Corelattions Between CD31, CD68, MMP-2 and MMP-9 Expression in Allograft Cardiac Rejection – Immunohistochemical Study

Abstract Introduction. The cardiac allograft rejections from the post-transplant period are attributable to the acute cellular rejection monitored by multiple endomyocardial biopsies. Compared to this, humoral rejection remains a matter of debate, with multiple therapeutic strategies, poor prognosis, and persisting uncertainty about diagnostic criteria. Acute allograft rejection is associated with significant modifications of the extracellular matrix compartment mainly regulated by matrix metalloproteinases (MMPs). In this context, the aim of this study was to evaluate the expression of MMP-2 and -9 and CD31, CD68 (endothelial and histiocytic markers) and the correlations between them using immunohistochemistry, in patients with cardiac allografts. Materials and methods. Tissue fragments were obtained by endomyocardial biopsy from 5 patients with allograft heart transplant, 2 in the medium post-transplant phase and 2 in late phase. After identifying and characterizing the morphological context the probes were processed by standard immunohistochemical technique using anti-MMP-2 and anti-MMP-9 antibodies (Santa Cruz Biotechnology, Inc.) and anti-CD31, anti-CD68 antibodies (Sigma). The samples were examined using the Olympus BX40 microscope with an Olympus E330 camera attached. Results and discussions. Sample examination revealed in all 4 cases the lack of IR (-) for CD31 and weak IR (+) for CD68 compared to MMPs, where we found moderate IR (++) for MMP-9 and weak IR (+) for MMP-2. These aspects complets the histological lesional aspects of these cases, indicating the lack of acute rejection. In conclusion, CD31 and CD68 IR correlated with MMPs IR (especially MMP-9) appear to represent predictive markers for cardiac allograft rejection and require further studies.

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P761The effect of extracorporeal photopheresis on cardiac allograft rejection and on lymphocyte subclasses

European Heart Journal ◽

10.1093/eurheartj/ehz747.0361 ◽

2019 ◽

Vol 40 (Supplement_1) ◽

Author(s):

T Teszak ◽

A Assabiny ◽

A Kiraly ◽

Z Tarjanyi ◽

N Parazs ◽

...

Keyword(s):

T Cells ◽

Regulatory T Cells ◽

B Lymphocytes ◽

Heart Transplant ◽

Allograft Rejection ◽

Cytotoxic T Cells ◽

Cardiac Allograft ◽

Extracorporeal Photopheresis ◽

Cardiac Allograft Rejection ◽

Cellular Rejection

Abstract Background Cardiac allograft rejection is known to have a profound impact on graft survival and mortality after heart transplant. Previous data on the efficacy of extracorporeal photopheresis (ECP) in the management of cardiac allograft rejection is encouraging. Though, clear evidence on the exact indication and data regarding its effect on distinct lymphocyte subtypes are still lacking. Based on their cytokine production, both helper and cytotoxic T cells can differentiate into either regulatory cells participating in the suppression of rejection or into effector cells responsible for its maintenance. Regulatory T cells are essential for the termination of rejection, while B lymphocytes and natural killer (NK) cells contribute to it. Purpose We aimed to investigate the anti-rejection efficacy and the impact of ECP on peripheral blood lymphocyte subclasses in adult heart transplant recipients. Methods In a retrospective analysis of 12 consecutive patients treated with ECP for cardiac allograft rejection between 2013 and 2019, we examined the grade of rejection in endomyocardial biopsies (EMB) based on the International Society for Heart and Lung Transplantation classification. We analysed the absolute counts and the percentages of helper, cytotoxic and regulatory T cells, B lymphocytes and NK cells with fluorescence activated cell sorting. Measurements were performed both before and after the ECP treatment period. Data values were given as either mean±standard deviation or median [min–max]. Results The patients underwent 26 [2–39] ECP treatments in addition to standard immunosuppressant therapy. Whereas grade 2R cellular rejection was detected in 83% of the cases prior to initiating ECP, none of the examined EMB specimen revealed rejection greater than grade 1R cellular rejection post ECP therapy. The average grade of cellular rejection improved significantly (1.25±0.45 vs. 0.50±0.53; p=0.022). The absolute count and the percentage of helper T cells increased significantly post ECP therapy (0.34 G/l±0.26 G/l vs. 0.51 G/l±0.39 G/l; p=0.018 and 3.43%±2.24% vs. 5.98%±3.64%; p=0.017, respectively). There was also a significant rise in the percentage of cytotoxic T cells (2.33%±1.46% vs. 4.16±2.98%; p=0.027). We noticed an almost significant twofold increase in the percentage of regulatory T cells on completion of the ECP therapy (0.20%±0.22% vs. 0.37%±0.20%; p=0.060). Neither B lymphocyte nor NK cell counts revealed any significant changes. Conclusion ECP was effective in reducing the severity of cardiac allograft rejection episodes. The significant decrease in rejection rates might be indicative of the predominance of anti-inflammatory helper and cytotoxic T cell subpopulations and the increase of regulatory T cell count post ECP therapy. However, discussion of the results are limited by small sample size and the effect of medical therapy on the lymphocytes.

