRO KATAYAMA AND KIYOSHI NISHIOKA Departmentof Dermatolcqy,Tokyo Medx?.l and Dental Univ. School of Medicine, Tokyo. MOuse transformedepidenw.1 cell line (Pam 212) cells generated the soluble rcediators(RAMCGF) to promote the mcwth of spleen-conditioned-medium dependent-mastcell iine (MC 9).in the pres_epce_yf all tra&-retinoic acid at the concentrationof IO -70 M. RAMCGF was non-aialyzable and eluted in between rrolecular weight of 45 and 6% on TSK 2000G column. Ciucmatofocusinganalysis revealed that this factor had pI range between 7.0 and 7.5. MC 9 cells respondedto Con A stimulatedspleen cell supematant, WMI-3 supzrnatantin addition to retinoic acid pretreated Pam212 supematant but not to several reccmbinantcytoklnes including ILlr,ILl,~,IL2,IL3 or IL4 at the concentration of 100 U/ml. Presence of correspondingantibodies did not abrogate the RAMm activity in the culture su~matant. In addition, F?AMcGFinduced sl~ll lymphocyte and large mast cell like colonies frcxntone marrow cells.
Basic fibroblast growth factor (bFGF) is an angiogenic factor and also a mitogen for the epidermal cell. In order to examine the role of bFGF in human skin we studied the distributionof bFGF in normal human skin and several skin diseases. bFGF was demonstratedby direct immunofluorescence staining of cryostat sections with a polyclonal anti bFGF antibody. In normal human skin, positive stainings for bFGF were observed in the basal cells. In psoriasis, the basal cells and several supra-basallayers at rate ridges were positively stained. Seborrheickeratosis and basal cell epitheliomashowed diffuse stainings in the basaloid cells of the tumor. These results indicate the importanceof bFGF for keratinocyteproliferation.
242
568
THE EFFECTS OF INTERLEUKIN 1 (IL-l) ALPHA AND BETA ON CULTURED NORMAL HUMAN EPIDERMAL KERATINCCYTES(NHEK)
ADDITIVE EFFECT OF ANTI ICAM- AND ANTI NAC 1 IN ThE INDUCTIONOF IdEUTROPHIL-IIEDIATED SCC CELL DISSOCIATION
NORIKO TAKAHASHI, YORO KAWA, SHINICHI TAKANASHI, TERUYOSHI TAKENAKA, EIKO ICHIKAWA, MASATO KASHIMA, SHINICHI WATANABE AND MASAKO MIZOGUCHI
EIROSHI KATAYAMA AND HIDE0 YAOITA Department of Dermatology,Jichi Medical School, Tochigi
Although IL-l alpha and beta are distinct gene products, they recognize the same receptor and share almost the same biologicalproperties.To find out the effects of both forms of IL-l on proliferation and differentiation of keratinocytes,NHEK were cultured with IL-l alpha, IL-1 beta or Ca2t. To estimate their proliferation, the number of cultured NHEK was counted and the 3H-thymidine uptake by NHEK was measured. To estimate their differentiation, epithelial keratins were studied by immunoblottingusing two monoclonal anticytokeratinantibodies (PKK-1 and MA-904), a monoclonal antihuman filaggrin antibody, and rabbit antiserum against involucrin. The results of these experiments show that only IL-l alpha, but not beta, suppresses the proliferation of NHEK and produces differentiationof them. These findings suggest that some biological properties may be different in IL-l alpha and beta.
In the iestructionof cancer cells by effector cells, direct contact of them is supposed to be necessary.l:owever, precise mechanism is unkonwn. We previouslyreported that direct contact of PMN (neutrophils)with SCC cells (Dj?l-1) evoked Dn4-1 dissociationin 48h in vitro. After that we found that addition of nreactivatedneutrwhils to DJ!4-1 culture in the presence'ofanti ICAX-1 ind;ced DJH-1 dissociationmuch earlier than nonactivatedones. This time we found that anti Mac 1 (CDllb)significantlyenhanced the dissociationin this system. Namely, addition af both anti Hat I and anti ICAEI-1to the reaction system of GMCSFpretreatedneutrophilswith upregulatedMac 1 on IFN -ipretreatedDJM-1 sheet with upregulatedICA?-1 enhanced DJM -1 dissociationthan the addition of anti ICAM- alone. Furthermoresuch dissociationwas partially inhibitedby anti laninin,which DJM-1 hai an its surface.
