PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jcinvestThe Journal of Clinical Investigation
 
J Clin Invest. 2010 April 1; 120(4): 1036–1039.
Published online 2010 March 24. doi:  10.1172/JCI42532
PMCID: PMC2846071

Prevention trumps treatment of antibody-mediated transplant rejection

Abstract

Belying the spectacular success of solid organ transplantation and improvements in immunosuppressive therapy is the reality that long-term graft survival rates remain relatively unchanged, in large part due to chronic and insidious alloantibody-mediated graft injury. Half of heart transplant recipients develop chronic rejection within 10 years — a daunting statistic, particularly for young patients expecting to achieve longevity by enduring the rigors of a transplant. The current immunosuppressive pharmacopeia is relatively ineffective in preventing late alloantibody-associated chronic rejection. In this issue of the JCI, Kelishadi et al. report that preemptive deletion of B cells prior to heart transplantation in cynomolgus monkeys, in addition to conventional posttransplant immunosuppressive therapy with cyclosporine, markedly attenuated not only acute graft rejection but also alloantibody elaboration and chronic graft rejection. The success of this preemptive strike implies a central role for B cells in graft rejection, and this approach may help to delay or prevent chronic rejection after solid organ transplantation.

Acute and chronic rejection

Newly transplanted organs are susceptible within a week to acute rejection, mediated dominantly by T cells, but are usually effectively protected from this form of inflammation and injury by currently used immunosuppressive agents such as calcineurin inhibitors, antiproliferative agents, mTOR inhibitors, and prophylactic therapy with T cell–specific antibodies. When acute rejection occurs, as it does in 5%–25% of solid organ recipients within the first year, it can typically be successfully treated with steroid therapy or, if needed, T cell–specific antibodies. However, the current immunosuppressive pharmacopeia is relatively ineffective in preventing or treating rejection mediated by B cells and the antibodies they produce. Antibody-mediated allograft injury, which occurs in 50% of heart transplant patients within 10 years, typically manifests more than a year after transplantation, more insidiously than T cell–mediated injury, and in a process characterized by complement deposition and microvascular obliteration that leads to tissue ischemia and eventually fibrosis with loss of graft function. Chronic graft rejection refers to this antibody-mediated process.

While factors contributing to chronic injury of organ transplants are multiple and include ischemia/reperfusion injury, preexisting donor disease, drug toxicities, and recurrence of original disease, the subtle development in the graft recipient of antibodies specific for the foreign donor tissue (alloantibodies) in the months and years following organ transplantation has been shown to be an accurate predictor of graft failure (1, 2). The magnitude of this problem is compounded by the practical difficulties in designing feasible clinical trials to evaluate methods for preventing alloantibody development and by the paucity of proven strategies to prevent alloantibody development in large animal models or humans. Nevertheless, data suggest that if preexisting alloantibody levels can be reduced, the risk of graft loss is lower (3).

B cell depletion as treatment for established antibody-mediated rejection

In the medical literature, organ transplant patients experiencing antibody-mediated rejection have been treated with rituximab (a CD20-specific monoclonal antibody that depletes the B cell population) or by targeting of their plasma cells (antibody-secreting differentiated B cells), and in most cases these patients possessed preexisting alloantibody or suffered from early antibody-mediated rejection (4, 5). As expected, it is difficult to reverse the damage done by alloantibody in the setting of an established B cell immune response, and the efficacy of targeting B cells with rituximab under these posttransplant circumstances has been difficult to clearly establish. The combination of B cell depletion with profound T cell immunosuppression may also be complicated by loss of protective immunity (6). In other words, infection or malignancy may ensue, especially when both T cell– and B cell–depleting antibodies are administered simultaneously or sequentially. Therefore, an alternative strategy, that being prevention as opposed to treatment of the B cell alloimmune response, even if resorting to B cell depletion, may be attractive.

