Humoral Allograft Rejection Research Articles

Significance of HLA-matching and anti-HLA antibodies in heart transplant patients receiving induction therapy?

ObjectivesThough Human Leukocyte Antigen (HLA) matching benefits are demonstrated in renal transplantation, evidence in heart transplantation is lacking, and its clinical feasibility is uncertain. Post-transplantation anti-HLA antibodies are being increasingly studied in organ transplantation, with diverging conclusions between transplantated organs. MethodsWe analyzed retrospectively the influence of HLA matching and anti-HLA antibodies on overall survival, acute rejection and chronic allograft vasculopathy in 309 patients receiving induction therapy and triple-drug immunosuppression. ResultsThe average number of HLA-A/B/DR mismatches between donor and recipient was 4.9 ± 1. The majority of mismatches was for Class I HLA-A/B with an average of 3.3, then for Class I HLA-DR with an average of 1.6. Overall, the HLA-A/-B/-DR mismatches had no influence on the cardiac allograft survival (p = 0.28). However, HLA-DR mismatches were negatively correlated to severe cellular and/or humoral allograft rejection (p = 0.04). Our analysis found anti-HLA antibodies in 27% of recipients, de novo anti-HLA antibodies in 16% of recipients, and donor-specific anti-HLA (DSA) antibodies in 8% of recipients. Furthermore, de novo DSA had no influence on the 5-year survival (78% with DSA vs. 92% without DSA; p = 0.49), which may be masked by the limited number of recipients in analysis By univariable analysis, anti-HLA antibodies (preexisting or de novo) unrelated or related to the donor had no influence on severe cellular and/or humoral rejection or on chronic allograft vasculopathy. ConclusionsHLA-DR mismatch was negatively correlated to severe cellular and/or humoral allograft rejection but had no influence on cardiac allograft survival. In this study, anti-HLA antibodies (preexisting or de novo) unrelated or related to the donor had no influence on cellular and/or humoral rejection or on chronic allograft vasculopathy. The results of this study add to the controversy on the impact of allo-antibodies in heart transplant recipients receiving induction therapy and contemporary immunosuppression.

Early Acute Graft Rejection in a Heart Transplanted Child with Dilated Cardiomyopathy

Abstract Dilated cardiomyopathy (DCM) is the most common type of cardiomyopathy in children. Heart transplantation is considered standard therapy in dilated cardiomyopathy with end-stage heart failure. We present a case of a 15-year-old patient diagnosed with DCM in the neonatal period, who underwent heart transplantation for end-stage heart failure. Despite the use of induction therapy, the endomyocardial biopsy performed at two weeks post-transplant revealed mixed moderate cellular (2R) and humoral (pAMR2) allograft rejection. Aggressive rejection treatment was initiated with good outcome. Besides endomyocardial biopsy, advanced echocardiography can also be a valuable noninvasive tool for rejection assessment.

Detrimental Effects of Donor Brain Death on Tolerance Induction May Be Eliminated by Delaying Mixed Chimerism in Non-Human Primates

Induction of tolerance in combined heart/kidney transplantation in non-human primates is possible by applying a mixed chimerism-based conditioning protocol which induces indefinite tolerance. However, donor brain death negatively impacts tolerance induction, leading to a higher incidence of humoral as well as cellular rejection. Here, we investigated whether delaying conditioning and donor bone marrow transplantation (DBMT) until four months after organ transplantation would diminish the detrimental effects of donor brain death and permit tolerance induction despite donor brain death. Nine cynomolgus monkeys underwent combined heart/kidney transplantation using organs from donors rendered brain-dead 4 hours prior to organ procurement. Six recipients underwent nonmyeloablative conditioning that included total body irradiation, thymic irradiation, horse anti-thymocyte globulin, anti-CD154 mAb, CyA and DBMT on the day of solid organ transplantation, three animals were kept on triple immunosuppression for four months after combined heart/kidney transplantation before undergoing the identical conditioning protocol prior to DBMT. All drugs were stopped 29 days after DBMT. Three brain-dead donor animals rejected their allografts on day 127, 131 and 383, respectively. Two brain dead donor animals were euthanized without signs of rejection on day 30 and day 114 after transplantation due to sepsis and PTLD, respectively. One brain dead donor animal underwent elective euthanasia on day 400 after transplantation without signs of rejection. One animal in the delayed conditioning group developed no detectable donor chimerism and rejected its allografts on day 135, another delayed animal was euthanized for sepsis without signs of graft rejection on day 59 after DBMT. A third animal in the same group is still ongoing without histological signs of cellular or humoral allograft rejection on day 420 after DBMT. Donor brain death negatively impacts tolerance induction after mixed chimerism conditioning leading to a higher incidence of allograft rejection. Delaying recipient conditioning and DBMT might eliminate the detrimental effects of donor brain death and its associated pro-inflammatory side-effects, thus increasing the chance of long-term tolerance.

Humoral Response in Liver Allograft Transplantation: A Review of the Role of Anti-Human Leukocyte Antigen (HLA) Antibodies.

Antibody-mediated rejection (AMR) is relatively uncommon in AB0-compatible liver grafts; however, recent data suggest that antibody-mediated mechanisms may play a role in the differential pathogenesis of liver graft rejection. In this sense, it has been shown that staining for diffuse C4d deposition (in endothelium or stroma of portal tracts or sinusoids) could be used as a tissue biomarker of humoral allograft rejection and fibrosis in biopsies obtained in post-transplantation period, in addition to the determination of donor specific antibodies (DSA). Therefore, more stringer criteria have to be established in order to define real antibody-mediated injury (acute rejection, chronic rejection or fibrosis development) in liver graft transplant. These criteria should intend to correlate allograft dysfunction associated with compatible histological findings, presence of DSAs and C4d positivity and should be clarified with respect to rejection injury frequency, its severity and its response to antibody-depleting immunosuppression in the future. This could improve our progressive understanding of the clinical-pathological characteristic and diagnostic approaches of AMR and fibrosis.

Decreased humoral antibody episodes of acute renal allograft rejection in recipients expressing the HLA-DQβ1*0202 allele

The present investigation was designed to show the effect of human leukocyte antigen (HLA) class II molecular allelic specificities in the recipient on the induction of humoral antibody rejection, identified by C4d peritubular capillary staining, as well as specific antibody identified by Luminex technology. Major histocompatibility complex (MHC) class II molecules are expressed on dendritic cells, macrophages, and B lymphocytes and they present antigenic peptides to CD4 positive T lymphocytes. Human renal peritubular and glomerular capillaries express class II MHC molecules upon activation. Expression of class II molecules on renal microvascular endothelial cells exposes them to possible interaction with specific circulating antibodies. We hypothesize that HLA-DQβ1*0202 expression in recipients decreases the likelihood of antibody-mediated renal allograft rejection. We found that 80% (=25) of DQ2 positive haplotype recipients failed to induce humoral antibody renal allograft rejection and 20% (n=25) of DQ2 positive haplotype recipients induced humoral antibody renal allograft rejection (p=0.008). By contrast, 48% (n=46) of DQ2 negative haplotype recipients failed to induce a humoral antibody component of renal allograft rejection and 52% (n=46) of DQ2 negative haplotype recipients induced humoral antibody-mediated renal allograft rejection. Our results suggest that recipients who express the DQβ1*0202 allele are less likely to induce a humoral antibody component of acute renal allograft rejection than are those expressing DQ1, DQ3, or DQ4 alleles. DQβ1*0202 allele expression in recipients could possibly be protective against acute humoral allograft rejection and might serve as a future criterion in recipient selection and in appropriate therapy for acute renal rejection episodes.

