J C Voltarelli

Northwestern University, Evanston, IL, United States

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Publications (151)417.13 Total impact

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    ABSTRACT: Autologous hematopoietic stem cell transplantation (AHSCT) has been experimented as a treatment in patients affected by severe forms of multiple sclerosis (MS) who failed to respond to standard immunotherapy. The rationale of AHSCT is to "reboot" the immune system and reconstitute a new adaptive immunity. The aim of our study was to identify through a robust and unbiased transcriptomic analysis any changes of gene expression in T cells potentially underlying the treatment effect in patients who underwent non-myeloablative AHSCT for treatment of MS. We evaluated by microarray DNA-chip technology the gene expression of peripheral CD4+ and CD8+ T cell subsets sorted from patients with MS patients before AHSCT, at 6 months, 1 year and 2 years after AHSCT and from healthy control subjects. Hierarchical clustering analysis revealed that reconstituted CD8+ T cells of MS patients at 2 years post-transplantation aggregated together with healthy controls, suggesting a normalization of gene expression in CD8+ cells post-therapy. When we compared the gene expression in MS patients before and after therapy we detected a large number of differentially expressed genes (DEG) in both CD8+ and CD4+ T cell subsets at all time-points after transplantation. We catalogued the biological function of DEG and we selected 27 genes known to be involved in immune function for accurate quantification of gene expression by real-time PCR. The analysis confirmed and extended with quantitative data a number of significant changes in both the CD4+ and CD8+ T cells subsets from MS post-transplant. Notably, CD8+ T cells revealed more extensive changes in the expression of genes involved in effector immune responses.
    Clinical science (London, England : 1979). 08/2014;
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    ABSTRACT: This study evaluated the influence of fludarabine on the pharmacokinetics of busulfan administered orally to patients receiving a conditioning regimen for hematopoietic allogeneic stem cell transplantation (HSCT). Twenty-six patients treated with oral busulfan (1 mg/kg/6 h for 4 days) were divided into two groups according to the concomitant administration of fludarabine (n = 11; 30 mg/m(2) for 5 days) or subsequent administration of cyclophosphamide (n = 15; 60 mg/kg for 2 days). Serial blood samples were collected on Day 4 of busulfan administration. Plasma busulfan concentrations were determined by HPLC-UV and the pharmacokinetic parameters were calculated using the WinNonlin program. Patients concomitantly treated with fludarabine showed reduced apparent clearance of busulfan (110.5 mL/h/kg vs. 157.4 mL/h/kg) and higher AUC0-6 (area under the plasma concentrations vs. time curve) than patients subsequently treated with cyclophosphamide (7.9 µg h/mL vs. 5.7 µg h/mL). No association was observed between busulfan AUC0-6 and clinical evolution of the patients. Although plasma busulfan concentrations were higher in patients receiving concomitant fludarabine, myelosuppression-related toxicity was less frequent than in patients treated with busulfan and cyclophosphamide. The results suggest that patients treated with fludarabine should receive 30% lower busulfan doses during conditioning protocols for HSCT.
    The Journal of Clinical Pharmacology 08/2013; · 2.84 Impact Factor
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    ABSTRACT: BACKGROUND: Autologous haemopoietic stem-cell transplantation (HSCT) benefits patients with systemic sclerosis but has been associated with significant treatment-related mortality and failure to improve diffusion capacity of carbon monoxide (DLCO). We aimed to assess efficacy of HSCT and use of rigorous cardiac screening in this group. METHODS: We assessed patients with diffuse systemic sclerosis or limited systemic sclerosis and interstitial lung disease who were treated with HSCT as part of a study or on a compassionate basis at Northwestern University (Chicago, IL, USA) or the University of São Paulo (Ribeirão Preto, Brazil). Unselected peripheral blood stem cells were harvested with cyclophosphamide (2 g/m(2)) and filgrastim. The transplant regimen was a non-myeloablative regimen of cyclophosphamide (200 mg/kg) and rabbit anti-thymocyte globulin (rATG; 4·5-6·5 mg/kg). We followed patients up to 5 years for overall survival, relapse-free survival, modified Rodnan skin score, and pulmonary function tests. FINDINGS: Five (6%) of 90 patients died from treatment-related causes. Despite standard guidelines that recommend echocardiogram for screening before transplantation, four treatment-related deaths occurred because of cardiovascular complications (one constrictive pericarditis, two right heart failures without underlying infection, and one heart failure during mobilisation), and one death was secondary to sepsis without documented underlying heart disease. Kaplan-Meier analysis showed survival was 78% at 5 years (after eight relapse-related deaths) and relapse-free survival was 70% at 5 years. Compared with baseline, we noted improvements after HSCT in modified Rodnan skin scores at 1 year (58 patients; p<0·0001), 2 years (42 patients; p<0·0001), and 3 years (27 patients; p<0·0001) and forced vital capacity at 1 year (58 patients; p=0·009), 2 years (40 patients; p=0·02), and 3 years (28 patients; p=0·004), but total lung capacity and DLCO were not improved significantly after HSCT. Overall mean DLCO was significantly improved in patients with normal baseline echocardiograms (p=0·005) or electrocardiographs (p=0·05). INTERPRETATION: Autologous HSCT with a non-myeloablative regimen of cyclophosphamide and rATG with a non-selected autograft results in sustained improvement in skin thickness and forced vital capacity. DLCO is affected by baseline cardiac function. Guidelines for cardiac screening of patients with systemic sclerosis to assess treatment-related risk from pulmonary artery hypertension, primary cardiac involvement, or pericardial disease should be reconsidered and updated. FUNDING: None.
