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Ontogeny and Ia antigen expression of the rat epidermal Langerhans cell lineage during perinatal period
119 A-30
IYYUNOHlSTOCHEYlCAL OBSERVATION ON THE KINETICS POSSESSING MITE ANTIGEN
Y. Tanaka
(Division
of
Dermatology.
Nagasaki
K. Yaeda.
OF LANGERHANS CELLS
Chuo National
K. Yamamoto. Y. Tanaka. H. Yoshida Nagasnki University School of Medicine) and
H. Morita, T. Kihara, M. Kuramoto, S. Minami, M. Yu, M. Yamagata, S. Sagami. Departmentof Dermatology,Hyogo College of Medicine.
Hospital).
(Department of Dermatology. A. Anan (Nagasaki City)
Recently. We haveobserved that eczematous reaction could be Induced by application of mire antigen after 48hrs in the majority of patients atopic dermatitis (AD). The percutaneous entry of mite antigen was
topical
with also
demonstrated
by
immunofluorescence
staining.
Furthermore.
it
was
of OKT6 positive cells which were distributed in the dermis by use of immune-double labeling technique. On the other hand. OUTimmunoelectron microscope study confirmed that mite antigen was trapped and phagacytosed by the dermal macrophages. To further elucidate the response to mite antigen. skin biopsieswere done at the sitesof patchtestingafterIhr. Ghrs. 24hrs and 48hrs and examined by immunohistochemical methods. AS the results. manyLangerhans cellspossessing miteantigenwere seenin the epidermis duringbeginning 6 hrs. But. after24 hours. mostof the cellswereobservedin the dermis. shoringapportionto lymphocytes. revealed
A -37
that
mite
antigen
was seen
on the
FLOW CYTOMETRICANALYSIS OF L3T4-POSITIVECELLS IN LYMPH NODE OF MURINE CONTACT HYPERSENSITIVITYREACTION
h-30
surface
ONTOGENY AND IA ANTIGEN EXPRESSIONOF THE RAT EPIDERMAL LANCERHANSCELL LINEAGE DURING PERINATAL PERIOD
L. Hsiao' ', M. Takeys', T. Arao' and K. Takahashi' 'Departmentof DermatoloXyand 2Second Departmentof Pathology, Kumamoto UniversityMedical School, Kumamoto,Japan A new mouse anti-ratmacrophage monoclonal antibody,TRPM-1, produced by Takeya et al. was reported to recognizeLangerhans cells and indeterminatedendritic cells (ID&) in rat epidermis. THPM-lnositive IDCs anneared in rat widerma sheets from the 17th day of Com&ing numbers of the TRPM-l-positivecells with.Iagestation. positive ones, only a few epidermaldendritic cells were positive for Ia antigen during fetal life. However, one day before birth the Ia-positivecells began to increase. After birth, Ia-positivecells birth when Birbeck increasedmarkedly. At about 5 days after granules began to appear in the Langerhanscells, the ratio of Iawsitive cells to TRPM-l--positive cells became constant and remained so afterwards. The Ia antigen of the Langerhanscell lineage is a significantdeterminantof its immunologicresponsiveness. These results disclosedthat Ia antigen expressionof this cell lineage was suppressedduring fetal life and that its maturationwas not completeduntil five days after birth.
At the 11th and 12th meetings of this society.we demonstrated that the Ia-positivedendritic (Ia-D) cells increasedaround the marginal sinus in the lymph node at 24-hours after induction,and then spread into the paracorticalarea. We also reported that L3T4 blast cells appear in the lymph nodes on the fifth day after induction, as proven by flow cytometricanalysis. This time, using monaclonal anti-L3T4antibodies,and monoclonal anti-IL-2 receptor (IL-2R) antibodies,cells showing positive reactionsto L3T4, IL-2R detected by the inrmunofluorescent technique were subjected to two-coloranalysis usinr!the EPICS V cell sorter. in the lymph node on the fou&h day of induction phase As a res;lt, of contact hvoersensitivitvreaction.L3T4 ILZ-RC cells were found to be more n&zrous comparid to the number of cells in untreated mice.