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Post transplant monitoring of soluble donor HLA antigens predicts the development of chronic humoral rejection of cardiac allografts

Human Immunology ◽

10.1016/0198-8859(95)92900-p ◽

1995 ◽

Vol 44 ◽

pp. 23

Keyword(s):

Hla Antigens ◽

Humoral Rejection ◽

Post Transplant ◽

Cardiac Allografts

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Circulating Sphingosine-1-Phosphate as A Non-Invasive Biomarker of Heart Transplant Rejection

Scientific Reports ◽

10.1038/s41598-019-50413-8 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Estefanía Tarazón ◽

Carolina Gil-Cayuela ◽

María García Manzanares ◽

Marta Roca ◽

Francisca Lago ◽

...

Keyword(s):

Heart Failure ◽

Heart Transplant ◽

Allograft Rejection ◽

Transplant Rejection ◽

Cardiac Allograft ◽

Cardiac Allograft Rejection ◽

Non Invasive ◽

Novel Approach ◽

Sphingosine 1 Phosphate ◽

Cardiac Rejection

Abstract Accumulating evidence has confirmed that the expression of sarcoplasmic reticulum calcium ATPase 2a (SERCA2a) is downregulated in heart failure and cardiac allograft rejection. Although many SERCA2a-related genes and proteins involved in the regulation of myocardial Ca2+ fluxes have been explored, its related metabolites remain poorly studied. Our main objective was to identify circulating SERCA2a-related metabolites altered in cardiac allograft rejection and to determine whether these could serve as non-invasive biomarkers. Sixty plasma samples from adult heart transplant were included in a metabolomic analysis. Sphingosine-1 phosphate (S1P), metabolite closely related with SERCA, were increased in patients with cardiac rejection (p < 0.0001). S1P discriminated between patients with and without rejection: normal grafts vs. all rejecting grafts (AUC = 0.911, p < 0.0001), normal grafts vs. Grade 1 R (AUC = 0.819, p < 0.01), Grade 2 R (AUC = 0.911, p < 0.0001), Grade 3 R (AUC = 0.996, p < 0.0001). In addition, we found changes in key enzymes and receptors of S1P pathway analysed on explanted hearts from heart failure patients. This preliminary study reveals that circulating S1P determination could be a novel approach to detect cardiac rejection, showing a robust capability for detection that improves gradually with the severity of rejection. These alterations could be relevant to better understand the involvement of calcium regulation on the pathophysiology of rejection.

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POST-TRANSPLANT DE NOVO AUTOIMMUNITY CONTRIBUTES TO CARDIAC ALLOGRAFT REJECTION AND IT IS MEDIATED VIA INDIRECT PRESENTATION OF DONOR MHC PEPTIDES AND ORGAN-SPECIFIC AUTOANTIGENS.