Preemptive B cell depletion

In their study in this issue of the JCI, Kelishadi et al. (7) show that preemptive treatment of cynomolgus monkeys transplanted with an allogeneic heart with rituximab on the day of the transplant substantially eliminated the injury attributable to B cells. In particular, infiltration of the graft by B cells was markedly reduced, as were intragraft levels of B cell–activating factor (BAFF; also known as B lymphocyte survival factor [BlyS]) and the B cell costimulatory molecules CD80 and CD86. In addition, the downstream effects of B cell activation were attenuated; for example, levels of alloantibody in the blood were reduced and less complement deposition in the graft was observed. Perhaps most importantly, these mechanistic changes were reflected by a substantial improvement in the microvascular integrity of the transplanted hearts (i.e., there was less chronic allograft vasculopathy) and by improved cardiac function, with four of four hearts beating well by 90 days compared with only three of seven in cynomolgus monkeys treated with cyclosporine alone. As seen from the control animals treated with cyclosporine alone, calcineurin inhibitors alone were unable to effectively prevent the injury inflicted by B cell–mediated rejection. Cotreatment with rituximab and cyclosporine also effectively prevented acute rejection compared with cyclosporine treatment alone, suggesting a role for B cells in acute rejection as well as chronic rejection.

Implications for human transplant patients

Therapeutic targeting of CD20 in transplantation may be appealing because of CD20’s stable expression primarily on B cells in the peripheral blood and its absence from plasma cells, pro-B cells, and hematopoietic stem cells, thus permitting maintenance of serum IgG levels and posttreatment recovery by spared pro-B and stem cells (8). For the same reasons, therapeutic targeting of CD20 may not be as effective in treating recipients known to have donor-specific alloantibody prior to transplantation, since memory B cells and plasma cells capable of producing antibody specific for the donor organ would already be primed. Since many T cell–mediated immune responses include a B cell component, the impact of B cell depletion may extend beyond suppression of measurable antibody (9), as is suggested by the observation in the current study that acute rejection was reduced from a 57% incidence in cynomolgus monkeys treated with cyclosporine alone to zero by addition of rituximab to the treatment regimen (7).

Nonhuman primate (NHP) models, such as the one used by Kelishadi et al. (7), are far closer, genetically, to the human condition than any rodent model might be, and thus the current report is expected to predict better than any rodent model of transplantation how humans might respond to B cell depletion. Nevertheless, it is worth noting that even observations in NHPs in the field of organ transplantation have sometimes been difficult to translate directly into the clinic (10, 11). By analogy, human heart transplant patients usually receive three or four simultaneous immunosuppressive agents to prevent T cell–mediated rejection, whereas the cynomolgus monkeys in the study by Kelishadi et al. received high-dose cyclosporine as their sole immunosuppressive agent (7). The applicability of the findings of the current study to human organ transplantation will therefore require rigorous testing in order to determine whether preemptive CD20 monoclonal antibody treatment in the setting of more intense T cell immunosuppression is accompanied by opportunistic infection.

Other B cell strategies for transplantation

Targeting B cell immunity without depleting these cells in order to prevent alloantibody development may also lead to opportunities to prevent allograft injury (Figure (Figure1).1). Such strategies include targeting complement pathway components (12) and B cell cytokines and/or chemokines such as BAFF and/or a proliferation-inducing ligand (APRIL), which may influence both B and T cell responses (13, 14). Other biologics being considered for development for the targeting of B cell responses in the setting of transplantation are those that affect the costimulatory pathways. Interactions between CD28 on CD4+ T cells and CD80/CD86 on B cells, as well as between CD40 ligand (CD40L; also known as CD154) on activated CD4+ T cells and CD40 on B cells have been shown to participate in providing T cell help to B cells (15). The CD40/CD40L interaction stimulates B cell proliferation and isotype switching in the appropriate cytokine milieu (16, 17). CD28 and CTLA4 expression have also been shown to be involved in germinal center formation (18).

Figure 1
B cell– and antibody-related biologics in transplantation.

Each of these potential therapies is under active investigation. It will be important to compare the relative safety and efficacy of such strategies with that of profound B cell depletion with rituximab. Additionally, it will be necessary to determine the durability and need for repeated application of B cell therapy in the setting of constant exposure to alloantigen, as is the case with an organ transplant. Nevertheless, the current report by Kelishadi et al. (7) offers clear experimental evidence in a large animal model that B cell targeting in parallel with T cell inhibition can prevent alloantibody development and lead to improved long-term graft histology and better small blood vessel patency. Prevention of chronic rejection would represent a major advance for the field of transplantation, and prevention of alloantibody development is more likely to succeed than are strategies to reverse ongoing antibody-mediated graft injury.

Acknowledgments

The authors are supported by NIH grant AI074635 (to S.J. Knechtle) and by Roche Organ Transplantation Research Foundation grant 962141545 (to N. Iwakoshi).

