Banas, R

Demetris, AJ

Dodson, F

Fontes, P

Fung, J

Jordan, M

Lombardozzi, S

Pavlick, M

Pham, S

Rao, AS

Shapiro, R

Starzl, TE

Trucco, M

Zeevi, A

English

PubMed

Name not available

Serial evaluation of immune profiles of simultaneous bone marrow and whole organ transplant recipients

D-Scholarship@Pitt

·Serial Evaluation of Immune Profiles of Simultaneous Bone Marrow and Whole Organ Transplant Recipients A. Zeevi. M. Pavlick. S. Lombardozzi. R. Banas. A.S. Rao. P. Fontes. A.J. Demetris. R. Shapiro. \ n S/0 M. Jordan. F. Dodson. S. Pham. M. Trucco. J. Fung, and T.E. Starzl • I BONE MARROW (BM) augmentation of donor-derived chimerism was achieved in recipients of kid-ney. liver. heart. and pancreas islet transplantation. l We have proposed that these donor-derived cells are essential for graft acceptance and the induction of donor-specific nonreactivity.2.3 The first 15 BM-augmented transplant recipients and 16 nonmarrow controls who were more than 120 days posttransplant underwent sequential in vitro im-munological evaluations to determine the development of donor-specific hyporeactivity.4 Based on proliferative re-sponses (pre- and posttransplant) of peripheral blood mononuclear cells (PBMCs) to mitogens and alloantigens. the recipients were classified into four categories: donor-specific hyporeactive. intermediate. responsive. and sup-pressed.4 Fifty-three percent of BM treated recipients (eight of 15) exhibited progressive modulation of antidonor responses. whereas only 12% of nonmarrow controls showed donor-specific hyporeactivity. However. the fol-low-up time in the nonmarrow control group was shorter (112 ::: 10 days) as compared with that of patients in the study group (282 ::t: 113 days).4 In this report. we reanalyzed the immune profile of BM-treated and nonmarrow controls 3 months later. and the results were compared with the earlier report. METHODS Case Material Since December 1992. 64 patients have received simultaneous 8M-positive whole-organ transplants. whereas 53 recipients of whole organs alone have been monitored as contemporaneous controls. However. the present study included 17 of 19 BM-augmented recipients (kidneys. n = 6: kidney plus islets. n = 2; livers. n = 7; liver plus islets. n = 1: and heart. n = 1) and 22 of 25 nonmarrow controls (kidneys. n = 6; livers. n = 14: and hearts, n .. 2) who were more than 120 days posttransplant and in whom in vitro immune monitoring was possible. Immunosuppression was aimilar in both groups and consisted of FK 506 and prednisone routinely used at our center. I Using HLA-specific markers and probes for Y chromosome (in male to female transplant recipi-ents), all patients were examined for donor chimerism by fluores-cence-activated cell sorting (FACS). fluorescence in situ hybridiza-tion (FISH), and polymerase chain reaction (PCR).I In Vitro Immune Monitoring Recipients' PBMCs were prepared via a method described previ-ously, were tested for proliferative responses induced by mitogens (phytohemagglutinin [PHAI and concanavalin A [Con A» and by aIIoantigens (donor and third-party panel cells) in mixed lympho-Cyte reaction (MLR) assays, the details of which were summarized eiscwhere.5." . Based on the proliferative responses of recipients' PBMCs pre-and posttransplantation. we have classified them into four catego-ries." Category r. Donor-specific hyporeactive: significant decrease (over 70%) in posttransplant versus pre transplant donor-specific MLR responses while maintaining adequate reactivity to control third party stimulator cells and mitogens. Category II. Donor-specific intermediate: reduction (40% to 70%) in posttransplant donor-specific alloreactivity as compared with pre transplant baseline control. with no attenuation of respon-siveness to third-party stimulators or mitogens. Category III. Reactive: minimal change in pre transplant versus posttransplant donor-specific reactivity. Category IV. Suppressed: global nonreactivity to all in vitro stimuli. RESULTS Distribution of Donor-Specific MLR Patterns in 8M-Augmented and Control Transplant Recipients The overall percentage of donor-specific hyporeactive/in-termediate responders in the BM-augmented and control groups remained similar to that reported 