Transplantation Journal ◽

10.1097/00007890-200004271-00005 ◽

2000 ◽

Vol 69 (Supplement) ◽

pp. S112 ◽

Cited By ~ 1

Author(s):

Eugenia V. Fedoseyeva ◽

Koji Kishimoto ◽

Hillary Rolls ◽

Mohamed H. Sayegh ◽

Gilles Benichou

Keyword(s):

Allograft Rejection ◽

De Novo ◽

Cardiac Allograft ◽

Cardiac Allograft Rejection ◽

Post Transplant ◽

Organ Specific

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Analysis of ED-A+ fibronectin expression after heterotopic rat heart transplantation: association to cardiac allograft rejection and implications for targeted post-transplant therapy

The Thoracic and Cardiovascular Surgeon ◽

10.1055/s-0029-1247072 ◽

2010 ◽

Vol 58 (S 01) ◽

Author(s):

M Franz ◽

A Berndt ◽

K Grün ◽

P Richter ◽

H Kosmehl ◽

...

Keyword(s):

Heart Transplantation ◽

Allograft Rejection ◽

Rat Heart ◽

Cardiac Allograft ◽

Cardiac Allograft Rejection ◽

Post Transplant ◽

Fibronectin Expression

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Abstract 1128: The Leukocyte Integrin Mac-1 Promotes Acute Cardiac Allograft Rejection

Circulation ◽

10.1161/circ.116.suppl_16.ii_227-a ◽

2007 ◽

Vol 116 (suppl_16) ◽

Author(s):

Koichi Shimizu ◽

Peter Libby ◽

Masashi Sakuma ◽

Yunmei Wang ◽

Richard N Mitchell ◽

...

Keyword(s):

Graft Survival ◽

Adoptive Transfer ◽

Allograft Rejection ◽

Arterial Disease ◽

Cardiac Allograft ◽

Post Transplantation ◽

Cardiac Allograft Rejection ◽

Cardiac Allografts ◽

Antigen Presenting ◽

Prolong Graft Survival

Background: In allograft rejection, donor endothelial cells (EC) are the principal early targets of host leukocytes and alloantibodies. Although the leukocyte integrin Mac-1(αmβ2, CD11b/ CD18) facilitates leukocyte-leukocyte as well as leukocyte-EC interactions, its roles in allograft survival and vasculopathy remains unclear. This study used murine cardiac transplantation to test the hypothesis that Mac-1 deficiency in the recipient reduces accumulation of graft infiltrating cells and attenuates cardiac allograft rejection. Results: In total-allomismatched murine cardiac allografts (BALB/c donor hearts and C57BL/6 recipients), survival averaged 13.8 ± 2.3 days (n=6) for allografts in Mac-1-deficient (Mac-1−/−) recipients compared to 8.3 ± 1.3 days (n = 18, p<0.0001) in wild-type (WT) recipients. At 7 d post-transplantation, allografts in Mac-1−/− recipients had significantly lower parenchymal rejection (PR) scores (2.3 ± 0.4) than WT recipients (3.0 ± 0.4, p<0.01). Allograft accumulation of neutrophils, T cells, and macrophages (Mϕ) significantly diminished in Mac-1−/− compared to WT recipients. To determine the cell type involved in acute graft dysfunction, we performed adoptive transfer experiments. Adoptive transfer of WT, but not Mac-1−/−, Mϕ exacerbated parenchymal rejection of 7-day allografts placed into Mac-1−/− recipients (PR score: 2.3 ± 0.4 for WT vs. 1.5 ± 0.5 for Mac1−/− Mϕ; p < 0.05), and significantly reduced graft survival (8.0 ± 1.4 vs. 13.5 ± 2.6 days, p<0.01). Neither WT nor Mac-1−/− neutrophil adoptive transfer affected graft survival in Mac-1−/− recipients. Mac-1−/− Mϕ co-cultured with allo-EC (BALB/c) expressed significantly lower levels of the co-stimulatory molecules CD40 and CD80, and mixed leukocyte reaction using allo-EC-primed Mac-1−/− Mϕ showed significantly lower antigen-presenting function than WT Mϕ. Despite attenuating acute rejection, recipient Mac-1-deficiency did not prevent graft arterial disease. Conclusion: These studies demonstrate that Mac-1 contributes to acute allograft rejection by modulating macrophage recruitment, antigen presentation, and T cell priming. These observations establish a molecular target for modulating recipient responses to prolong graft survival.

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