Footnotes

Conflict of interest: Stuart J. Knechtle acknowledges stock ownership in Bristol-Myers Squibb.

Citation for this article: J Clin Invest. 2010;120(4):1036–1309. doi:10.1172/JCI42532

See the related article beginning on page 1275.

References

1. Terasaki PI. Humoral theory of transplantation. Am J Transplant. 2003;3(6):665–673. doi: 10.1034/j.1600-6143.2003.00135.x. [PubMed] [Cross Ref]
2. Terasaki PI, Cai J. Human leukocyte antigen antibodies and chronic rejection: from association to causation. Transplantation. 2008;86(3):377–383. [PubMed]
3. Everly MJ, et al. Reducing de novo donor-specific antibody levels during acute rejection diminishes renal allograft loss. Am J Transplant. 2009;9(5):1063–1071. doi: 10.1111/j.1600-6143.2009.02577.x. [PubMed] [Cross Ref]
4. Ramos EJ, et al. The effect of desensitization protocols on human splenic B-cell populations in vivo. Am J Transplant. 2007;7(2):402–407. doi: 10.1111/j.1600-6143.2006.01632.x. [PubMed] [Cross Ref]
5. Zarkhin V, et al. A randomized, prospective trial of rituximab for acute rejection in pediatric renal transplantation. Am J Transplant. 2008;8(12):2607–2617. doi: 10.1111/j.1600-6143.2008.02411.x. [PubMed] [Cross Ref]
6. Kamar N, et al. Incidence and predictive factors for infectious disease after rituximab therapy in kidney-transplant patients. Am J Transplant. 2010;10(1):89–98. [PubMed]
7. Kelishadi SS, et al. Preemptive CD20+ B cell depletion attenuates cardiac allograft vasculopathy in cyclosporine-treated monkeys. . J Clin Invest. 2010;120(4):1275–1284. [PMC free article] [PubMed]
8. Teeling JL, et al. The biological activity of human CD20 monoclonal antibodies is linked to unique epitopes on CD20. J Immunol. 2006;177(1):362–371. [PubMed]
9. Pescovitz MD, et al. Rituximab, B-lymphocyte depletion, and preservation of beta-cell function. N Engl J Med. 2009;361(22):2143–2152. doi: 10.1056/NEJMoa0904452. [PubMed] [Cross Ref]
10. Hale DA, Dhanireddy K, Bruno D, Kirk AD. Induction of transplantation tolerance in non-human primate preclinical models. Philos Trans R Soc Lond B Biol Sci. 2005;360(1461):1723–1737. [PMC free article] [PubMed]
11. Knechtle SJ, Hamawy MM, Hu H, Fechner JH, Jr, Cho CS. Tolerance and near-tolerance strategies in monkeys and their application to human renal transplantation. Immunol Rev. 2001;183:205–213. doi: 10.1034/j.1600-065x.2001.1830116.x. [PubMed] [Cross Ref]
12. Sacks S, Lee Q, Wong W, Zhou W. The role of complement in regulating the alloresponse. Curr Opin Organ Transplant. 2009;14(1):10–15. [PubMed]
13. Walters S, et al. Increased CD4+Foxp3+ T cells in BAFF-transgenic mice suppress T cell effector responses. J Immunol. 2009;182(2):793–801. [PubMed]
14. Bloom D, et al. BAFF is increased in renal transplant patients following treatment with alemtuzumab. Am J Transplant. 2009;9(8):1835–1845. doi: 10.1111/j.1600-6143.2009.02710.x. [PubMed] [Cross Ref]
15. Larsen CP, et al. Rational development of LEA29Y (belatacept), a high-affinity variant of CTLA4-Ig with potent immunosuppressive properties. Am J Transplant. 2005;5(3):443–453. [PubMed]
16. Li Y, Ma L, Yin D, Shen J, Chong AS. Long-term control of alloreactive B cell responses by the suppression of T cell help. J Immunol. 2008;180(9):6077–6084. [PMC free article] [PubMed]
17. Quezada SA, Jarvinen LZ, Lind EF, Noelle RJ. CD40/CD154 interactions at the interface of tolerance and immunity. Annu Rev Immunol. 2004;22:307–328. doi: 10.1146/annurev.immunol.22.012703.104533. [PubMed] [Cross Ref]
18. Ferguson SE, Han S, Kelsoe G, Thompson CB. CD28 is required for germinal center formation. . J Immunol. 1996;156(12):4576–4581. [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation