Paola Pontrelli | University of Bari Italy (original) (raw)

Papers by Paola Pontrelli

International Journal of Molecular Sciences

CD40 crosslinking plays an important role in regulating cell migration, adhesion and proliferatio... more CD40 crosslinking plays an important role in regulating cell migration, adhesion and proliferation in renal cell carcinoma (RCC). CD40/CD40L interaction on RCC cells activates different intracellular pathways but the molecular mechanisms leading to cell scattering are not yet clearly defined. Aim of our study was to investigate the main intracellular pathways activated by CD40 ligation and their specific involvement in RCC cell migration. CD40 ligation increased the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun NH (2)-terminal kinase (JNK) and p38 MAPK. Furthermore, CD40 crosslinking activated different transcriptional factors on RCC cell lines: AP-1, NFkB and some members of the Nuclear Factor of Activated T cells (NFAT) family. Interestingly, the specific inhibition of NFAT factors by cyclosporine A, completely blocked RCC cell motility induced by CD40 ligation. In tumor tissue, we observed a higher expression of NFAT factors and in particular an increased ...

Aging, Apr 20, 2021

Pentraxins are a family of evolutionarily conserved pattern recognition molecules with pivotal ro... more Pentraxins are a family of evolutionarily conserved pattern recognition molecules with pivotal roles in innate immunity and inflammation, such as opsonization of pathogens during bacterial and viral infections. In particular, the long Pentraxin 3 (PTX3) has been shown to regulate several aspects of vascular and tissue inflammation during solid organ transplantation. Our study investigated the role of PTX3 as possible modulator of Complement activation in a swine model of renal ischemia/reperfusion (I/R) injury. We demonstrated that I/R injury induced early PTX3 deposits at peritubular and glomerular capillary levels. Confocal laser scanning microscopy revealed PTX3 deposits co-localizing with CD31+ endothelial cells. In addition, PTX3 was associated with infiltrating macrophages (CD163), dendritic cells (SWC3a) and myofibroblasts (FSP1). In particular, we demonstrated a significant PTX3-mediated activation of classical (C1q-mediated) and lectin (MBL-mediated) pathways of Complement. Interestingly, PTX3 deposits co-localized with activation of the terminal Complement complex (C5b-9) on endothelial cells, indicating that PTX3-mediated Complement activation occurred mainly at the renal vascular level. In conclusion, these data indicate that PTX3 might be a potential therapeutic target to prevent Complement-induced I/R injury.

The FASEB Journal, Nov 17, 2022

Kidney International Reports, Jun 1, 2023

Nephrology Dialysis Transplantation, Jun 1, 2023

Association between QT intervals and death. The model was adjusted for age, gender, cause of end-... more Association between QT intervals and death. The model was adjusted for age, gender, cause of end-stage renal failure, serum concentrations of iron, potassium, magnesium, calcium, and phosphorus, history of dialysis, and history of cardiovascular disease.

Biomedicines

Hypoxia activates hypoxia-related signaling pathways controlled by hypoxia-inducible factors (HIF... more Hypoxia activates hypoxia-related signaling pathways controlled by hypoxia-inducible factors (HIFs). HIFs represent a quick and effective detection system involved in the cellular response to insufficient oxygen concentration. Activation of HIF signaling pathways is involved in improving the oxygen supply, promoting cell survival through anaerobic ATP generation, and adapting energy metabolism to meet cell demands. Hypoxia can also contribute to the development of the aging process, leading to aging-related degenerative diseases; among these, the aging of the immune system under hypoxic conditions can play a role in many different immune-mediated diseases. Thus, in this review we aim to discuss the role of HIF signaling pathways following cellular hypoxia and their effects on the mechanisms driving immune system senescence.

Journal of Nephrology

Background COVID-19 in kidney transplant recipients is associated with high morbidity and mortali... more Background COVID-19 in kidney transplant recipients is associated with high morbidity and mortality. In this study we aimed to evaluate: (i) the seroconversion rate after BNT162b2 (Pfizer-BioNTech) SARS-CoV-2 vaccine, (ii) factors associated with humoral response, (iii) clinical outcome of COVID-19 in kidney transplanted patients. Methods We enrolled a cohort of 743 kidney transplant recipients followed up from March 2020 until April 2022. A subset of 336 patients, who received three-doses of SARS-CoV-2 vaccine, was analyzed in terms of kinetics of humoral immune response and compared to a control group of 94 healthcare workers. Antibody response was tested before vaccination (T0), 15 and 90 days after the second dose (T1 and T2), on the day of the third dose (T3) and one month after the third dose (T4). Results We observed that 66 out of 743 subjects had COVID-19 infection pre-vaccination: 65.2% had severe symptoms, 27.3% were hospitalized (9 deaths), none were asymptomatic. After ...

