Integrative analysis correlates donor transcripts to recipient autoantibodies in primary graft dysfunction after lung transplantation

Up to one in four lung-transplanted patients develop pulmonary infiltrates and impaired oxygenation within the first days after lung transplantation. Known as primary graft dysfunction (PGD), this condition increases mortality significantly. Complex interactions between donor lung and recipient immune system are the suspected cause. We took an integrative, systems-level approach by first exploring whether the recipient''s immune response to PGD includes the development of long-lasting autoreactivity. We next explored whether proteins displaying such differential autoreactivity also display differential gene expression in donor lungs that later develop PGD compared with those that did not. We evaluated 39 patients from whom autoantibody profiles were already available for PGD based on chest radiographs and oxygenation data. An additional nine patients were evaluated for PGD based on their medical records and set aside for validation. From two recent donor lung gene expression studies, we reanalysed and paired gene profiles with autoantibody profiles. Primary graft dysfunction can be distinguished by a profile of differentially reactive autoantibodies binding to 17 proteins. Functional analysis showed that 12 of these proteins are part of a protein–protein interaction network (P=3 × 10⁻⁶) involved in proliferative processes. A nearest centroid classifier assigned correct PGD grades to eight out of the nine patients in the validation cohort (P=0·048). We observed significant positive correlation (r=0·63, P=0·011) between differences in IgM reactivity and differences in gene expression levels. This connection between donor lung gene expression and long-lasting recipient IgM autoantibodies towards a specific set of proteins suggests a mechanism for the development of autoimmunity in PGD.     

Endomyocardial biopsy of normal donor hearts before cardiac transplantation. A morphological and morphometrical study in 97 cases.

Endomyocardial biopsies from 97 normal donor hearts were examined. Morphometric analysis showed: mean myocyte diameter 22.21 +/- 6.93 mu, mean nuclear dimension 7.32 +/- 2.33 mu, mean nuclear/sarcoplasmic ratio 0.33 +/- 0.02. 31 biopsies showed enlarged myocytes (mean diameter 31.65 +/- 3.98 mu) with increased nuclear size (mean 10.45 +/- 1.39 mu), but preserved nuclear/sarcoplasmic ratio (mean 0.33 +/- 0.01). The mean age of these latter subjects was significantly higher. Endocardial thickness mean value was 17.73 +/- 4.58 mu, but in 28 cases the value exceeded the considered upper normal limit of 20 mu. Interstitial mononuclear cells were rare and randomly present. Interstitial fibrosis was observed in 15% and focal fibrosis in 27% of cases. Our results show that histology of biopsies from clinically normal hearts can widely vary, sometime overriding the pathologic boundaries. These apparently "benign" abnormalities should be kept in mind when specific pathologic substrates of cardiac diseases have to be defined.     

Lectin complement pathway gene profile of the donor and recipient does not influence graft outcome after kidney transplantation

In kidney transplantation, complement activation was found to be induced by donor brain death, renal ischemia-reperfusion injury and allograft rejection. There are three known pathways of complement activation: the classical, lectin and the alternative pathway. The lectin complement pathway can be activated upon pattern recognition by mannan binding lectin (MBL) or ficolins (FCN). Single nucleotide polymorphisms (SNPs) in the genes encoding the lectin pathway proteins determine their functional activity and serum levels. The aim of this study was to investigate the role of the lectin gene profile of the donor and recipient on post-transplant outcome. A total of 12 functional SNPs in the MBL2, FCN2 and MBL-associated serine proteases 2 (MASP2) genes of 1271 donor-recipient pairs were determined. Lectin genotypic variants were analyzed for association with primary non-function (PNF), delayed graft function (DGF), biopsy proven acute rejection, death-censored graft survival and patient survival. Multivariate analyses found no association of donor and recipient MBL2 and MASP2 genotype with allograft outcome. Analysis of separate functional SNPs and haplotypes in the FCN2 gene of the donor and recipient did not reveal an association with transplant outcome. Also, the joint effect of the MBL2 and FCN2 genotype was not associated with allograft outcome.This study shows that the genetic profile of the lectin pathway of complement activation of the donor and recipient is not associated with allograft outcome after kidney transplantation.     

Refining murine heterotopic heart transplantation: A model to study ischemia and reperfusion injury in donation after circulatory death hearts

Heart transplantation is a lifesaving procedure, which is limited by the availability of donor hearts. Using hearts from donors after circulatory death, which have sustained global ischemia, requires thorough studies on reliable and reproducible models that developing researchers may not have mastered. By combining the most recent literature and our recommendations based on observations and trials and errors, the methods here detail a sound in vivo heterotopic heart transplantation model for rats in which protective interventions on the ischemic heart can be studied, and thus allowing the scientific community to advance organ preservation research. Knowledge gathered from reproducible animal models allow for successful translation to clinical studies.     

Recurrence of bronchioloalveolar carcinoma in donor lung after lung transplantation: microsatellite analysis demonstrates a recipient origin

Bronchioloalveolar carcinoma is a distinctive subtype of pulmonary adenocarcinoma, without effective therapy, although there have recently been some attempts to use lung transplantation. However, a high post-transplantation local recurrence rate is described with some controversy regarding the possible involved mechanisms, the main possibilities being the lymphatic spread and aerosolization. Presented herein is a case of a bilateral lung transplantation for a bilateral and pneumonic form of non-mucinous bronchioloalveolar carcinoma in a 43-year-old woman. The histological analysis of mediastinal lymph nodes during surgery did not show neoplastic cells. Thirty-five months after transplantation several nodular opacities in donor lungs were detected. Three pulmonary wedge resections were performed showing a non-mucinous bronchioloalveolar carcinoma with the same histological characteristics as the primary. Again, the mediastinal lymph nodes were tumor free. A complete microsatellites molecular analysis was performed to compare the primary and recurrent carcinoma using capillary electrophoresis, showing that the recurrent tumor was generated in a recipient cellular clone. The absence of lymph node metastasis and the molecular evidence of the recipient origin of the neoplasm supports the contamination of the new lungs at the time of implantation as being the reason for the high incidence of recurrence after lung transplantation in this kind of disease.     

热缺血20 min猪心脏死亡供体心冠状动脉内皮功能的变化

BACKGROUND: Myocardial and coronary endothelial injury occurs in donor hearts due to warm ischemia during cardiac transplantation. Coronary endothelial structure and function play a critical role in long-term outcomes for patients after cardiac transplantation.  OBJECTIVE: To study the effect of hypoxia-induced warm ischemia (20 minutes) on coronary endothelial function of porcine donor hearts after cardiac death (DCD). METHODS: Sixteen healthy Swedish domestic pigs were randomized into control (n=6), DCD (n=5), and DCD plus cold storage (n=5) groups, respectively. A DCD model in pigs was established using the method of hypoxia-induced 20-minute warm ischemia in the DCD and DCD plus cold storage groups. Isolation of the heart left anterior descending coronary artery or combined with heart preservation pretreatment for 4 hours was performed in the DCD and DCD plus cold storage groups. The maximum coronary endothelium-dependent relaxation was determined in the three groups. RESULTS AND CONCLUSION:There were no significant differences in the maximum coronary endothelium-dependent relaxation and the minus logarithmic of substance concentration induced 50%maximal relaxation among three groups (P > 0.05). These results indicate that 20-minute warm ischemia cannot lead to obvious coronary endothelial dysfunction. In addition, DCD combined with 4-hour cold storage does not affect coronary endothelial function. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Malignant transformation of thymoma in recipient rats by heterotopic thymus transplantation from HTLV-I transgenic rats

Transgenic rats expressing the pX gene of human T lymphocyte virus type-I (HTLV-I) under control of the rat lymphocyte-specific protein tyrosine kinase type-I promoter (lck-pX rats) developed benign epithelial thymomas. When the thymuses of newborn lck-pX rats were transplanted into the subcapsular space of the kidney in other thymectomized lck-pX rats, similar tumors developed in the transplanted thymuses. Following the tumor growth, dissemination in the abdominal cavity and distant metastasis occurred. The tumors were histopathologically similar to the original thymomas, but prominent nuclear atypia and high mitotic activity were present. The Ki-67 index was twice as high as that in the originals. The tumors were transplantable into the subcutis of lck-pX rats, although transplantation of the originals never succeeded. All evidence indicated that malignant transformation of thymoma was induced by the heterotopic transplantation. Expression of the pX transgene in the transformed tumors were significantly reduced. Among host genes, the expression of p16ink4a/ARF, which was significantly upregulated in the originals, was never detected in the transformed tumors. Genomic Southern blots and PCR suggest that homozygous deletion of the p16ink4a/ARF gene may play important roles in malignant transformation in this model. Our model described here is a useful unique model for in vivo malignant transformation.     

Toxoplasmosis after renal transplantation: implications of a missed diagnosis

We describe a renal transplant patient with a primary Toxoplasma gondii infection presenting as pneumonitis, with subsequent chorioretinitis and encephalitis. The diagnostic challenges of T. gondii infection in immunocompromised patients are discussed.     

First case of Mycobacterium tuberculosis transmission by heart transplantation from donor to recipient

We report the first documented case of a Mycobacterium tuberculosis transmission by an orthotopic heart transplantation from the donor to the recipient. Mycobacterium tuberculosis positive blood culture showed systemic prevalence of the Mycobacteria, however, prophylactic therapy was able to prevent a clinical manifestation of tuberculosis in the recipient.     

The effect of recipient cytokine gene polymorphism on cardiac transplantation outcome

We determined the association between clinical outcomes after heart transplantation and gene polymorphism in five cytokines (tumor necrosis factor-alpha, transforming growth factor (TGF)-beta, interleukin-10, interleukin-6, and interferon-gamma) reported to influence expression in vitro. Ninety-five patients were studied. Cytokine genotyping was performed by sequence specific priming polymerase chain reaction. Clinical outcomes studied in the first posttransplant year included: (1) documented viral, bacterial, or fungal infection; (2) cytomegalovirus infection; (3) acute cellular rejection (International Society for Heart and Lung Transplantation > or = grade IIIA); (4) time to first rejection episode; and (5) the development of allograft vasculopathy. Patients with the TGF-beta genotype 10 T/T 25 G/G or 10 T/C 25 G/G had a longer time to first rejection (median time to first rejection episode 321 days) than those with the TGF-beta genotype 10C/C 25 G/C or 10 C/C 25 C/C (median time to first rejection 88 days). There was a trend toward a higher frequency of the tumor necrosis factor-alpha genotype -308 G/A or A/A in patients without infection (19/59, 32%) as compared with patients with infection (5/31, 16%). In both cases, these differences failed to reach significance when adjusted for multiple comparisons. No other significant association was found with clinical outcomes and polymorphisms in the five cytokine genes studied in this population.     

Fluorescence in situ hybridization for the Y-chromosome can be used to detect cells of recipient origin in allografted hearts following cardiac transplantation.

The purpose of this study was to determine if fluorescence in situ hybridization for the Y-chromosome can be used to detect cells of recipient origin in allografted hearts following cardiac transplantation. Formalin-fixed, paraffin-embedded tissue sections of coronary arteries from two hearts surgically explanted from heart transplant recipients undergoing retransplantation because of accelerated arteriosclerosis were examined by fluorescence in situ hybridization for the presence of cells containing the Y-chromosome using a biotinylated Y-chromosome cocktail probe. In both cases, the recipients were male and the original donor hearts were obtained from female donors. Hybridization was detected in cells morphologically recognizable as infiltrating lymphocytes, macrophages, and mast cells, establishing that these cells in the donor hearts were of recipient origin. In contrast, hybridization was not detected in cardiac myocytes, in vascular smooth muscle cells, or in the majority (>95%) of endothelial cells, suggesting that these cells were of donor origin. Although hybridization was detected in rare flattened cells lining vascular lumina, these cells did not stain for factor VIII, suggesting that they were, in fact, flattened inflammatory cells and not endothelial cells. These results demonstrate that, when the recipient and donor are of the opposite sex, fluorescence in situ hybridization for the Y-chromosome can be used to detect graft chimerism in transplanted hearts.     

Massive haemoptysis after living donor liver transplantation

A 27 year old man with hereditary haemorrhagic telangiectasia who developed progressive liver dysfunction underwent living related right lobe transplantation. Pulmonary arteriography did not reveal arteriovenous malformation or abnormal intrapulmonary venous channels. The postoperative course was characterised by persistent hypoxaemia and respiratory failure developed. On day 6, a massive haemoptysis developed and the patient died shortly thereafter. The native liver showed a nodular pseudocirrhotic transformation, with highly dilated and irregularly interconnected vein-like or arterial-like structures in the fibrous septa. Pathological examination of both lungs showed irregular thickening of the wall of the arteries, secondary to eccentric and/or concentric myointimal hyperplasia. This case suggests that massive haemoptysis can develop even when arteriovenous malformations are undetectable by pulmonary arteriography, and it questions the role and the appropriateness of living donor liver transplantation in high risk patients.     

No increased mortality from donor or recipient hepatitis B- and/or hepatitis C-positive serostatus after related-donor allogeneic hematopoietic cell transplantation

Limited data exist on allogeneic transplant outcomes in recipients receiving hematopoietic cells from donors with prior or current hepatitis B (HBV) or C virus (HCV) infection (seropositive donors), or for recipients with prior or current HBV or HCV infection (seropositive recipients). Transplant outcomes are reported for 416 recipients from 121 centers, who received a human leukocyte antigen-identical related-donor allogeneic transplant for hematologic malignancies between 1995 and 2003. Of these, 33 seronegative recipients received grafts from seropositive donors and 128 recipients were seropositive. The remaining 256 patients served as controls. With comparable median follow-up (cases, 5.9?years; controls, 6.7?years), the incidence of treatment-related mortality, survival, graft-versus-host disease, and hepatic toxicity, appears similar in all cohorts. The frequencies of hepatic toxicities as well as causes of death between cases and controls were similar. Prior exposure to HBV or HCV in either the donor or the recipient should not be considered an absolute contraindication to transplant.     

Donor and recipient origin of herpesvirus-reactive lymphocytes after bone marrow transplantation

Summary Cellular reactivity to herpesviruses was studied after bone marrow transplantation (BMT). In 3 patients all virus-stimulated lymphocytes were of donor type. In the fourth patient, 14 per cent of the HSV stimulated cells were of recipient origin at 6 months after BMT.With 1 Figure