569
570
GENERATION MICE AFTER
OF H-2 CLASS II-REACTIVE CD8+ CELLS IN CLASS II-DISPARATE SKIN GRAFT REJECTION
A histopathological,immunohistochemical, and ultrastructuralstudy.
KAZUHIRO KAWAI, MASAAKI IT0 AND YOSHIO SAT0 Department of Dermatology, Niigata University School of Medicine, Niigata
MASA IWASAKI1' *, SHIN-ICHIRO
NISHIYAMA
1 Department
of
TAKEZRKIl AND SHIGEO of LJermatology, Kltaaato Univ. School
Medicine, Sagamihara 2 Departmentof Dermatology,Sakurai Hospital,Mlzusawa
Primary rejection of H-2 class II-dispar+atE,;:,in grafts is known to be mediated by CD4 predominant T cell subset involved in HOWeVer, second-set rejection of class II-disparate skin In the grafts has not been well documented. of present study, we have shown that depletion CD4+ cells by in viva monoclonal antibody treatment results in prolongation of isolated class II-disparate skin graft survival in naive mice but CD8+ cells from these not in sensitized mice. sensitized mice depleted of CD4+ cells expressed These results class II reactivity in vitro. indicate the possibility that class II-reactive CD8+ cells generated in primary rejection of class II-disparate skin grafts could mediate second-set rejection in sensitized mice without CD4+ cells.
Lymphomatoidpapulosis (LyP) was studied by immunohistochemistry,and light and electron microscopy in four cases. According to histologic types of LyP (Willemzeet al. 1982), 2 cases were classifiedas type A. 1 as type B, and 1 as atypical large transitionalfarm. ~mmunoh~stochemically, cells expressedT cell antigens in all cases. However, in two cases these cells lacked expressionof some T cell antigens. In all casesthese cells had several activated antigen (CD25,~~30, CD71, HLA DR), hut in one case did not have EMA (epithelialmembrane antigen). Electron microscopy showed that atypical lymphoid cells were admixed with numerous macrophaqes.
571
J72
IMNUNOHISTOCHEMICALLOCALIZATIONOF BASIC FIBROBLASTGROWTH FACTOR IN SEVERAL SKIN DISEASES
RETINOIC ACID STI!.%X,ATED MXSE TRANSFORNEDEPIDERMAL CELL LINE(PAM212)INWCED MAST CELL LIKE COLONIES FROM BONE MARROWCELLS
HITOSHI YAGUCHI, RYOJI TSUBOI, RIE "EKI AND HIDEOKI OGAWA Departmentof Dermatology,Juntendo Univ. School of Medicine, Tokyo
ICHIRO KATAYAMA AND KIYOSHI NISHIOKA Departmentof Dermatolcqy,Tokyo Medx?.l and Dental Univ. School of Medicine, Tokyo. MOuse transformedepidenw.1 cell line (Pam 212) cells generated the soluble rcediators(RAMCGF) to promote the mcwth of spleen-conditioned-medium dependent-mastcell iine (MC 9).in the pres_epce_yf all tra&-retinoic acid at the concentrationof IO -70 M. RAMCGF was non-aialyzable and eluted in between rrolecular weight of 45 and 6% on TSK 2000G column. Ciucmatofocusinganalysis revealed that this factor had pI range between 7.0 and 7.5. MC 9 cells respondedto Con A stimulatedspleen cell supematant, WMI-3 supzrnatantin addition to retinoic acid pretreated Pam212 supematant but not to several reccmbinantcytoklnes including ILlr,ILl,~,IL2,IL3 or IL4 at the concentration of 100 U/ml. Presence of correspondingantibodies did not abrogate the RAMm activity in the culture su~matant. In addition, F?AMcGFinduced sl~ll lymphocyte and large mast cell like colonies frcxntone marrow cells.
Basic fibroblast growth factor (bFGF) is an angiogenic factor and also a mitogen for the epidermal cell. In order to examine the role of bFGF in human skin we studied the distributionof bFGF in normal human skin and several skin diseases. bFGF was demonstratedby direct immunofluorescence staining of cryostat sections with a polyclonal anti bFGF antibody. In normal human skin, positive stainings for bFGF were observed in the basal cells. In psoriasis, the basal cells and several supra-basallayers at rate ridges were positively stained. Seborrheickeratosis and basal cell epitheliomashowed diffuse stainings in the basaloid cells of the tumor. These results indicate the importanceof bFGF for keratinocyteproliferation.
242