Frontiers in Endocrinology

Several insults can lead to acute kidney injury (AKI) in native kidney and transplant patients, w... more Several insults can lead to acute kidney injury (AKI) in native kidney and transplant patients, with diabetes critically contributing as pivotal risk factor. High glucose per se can disrupt several signaling pathways within the kidney that, if not restored, can favor the instauration of mechanisms of maladaptive repair, altering kidney homeostasis and proper function. Diabetic kidneys frequently show reduced oxygenation, vascular damage and enhanced inflammatory response, features that increase the kidney vulnerability to hypoxia. Importantly, epidemiologic data shows that previous episodes of AKI increase susceptibility to diabetic kidney disease (DKD), and that patients with DKD and history of AKI have a generally worse prognosis compared to DKD patients without AKI; it is therefore crucial to monitor diabetic patients for AKI. In the present review, we will describe the causes that contribute to increased susceptibility to AKI in diabetes, with focus on the molecular mechanisms t...

Journal of Nephrology

Background Hemodialysis patients present a dramatic increase in cardiovascular morbidity/mortalit... more Background Hemodialysis patients present a dramatic increase in cardiovascular morbidity/mortality. Circulating immune cells, activated by both uremic milieu and dialysis, play a key role in the pathogenesis of dialysis-related vascular disease. The aim of our study was to identify, through a high-throughput approach, differences in gene expression profiles in the peripheral blood mononuclear cells (PBMCs) of patients treated with on-line hemodiafiltration and bicarbonate hemodialysis. Methods The transcriptomic profile was investigated in PBMCs isolated from eight patients on on-line hemodiafiltration and eight patients on bicarbonate hemodialysis by microarray analysis. The results were evaluated by statistical and functional pathway analysis and validated by real time PCR (qPCR) in an independent cohort of patients (on-line hemodiafiltration N = 20, bicarbonate hemodialysis n = 20). Results Eight hundred and forty-seven genes were differentially expressed in patients treated with...

Frontiers in Immunology

Atypical hemolytic–uremic syndrome (aHUS) is a severe thrombotic microangiopathy in which kidney ... more Atypical hemolytic–uremic syndrome (aHUS) is a severe thrombotic microangiopathy in which kidney involvement is common. aHUS can be due to either genetic or acquired abnormalities, with most abnormalities affecting the alternative complement pathway. Several genetic factors/alterations can drive the clinical presentation, therapeutic response, and risk of recurrence, especially recurrence following kidney transplantation. We report here the case of a 22-year-old man who developed a severe form of aHUS. Renal biopsy revealed thrombotic microangiopathy and features of chronic renal damage. Despite two eculizumab infusions, the patient remained dialysis dependent. Two novel rare variants, c.109G>A (p.E37K) and c.159 C>A (p.Y53*), were identified in the factor H-related 2 (FHR2) gene, and western blot analysis revealed a significant reduction in the level of FHR2 protein in the patient’s serum. Although FHR2 involvement in complement 3 glomerulopathy has been reported previously, ...

American Journal of Transplantation, 2022

Kidney transplant recipients (KTR) have been considered as patients at higher risk of SARS-CoV-2-... more Kidney transplant recipients (KTR) have been considered as patients at higher risk of SARS-CoV-2-related disease severity, thus COVID-19 vaccination was highly recommended. However possible interferences of different immunosuppression with development of both humoral and T-cell-mediated immune response to COVID-19 vaccination has not been determined. Here we evaluated the association between mTOR-inhibitors (mTOR-I) and immune response to mRNA BNT162b2 (Pfizer-Biontech) vaccine in KTR. To this aim 132 consecutive KTR vaccinated against COVID-19 in the early 2021 were enrolled, and humoral and T-cell-mediated immune response were assessed after 4-5 weeks. Patients treated with mTOR-I showed significantly higher anti-SARS-CoV-2 IgG titer (p=0.003) and higher percentages of anti-SARS-CoV-2 S1/RBD Ig (p=0.024), than those without. Moreover, SARS-CoV-2-specific T-cell-derived IFNγ release was significantly increased in patients treated with mTOR-I (p<0.001), than in those without. Multivariate analysis confirmed that therapy with mTOR-I gained better humoral (p=0.005) and T-cell-mediated immune response (p=0.005) in KTR. The presence of mTOR-I is associated to a better immune response to COVID-19 vaccine in KTR compared to therapy without mTOR-I, not only by increasing vaccine-induced antibodies but also stimulating anti-SARS-CoV-2 T-cell response. These finding are consistent with a potential beneficial role of mTOR-I as modulators of immune response to COVID-19 vaccine in KTR.