A-40
GENETIC
FACTOR
RESTRICTION
IN MURINE
CONTACT
Y. Tokura; M.Takigawa:
OF UVB-INDUCED
T-CELL-SUPPRESSOR
PHOTOSENSITIVITY * =partrnent of Medicine, Hanamatsu,
T. Sato**and M. Yamada:
ofCern!atology,HamyMtsu UniversitySchool and **TokyoMedical and Dental University
Inmurine oontxtpbotaensitivity (CPS) to TZSA,wehave reported that antigen-spxific suppressorT cells and factor (TsF) are inducedby preexposureof the photosensitizingsite to low doses of LWB. The TsF is a single chain factor baring both antigen-binding site and I-J determinants.In this repxt,we exanined the genetic restrictionof the factor in terms of both H-2 and Igh-asscciated genes. 'IheCPS respvlsesof BALB/c (H-2:Igh-1') and 5UB.B (H-2: Igh-1') but not DBA/I (H-2: Igh-p) were suppressedby the injection of the B&B/c TsF. Fnrthznn~e,~the faapr significantlysuppressed the CPS reaction in BAB-14 (H-2, Igh-v"C ) but rot C.B-20 (H-2: Igh-lb) or C.&Z0 (H-2$Igh-l*).'Ihisindicatedthat identity at the Igh-V locus, but not at the MHC, of the strain prcducinq the factor and the recipientwas r-red for suppression.$cause of the single chain natureof the factor,its-that the I-J molecule present in our TsF is closely related to wt only recognitionbut also 19-h-V restrictionfunctions.
SUSCEPTIBILITYTO NC ACTIVITY AND MIXED SKIN CELL LYMPNOCYTE REACTION (MSLR) IN FRACTIONEDEPIDERMAL CELLS M. Kashitani,S. Tanaka, K. Takahashi,H. Okada, N. Tomioka, K. Orita DePartmentof 1st Surgery, Okayama Univ. Medical School, OkayPreviouslywe reported that the rat epidermal cells were susceptible in natural cytotoxic (NC) activity of syngeneicand allogeneic spleen cells, and that ehey induced syngeneic and allogeneicT cell proliferationin mixed skin cell lymphocytereaction (MSLR). Here we have examinedwhether there were potent subpopulationsof epidermal tells in these immunologicalresposes. 6 subpopulationsof epidenoal cellswereobtainedbyPercol1 gradient sedimentation. The fraction of intermediate gravity was more susceptible in NC activity of syngeThe neic and allogeneic spleen cells than whole epidermal cells was. other fractions were less or not susceptible in NC activity. On the other hand, the lightest fractionwas more seimulatoryinMSLR of
syngeneicand allcgeneic combinationthan whole epidermal cells was. The other fractionswere less stimularoryin MSLR, and the lighter the fractionwas, the higher the stimulatingcapabilitywas. Thus the responsiblecells for SCMC and MSLR were different from each ocher, and there must be something different from the other epidermalcells in the intermediatefraction.
A-41
PARTIAL ~IZATICN PEMPHIGOIDANTT~
AN!JGRaTIc MPRESSICN OF l?aJLuxs SYN'IWSIZEEBY PAM CELL!?
S. Miyakawa, S. Taji.naard T. Nishikawa. Departmentof I)ennatolcgy, Keio UniversitySchcol of Medicine,Tokyo wlllous pemphigoid (B.P.)antigen produced by Pam cells (rwuse epiderwl cell lines) was studied by inxnunoprecipitation ~rcceduredescribed by Stanley et al. The protein of ms1ecula.r weight 22OKD which wai reportedby Stanley et al was identified. Pulse-chaseexperimentshcnvedthat this protein was relatively st&le. l%e protein treated by Tunicamycinwas shown to react with B.P. antiserum. Fran this exceriment. the suaar nwietv was considerednot to be involved in the reaction cd&d by this nrotein and B.P. antisenmn. RNA was isolated fran Pan cells and Lanslated proteins by cell free translationin the rabbit reticulccytelysate were imnunoprecipitated with B.P. antiserwn. Slightly higher n~&~~lar protein was identified. 'Ihisresult suggested that B.P. antigen was genetically expressed by Pam cells.
