Fung, JJ

Pan, F

Park, IY

Rao, AS

Starzl, TE

Sun, H

Tsugita, M

Valdivia, LA

Wakizaka, Y

PubMed

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Effect of Tacrolimus and Splenectomy on' Engraftment and GVHD After Bone Marrow Xenotransplantation in· the~ Reciprocal Hamster to Rat Animal Models F. Pan, M. Tsugita, A.S. Rao, Y. Wakizaka, H. Sun, I.Y. Park, J.J. Fung, T.E. Starzl, and L.A. Valdivia BONE marrow transplantation (BMTx) following lethal irradiation is an established modality to induce donor-specific tolerance in the rat to mouse xenogeneic combina-tion. 1 The induction of donor-specific tolerance in this model is accompanied by a distinct absence of graft-versus-host disease (GVHD). The latter outcome, however, is not witnessed when BMTx is performed in hamster to rat strain combinations in which, despite splenectomy (Spx), the recipients invariably succumb to lethal GVHO.2 The advent of tacrolimus (TCL. FK 506) as a potent immunosuppres-sive agent for the prevention and treatment of GVHO in allogeneic BMT~.4 prompted us to assess its efficacy on engraftment and GVHO after BMTx in the reciprocal hamster to rat xenograft combination. MATERIALS AND METHODS Animals. Syrian hamsters (100 to 120 g) and LEW rats (180 to 220 g) were used as donors and recipients. respectively. Bone Marrow Transplantation. LEW recipients were splenecto-mized 7 days before BMTx and injected with 3 x IO~ unfraction-ated hamster bone marrow cells (BMC) following irradiation (10.5 Gy). The hamster recipients received a similar quantity of LEW BMC and irradiation dose but were not subjected to Spx. Failure of BM to engraft was defined as recipient death within 14 days postinfusion without recovery of peripheral white blood cell count. Experimental groups are depicted in Table l. Tacrolimus. FK 506 (1 mgikg/d: 1M) was given daily to the recipients from days 0 to 30. DetectIOn of Chimensm. The presence of donor cells in the recipient's peripheral hlood was determined hy flow (.-ytometry \Ising LEW antihamster or hamster anti-LEW IgG antihodies. \\hich were prepared hy immunization of the appropriate animals with splenocytes. The IgG fraction was obtained by purification using gel chromatography. The primary antibody was biotinylated and PE-labeled streptavidin was used for its identification. GVHD. Oinica1 cutaneous and/or hepatic manifestations of GVHD were confirmed by histopathological examination and immunostaining. RESULTS AND DISCUSSION Failure of engraftment (FE) was witnessed in all rats preconditioned with irradiation followed by hamster BMTx (Table 1, group II) with the addition of TCL having no beneficial effect (group III). Interestingly the use of a similar regimen in the reciprocal rat to hamster strain combination resulted in much lower FE and markedly prolonged animal survival with no evidence of GVHO at day 30 (group VIII). Splenectomy prior to irradiation and BMTx in the hamster to rat model also resulted in a much lower FE but with obvious manifestations of cutaneous From the Pittsburgh Transplantation Institute and the Depart-ments of Surgery (F.P., A.S.R., Y.W., H.S .• l.Y.P., J.J.F., T.E.S •• L.A.V.) and Pathology (A.S.R.), University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; and the Institute of Gastroen-terology (F.P., A.S.R., Y.W .• H.S., l.Y.P., J.J.F .• T.E.S., L.A.V.), Tokyo Women's Medical College, Tokyo, Japan. Aided by Project Grant No. DK29961 from the National Insti-tutes of Health, Bethesda, MD. Address reprint requests to Luis A. Valdivia. MD. PhD. Pitts-burgh Transplantation Institute. E1546 Biomedical Science Tower. 200 Lothrop Street. Pittsburgh, PA 15213. '9 1996 by Appleton & Lange 0041-1345/96/$3.00/ + 0 Table 1. Failure of Engraftment (FE), GVHD. and Survival of Recipients After BMTx in the ReCiprocal Hamster to Rat Xenograft Combination Group Treatment n FE(%) A. Hamster to rat I Radiation (Rad)' alone 5 100 II Rad + BMTx* 6 100 III Rad + BMTx ~ TCl§ 7 100 IV Rad + BMTx + Spx 22 100 V Rad + BMTx T Spx + TCl 10 0 B. Rat to hamster N/A ~ not applicable . . Postoperative day. "0.5 Gy. VI Rad alone VII Rad ... BMTx VIII Rad + BMTx ,. TCl '3 '. '0' untracllOnated BM cells. ';, mg/kgld 1M: 0-30 days. 5 100 39 49 10 20 GVHD(%) Survival (days) (at POD' 30\ Max Median Min N/A 9 8 6 N/A 20 13 11 N/A 15 10 5 27 59 21 14 0 44 40 34 N/A 8 8 6 28 112 20 7 0 131 91 9 736 Transplantation Proceedings. Vol 28. No 2 (April), 1996: pp 736-737 EFFECT OF TACROLIMUS AND SPLENECTOMY GVHD (group IV). However, the administration of TCL to group IV animals resulted in complete abrogation of FE and of cutaneous GVHD, which markedly prolonged their survival. The animals in groups IV and V nevertheless succumbed to lethal GVHD, which was essentially limited to the liver. In splenectomized TeL-treated rats, which enjoyed mod-erately prolonged survival (group V) following hamster BMTx, 80% of peripheral blood cells at 14 days postinfu-sion were of hamster origin. Interestingly, reconstitution with hamster RBCs was only possible in animals in group V. When tested in CDC assays, hamster serum (unlike that from naive LEW rats) had no detectable titers of antirat preformed antibodies, which may provide an explanation for the relative ease with which engraftment could be achieved in the rat to hamster strain combination. Taken together, these data support earlier observations that nat-737 ural (preformed) antibodies are the main barrier to xeno-geneic bone marrow engraftment.5 Furthermore, Spx prior to BMTx across xenogeneic barriers enhances engraftment, an effect further supplemented by TCL, which also attenu-ates cutaneous (but not hepatic) manifestations of GVHD. REFERENCES 1. I1dstad ST, Wren SM, Boggs SS, et al: 1 Exp Med 174:467, 1991 2. Patijn GA, Valdivia LA, Pan F, et al: Transplantation 57: 1528,1994 3. Valdivia LA, Demetris AJ, Fung II, et al: Transplantation 55:659, 1993 4. Markus PM, Cai X, Ming W, et al: Transplantation 52:590, 1991 5. Aksentijevich I, Sachs DH, Sykes M: 1 Immuno1147:79, 1991 _ .. -,--_ .......... _------

Effect of tacrolimus and splenectomy on engraftment and GVHD after bone marrow xenotransplantation in the reciprocal hamster to rat animal models

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Effect of tacrolimus and splenectomy on engraftment and GVHD after bone marrow xenotransplantation in the reciprocal hamster to rat animal models

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