Nephrology Dialysis Transplantation, 2018

To analyze the differences in central nervous system structure between patients on haemodialysis ... more To analyze the differences in central nervous system structure between patients on haemodialysis and non-dialysis dependent CKD patients (NDD-CKD). METHODS: Cross section of KIDBRAIN study that includes 100 patients: 68 on haemodialysis and 32 NDD-CKD (stage 4 and 5). We analyzed grey matter volume (cortical and subcortical), white matter volume and axonal functional integrity evaluated by anisotropy fraction (FA) obtained by brain MRI; also, the influence of the 24 hoursurine creatinine clearance over the structures of the CNS including models adjusted to age, sex, DM, cardiovascular history disease, serum cystatine C and systolic blood pressure. The CNS structural differences between NDD-CKD patients and those on haemodialysis were studied in two models: 1.-including all the patients on haemodialysis (68) and 2.-excluding those with residual renal function (18). RESULTS: Mean age: 60,4615,7 years, male 68%, DM 37,5%, cardiovascular history disease 18,7%, serum cystatine C 3,561,0 mg/dl, systolic blood pressure 139,8619,6 mmHg. We found an independent correlation (Beta: 0,486, p:0,021) in the adjusted model between 24-hours-creatinine clearance (19,66 6 ml/min) and white matter volume (r:0,499, p:0,004). There is no assocciation between 24-hours-creatinine clearance and grey matter volume, anisotropy fraction in NDD-CKD.Globally, there is no differences in grey and white matter volumes and anisotropy fraction in haemodialysis and NDD-CKD patients. However, when we excluded the patients with residual renal function [700ml (300-1050ml)] there is differences in grey matter volume (p:0,038) and white matter volume (p:0,024) but no differences in anisotropy fraction (p:0,16). Multivariable analysis confirmed that haemodialysis patients have a less grey matter volume (p:0,015) independently. On the opposite, white matter volumen is dependent to age, sex and DM. CONCLUSIONS: The loss of renal function is accompanied with structural changes in central nervous system at the expense of less grey and white matter volume. This effect is independent of other cardiovascular risk factors. These changes get worse with the dialysis, especially when the residual renal function is lost.

Background: Lung transplantation is still the best therapeutic option for cystic fibrosis (CF) pa... more Background: Lung transplantation is still the best therapeutic option for cystic fibrosis (CF) patients, but, unfortunately, immunosuppressive therapies, often employed at high dosages to avoid acute rejection, may induce severe complications. In particular, patients treated with high dose of mammalian target of rapamycin inhibitors (mTOR-Is) may experience lung fibrosis (including bronchiolitis obliterans-organizing pneumonia). Although epithelial to mesenchymal transition (EMT) of airway cells has a central role in this process, the complete biological machinery is not completely clarified.Methods: In order to improve our knowledge on this process, primary bronchial epithelial cells carrying F508del mutation were treated with 5 and 100 nM everolimus (EVE) for 24 hours. Subsequently, RNA was hybridized to the Human HT-12 v3 Expression BeadChip (Illumina). Real-Time PCR was, then, used to validate microarray results and to measure major EMT biomarkers. Trans-epithelial resistance wa...

Nephrology Dialysis Transplantation, 2018

Nephrology Dialysis Transplantation, 2020

Background and Aims Chronic Kidney Disease (CKD) is strongly associated with increased circulator... more Background and Aims Chronic Kidney Disease (CKD) is strongly associated with increased circulatory complications that can lead to accelerated cardiovascular aging. During CKD, persistent oxidative stress triggered by mitochondrial dysfunction can affect Klotho protein levels together with circulating FGF23. mTOR pathway is involved in pro-survival mechanism and can be differently modulated by several available therapeutics, therefore exerting central function in senescence, autophagy and cellular regeneration. The aim of this study was to evaluate the beneficial effect of mTOR inhibition on cardiovascular aging in transplanted patient with Chronic Allograft Dysfunction. Method In this study, 250 transplanted patients with calcineurin inhibitory (CNI) therapy and graft survival higher than 6 months and lower than 60 months were enrolled. The patients were then divided in three groups: (MMF group) patients undergoing CNI therapy with CS (Ciclosporin) and MMF, (mTOR Inhibitors group) p...