3 months earlier (53% and 12% versus 41 % and 18%. respectively) (Table 1). In the current analysis. five of eight BM/liver recipients (62%) exhibited modulation of donor-specific reactivity (four hyporeactive and one intermediate), whereas in the control group only three of 14 patients (21%) showed a reduction in antidonor reactivity (three intermediate). BMI kidney and kidney control recipients had a similar distribu-tion of donor-specific nonreactivity (25% and 16%, respec-tively). Furthermore. all control recipients who previously exhibited global nonreactivity (kidneys. n = 2; livers. n = 2) regained their in vitro immune responsiveness and became reactive to donor cells (Table 1). From the Pittsburgh Transplantation Institute and the Depart-ments of Pathology (A.Z .• M.P .. A.B .• A.S.A., A.J.D.), Pediatrics (S.L., M.T.), and Surgery (A.S.A.. P.F .• R.S., M.J .• F.D., S.P .• J.F., T.E.S.I, University of Pittsburgh Medical Center. Pittsburgh, Pennsylvania. Supported by Project Grant No. DK 29961 from National Institutes of Health. Bethesda, Maryland. and the Pathology Education and Research Fund. Address reprint requests to Adriana Zeevi, Ph.D., Department of Pathology, Biomedical Science Tower. Room W1551, Univer-sity of Pittsburgh, Pittsburgh, PA 15261. © 1995 by Appleton & Lange 0041-1345/95/$3.00/ + 0 213 . Tflllsplantation Proceedings. Vol 27. No 1 (February). 1995: pp 213-215 214 ZEEVI. PAVUCK. LOMBARDOZZI ET Al Table 1. Donor-8pecific MLR Patterns in BM-Augmented and Control Transplant Recipients Report Paned April 1994 July 1994 Groups (POD:!: SO)" nt With 8M 15 (150 :: 60) Without 8M 16 (112 :: 10) With 8M 17 (2B5 = 119) Without 8M 22 (153 = 74) 'Postoperatlve day when last sample was tested. t Number of recIpIents per group. Hyporeactlve 20 (1K.2l)t 12 (1K.1l)i 29 (1 K. 4l)i 0 iNumber of transplant recIpIents for each organ: K. kidney; L hver. H. heart. Immune Reactivity. Immunosuppression, and Chimerism As shown in Tables 2 and 3, the number of patients off steroids was higher in the kidney and liver (66% and 75%, respectively) transplant recipients who also exhibited di-minished donor-specific MLR responses than in the donor-reactive patients (0% and 43% respectively) in both BM-augmented and nonaugmented groups. Chimerism in the BM-augmented group was detectable by flow cytometry for up to 18 months after transplantation and ranged between 0.5% and 2.6% in both the hyporeac-tive and donor-reactive recipients. whereas in the majority of the control recipients detection of donor cells by FACS analysis yielded negative results 30 to 60 days posttransplant (donor-derived cells accounted for less than 0.5%) (Tables 2 and 3). Shift in Donor-Specific Reactivity Four BM-treated recipients (two livers. one with pancreatic islets; and two kidneys. one with islets) who exhibited early evidence of donor-specific hyporeactive or intermediate reactivity reverted into reactive against the donor when last tested. In three of four patients. we also have evidence of an increase (three- to lO-fold) in circulating donor-specific helper T cells (data not shown). All four recipients experi-enced late rejection episodes. Heightened donor-specific Table 2. Correlation Between Immune Reactivity, Immunosuppression, and Chimerism: Kidney Transplant Recipients Immunosuppression Chimensm (%) MLR Reactivity Profile FK Level (ng/mL) Hyporesponslve/intermediate 01! PrednIsone Study (n = 2) 0.1, O.B 1/2 Control (n = 1) 0.9 1/1 ResponSive Study (n = 6) Control (n = 5) 1.0 = 0.5 0.9:!: 0.4 'Level of circulating dOnot' cells. T Percentage of patIents positIve. NF. not feeslble. 