IYYUNOHlSTOCHEYlCAL OBSERVATION ON THE KINETICS POSSESSING MITE ANTIGEN
Y. Tanaka
(Division
of
Dermatology.
Nagasaki
K. Yaeda.
OF LANGERHANS CELLS
Chuo National
K. Yamamoto. Y. Tanaka. H. Yoshida Nagasnki University School of Medicine) and
H. Morita, T. Kihara, M. Kuramoto, S. Minami, M. Yu, M. Yamagata, S. Sagami. Departmentof Dermatology,Hyogo College of Medicine.
Hospital).
(Department of Dermatology. A. Anan (Nagasaki City)
Recently. We haveobserved that eczematous reaction could be Induced by application of mire antigen after 48hrs in the majority of patients atopic dermatitis (AD). The percutaneous entry of mite antigen was
topical
with also
demonstrated
by
immunofluorescence
staining.
Furthermore.
it
was
of OKT6 positive cells which were distributed in the dermis by use of immune-double labeling technique. On the other hand. OUTimmunoelectron microscope study confirmed that mite antigen was trapped and phagacytosed by the dermal macrophages. To further elucidate the response to mite antigen. skin biopsieswere done at the sitesof patchtestingafterIhr. Ghrs. 24hrs and 48hrs and examined by immunohistochemical methods. AS the results. manyLangerhans cellspossessing miteantigenwere seenin the epidermis duringbeginning 6 hrs. But. after24 hours. mostof the cellswereobservedin the dermis. shoringapportionto lymphocytes. revealed
A -37
that
mite
antigen
was seen
on the
FLOW CYTOMETRICANALYSIS OF L3T4-POSITIVECELLS IN LYMPH NODE OF MURINE CONTACT HYPERSENSITIVITYREACTION
h-30
surface
ONTOGENY AND IA ANTIGEN EXPRESSIONOF THE RAT EPIDERMAL LANCERHANSCELL LINEAGE DURING PERINATAL PERIOD
L. Hsiao' ', M. Takeys', T. Arao' and K. Takahashi' 'Departmentof DermatoloXyand 2Second Departmentof Pathology, Kumamoto UniversityMedical School, Kumamoto,Japan A new mouse anti-ratmacrophage monoclonal antibody,TRPM-1, produced by Takeya et al. was reported to recognizeLangerhans cells and indeterminatedendritic cells (ID&) in rat epidermis. THPM-lnositive IDCs anneared in rat widerma sheets from the 17th day of Com&ing numbers of the TRPM-l-positivecells with.Iagestation. positive ones, only a few epidermaldendritic cells were positive for Ia antigen during fetal life. However, one day before birth the Ia-positivecells began to increase. After birth, Ia-positivecells birth when Birbeck increasedmarkedly. At about 5 days after granules began to appear in the Langerhanscells, the ratio of Iawsitive cells to TRPM-l--positive cells became constant and remained so afterwards. The Ia antigen of the Langerhanscell lineage is a significantdeterminantof its immunologicresponsiveness. These results disclosedthat Ia antigen expressionof this cell lineage was suppressedduring fetal life and that its maturationwas not completeduntil five days after birth.
At the 11th and 12th meetings of this society.we demonstrated that the Ia-positivedendritic (Ia-D) cells increasedaround the marginal sinus in the lymph node at 24-hours after induction,and then spread into the paracorticalarea. We also reported that L3T4 blast cells appear in the lymph nodes on the fifth day after induction, as proven by flow cytometricanalysis. This time, using monaclonal anti-L3T4antibodies,and monoclonal anti-IL-2 receptor (IL-2R) antibodies,cells showing positive reactionsto L3T4, IL-2R detected by the inrmunofluorescent technique were subjected to two-coloranalysis usinr!the EPICS V cell sorter. in the lymph node on the fou&h day of induction phase As a res;lt, of contact hvoersensitivitvreaction.L3T4 ILZ-RC cells were found to be more n&zrous comparid to the number of cells in untreated mice.
A-40
GENETIC
FACTOR
RESTRICTION
IN MURINE
CONTACT
Y. Tokura; M.Takigawa:
OF UVB-INDUCED
T-CELL-SUPPRESSOR
PHOTOSENSITIVITY * =partrnent of Medicine, Hanamatsu,
T. Sato**and M. Yamada:
ofCern!atology,HamyMtsu UniversitySchool and **TokyoMedical and Dental University
Inmurine oontxtpbotaensitivity (CPS) to TZSA,wehave reported that antigen-spxific suppressorT cells and factor (TsF) are inducedby preexposureof the photosensitizingsite to low doses of LWB. The TsF is a single chain factor baring both antigen-binding site and I-J determinants.In this repxt,we exanined the genetic restrictionof the factor in terms of both H-2 and Igh-asscciated genes. 'IheCPS respvlsesof BALB/c (H-2:Igh-1') and 5UB.B (H-2: Igh-1') but not DBA/I (H-2: Igh-p) were suppressedby the injection of the B&B/c TsF. Fnrthznn~e,~the faapr significantlysuppressed the CPS reaction in BAB-14 (H-2, Igh-v"C ) but rot C.B-20 (H-2: Igh-lb) or C.&Z0 (H-2$Igh-l*).'Ihisindicatedthat identity at the Igh-V locus, but not at the MHC, of the strain prcducinq the factor and the recipientwas r-red for suppression.$cause of the single chain natureof the factor,its-that the I-J molecule present in our TsF is closely related to wt only recognitionbut also 19-h-V restrictionfunctions.
SUSCEPTIBILITYTO NC ACTIVITY AND MIXED SKIN CELL LYMPNOCYTE REACTION (MSLR) IN FRACTIONEDEPIDERMAL CELLS M. Kashitani,S. Tanaka, K. Takahashi,H. Okada, N. Tomioka, K. Orita DePartmentof 1st Surgery, Okayama Univ. Medical School, OkayPreviouslywe reported that the rat epidermal cells were susceptible in natural cytotoxic (NC) activity of syngeneicand allogeneic spleen cells, and that ehey induced syngeneic and allogeneicT cell proliferationin mixed skin cell lymphocytereaction (MSLR). Here we have examinedwhether there were potent subpopulationsof epidermal tells in these immunologicalresposes. 6 subpopulationsof epidenoal cellswereobtainedbyPercol1 gradient sedimentation. The fraction of intermediate gravity was more susceptible in NC activity of syngeThe neic and allogeneic spleen cells than whole epidermal cells was. other fractions were less or not susceptible in NC activity. On the other hand, the lightest fractionwas more seimulatoryinMSLR of
syngeneicand allcgeneic combinationthan whole epidermal cells was. The other fractionswere less stimularoryin MSLR, and the lighter the fractionwas, the higher the stimulatingcapabilitywas. Thus the responsiblecells for SCMC and MSLR were different from each ocher, and there must be something different from the other epidermalcells in the intermediatefraction.
A-41
PARTIAL ~IZATICN PEMPHIGOIDANTT~
AN!JGRaTIc MPRESSICN OF l?aJLuxs SYN'IWSIZEEBY PAM CELL!?
S. Miyakawa, S. Taji.naard T. Nishikawa. Departmentof I)ennatolcgy, Keio UniversitySchcol of Medicine,Tokyo wlllous pemphigoid (B.P.)antigen produced by Pam cells (rwuse epiderwl cell lines) was studied by inxnunoprecipitation ~rcceduredescribed by Stanley et al. The protein of ms1ecula.r weight 22OKD which wai reportedby Stanley et al was identified. Pulse-chaseexperimentshcnvedthat this protein was relatively st&le. l%e protein treated by Tunicamycinwas shown to react with B.P. antiserum. Fran this exceriment. the suaar nwietv was considerednot to be involved in the reaction cd&d by this nrotein and B.P. antisenmn. RNA was isolated fran Pan cells and Lanslated proteins by cell free translationin the rabbit reticulccytelysate were imnunoprecipitated with B.P. antiserwn. Slightly higher n~&~~lar protein was identified. 'Ihisresult suggested that B.P. antigen was genetically expressed by Pam cells.