Long-Term Outcomes Using Deceased Donor Kidneys From Cobra Bite Brain Dead Victims

Long-Term Outcomes Using Deceased Donor Kidneys From Cobra Bite Brain Dead Victims

Successful Treatment of Refractory Acute Humoral Allograft Rejection with Single Dose Rituximab

Successful Treatment of Refractory Acute Humoral Allograft Rejection with Single Dose Rituximab

52-P: Anti-GSTT1 antibodies in patients with humoral kidney allograft rejection; its association with C4d deposition on renal biopsies

52-P: Anti-GSTT1 antibodies in patients with humoral kidney allograft rejection; its association with C4d deposition on renal biopsies

The Cellular Lesion of Humoral Rejection: Predominant Recruitment of Monocytes to Peritubular and Glomerular Capillaries

Accumulation of inflammatory cells within capillaries is a common morphologic feature of humoral renal allograft rejection and is most easily appreciated if it occurs in glomeruli. The aim of our study was to determine the amount and composition of immune cells within glomeruli and peritubular capillaries (PTC) in cellular and humoral allograft rejection. Immunofluorescent double-labeling for CD31 and CD3 or CD68 was used for phenotyping and enumerating immune cells within glomeruli and PTC. The major findings are: (1) accumulation of immune cells in PTC is far more common than it would be anticipated based on the assessment by conventional histology; (2) it is not the absolute number of immune cells accumulating within capillaries, but rather the composition of the intracapillary cell population that distinguishes humoral rejection from cellular rejection and (3) in C4d positive biopsies a predominantly monocytic cell population accumulates not only within glomeruli but also within PTC. The median value of monocyte/T-cell ratio within PTC was 2.3 in C4d positive biopsies but only 1 (p = 0.0008) in C4d negative biopsies. Given their prominent presence within capillaries and their extensive biological versatility monocytes might contribute to the capillary damage observed in acute and chronic allograft rejection.

Prolonged Cardiac Allograft Survival in Presensitized Rats After a High Activity Yunnan-Cobra Venom Factor Therapy

Complement-dependent antibody-mediated acute humoral rejection is the major obstacle of clinical transplantation across ABO incompatibility and human leukocyte antigen presensitization. We previously demonstrated that Yunnan-cobra venom factor (Y-CVF) could almost completely abrogate complement activity and successfully prevent hyperacute rejection in some xenotransplant models without any obvious toxicity. In this study we investigated whether depletion of complement by Y-CVF prevented acute humoral allograft rejection in presensitized rats thereby prolonging graft survival. Methods Presensitization was achieved in Lewis rats by sequential grafting of three full-thickness skin pieces from Brown Norway rats. Serum cytotoxic alloantibody titers were determined by a modified in vitro complement-dependent microcytotoxicity assay. After presensitization, each Lewis rat received a heterotopic Brown Norway cardiac allograft. Fifteen recipients were divided into two groups: (1) no treatment control ( n = 7); (2) Y-CVF therapy group (86 u/kg, IV, day −1) ( n = 8). After cessation of the heart beat, allograft rejection was confirmed by pathologic as well as IgG and C3 immunohistochemical examinations. Results The mean graft survival time was significantly prolonged to 99.50 ± 38.72 hours among rats that received Y-CVF vs 12.71 ± 13.94 hours in nontreated controls ( P < .001). Upon pathological and immunohistochemical examination, acute humoral rejection was mainly exhibited in the control group, whereas acute cellular rejection was mainly displayed in the Y-CVF therapy group. Conclusions Our study demonstrated that complement depletion by Y-CVF significantly inhibited acute humoral allograft rejection in presensitized rats. As a therapeutic immunointervention tool for complement, Y-CVF has shown potential efficacy across ABO incompatible and positive cross-match barriers.

C3d and the Septal Microvasculature as a Predictor of Chronic Lung Allograft Dysfunction

Studies have shown a potential role for humoral rejection in the evolution of lung graft dysfunction, apparently based on antibodies without human leukocyte antigen specificity. The correlation between extent of C4d deposition with clinical status further illustrates the importance of humoral immunity. Our study examines the potential value of C3d as a further diagnostic adjunct. C3d deposition was examined in lung allograft specimens using frozen tissue indirect direct immunofluorescence (IIF) and avidin biotin immunohistochemical applied to paraffin embedded tissue. Intermediate/extensive amounts of C3d using IIF and immunohistochemical (IH) methodologies correlated with chronic graft dysfunction; IIF C3d deposition was associated with septal and bronchial wall fibrosis ( p < 0.0001). Weak/absent amounts of IIF and IH C3d correlated with clinical stability ( p < 0.0001). Higher levels of C3d by IH were more sensitive than by IIF as a bronchiolitis obliterans syndrome determinant. C3d and C4d deposition using immunofluorescence and IH were correlated ( p < 0.00001). C3d deposition appears prognostically significant. Higher tissue expression of C3d mark chronic graft dysfunction/persistent graft failure following transplantation. The close correlation between C3d and C4d lends credence to the role of humoral allograft rejection as a pulmonary graft dysfunction contributing factor. C3d by IH manifests higher sensitivity but similar specificity compared to C3d by IIF.

Ultrastructure as a Diagnostic Adjunct in the Evaluation of Lung Allograft Biopsies

Humoral immunity contributes significantly to lung graft dysfunction. Recognizing a role of ultrastructural studies in the evaluation and diagnosis of chronic humoral allograft rejection in the kidney, the authors sought to explore its utility as a diagnostic adjunct in lung allograft biopsies. Ultrastructural studies were conducted on 44 biopsies from 26 lung transplant recipients. Endothelial cell activation and necrosis were seen in the setting of acute humoral allograft rejection. Septal chronic vasculopathic changes of thickening and lamellation of the basement membrane zone (BMZ) and BMZ collagen deposition were correlated with greater numbers of humoral allograft rejection episodes and with the development of chronic graft dysfunction/bronchiolitis obliterans syndrome. There was a positive correlation between the extent of septal fibrosis and certain chronic vasculopathic changes, namely collagen deposition in the BMZ and BMZ wrinkling. Patients with chronic graft dysfunction and multiple rejection episodes manifested low diffusion capacities (less than 50% predicted). The results indicate that ultrastructural analysis is useful in identification of septal fibrosis and chronic vasculopathy of the septal microvasculature, correlating with chronic graft dysfunction, encompassing not only fibrotic sequelae of the bronchial wall but also irreversible terminal lung parenchymal changes, the latter associated with repeated episodes of humoral rejection.

Use of C4d as a Diagnostic Adjunct in Lung Allograft Biopsies

Purpose: Humoral allograft rejection is a defined mechanism for cardiac and renal graft dysfunction; C4d deposition, a stable component of complement activation, inversely correlates with graft survival. With the recent recognition of humoral rejection in lung grafts, we examined C4d's role as a prognostic adjunct in lung allografts. Material and Methods: Twenty‐three lung recipients underwent biopsies for deterioration in clinical status or routine surveillance. Clinically unwell patients possessed acute rejection or bronchiolitis obliterans syndrome (BOS). Biopsies attributable to infection were excluded from the study. In addition to routine light microscopy, an attempt was made to correlate the clinical status and morphologic findings with the pattern of C4d deposition and also to compare these clinical and morphologic parameters with the other assessed immunoreactants. Panel reactive antibody testing was also carried out at various points in their post transplantation course whereby in 6 of the cases the samples were procured at exactly the same time as the tissue samples. Results: The patients were segregated into two groups: those patients with recurrent acute rejection and those with BOS. In those patients with symptomatic acute rejection, all biopsies showed light microscopic and immunofluorescent evidence of humoral allograft rejection. The level of C4d was positively correlated with the degree of parenchymal injury, the hallmark being one of septal capillary necrosis. In addition, high and intermediate levels of C4d correlated with a clinical diagnosis of acute rejection. C4d was the strongest predictor of parenchymal injury and of the clinical status (p < .0001) compared to other the immunoreactants C1q, C5b‐9 and immunoglobulin. There was no specific correlation between C4d deposition and the presence of acute cellular rejection. In those patients fulfilling clinical criteria of BOS, deposits of C4d as well as other immunoreactants were found in the bronchial wall as opposed to the rarity of this finding in bon‐BOS patients. However the only statistically significant predictor of BOS was bronchial wall deposition of C1q. In no case were panel reactive antibodies at significant levels discovered post transplantation. Conclusions: In the context of acute rejection, C4d deposition correlates with clinical evidence of rejection and the degree of humoral rejection assessed pathologically; there is no association with the presence of histocompatibility related antibodies. It is a more specific predictor of allograft status compared to other immunoreactants. C4d deposition within the bronchial wall is a feature of BOS and hence may be used as a marker of chronic graft dysfunction. The antigenic target resulting in C4d deposition may not be histocompatibility related.

Non-Invasive Monitoring of Renal Transplant Recipients: Urinary Excretion of Soluble Adhesion Molecules and of the Complement-Split Product C4d

Background: The number of inducible adhesion molecules known to be involved in cell-mediated allograft rejection is still increasing. In addition, recent data describe complement activation during acute humoral allograft rejection. The aim of this study was to assess whether specific molecules from either pathway are excreted into urine and whether they can provide useful diagnostic tools for the monitoring of renal transplant recipients. Methods: Urinary concentrations of soluble adhesion molecules (sICAM-1, sVCAM-1) and of the complement degradation product C4d were determined by standardized ELISA technique in 75 recipients of renal allografts and 29 healthy controls. Patient samples were assigned to four categories according to clinical criteria: group 1: acute steroid-sensitive rejection (ASSR, n = 14), group 2: acute steroid-resistant rejection (ASRR, n = 12), group 3: chronic allograft dysfunction (CAD, n = 20) and group 4: stable graft function (SGF, n = 29). Results: All patients with rejection episodes (groups 1–3) had significantly higher values of urinary sC4d compared with healthy controls and patients with stable graft function (p < 0.05). The urinary levels of sVCAM-1 were significantly higher in group 2 (ASRR) compared with all other groups (p < 0.001). Uniformly low amounts of s-VCAM-1 and complement-split product C4d were excreted by healthy controls (group 0). In contrast, urinary sICAM-1 concentration in healthy controls was almost as high as in group 2 (ASRR) whereas patients with a stable functioning graft (group 4) excreted significantly less sICAM-1 (p < 0.05). Conclusion: The evaluation of sVCAM-1 and sC4d excretion in urine can provide a valuable tool with regard to the severity and type of allograft rejection. With respect to long-term allograft survival, serial measurements of these markers should have the potential to detect rejection episodes and prompt immediate treatment.

When renal allografts turn DARC.

The Duffy antigen-receptor for chemokines (DARC) is a chemokine-binding protein that is up-regulated on peritubular capillaries (PTC) during cellular renal allograft rejection. C4d deposition and accumulation of inflammatory cells in PTC are indicators of humoral renal allograft rejection. Because DARC is expressed at the site of C4d deposition and might be involved in inflammatory cell recruitment, the authors evaluated the expression of DARC in different forms of human renal allograft rejection. Deposition of C4d and DARC expression were evaluated by immunohistochemistry in 42 renal transplant biopsy specimens. Biopsy specimens were subdivided according to histologic and immunohistochemical results, that is, C4d-negative biopsy specimens with (Banff 1, n=8) or without signs of cellular rejection (n=16), and C4d-positive biopsies (humoral rejection) with (Banff 1 rejection, n=7) or without cellular rejection (n=11). DARC expression was found on a small number of PTC and veins in patients without rejection. Cellular and humoral rejection led to a comparable increase in the number of DARC-positive PTC (9.7 and 8.7 vs. 2.6 vessels per high-power field [HPF], respectively). The highest numbers were found in biopsy specimens with signs of both humoral and cellular rejection (17.5 vessels per HPF). This is the first study that demonstrates an induction of a chemokine-binding protein at the site of C4d deposition in humoral allograft rejection. The additive effect of humoral and cellular rejection on DARC expression might imply different pathways of DARC induction for different forms of allograft rejection.

Direct and indirect immunofluorescence as a diagnostic adjunct in the interpretation of nonneoplastic medical lung disease.

Fresh open lung biopsy material from 57 patients was incubated with fluoresceinated complement and immunoglobulin antisera. An indirect immunofluorescent assay using neonatal lung as substrate was conducted as well. Direct immunofluorescent patterns could be categorized into interalveolar septal capillary deposition, large vessel wall localization, alveolar basement membrane localization, or a pauci-immune immunofluorescence pattern. With respect to the septal capillary pattern, endothelial cell decoration was seen with scleroderma, mixed connective tissue disease, anti-Ro-associated lupus erythematosus, dermatomyositis, humoral allograft rejection, and patients with isolated pulmonary fibrosis in whom autoantibodies were established, including antiphospholipid antibodies. A similar pattern of endothelial cell staining was seen in these cases via the indirect assay. Granular mural septal capillary deposition was seen in the aforesaid settings along with rheumatoid factor-positive rheumatoid arthritis, type II cryofibrinogenemia, and mixed cryoglobulinemia and, in some cases, light microscopically corresponded to a neutrophilic capillaritis. Isolated vascular IgA corresponded with rheumatoid arthritis corresponding to IgA-specific antiendothelial cell antibodies, celiac disease-associated pulmonary hemorrhage, Schönlein-Henoch purpura and with IgA antiphospholipid antibodies. Alveolar wall deposition was seen with anti-glomerular basement membrane disease.

Direct and Indirect Immunofluorescence as a Diagnostic Adjunct in the Interpretation of Nonneoplastic Medical Lung Disease

Fresh open lung biopsy material from 57 patients was incubated with fluoresceinated complement and immunoglobulin antisera. An indirect immunofluorescent assay using neonatal lung as substrate was conducted as well. Direct immunofluorescent patterns could be categorized into interalveolar septal capillary deposition, large vessel wall localization, alveolar basement membrane localization, or a pauci-immune immunofluorescence pattern. With respect to the septal capillary pattern, endothelial cell decoration was seen with scleroderma, mixed connective tissue disease, anti-Ro-associated lupus erythematosus, dermatomyositis, humoral allograft rejection, and patients with isolated pulmonary fibrosis in whom autoantibodies were established, including antiphospholipid antibodies. A similar pattern of endothelial cell staining was seen in these cases via the indirect assay. Granular mural septal capillary deposition was seen in the aforesaid settings along with rheumatoid factor-positive rheumatoid arthritis, type II cryofibrinogenemia, and mixed cryoglobulinemia and, in some cases, light microscopically corresponded to a neutrophilic capillaritis. Isolated vascular IgA corresponded with rheumatoid arthritis corresponding to IgA-specific antiendothelial cell antibodies, celiac disease-associated pulmonary hemorrhage, Schonlein-Henoch purpura and with IgA antiphospholipid antibodies. Alveolar wall deposition was seen with anti-glomerular basement membrane disease.

Immunological characterization of anti-endothelial cell antibodies induced by cytomegalovirus infection.