Transplantation, 2014

CAMR is the main cause of chronic graft injury and loss, but its pathogenesis is still largely un... more CAMR is the main cause of chronic graft injury and loss, but its pathogenesis is still largely unclear. The aim of the present study was to investigate the molecular mechanisms underlying the development of CAMR by the analysis of gene expression profi les of both total peripheral lymphomonocytes (PLM) and isolated CD4+ and CD8+ T lymphocytes. We enrolled 18 patients with biopsy-proven CAMR and 18 transplant recipients with normal graft histology/function (control group). Gene expression profi le of PLM and CD4+ and CD8+ T lymphocytes isolated from both groups (n=8/group) was assessed by Agilent microarrays, evaluated by standard statistical and functional pathway analysis (Ingenuity Pathway Analysis, IPA) and validated by quantitative RT-PCR in an independent set of patients (n=6/group). We identifi ed 67 probesets, corresponding to 53 genes, differentially expressed in the 2 groups (FDR<5%, fold-change ≥2). The main canonical pathway identifi ed by these genes was the IFN-alpha pathway (p=6.0*10-5), while network analysis showed that 22/53 of the differentially-expressed genes regulate or are regulated by IFN-alpha (IPA score=52, p≤.01). Quantitative RT-PCR on the main differentially expressed genes (GBP1, CXCL9, Serping1, C1QA, SLC1A7) confirmed the results obtained by microarrays. We, then, analyzed the gene expression profi le of CD4+ and CD8+T lymphocytes isolated from an independent cohort of 4 CAMR and 4 control subjects. Only one gene (TNFSF10) was differentially expressed in CAMR patients in both cell population. CD4+T cells showed a specifi c IFN-alpha signature (p=5.06*10-11), while CD8+T were characterized by several differentially regulated pathways (Tryacylglicerol biosynthesis p=1.4*10-3; TREM1 signaling p=5.8*10-2; Toll like receptor signaling p=5.2*10-2), but IFN-alpha signaling. Immunofl uorescence analysis of graft biopsies demonstrated a statistically signifi cant increased expression of MXA, an IFN-alpha-dependent protein, in CAMR patients (CAMR 1471.6±28.9 pixels vs control 689±44 pixels, p=.02). Our data suggest a key role for IFN-alpha in modulating CD4+T cell response during CAMR. This observation may open new perspectives for early non-invasive diagnosis of CAMR.

Transplantation, 2014

Conclusion: A panel of 3 urinary proteins, mined by high throughput proteomics, and validated by ... more Conclusion: A panel of 3 urinary proteins, mined by high throughput proteomics, and validated by customized ELISA, allows for non-invasive diagnosis of AR, not confounded by BKVN. This urinary assay requires no local sample processing, is robust and can be used as a sensitive assay for outpatient renal transplant injury surveillance.

Nephrology Dialysis Transplantation, 2018

Background. Malignancies represent the third leading cause of post-transplant mortality worldwide... more Background. Malignancies represent the third leading cause of post-transplant mortality worldwide. The main challenge for transplant physicians is a timely diagnosis of this condition. The aim of the study was to identify a soluble diagnostic marker for monitoring the development of post-transplant malignancies. Methods. This is a multicentre, observational, perspective, case-control study. We enrolled 47 patients with posttransplant solid neoplasia. As a control group we employed 106 transplant recipients without a history of neoplasia and matched them with cases for the main demographic and clinical features. We investigated the transcriptomic profiles of peripheral blood mononuclear cells from kidney graft recipients with and without post-transplant malignancies enrolled in two of the participating centres, randomly selected from the whole study population. Microarray results were confirmed by quantitative polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) in the remaining patients from the same transplant centres and validated in a further independent group enrolled in two different transplant centres. Results. We identified 535 differentially expressed genes comparing patients with and without post-transplant malignancies (fold change !2.5; false discovery rate <5%). The cancer pathway was closely related to gene expression data, and one of the most down-regulated genes in this pathway was interleukin-27 (IL-27), a cytokine regulating anti-tumour immunity. Quantitative PCR and ELISA confirmed the microarray data. Interestingly, IL-27 plasma levels were able to discriminate patients with post-transplant neoplasia with a specificity of 80% and a sensitivity of 81%. This observation was confirmed in an independent set of patients from two different transplant centres. Conclusions. Our data suggest that IL-27 may represent a potential immunological marker for the timely identification of post-transplant neoplasia.

