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-91- Immunology(3) 9/5 Room-A
REGULATORY FUNCTION OF M-CSF, GM-CSF AND TNF-a ON THE EXPRESSION OF COSTIMULATORY MOLECULES OF MURINE EPIDERMAL LANGERHANS CELLS (LC).
Claudio Salgado1, Koichiro Nakamura1, Yoichi Koyama2, Sinkichi Irie2, Kunihiko Tamaki1.
1Department of Dermatology, University of Tokyo, 2Nippi Research Institute of Biomatrix, Tokyo, Japan.
LC are bone marrow derived antigen presenting cells and activate naive T cells. In order to clarify the capacity of M-CSF, GM-CSF and TNF-a on the expression of costimulatory molecules (CD40, CD80 and CD86) of LC, LC were purified using panning method and cultured in combination with these cytokines. After 12 hours culture, the expression was examined by FACS. M-CSF and GM-CSF (1ng/ml) modulated CD80 and CD86 expression in different patterns. Compared to medium only cultured LC (100% MFI), M-CSF up-regulated CD80 to 188% and CD86 to 137%, GM-CSF up-regulated CD80 to 149% and CD86 to 169%. TNF-a strikingly up-regulated CD40 (296%) at 10 ng/ml, while GM-CSF up-regulated to 135% and M-CSF did not have any effect after 12 hours culture. These results suggest that the expression of costimulatory molecules in LC induced by M-CSF, GM-CSF and TNF-a may be regulated by different mechanisms.
-92- Immunology(3) 9/5 Room-A
DENDRITIC CELLS IN CORNAL LIMBUS REDUCES WITH STORAGE TIME.
Naoshi Shinozaki, Shigeto Shimmura, Jun Shimazaki, Kazuo Tsubota.
Tokyo Dental College, Ichikawa General Hospital, Department of Ophthalmology, Chiba, Japan.
In cases of Stevens-Johnson syndrome or chemical burn, there is a high rate of recurrence after simple corneal transplantation resulting in decreased visual acuity. Due to the lack of corneal stem cells in these diseases, the cornea is unable to re-epithelialize. Limbal transplantation is the implantation of the donor corneal epithelial stem cells to the recipient. But the high density of dendritic cells in the limbus results in a higher rate of rejection after surgery. 36 cases of limbal transplantation were carried out at Corneal Transplant Center, Tokyo Dental College from 1994 to 1996; the rejection rate was only 30.6%, which is equivalent to that of simple corneal transplantation. In this study, we measured the density of dendritic cells in donor limbal tissue after enucleation, using whole-mount immunohistochemical technique with anti-S-100, anti-CD1a, anti-HLA-DR antibody. The results were 283.8¡Þ38.9¡¢112.5¡Þ25.3¡¢39.8¡Þ26.1¡¢8.5¡Þ6.2¡¢2.2¡Þ0.7¡¢0¡Þ0 mm2 after preservation in conventional corneo-screlal storage medium (Optisol GSR) for 0,1,2,3,5,7 days respectively. The number of immunoreactive dendritic cells decreases during storage time which suggests that the reduced number of antigen presenting cells in a graft can lead to a lower rate of rejection after limbal transplantation.
-93- Immunology(3) 9/5 Room-A
ONTOGENY OF TCR ab+ AND gd+ CELLS IN THE MURINE FETAL SKIN.
Koichiro Nakamura1, Nami Yasaka2, Chung Hsing Chang1, Masutaka Furue3, and Kunihiko Tamaki1.
1Department of Dermatology, University of Tokyo,2University of Teikyo,3University of Tokyo, Tokyo Branch Hospital, Tokyo, Japan.
The murine epidermis contains Thy-1+ dendritic epidrmal T cells expressing TCR Vg3 / Vd1. However TCR Vg2 or Vd4 + cells are identified in the epidermis of nude mice, suggesting extrathymic pathway into the skin. In order to examine the character of T cells in the murine fetal skin, we analyzed the population of T cells using FACS. TCR gd+ T cells and TCR ab+ T cells were identified in the fetal epidermis and dermis. TCR gd+ T cells represent TCR Vg3 and Vg2. At 16 fetal day the numbers of these T cells in the dermis reached at the peak and then declined in the new born mice. Immunohistochemical analysis of epidermal and dermal sheets also confirmed the existence TCR ab and TCR gd+ cells in the fetal skin. We presume that these T cells have the capacity to reside in the skin and further investigate the possibility of development of T cells in the fetal skin.
-94- Immunology(3) 9/5 Room-A
POSSIBLE INVOLVEMENT OF DENDRITIC EPIDERMAL T CELLS IN THE RESISTANCE TO CUTANEOUS GVHD IN EUTHYMIC MICE.
Katsuhiro Tomiyama, Masahiro Minagawa, Akiko Ito, Kazuhiro Kawai, Masaaki Ito.
Department of Dermatology, Niigata University School of Medicine, Niigata, Japan.
While experimental GVHD in euthymic (+/+) mice is easily induced, it is difficult to find cutaneous manifestation except for in the irradiated bone marrow chimeras. Previously, we have reported that athymic nude (nu/nu) mice transferred MHC class II-disparate T cells show systemic GVHD with obvious cutaneous lesions. The differential susceptibility to cutaneous GVHD between +/+ and nu/nu mice may be regulated by factors present in the skin. One candidate for such a factor is dendritic epidermal T cells (DETC) that reside in +/+ mice but not in nu/nu mice. To determine this possibility, systemic GVHD was induced and cutaneous lesions were examined in the recipient mice engrafted with the skin from +/+, nu/nu, or SCID mice. Skin grafts from nu/nu mice but not +/+ mice showed pathological findings compatible with GVHD. Interestingly, the skin grafts from SCID mice also showed cutaneous GVHD. These results demonstrated that susceptibility to cutaneous GVHD was dependent on cutaneous environment. Thus, DETC might provide the condition resistant to epidermal infiltration of allo-reactive T cells, possibly by regulating the expression of some molecules such as MHC class II on the surrounding keratinocytes.
-95- Immunology(3) 9/5 Room-A
EFFECT OF REPEATED ANTIGENIC STIMULATIONS ON CYTOKINE PROFILE OF REGIONAL LYMPH NODE CELLS IN MURINE CONTACT HYPERSENSITIVITY.
Masaru Natsuaki,Kayoko Abe,Yukio Kitano.
Department of Dermatology,Hyogo College of Medicine,Hyogo,Japan.
We investigated a cytokine profile of regional lymph node cells(LNC) in murine contact hypersensitivity(CH). BALB/c mice were sensitized with dinitrofluorobenzene(DNFB) and challenged repeatedly with DNFB. The regional LNC were cultured with a specific hapten and several cytokines in the culture supernatant were measured. IFN-¦Ã was mainly produced by LNC in sensitized mice. However, IL-10 as well as IFN-¦Ã was produced in repeatedly challenged mice. The result indicates that regional LNC in CH has a nature as Th1 and the LNC in repeatedly challenged mice has both Th1 and Th2 nature.
-96- Immunology(3) 9/5 Room-A
ANALYSIS OF CONTACT PHOTOSENSITIVITY REACTION ON MICE BY IMMUNOHISTOCHEMISTORY AND CYTOKINES.
Keiko Suzuki, Syunsuke Yamazaki, Rikako Miyake, Hiroshi Kakishima.
Cosmetics Laboratory, Kanebo Ltd., Kanagawa, Japan.
We were performed T cell participation of contact photosensitivity reaction (CPSR) and kinetics of cytokines using RT-PCR method.In this study,Balb/c mice (female,6-week-old) and tetrachlorosalicylanilide(TCSA) were used. We examined immunohistochemistory of T lymphocyte population and kinetics of cytokines by the RT-PCR method using skin tissues collected at regular intervals in the challenge phase. It was found that after the challenge on reaction sites, phenotype of lymph node cells were notable expressed. As an hour progress advances after the challenge ,IL-4,IL-6,IL-10 and IL-2 mRNA were observed. IL-4 mRNA was particular observed.T lymphocytes at the CPSR site were found to play important roles. These date suggested that the expression of IL-4 among several Th2 types of cytokines is especially important for the formation of CPSR.
-97- Immunology(3) 9/5 Room-A
UVA RADIATION MODULATES THE ANTIGEN-PRESENTING FUNCTION OF LANGERHANS CELLS IN VITRO.
Ichiro Iwai, Masato Hatao, Eiichirou Yagi, Kenji Yamaguchi, Masako Naganuma.
Shiseido Research Center 1, Yokohama, Japan.
Effects of UVB on immune functions of the skin have long been studied from various aspects and have shown to induce a local immune suppression at the irradiated site or systemic immune suppression depending on the amount of UV radiation. However, only a few studies have been reported regarding an immune suppression induced by UVA. In this study, we investigated an immune modulation of Langerhans cells (LC) by in vitro UVA radiation. Epidermal cell suspension prepared from C3H/HeN female mice was exposed to UVA. After pulsed with trinitrobenzene sulfonic acid, the suspension was co-cultured with T cells prepared from syngeneic mice sensitized with 1,3,5-trinitrochlorobenzene. An antigen presenting function was evaluated with 3[H]-methylthymidine incorporation by T cells in a mixed epidermal-cell lymphocyte reaction. As a result, UVA radiation dose-dependently suppressed the antigen presenting function of LC accompanied by inhibition of the surface expression of accessory molecules, ICAM-1, B7-1 and B7-2. It is suggested that UVA suppressed the antigen-presenting function of LC , at least partially, by affecting the synthetic pathway of co-stimulatory molecules.