0/6 0/5 FACS' PCRt 1.9,0.5 100 0.83 NF 0.5-2.6 100 <0.5 60 DistributIon of MLR Profiles (%) IntermedIate ReactIve SuppreSSed 33 47 0 (2K 3l)t (4K. 2L. 1 H)t 0 63 25 (2K.8l)t (2K.2l)i 12 59 0 (1 K. 1L)t (6K. 3L. 1 H)t 1B B2 0 (1 K. 3L)t (5K. 11 L. 2H)t reactivity was maintained in three of four (liver plus islet. n = 1; kidney, n = 1; and kidney plus islet. n = 1) recipients after resolution of their ongoing rejection. whereas one liver patient reverted to being hyporeactive to the donor. The kidney recipient who was treated for recurrent latl: rejection episode exhibited antidonor alloreactivity before initiation of antirejection and after steroid recycle. The specificity of this response was confirmed when lymphocyte cultures propagated from pre- and posttherapy samples in bulk MLRs also exhibited vigorous proliferation to the donor-mismatched HLA class II DR! antigen (data not shown). Furthermore. the prerejection lymphocyte culture exhibited cell-mediated lymphocylolOXicity against donor lymphoblastoid B cells (data not shown). DISCUSSION The recognition that after whole-organ transplantation resident bone-marrow derived cells migrate out of the graft into the recipient and persist has led to the concept that establishment of chimerism is a seminal event in allograft acceptance and subsequent induction of donor-specific hyporeactivity. :.3 This spontaneous chimerism was safely augmented by simultaneous infusion of bone-marrow along with whole-organ transplants. I The in vitro immune reac-tivity profile of patients in the BM-augmented group was reminiscent of earlier observations in recipients of long-Table 3. Correlation Between Immune Reactivity, Immunosupprassion, and Chimerism: Liver Transplant Recipients ImmunosuppressIon ChimerIsm % MLR Reactivity FK Level 01! Profile (nglmL) Prednisone FACS' PCRt Hyporesponslvelintermediate Study (n = 5) 0.4 = 0.3 3/5 0.5-2.5 100 Control (n = 3) 0.4=0.1 3/3 NF 66 Responsive Study (n = 3) 0.3 = 0.1 1/3 0.5-2.3 100 Control (n = 11) 0.7 = 0.2 5/11 1.3. <0.5 56 'Level of circulating donor cells. 'Percentage of patIents posItIve. NF. not feasible. J islet, Jients . one oor. t late dore The ocyte les in I the I not Iiture lonor ation graft : that 19raft ecific ;afely. along reac-lWas long-1% 100 66 100 56 SERIAL EVALUATION OF IMMUNE PROFILES term functioning allografts who exhibited donor-specific hyporeactivity.7-" However. in the 8M-treated recipients these changes occurred more frequently and earlier than in Ihe contemporaneous controls. 4 \on!!cr follow-up of both 8M- and non-marrow-treated Iln:r recipients demonstrated that the 8M augmentation ILl' conhrmed an advantage because five of eight (62'i~) pro!!ressed to stable donor-specific hypo reactive status, whereas only 21 'ic of recipients of liver allograft alone exhihited donor-specific intermediate responses. These re-sult~ suggest that the inherent tolerogenecity of liver ai-1(J~rafts can be further enhanced by simultaneous 8M infusion. I n contrast to liver recipients, BM-enriched and control kidney rccipients had a similar pattern of donor-specific hyporeactivity that was maintained over the course of our follow-up. Nevertheless. the graft function of marrow-au!!mented kidney recipients was significantly better as l'Ompared with that of the controls. lo OJ nole is that none of the BM-treated or control rl'l'lpil'nts when last tested exhibited universal nonreactivity ;lIld all of the previous recipients in the control group who II L'rL' classified as suppressed reverted to become donor reactIve. 4 In summary. BM infusion with whole-organ transplanta-215 tion proved to be a safe procedure to augment donor chimerism in transplant recipients. l Serial immunologic examinations of these recipients may help us \0 better understand the role of both immune systems (donor and recipient) in establishing allograft acceptance and lolcr-ance. REFERENCES 1. Fontes P. Rao AS. Demetris AJ. ct a1: Lancet :-44: 151. 1 '1'14 2. Starzl TE. Demetris AJ. Murase N. ct al: Lancet 339:1579. 1992 3. Starzl TE. Dcmetris AJ. Trucco M. ct a1: Hep.nologv 17:11~7. 1993 4. Zeevi A. Pavlick M. Lombardozzi S. ct <11: Tran,plantation (in press) 5. Eiras G, Shimizu Y. Seventcr GA. ct al: Transplant Prot' 23:236. 1991 6. Zccvi A. Venkataramanan R. Burckart G. ct al: Ilum Immu-nol 21:143. 1981' 7. Thomas J. Thoma, F. MendeZ-Picon 1. ct al: Sur!!cry 1\ 1 :12:;. 1977 S. Reinsmoen NL. Kaufman D. Matas A. et al: Transplantation 50:783. 1990 9. Reinsmoen NL. Matas AJ: Transplantation 55:]017. 1993 10. Rao A. Fontes P. Zeevi A. ct al: Transplant Proc (in press) 

http://d-scholarship.pitt.edu/5141/1/31735062126127.pdf

Serial evaluation of immune profiles of simultaneous bone marrow and whole organ transplant recipients

Abstract

Similar works

Full text

Available Versions

Name not available

D-Scholarship@Pitt

Name not available