Nephrology Dialysis Transplantation, 2016

International Journal of Molecular Sciences

CD40 crosslinking plays an important role in regulating cell migration, adhesion and proliferatio... more CD40 crosslinking plays an important role in regulating cell migration, adhesion and proliferation in renal cell carcinoma (RCC). CD40/CD40L interaction on RCC cells activates different intracellular pathways but the molecular mechanisms leading to cell scattering are not yet clearly defined. Aim of our study was to investigate the main intracellular pathways activated by CD40 ligation and their specific involvement in RCC cell migration. CD40 ligation increased the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun NH (2)-terminal kinase (JNK) and p38 MAPK. Furthermore, CD40 crosslinking activated different transcriptional factors on RCC cell lines: AP-1, NFkB and some members of the Nuclear Factor of Activated T cells (NFAT) family. Interestingly, the specific inhibition of NFAT factors by cyclosporine A, completely blocked RCC cell motility induced by CD40 ligation. In tumor tissue, we observed a higher expression of NFAT factors and in particular an increased ...

Aging, Apr 20, 2021

Pentraxins are a family of evolutionarily conserved pattern recognition molecules with pivotal ro... more Pentraxins are a family of evolutionarily conserved pattern recognition molecules with pivotal roles in innate immunity and inflammation, such as opsonization of pathogens during bacterial and viral infections. In particular, the long Pentraxin 3 (PTX3) has been shown to regulate several aspects of vascular and tissue inflammation during solid organ transplantation. Our study investigated the role of PTX3 as possible modulator of Complement activation in a swine model of renal ischemia/reperfusion (I/R) injury. We demonstrated that I/R injury induced early PTX3 deposits at peritubular and glomerular capillary levels. Confocal laser scanning microscopy revealed PTX3 deposits co-localizing with CD31+ endothelial cells. In addition, PTX3 was associated with infiltrating macrophages (CD163), dendritic cells (SWC3a) and myofibroblasts (FSP1). In particular, we demonstrated a significant PTX3-mediated activation of classical (C1q-mediated) and lectin (MBL-mediated) pathways of Complement. Interestingly, PTX3 deposits co-localized with activation of the terminal Complement complex (C5b-9) on endothelial cells, indicating that PTX3-mediated Complement activation occurred mainly at the renal vascular level. In conclusion, these data indicate that PTX3 might be a potential therapeutic target to prevent Complement-induced I/R injury.

The FASEB Journal, Nov 17, 2022

Kidney International Reports, Jun 1, 2023

Nephrology Dialysis Transplantation, Jun 1, 2023

Association between QT intervals and death. The model was adjusted for age, gender, cause of end-... more Association between QT intervals and death. The model was adjusted for age, gender, cause of end-stage renal failure, serum concentrations of iron, potassium, magnesium, calcium, and phosphorus, history of dialysis, and history of cardiovascular disease.

Biomedicines

Hypoxia activates hypoxia-related signaling pathways controlled by hypoxia-inducible factors (HIF... more Hypoxia activates hypoxia-related signaling pathways controlled by hypoxia-inducible factors (HIFs). HIFs represent a quick and effective detection system involved in the cellular response to insufficient oxygen concentration. Activation of HIF signaling pathways is involved in improving the oxygen supply, promoting cell survival through anaerobic ATP generation, and adapting energy metabolism to meet cell demands. Hypoxia can also contribute to the development of the aging process, leading to aging-related degenerative diseases; among these, the aging of the immune system under hypoxic conditions can play a role in many different immune-mediated diseases. Thus, in this review we aim to discuss the role of HIF signaling pathways following cellular hypoxia and their effects on the mechanisms driving immune system senescence.

Journal of Nephrology

Background COVID-19 in kidney transplant recipients is associated with high morbidity and mortali... more Background COVID-19 in kidney transplant recipients is associated with high morbidity and mortality. In this study we aimed to evaluate: (i) the seroconversion rate after BNT162b2 (Pfizer-BioNTech) SARS-CoV-2 vaccine, (ii) factors associated with humoral response, (iii) clinical outcome of COVID-19 in kidney transplanted patients. Methods We enrolled a cohort of 743 kidney transplant recipients followed up from March 2020 until April 2022. A subset of 336 patients, who received three-doses of SARS-CoV-2 vaccine, was analyzed in terms of kinetics of humoral immune response and compared to a control group of 94 healthcare workers. Antibody response was tested before vaccination (T0), 15 and 90 days after the second dose (T1 and T2), on the day of the third dose (T3) and one month after the third dose (T4). Results We observed that 66 out of 743 subjects had COVID-19 infection pre-vaccination: 65.2% had severe symptoms, 27.3% were hospitalized (9 deaths), none were asymptomatic. After ...

Frontiers in Endocrinology

Several insults can lead to acute kidney injury (AKI) in native kidney and transplant patients, w... more Several insults can lead to acute kidney injury (AKI) in native kidney and transplant patients, with diabetes critically contributing as pivotal risk factor. High glucose per se can disrupt several signaling pathways within the kidney that, if not restored, can favor the instauration of mechanisms of maladaptive repair, altering kidney homeostasis and proper function. Diabetic kidneys frequently show reduced oxygenation, vascular damage and enhanced inflammatory response, features that increase the kidney vulnerability to hypoxia. Importantly, epidemiologic data shows that previous episodes of AKI increase susceptibility to diabetic kidney disease (DKD), and that patients with DKD and history of AKI have a generally worse prognosis compared to DKD patients without AKI; it is therefore crucial to monitor diabetic patients for AKI. In the present review, we will describe the causes that contribute to increased susceptibility to AKI in diabetes, with focus on the molecular mechanisms t...

Journal of Nephrology

Background Hemodialysis patients present a dramatic increase in cardiovascular morbidity/mortalit... more Background Hemodialysis patients present a dramatic increase in cardiovascular morbidity/mortality. Circulating immune cells, activated by both uremic milieu and dialysis, play a key role in the pathogenesis of dialysis-related vascular disease. The aim of our study was to identify, through a high-throughput approach, differences in gene expression profiles in the peripheral blood mononuclear cells (PBMCs) of patients treated with on-line hemodiafiltration and bicarbonate hemodialysis. Methods The transcriptomic profile was investigated in PBMCs isolated from eight patients on on-line hemodiafiltration and eight patients on bicarbonate hemodialysis by microarray analysis. The results were evaluated by statistical and functional pathway analysis and validated by real time PCR (qPCR) in an independent cohort of patients (on-line hemodiafiltration N = 20, bicarbonate hemodialysis n = 20). Results Eight hundred and forty-seven genes were differentially expressed in patients treated with...

Frontiers in Immunology

Atypical hemolytic–uremic syndrome (aHUS) is a severe thrombotic microangiopathy in which kidney ... more Atypical hemolytic–uremic syndrome (aHUS) is a severe thrombotic microangiopathy in which kidney involvement is common. aHUS can be due to either genetic or acquired abnormalities, with most abnormalities affecting the alternative complement pathway. Several genetic factors/alterations can drive the clinical presentation, therapeutic response, and risk of recurrence, especially recurrence following kidney transplantation. We report here the case of a 22-year-old man who developed a severe form of aHUS. Renal biopsy revealed thrombotic microangiopathy and features of chronic renal damage. Despite two eculizumab infusions, the patient remained dialysis dependent. Two novel rare variants, c.109G>A (p.E37K) and c.159 C>A (p.Y53*), were identified in the factor H-related 2 (FHR2) gene, and western blot analysis revealed a significant reduction in the level of FHR2 protein in the patient’s serum. Although FHR2 involvement in complement 3 glomerulopathy has been reported previously, ...

American Journal of Transplantation, 2022

Kidney transplant recipients (KTR) have been considered as patients at higher risk of SARS-CoV-2-... more Kidney transplant recipients (KTR) have been considered as patients at higher risk of SARS-CoV-2-related disease severity, thus COVID-19 vaccination was highly recommended. However possible interferences of different immunosuppression with development of both humoral and T-cell-mediated immune response to COVID-19 vaccination has not been determined. Here we evaluated the association between mTOR-inhibitors (mTOR-I) and immune response to mRNA BNT162b2 (Pfizer-Biontech) vaccine in KTR. To this aim 132 consecutive KTR vaccinated against COVID-19 in the early 2021 were enrolled, and humoral and T-cell-mediated immune response were assessed after 4-5 weeks. Patients treated with mTOR-I showed significantly higher anti-SARS-CoV-2 IgG titer (p=0.003) and higher percentages of anti-SARS-CoV-2 S1/RBD Ig (p=0.024), than those without. Moreover, SARS-CoV-2-specific T-cell-derived IFNγ release was significantly increased in patients treated with mTOR-I (p<0.001), than in those without. Multivariate analysis confirmed that therapy with mTOR-I gained better humoral (p=0.005) and T-cell-mediated immune response (p=0.005) in KTR. The presence of mTOR-I is associated to a better immune response to COVID-19 vaccine in KTR compared to therapy without mTOR-I, not only by increasing vaccine-induced antibodies but also stimulating anti-SARS-CoV-2 T-cell response. These finding are consistent with a potential beneficial role of mTOR-I as modulators of immune response to COVID-19 vaccine in KTR.

Nephrology Dialysis Transplantation, 2018

To analyze the differences in central nervous system structure between patients on haemodialysis ... more To analyze the differences in central nervous system structure between patients on haemodialysis and non-dialysis dependent CKD patients (NDD-CKD). METHODS: Cross section of KIDBRAIN study that includes 100 patients: 68 on haemodialysis and 32 NDD-CKD (stage 4 and 5). We analyzed grey matter volume (cortical and subcortical), white matter volume and axonal functional integrity evaluated by anisotropy fraction (FA) obtained by brain MRI; also, the influence of the 24 hoursurine creatinine clearance over the structures of the CNS including models adjusted to age, sex, DM, cardiovascular history disease, serum cystatine C and systolic blood pressure. The CNS structural differences between NDD-CKD patients and those on haemodialysis were studied in two models: 1.-including all the patients on haemodialysis (68) and 2.-excluding those with residual renal function (18). RESULTS: Mean age: 60,4615,7 years, male 68%, DM 37,5%, cardiovascular history disease 18,7%, serum cystatine C 3,561,0 mg/dl, systolic blood pressure 139,8619,6 mmHg. We found an independent correlation (Beta: 0,486, p:0,021) in the adjusted model between 24-hours-creatinine clearance (19,66 6 ml/min) and white matter volume (r:0,499, p:0,004). There is no assocciation between 24-hours-creatinine clearance and grey matter volume, anisotropy fraction in NDD-CKD.Globally, there is no differences in grey and white matter volumes and anisotropy fraction in haemodialysis and NDD-CKD patients. However, when we excluded the patients with residual renal function [700ml (300-1050ml)] there is differences in grey matter volume (p:0,038) and white matter volume (p:0,024) but no differences in anisotropy fraction (p:0,16). Multivariable analysis confirmed that haemodialysis patients have a less grey matter volume (p:0,015) independently. On the opposite, white matter volumen is dependent to age, sex and DM. CONCLUSIONS: The loss of renal function is accompanied with structural changes in central nervous system at the expense of less grey and white matter volume. This effect is independent of other cardiovascular risk factors. These changes get worse with the dialysis, especially when the residual renal function is lost.

Background: Lung transplantation is still the best therapeutic option for cystic fibrosis (CF) pa... more Background: Lung transplantation is still the best therapeutic option for cystic fibrosis (CF) patients, but, unfortunately, immunosuppressive therapies, often employed at high dosages to avoid acute rejection, may induce severe complications. In particular, patients treated with high dose of mammalian target of rapamycin inhibitors (mTOR-Is) may experience lung fibrosis (including bronchiolitis obliterans-organizing pneumonia). Although epithelial to mesenchymal transition (EMT) of airway cells has a central role in this process, the complete biological machinery is not completely clarified.Methods: In order to improve our knowledge on this process, primary bronchial epithelial cells carrying F508del mutation were treated with 5 and 100 nM everolimus (EVE) for 24 hours. Subsequently, RNA was hybridized to the Human HT-12 v3 Expression BeadChip (Illumina). Real-Time PCR was, then, used to validate microarray results and to measure major EMT biomarkers. Trans-epithelial resistance wa...

Nephrology Dialysis Transplantation, 2018

Nephrology Dialysis Transplantation, 2020

Background and Aims Chronic Kidney Disease (CKD) is strongly associated with increased circulator... more Background and Aims Chronic Kidney Disease (CKD) is strongly associated with increased circulatory complications that can lead to accelerated cardiovascular aging. During CKD, persistent oxidative stress triggered by mitochondrial dysfunction can affect Klotho protein levels together with circulating FGF23. mTOR pathway is involved in pro-survival mechanism and can be differently modulated by several available therapeutics, therefore exerting central function in senescence, autophagy and cellular regeneration. The aim of this study was to evaluate the beneficial effect of mTOR inhibition on cardiovascular aging in transplanted patient with Chronic Allograft Dysfunction. Method In this study, 250 transplanted patients with calcineurin inhibitory (CNI) therapy and graft survival higher than 6 months and lower than 60 months were enrolled. The patients were then divided in three groups: (MMF group) patients undergoing CNI therapy with CS (Ciclosporin) and MMF, (mTOR Inhibitors group) p...

