Cyclosporine and methylprednisolone for prophylaxis of acute graft-versus-host disease

Twenty-eight consecutive HLA matched patients undergoing allogeneic bone marrow transplantation received prophylaxis for acute graft-versus-host disease with combined cyclosporine and methylprednisolone. The incidence of grades II-IV acute GVHD was 28.5%, a figure similar to that reported for two other drug regimens. The incidence of chronic GVHD in patients surviving longer than 150 days was 73%. Toxicity, especially renal, was acceptable and a number of potential problems associated with the use of methotrexate were avoided. While this regimen and similar ones have reduced the incidence and severity of acute GVHD the problem remains formidable and newer approaches are clearly needed.     

ATG plus corticosteroid therapy for acute graft-versus-host disease: predictors of response and survival

 Innovative treatment strategies for acute graft-versus-host disease (aGVHD) have not replaced corticosteroids as the primary therapy. We retrospectively reviewed 74 patients who received equine antithymocyte globulin (ATG) in addition to corticosteroids as therapy for GVHD, 21 who received primary therapy and 53 who received ATG after progressing or failing to improve with corticosteroids alone. The groups were comparable in clinical characteristics and in timing and severity of GVHD. After primary therapy with ATG 67% of patients' GVHD symptoms were stable or improved by 28 days versus 56% in those receiving secondary ATG (p=0.57). In univariate analysis the absence of multiple organ, GI, and liver aGVHD and a clinical stage score ≤4 were predictive of a favorable response, while in a multivariate logistic regression model only a clinical stage score ≤4 was independently associated with a favorable response (odds ratio 0.08, 95% CI 0.02–0.32, p=0.003). ATG response rates and 6-month survival (38 vs. 40%, p=0.89) were similar following primary and secondary ATG. Patients stable or improved 28 days after ATG therapy had a significantly better 6-month survival than those whose aGVHD had progressed (50 vs. 30%, p=0.02). Further study is required to assess whether some initial presentations of aGVHD would predictably fail corticosteroid therapy and may thus suggest a role for ATG in the primary management of aGVHD. For this determination, formal prospective comparative trials are needed. Received: 11 March 1997/Accepted: 16 June 1997     

Immunobiology of acute graft-versus-host disease

Graft-versus-host disease (GVHD) has been the primary limitation to the wider application of allogeneic bone marrow transplantation (BMT). The immunobiology of acute GVHD is complex and can be conceptualized to be a three-step process. In step 1, the conditioning regimen (irradiation and/or chemotherapy) leads to the damage and activation of host tissues and induces the secretion of inflammatory cytokines TNF-alpha and IL-1. As a consequence expression of MHC antigens and adhesion molecules is increased, thus enhancing the recognition of host alloantigens by donor T cells. Donor T-cell activation in step 2 is characterized by donor T-cell interaction with host APCs and subsequent proliferation, differentiation, and secretion of cytokines. Cytokines such as IL-2 and IFN-gamma enhance T-cell expansion, induce cytotoxic T cells (CTL) and natural killer (NK) cell responses, and prime additional mononuclear phagocytes to produce TNF-alpha and IL-1. These inflammatory cytokines in turn stimulate production of inflammatory chemokines, thus recruiting effector cells into target organs. In step 3, effector functions of mononuclear phagocytes are triggered via a secondary signal provided by lipopolysaccharide (LPS) that leaks through the intestinal mucosa damaged during step 1. This mechanism may result in the amplification of local tissue injury and further promotion of an inflammatory response, which, together with the CTL and NK components, leads to target tissue destruction in the transplant host.     

Management of acute graft-versus-host disease

Current graft-versus-host disease (GVHD) prophylaxis is not uniformly successful. Since the development of GVHD has a profound impact on transplant success, treatment is required. The mainstay of therapy has been glucocorticoids (steroids), along with anti-thymocyte globulin, cyclosporine, monoclonal antibodies, and aggressive supportive care, resulting in response rates of 30-50%, dependent upon severity of the disease, type of transplant, underlying diagnosis, prophylaxis given and other factors. Dose intensification or combinations of agents may increase response rates (60-80%), however, without necessarily improving survival, due to intervening complications. Nevertheless, patients with complete responses have a higher probability of survival, and a subgroup of patients will not develop chronic GVHD. As our understanding of cell recruitment, cell interactions and cytokine networks improves, new strategies are likely to be developed. The sequential use of antibodies directed at different cell populations or cytokines, along with anti-inflammatory measures, may be helpful. Prevention of GVHD has to remain our objective; however, efforts at therapy must continue until that goal is achieved.     

Management of acute graft-versus-host disease

Acute graft-versus-host disease (GvHD) is a frequent complication of allogeneic haemopoietic stem cell transplantation (HSCT) and donor lymphocyte infusions (DLI). Its incidence and severity depends on several factors, such as prophylaxis method, donor/recipient matching, intensity of the conditioning regimen and composition of the graft. Significant progress has been made in recent years in understanding the pathogenesis of the disease, and some of these advances have been translated into clinical trials. First-line treatment of acute GvHD is based on corticosteroids, and produce sustained responses in 50-80% of patients depending on the initial severity. Non-responders are offered second-line therapy, with combinations of immunosuppressive agents, but 1-year survival is 30% in most large trials. New strategies explored include infusion of expanded mesenchymal stem cells (MSC), down regulation of antigen-presenting cells (APC) and suicide gene transduced T cells. Acute GvHD is complicated by severe immunodeficiency causing life-threatening infections. To date, GvHD has not been differentiated from the graft-versus-leukaemia effect. The present review will discuss some of these aspects.     

Cyclosporine and methylprednisolone after allogeneic marrow transplantation: association between low cyclosporine concentration and risk of acute graft-versus-host disease

A combination of cyclosporine (CSA) and methylprednisolone (MP) was used as graft-versus-host disease (GVHD) prophylaxis in 25 patients age 11-47 years (median 27 years) who received HLA-compatible sibling marrow transplants after myeloablative therapy for leukemia, myelodysplasia or lymphoma. CSA was initiated at 3 mg/kg/day in two divided doses, and the dose was adjusted to maintain a trough whole blood h.p.l.c. concentration between 200 and 800 ng/ml. While on i.v. CSA, the dose of CSA was increased for 10 of the 25 patients. The actuarial rate of grades II-IV acute GVHD was 37%. Those patients who developed moderate to severe GVHD had a significantly higher early mortality than those who did not (56% vs 12%, p = 0.02). There was a significant association between the development of acute GVHD and a mean week 2 CSA trough concentration less than 250 ng/ml. Life threatening regimen-related toxicities in the first 100 days included capillary leak syndrome, acute pancreatitis and small bowel perforation. Although the combination of CSA and MP in this dosing schedule was active in preventing acute GVHD, nephrotoxicity remained a problem, and outcome was limited by the inability to achieve the target CSA trough concentration in a substantial proportion of patients.     

Diagnosis and management of acute graft-versus-host disease

A joint working group established by the Haemato-oncology subgroup of the British Committee for Standards in Haematology (BCSH) and the British Society for Bone Marrow Transplantation (BSBMT) has reviewed the available literature and made recommendations for the diagnosis and management of acute graft-versus-host disease. This guideline includes recommendations for the diagnosis and grading of acute graft-versus-host disease as well as primary treatment and options for patients with steroid-refractory disease. The goal of treatment should be effective control of graft-versus-host disease while minimizing risk of toxicity and relapse.     

Prophylaxis and treatment of acute graft-versus-host disease

Acute graft-versus-host disease (GVHD) remains a major obstacle to successful allogeneic hematopoietic stem cell transplantation (HSCT). The ability to prevent GVHD--the application of successful prophylaxis--is crucial as treatment when prophylaxis fails or remains suboptimal. A calcineurin inhibitor in combination with methotrexate is still the mainstream regimen for prophylaxis of GVHD. Despite a steady increase in the repertoire of available drugs, corticosteroids remain the first-line therapy for patients who fail prevention and develop GVHD. Pan T-cell depletion studies suggest that success in prophylaxis and treatment of GVHD will depend on whether GVHD can be prevented without losing anti-malignancy and anti-infectious effects. Better understanding of the allogeneic response that is responsible for GVHD will facilitate the development of such an approach.     

Treatment of gastrointestinal acute graft-versus-host disease

Opinion statement Therapy of acute graft-versus-host disease (GVHD) aims to selectively alter the grafthost interactions to foster antitumor effect and minimize antihost effects. The immunosuppression produced by the various therapies ranges from broad, nonselective effects to relatively narrow targeted impact. Despite advances in understanding the pathophysiology of GVHD, newer agents with more selective effects have not yet produced therapeutic advances. The newer targeted agents continue to produce a degree of immunosuppression in which infection and relapse of malignancy are all too common. High-dose systemic steroids remain, as they have for two decades, the initial treatment of choice. Patients failing to respond to steroids continue to represent a challenge, as no second-line therapy is clearly superior to the others. However, some of the new agents appear to be particularly effective in certain organs involved with acute GVHD. For those patients with steroid-refractory GVHD involving primarily the gut, we favor infliximab with concomitant antifungal therapy. For those with primarily skin or liver disease, we favor extracorporeal photochemotherapy.     

Cytokine dysregulation and acute graft-versus-host disease.

We suggest that acute GVHD after marrow transplantation reflects (1) host injury due to the conditioning regimen followed by the production of inflammatory cytokines; (2) stimulation of mature donor T cells in the milieu of increased cell surface expression of leukocyte adhesion molecules and HLA molecules, followed by the autocrine production of IL-2; and, finally, (3) recruitment and activation of additional mononuclear effector cells from donor marrow progenitors, which produce additional inflammatory cytokines, thus sustaining the response. The second step is critical for the amplification of the systemic inflammatory response, and it is absence in autologous, syngeneic, and T-cell-depleted transplants. These T cells may also contribute to the inflammatory cytokine network. Acute GVHD can occur in the absence of primary tissue injury in such settings as transfusion-related GVHD; however, it is likely that a greater HLA disparity between donor and host is required. We propose that inflammatory cytokine production is the final common pathway of acute GVHD. If this model is correct, control of cytokine dysregulation at any of several points should control GVHD. Further studies of GVHD and investigations of cytokine antagonists (eg, IL-4 or IL-10) or combinations of antagonists such as IL-1ra and soluble TNF receptor or pentoxifylline will allow us to determine the validity of this hypothesis.     

G-CSF-treated granulocytes inhibit acute graft-versus-host disease

It has been shown that in vivo and in vitro treatment with G-CSF induces the generation of low-density granulocytes (LDGs), which copurify with PBMCs and inhibit IFN-gamma production by human T cells. These results prompted us to postulate an immunomodulatory role for LDGs in acute graft-versus-host disease (aGVHD). Here it is shown that in the mouse experimental model, in vivo and in vitro G-CSF treatment generates LDGs capable of inhibiting 80% of T-cell IFN-gamma production. To assess the role of these LDGs in aGVHD, lethally irradiated (C57BL/6 x BALB/c) F1 hosts were reconstituted with T cell-depleted bone marrow cells plus nylon wool-purified spleen cells from G-CSF-treated (G-NWS) or -nontreated (NWS) C57BL/6 donors. Recipients of G-NWS had a 75% survival rate in contrast to a rate of 25% in the NWS recipients. The protective effect was completely abolished, and the mortality rate was 100% if donor-cell infusion was treated with anti-Gr1. Moreover, if LDGs were infused with NWS, full protection of aGVHD was observed, and no signs of disease were evidenced by mortality rate, weight loss, or histopathology of target organs. These results revealed the unexpected immunosuppressive capacity of G-CSF based on the generation of LDGs, leading to the possibility of using these cells as inhibitors of aGVHD.     

Risk factors for acute graft-versus-host disease

Acute graft-versus-host disease (GvHD) is an important complication of bone marrow transplantation in humans. Risk factors are imprecisely defined and controversial. We analysed data from 2036 recipients of HLA-identical sibling transplants for leukaemia or aplastic anaemia to identify risk factors for GvHD. Analyses indicate that grading of GvHD can be reproducibly divided into absent or mild versus moderate to severe; 2-year actuarial probability was 54% (95% confidence interval 52-56%) for absent or mild and 46% (44-48%) for moderate to severe. Factors predictive of development of moderate to severe GvHD include donor/recipient sex-match (female----male greater than others, relative risk 2.0, P less than 0.001). This risk was markedly increased if female donors for male recipients were previously pregnant or transfused (relative risk 2.9, P less than 0.0001). Older patients were at increased risk of GvHD (relative risk 1.6, P less than 0.001), but the age gradient was modest, even the youngest patients had a substantial risk of GvHD and, if parous or transfused female----male transplants were excluded, age was not a significant risk factor. Cyclosporine or methotrexate were equally effective at preventing GvHD and were superior to no prophylaxis (relative risk 2.3, P less than 0.01). These data should be useful in estimating the risk of acute GvHD in an individual patient and in designing clinical trials to investigate methods to modify or prevent GvHD.     

Cyclosporine as prophylaxis for graft-versus-host disease: a randomized study in patients undergoing marrow transplantation for acute nonlymphoblastic leukemia

Seventy-five patients, 13 to 49 years of age, with acute nonlymphoblastic leukemia in first remission were treated with cyclophosphamide, fractionated total body irradiation, and marrow transplantation from an HLA-identical sibling and randomized to receive either cyclosporine (CSP) (n = 36) or methotrexate (MTX) (n = 39) as prophylaxis for graft-v-host disease (GVHD). All patients engrafted, and 22 who were given CSP and 21 who were given MTX, are alive at 20 to 47 (median, 35) months (P = .5). Engraftment as assessed by granulocyte recovery (P less than .0005) and platelet transfusion requirement (P = .01) was faster in patients on CSP. Twelve patients (33%) on CSP and 22 (56%) on MTX developed acute GVHD of grades II through IV (P = .07) and 15 of 30 on CSP and 14 of 32 on MTX that were at risk developed chronic GVHD. The most frequent causes of death were interstitial pneumonitis and marrow relapse of leukemia, which occurred with similar frequency in both groups. Beneficial effects observed in patients on CSP included less severe mucositis and shorter duration of hospitalization; adverse effects included renal function impairment and hypertension. These data confirm that CSP is a useful immunosuppressant in patients undergoing marrow transplantation but fail to show a significant improvement in survival as compared with the standard regimen of MTX.     

The clinical spectrum of acute graft-versus-host disease

Acute graft-versus-host disease (GVHD), initiated by the reaction of donor T lymphocytes against nonshared recipient antigens, typically leads to a clinical syndrome characterized by cutaneous eruptions and intestinal and hepatic dysfunction. These three organ systems are considered in the clinical grading of acute GVHD. However, other targets may be involved. With conventional transplant conditioning regimens and in vivo prophylaxis, GVHD becomes clinically manifest within 2 to 4 weeks. With reduced-intensity conditioning, the onset of acute GVHD may be delayed until 2 to 3 months after transplantation. Hyperacute GVHD may occur within a week of transplantation after severely human leukocyte antigen (HLA)-mismatched transplants or transplants without GVHD prophylaxis. There is no reliable laboratory test for acute GVHD, and the diagnosis is based on clinical assessment.     

Endothelial damage and dysfunction in acute graft-versus-host disease

Clinical studies have suggested a potential involvement of endothelial dysfunction and damage in the development and severity of acute graft-versus-host disease (aGvHD). Accordingly, we found an increased percentage of apoptotic caspase 3 positive blood vessels in duodenal and colonic mucosa biopsies of patients with severe aGvHD. In murine experimental aGvHD, we detected severe microstructural endothelial damage and reduced endothelial pericyte coverage accompanied by reduced expression of endothelial tight junction proteins leading to increased endothelial leakage in aGvHD target organs. During intestinal aGvHD, colonic vasculature structurally changed, reflected by increased vessel branching and vessel diameter. As recent data demonstrated an association of endothelium-related factors and steroid refractory aGvHD (SR-aGvHD), we analyzed human biopsies and murine tissues from SR-aGvHD. We found extensive tissue damage but low levels of alloreactive T-cell infiltration in target organs, providing the rationale for T-cell independent SR-aGvHD treatment strategies. Consequently, we tested the endothelium-protective PDE5 inhibitor sildenafil, which reduced apoptosis and improved metabolic activity of endothelial cells in vitro. Accordingly, sildenafil treatment improved survival and reduced target organ damage during experimental SR-aGvHD. Our results demonstrate extensive damage, structural changes, and dysfunction of the vasculature during aGvHD. Therapeutic intervention by endothelium- protecting agents is an attractive approach for SR-aGvHD complementing current anti-inflammatory treatment options.     

Novel strategies for steroid-refractory acute graft-versus-host disease

PURPOSE OF REVIEW: Graft-versus-host disease is one of the commonest complications of allogeneic bone marrow or peripheral blood stem cell transplantation. This review will cover advances in the pathophysiology of graft-versus-host disease and new agents under investigation for the treatment of this disorder. Patients developing graft-versus-host disease who fail to respond to steroids have a poor prognosis. In this group of people, morbidity and mortality are very high. RECENT FINDINGS: Novel agents are currently under investigation for the treatment of such devastating disorders. Pentostatin, denileukin diftitox, mycophenolate mofetil, extracorporeal photopheresis, and several monoclonal antibodies have been used, some of them with encouraging results. SUMMARY: As supportive care improves and new agents are added to the armamentarium against steroid-refractory acute graft-versus-host disease, the prognosis of this entity may start to change. Patients with this complication after transplantation should be enrolled, whenever possible, in clinical trials to find effective therapies.     

A biomarker panel for acute graft-versus-host disease

No validated biomarkers exist for acute graft-versus-host disease (GVHD). We screened plasma with antibody microarrays for 120 proteins in a discovery set of 42 patients who underwent transplantation that revealed 8 potential biomarkers for diagnostic of GVHD. We then measured by enzyme-linked immunosorbent assay (ELISA) the levels of these biomarkers in samples from 424 patients who underwent transplantation randomly divided into training (n = 282) and validation (n = 142) sets. Logistic regression analysis of these 8 proteins determined a composite biomarker panel of 4 proteins (interleukin-2-receptor-alpha, tumor-necrosis-factor-receptor-1, interleukin-8, and hepatocyte growth factor) that optimally discriminated patients with and without GVHD. The area under the receiver operating characteristic curve distinguishing these 2 groups in the training set was 0.91 (95% confidence interval, 0.87-0.94) and 0.86 (95% confidence interval, 0.79-0.92) in the validation set. In patients with GVHD, Cox regression analysis revealed that the biomarker panel predicted survival independently of GVHD severity. A panel of 4 biomarkers can confirm the diagnosis of GVHD in patients at onset of clinical symptoms of GVHD and provide prognostic information independent of GVHD severity.