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New immunosuppressive reagent, FTY 720, spares immunologic memory
New Immunosuppressive Reagent, FTY 720, Spares Immunologic Memory G. Lei, H. Amemiya, S. Suzuki, T. Goto, T. Kokubo, M. Miyamoto, and H. Kimura
I
T HAS been known for some time that when a weak minor histocompatibility antigen such as sex-linked minor histocompatibility antigen H-Y difference prevails, unless presensitization is performed, immune responses represented by rejection of male isograft skin in vivo or cell-mediated lympholysis (CML) in vitro are not observed. Thus, under appropriate sensitization protocols, immunologic memory verified by test skin isograft rejection or MHC-restricted CML reactivity, has been well established in H-Y incompatible syngeneic animal models. In this regard, it is quite interesting to determine whether FTY 720, a new immunosuppressive reagent that has been recognized as a unique reagent to render lymphocytes apoptosis, jeopardize acquired immunologic memory in the systems. We provide evidence that pretreatment of H-Y antigen sensitized animals with a single dose of FTY 720 (10 mg/kg) which is considered to be relatively high dose for clinical setting, spared an acquired memory.
MATERIALS AND METHODS ANIMALS Adult male and female rats that include LEW (RT11) as well as congeneic recombinant strain of LEW.1A (RT1a) and LEW.1W (RT1u) were originally obtained from Zentralinstitute fu ¨r Versuchsiterzucht (Hannover, Germany) and are maintained in our animal facility under the specific pathogen free.
intravenous injection. Following the sensitization (2 months later) H-Y antigen-sensitized recipients (females) were divided into two groups: group (I) were challenged with syngeneic male skin grafts, and, likewise, group (II) that had been presensitized with H-Y antigen by DC, were challenged with male isografts following FTY 720 (10 mg/kg/rat) treatment. The FTY 720 were given at 4 weeks after DC administration to sensitize the female rats.
RESULTS AND DISCUSSIONS
A strain of inbred female LEW rats accepted males isograft (29/32: acceptors/recipients mean survival time [MST] 300 days) without sensitization. However, when they were once sensitized toward H-Y antigen under the conditions, they restored an ability to reject subsequent H-Y incompatible skin isografts (2/18 MST: 15.4 ⫾ 5.3 days) (group I). This was also the case where the H-Y antigen-sensitized recipients were pretreated with immunosupressive reagent FTY 720 (group II 4/15 MST: 33.9 ⫾ 11.9 days). Thus a state of immunological memory appears to be well preserved following FTY 720 treatment. Our data suggested that FTY 720 renders naive rather than memory type of lymphocytes more apoptotic cell death. REFERENCES 1. Billingham RE, Silvers WK: Transplantation of Tissues and Cells. Philadelphia: The Wistar Institute Press, 1961 2. Miyagi T, Kimura H: Transplant Proc 27:1568, 1995
Cell Preparation Suspensions of spleen and BMC were prepared according to standard procedures.1
Dendritic Cell Preparation Dendritic cells (DC) were prepared as was described previously.2
Experimental Groups Inbred strain of female LEW (RT11) rats were either presensitized toward H-Y antigen employing 50 ⫻ 106 syngeneic male bone marrow cells (BMC) or 105 male BMC culture-derived DC by
0041-1345/00/$–see front matter PII S0041-1345(00)01443-3 1628
From the Department of Research Surgery, National Children’s MRC (G.L., H.A., S.S., H.K.), Medicinal Biology Research Laboratories, Fujisawa Pharmaceutical Co, Ltd (T.G.), Research Center, Bayer Yakuhin Ltd (T.K.). Supported by a Research Grant from the Ministry of Health and Welfare of Japan, a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan, and the Human Science Foundation of Japan. Address reprint requests to Hiromitsu Kimura, MD, Department of Research Surgery, National Children’s Medical Research Center, 3-35-31, Taishido, Setagaya-ku, Tokyo, 1548509 Japan.
© 2000 by Elsevier Science Inc. 655 Avenue of the Americas, New York, NY 10010 Transplantation Proceedings, 32, 1628 (2000)
I
T HAS been known for some time that when a weak minor histocompatibility antigen such as sex-linked minor histocompatibility antigen H-Y difference prevails, unless presensitization is performed, immune responses represented by rejection of male isograft skin in vivo or cell-mediated lympholysis (CML) in vitro are not observed. Thus, under appropriate sensitization protocols, immunologic memory verified by test skin isograft rejection or MHC-restricted CML reactivity, has been well established in H-Y incompatible syngeneic animal models. In this regard, it is quite interesting to determine whether FTY 720, a new immunosuppressive reagent that has been recognized as a unique reagent to render lymphocytes apoptosis, jeopardize acquired immunologic memory in the systems. We provide evidence that pretreatment of H-Y antigen sensitized animals with a single dose of FTY 720 (10 mg/kg) which is considered to be relatively high dose for clinical setting, spared an acquired memory.
MATERIALS AND METHODS ANIMALS Adult male and female rats that include LEW (RT11) as well as congeneic recombinant strain of LEW.1A (RT1a) and LEW.1W (RT1u) were originally obtained from Zentralinstitute fu ¨r Versuchsiterzucht (Hannover, Germany) and are maintained in our animal facility under the specific pathogen free.
intravenous injection. Following the sensitization (2 months later) H-Y antigen-sensitized recipients (females) were divided into two groups: group (I) were challenged with syngeneic male skin grafts, and, likewise, group (II) that had been presensitized with H-Y antigen by DC, were challenged with male isografts following FTY 720 (10 mg/kg/rat) treatment. The FTY 720 were given at 4 weeks after DC administration to sensitize the female rats.
RESULTS AND DISCUSSIONS
A strain of inbred female LEW rats accepted males isograft (29/32: acceptors/recipients mean survival time [MST] 300 days) without sensitization. However, when they were once sensitized toward H-Y antigen under the conditions, they restored an ability to reject subsequent H-Y incompatible skin isografts (2/18 MST: 15.4 ⫾ 5.3 days) (group I). This was also the case where the H-Y antigen-sensitized recipients were pretreated with immunosupressive reagent FTY 720 (group II 4/15 MST: 33.9 ⫾ 11.9 days). Thus a state of immunological memory appears to be well preserved following FTY 720 treatment. Our data suggested that FTY 720 renders naive rather than memory type of lymphocytes more apoptotic cell death. REFERENCES 1. Billingham RE, Silvers WK: Transplantation of Tissues and Cells. Philadelphia: The Wistar Institute Press, 1961 2. Miyagi T, Kimura H: Transplant Proc 27:1568, 1995
Cell Preparation Suspensions of spleen and BMC were prepared according to standard procedures.1
Dendritic Cell Preparation Dendritic cells (DC) were prepared as was described previously.2
Experimental Groups Inbred strain of female LEW (RT11) rats were either presensitized toward H-Y antigen employing 50 ⫻ 106 syngeneic male bone marrow cells (BMC) or 105 male BMC culture-derived DC by
0041-1345/00/$–see front matter PII S0041-1345(00)01443-3 1628
From the Department of Research Surgery, National Children’s MRC (G.L., H.A., S.S., H.K.), Medicinal Biology Research Laboratories, Fujisawa Pharmaceutical Co, Ltd (T.G.), Research Center, Bayer Yakuhin Ltd (T.K.). Supported by a Research Grant from the Ministry of Health and Welfare of Japan, a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan, and the Human Science Foundation of Japan. Address reprint requests to Hiromitsu Kimura, MD, Department of Research Surgery, National Children’s Medical Research Center, 3-35-31, Taishido, Setagaya-ku, Tokyo, 1548509 Japan.
© 2000 by Elsevier Science Inc. 655 Avenue of the Americas, New York, NY 10010 Transplantation Proceedings, 32, 1628 (2000)