Transplantation, 2014

CAMR is the main cause of chronic graft injury and loss, but its pathogenesis is still largely un... more CAMR is the main cause of chronic graft injury and loss, but its pathogenesis is still largely unclear. The aim of the present study was to investigate the molecular mechanisms underlying the development of CAMR by the analysis of gene expression profi les of both total peripheral lymphomonocytes (PLM) and isolated CD4+ and CD8+ T lymphocytes. We enrolled 18 patients with biopsy-proven CAMR and 18 transplant recipients with normal graft histology/function (control group). Gene expression profi le of PLM and CD4+ and CD8+ T lymphocytes isolated from both groups (n=8/group) was assessed by Agilent microarrays, evaluated by standard statistical and functional pathway analysis (Ingenuity Pathway Analysis, IPA) and validated by quantitative RT-PCR in an independent set of patients (n=6/group). We identifi ed 67 probesets, corresponding to 53 genes, differentially expressed in the 2 groups (FDR<5%, fold-change ≥2). The main canonical pathway identifi ed by these genes was the IFN-alpha pathway (p=6.0*10-5), while network analysis showed that 22/53 of the differentially-expressed genes regulate or are regulated by IFN-alpha (IPA score=52, p≤.01). Quantitative RT-PCR on the main differentially expressed genes (GBP1, CXCL9, Serping1, C1QA, SLC1A7) confirmed the results obtained by microarrays. We, then, analyzed the gene expression profi le of CD4+ and CD8+T lymphocytes isolated from an independent cohort of 4 CAMR and 4 control subjects. Only one gene (TNFSF10) was differentially expressed in CAMR patients in both cell population. CD4+T cells showed a specifi c IFN-alpha signature (p=5.06*10-11), while CD8+T were characterized by several differentially regulated pathways (Tryacylglicerol biosynthesis p=1.4*10-3; TREM1 signaling p=5.8*10-2; Toll like receptor signaling p=5.2*10-2), but IFN-alpha signaling. Immunofl uorescence analysis of graft biopsies demonstrated a statistically signifi cant increased expression of MXA, an IFN-alpha-dependent protein, in CAMR patients (CAMR 1471.6±28.9 pixels vs control 689±44 pixels, p=.02). Our data suggest a key role for IFN-alpha in modulating CD4+T cell response during CAMR. This observation may open new perspectives for early non-invasive diagnosis of CAMR.

Transplantation, 2014

Conclusion: A panel of 3 urinary proteins, mined by high throughput proteomics, and validated by ... more Conclusion: A panel of 3 urinary proteins, mined by high throughput proteomics, and validated by customized ELISA, allows for non-invasive diagnosis of AR, not confounded by BKVN. This urinary assay requires no local sample processing, is robust and can be used as a sensitive assay for outpatient renal transplant injury surveillance.

Nephrology Dialysis Transplantation, 2018

Background. Malignancies represent the third leading cause of post-transplant mortality worldwide... more Background. Malignancies represent the third leading cause of post-transplant mortality worldwide. The main challenge for transplant physicians is a timely diagnosis of this condition. The aim of the study was to identify a soluble diagnostic marker for monitoring the development of post-transplant malignancies. Methods. This is a multicentre, observational, perspective, case-control study. We enrolled 47 patients with posttransplant solid neoplasia. As a control group we employed 106 transplant recipients without a history of neoplasia and matched them with cases for the main demographic and clinical features. We investigated the transcriptomic profiles of peripheral blood mononuclear cells from kidney graft recipients with and without post-transplant malignancies enrolled in two of the participating centres, randomly selected from the whole study population. Microarray results were confirmed by quantitative polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) in the remaining patients from the same transplant centres and validated in a further independent group enrolled in two different transplant centres. Results. We identified 535 differentially expressed genes comparing patients with and without post-transplant malignancies (fold change !2.5; false discovery rate <5%). The cancer pathway was closely related to gene expression data, and one of the most down-regulated genes in this pathway was interleukin-27 (IL-27), a cytokine regulating anti-tumour immunity. Quantitative PCR and ELISA confirmed the microarray data. Interestingly, IL-27 plasma levels were able to discriminate patients with post-transplant neoplasia with a specificity of 80% and a sensitivity of 81%. This observation was confirmed in an independent set of patients from two different transplant centres. Conclusions. Our data suggest that IL-27 may represent a potential immunological marker for the timely identification of post-transplant neoplasia.

Nephrology Dialysis Transplantation, 2016