We have previously shown that the levels of anti-endothelial cell antibodies (AECA) determined by an enzyme immunoassay are elevated during cytomegalovirus (CMV) infection in cardiac and renal transplant recipients. In a separate study, high levels of AECA are associated with higher frequency of humoral allograft rejection (AR), chronic AR and lower 2 year allograft survival in cardiac transplant recipients. These results suggests that high levels of AECA produced during CMV infection may have a pathogenic role or be a risk factor for humoral AR, chronic AR and decreased allograft survival. Here we examined the reactivity of AECA against endothelial cells and other tissues to further characterize AECA induced by CMV infection. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/Western blot analysis was performed. The number and intensity of bands reactive with human umbilical vein ECs (HUVECs) increased during and after CMV infection. AECA(+) plasma reacted with multiple antigens expressed not only on endothelial cells but also on human fibroblasts, keratinocytes, platelets (PLs), peripheral blood mononuclear cells (PBMCs), Raji cells and THP-1 cells. Each individual's AECA(+) plasma showed different patterns of reactivity against these cells, whereas each plasma showed similar patterns of reactivity against ECs, PLs or peripheral blood mononuclear cells obtained from different individuals. AECA(+) plasma also showed a similar pattern of reactivity against HUVECs pretreated with/without interferon-gamma/tumor necrosis factor-alpha. The reactivity of preabsorbed sera with PLs significantly decreased against most reactive antigens expressed on PLs and other cell types. (1) Antibodies induced by CMV infection are not specific to endothelial cells and appear to react with multiple cell types, (2) AECA (+) plasma react with multiple antigens expressed on various cell types that are primarily constitutively expressed on these cells and are not individual specific, (3) CMV-induced antibodies in AECA (+) plasma are primarily autoantibodies. These results suggest that the elevated AECA levels seen in CMV-infected transplant recipients may represent a polyclonal activation of humoral immune responses induced by CMV, which is of uncertain pathogenic significance.

Allograft conduit wall calcification in a model of chronic arterial graft rejection.

Early allograft vascular wall degeneration has emerged as a major important complication in young patients. To explain this mechanism, we reviewed studies on explants of allograft valved conduits implanted heterotopically into the infrarenal aorta in inbred rats (LEW; RT1I and CAP-RT1C). The following strain combinations (isografts and allografts) were used: syngeneic, LEW- > LEW, strongly allogeneic, and CAP > LEW (RT1- and non-RT1-incompatible). Second-set skin grafting was performed 3 weeks after the heterotrophic implant to test for immunogenicity and presensitization. The animals (LEW) were sacrificed serially on days 20, 30, 50, and 100 for immunofluorescence and SEM studies. Endothelial disruption was observed on day 30, while valve leaflets appeared normal. Humoral allograft rejection was demonstrated and associated with production of antibodies (IgG) against the endothelial cells and around the smooth muscle cells, and in areas of smooth cell necrosis, through 100 days. Neointimal repopulation by host cells and migrated smooth muscle cells was also observed in both viable and allovital grafts. Allovital grafts demonstrated more disorganized collagen and elastic fibers, as well as calcific degeneration in the media and neointima on day 50; the viable conduits showed such structural changes on day 100. In conclusion, vascular walls of allovital conduits calcified earlier than the viable conduits without discernible calcification of the valves. There is therefore evidence to prove causative relationships between cellular viability, immune response, and fibroproliferative calcific degeneration in allograft vascular conduits.

Role of Endothelial Cells in Transplantation (Part 2 of 2)

Endothelial cell activation with accompanying vascular inflammatory changes is considered central to the experimental manifestations of both hyperacute and delayed xenograft rejection responses. Natural xenoreactive antibodies directed at &#913;-galactosyl residues of xenogeneic glycoproteins and glycolipids, with associated complement activation via the classical pathway, are considered major immediate mediators of graft endothelial cell injury in the clinically relevant discordant swine to primate combinations. In delayed xenograft rejection processes, where recipients are treated prophylactically to ameliorate these initial events, activation of infiltrating mononuclear phagocytes and natural killer cells are associated with ongoing endothelial cell activation processes, procoagulant generation and vascular thrombosis. Allograft hyperacute rejection is observed when vascularised organs are transplanted to sensitized individuals with high levels of cytotoxic antibodies. Less dramatic forms of humoral allograft rejection (termed accelerated or vascular rejection) and the more common cell-mediated endothelialitis are associated with significant graft damage. Endothelial cell activation is also linked with graft preservation injury, forms of chronic rejection and delayed graft loss. Experimental work is currently being directed at the control of hyperacute rejection, the close understanding of endothelial cell thromboregulation in both transplanted xeno- and allografts and the development of novel therapeutic agents including gene therapy and the possible use of organs from transgenic animals.