    The Lancet 01/2013; · 39.21 Impact Factor
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    Revista Brasileira de Hematologia e Hemoterapia 01/2013; 35(2):134-43.
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    ABSTRACT: Under many circumstances, the host constituents that are found in the tumor microenvironment support a malignancy network and provide the cancer cells with advantages in proliferation, invasiveness and metastasis establishment at remote organs. It is known that Toll like receptors (TLRs) are expressed not only on immune cells but also on cancer cells and it has suggested a deleterious role for TLR3 in inflammatory disease. Hypothesizing that altered IFNγ signaling may be a key mechanism of immune dysfunction common to cancer as well CXCR4 is overexpressed among breast cancer patients, the mRNA expression of TLR3, CXCR4 and IFNγ in breast cancer tumor tissues was investigated. No statistically significant differences in the expression of CXCR4 mRNA, IFNγ and TLR3 between healthy and tumor tissues was observed, however, it was verified a positive correlation between mRNA relative expression of TLR3 and CXCR4 (p < 0.001), and mRNA relative expression of TLR3 was significantly increased in breast cancer tumor tissue when compared to healthy mammary gland tissue among patients expressing high IFNγ (p = 0.001). Since the tumor microenvironment plays important roles in cancer initiation, growth, progression, invasion and metastasis, it is possible to propose that an overexpression of IFNγ mRNA due to the pro-inflammatory microenvironment can lead to an up-regulation of CXCR4 mRNA and consequently to an increased TLR3 mRNA expression even among nodal negative patients. In the future, a comprehensive study of TLR3, CXCR4 and IFNγ axis in primary breast tumors and corresponding healthy tissues will be crucial to further understanding of the cancer network.
    Molecular Biology Reports 10/2012; · 2.51 Impact Factor
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    ABSTRACT: Three clinical trials are being conducted in Brazil by our research group on the use of autologous, BM-derived, hematopoietic SCT (auto-BMHSCT) for the treatment of retinitis pigmentosa (RP), dry age-related macular degeneration and ischaemic retinopathy (including diabetic retinopathy with macular ischaemia). These studies are registered with ClinicalTrials.gov, numbers NCT01068561, NCT01518127 and NCT01518842, respectively. 1 We published the results of phase I of the clinical trial using an intravitreal injection of 0.1 mL of a cell suspension containing 0.92 Â 10 4 –2.91 Â 10 4 (mean, 1.68 Â 10 4) BM-derived hemato-poietic stem cells (CD34 þ) for retinal dystrophy. The second phase of this study (NCT01560715) has already begun, and now we describe the case of a participant in this study who had cystoid macular oedema (CMO) associated with RP. After this, patient underwent treatment with an intraocular injection (intravitreal) of auto-BMHSCT in which the patient showed complete resolution of the oedema 7 days after injection, and this effect remained for 1 month of follow-up, which was observed through examination using optical coherence tomography (Figure 1). 2, 3 The improvement of the oedema led to an improvement in visual acuity (20/50–20/32) and an improvement in macular sensitivity, as measured by the microperimetry test (Figure 2). This patient had macular oedema associated with RP that started 2 years ago and had already undergone treatment with systemic carbonic anhydrase inhibitors (acetazolamide) 250 mg three times daily for 3 months and dorzolamide eye drops three times daily for 1 year without any sign of improvement in macular oedema. This group of drugs is commonly used to treat macular oedema associated with RP. 4–6 Acetazolamide increases fluid transport through the retinal pigment epithelium (RPE) by active ion transport and acidification of the RPE. 7 Macular oedema owing to low-grade inflammation may respond better to this mechanism than the diffuse loss of endothelial tight junctions that occurs in diabetes. Topical carbonic anhydrase inhibitors have a lower rate of systemic side effects than acetazolamide but are less efficacious. 4 CME that is refractory to acetazolamide treatment usually leads to permanent visual acuity loss. 5,6 Thus, the short time of response after the injection of auto-BMHSCT is strong evidence for the action of this therapy. The pathophysiology of macular oedema associated with retinitis is still controversial, but it may possibly be due to the blood–retinal barrier breakdown. However, the mechanism by which auto-BMHSCT led to improvement of the oedema is still Baseline
    Bone marrow transplantation 09/2012; · 3.00 Impact Factor
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    ABSTRACT: Aims and objectives.  To identify differences in cyclosporine levels between blood samples collected from a peripheral venous access, catheter line used for drug infusion and catheter line not used for drug infusion in adult patients receiving allogeneic haematopoietic stem cell transplantation. Background.  Cyclosporine is an immunosuppressant that prevents graft-versus-host disease, has a narrow therapeutic window and causes nephrotoxicity. For cyclosporine infusion, a tunnelled central venous access device is used; however, because of the lipophilic properties of the drug, it can adsorb to the catheter surface and falsely raise cyclosporine concentrations in blood specimens. Design.  Prospective observational study. Methods.  The study collected 135 blood samples from 16 patients. In 13 subjects, samples were obtained from the three lines at three time points (1, 7 and 14 days after the start of cyclosporine infusion), and for three subjects, samples were only obtained at 1 and 7 days after the start of infusion. The 5-ml blood discard method was used for samples collected from the catheter. Using this procedure, the catheter line was washed with saline solution, 5 ml of blood and saline solution were aspirated from the catheter line and discarded, and then sample blood used for the test was collected. The paired t-test with the Bonferroni correction was used to analyse the differences in cyclosporine serum levels. Results.  Significant differences were observed when the drug serum levels obtained in the line used for drug infusion were compared with the levels obtained in the line not used for infusion or the peripheral venous line. No differences in drug levels were identified in blood collected from the peripheral venous line and the line not used for drug infusion. Conclusion.  Drug adsorption occurs in the line used for infusion. Therefore, the blood sample collected from the line not used for cyclosporine infusion can be considered reliable for drug concentration determination. Relevance to clinical practice.  Nurses should standardise one line of the tunnelled central venous access device for cyclosporine infusion, which avoids the need for evasive procedures and provides patients with more comfort.
    Journal of Clinical Nursing 07/2012; · 1.32 Impact Factor
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    ABSTRACT: Hematopoietic cell transplantation (HCT) is an emerging therapy for patients with severe autoimmune diseases (AID). We report data on 368 patients with AID who underwent HCT in 64 North and South American transplantation centers reported to the Center for International Blood and Marrow Transplant Research between 1996 and 2009. Most of the HCTs involved autologous grafts (n = 339); allogeneic HCT (n = 29) was done mostly in children. The most common indications for HCT were multiple sclerosis, systemic sclerosis, and systemic lupus erythematosus. The median age at transplantation was 38 years for autologous HCT and 25 years for allogeneic HCT. The corresponding times from diagnosis to HCT were 35 months and 24 months. Three-year overall survival after autologous HCT was 86% (95% confidence interval [CI], 81%-91%). Median follow-up of survivors was 31 months (range, 1-144 months). The most common causes of death were AID progression, infections, and organ failure. On multivariate analysis, the risk of death was higher in patients at centers that performed fewer than 5 autologous HCTs (relative risk, 3.5; 95% CI, 1.1-11.1; P = .03) and those that performed 5 to 15 autologous HCTs for AID during the study period (relative risk, 4.2; 95% CI, 1.5-11.7; P = .006) compared with patients at centers that performed more than 15 autologous HCTs for AID during the study period. AID is an emerging indication for HCT in the region. Collaboration of hematologists and other disease specialists with an outcomes database is important to promote optimal patient selection, analysis of the impact of prognostic variables and long-term outcomes, and development of clinical trials.
    Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 06/2012; 18(10):1471-8. · 3.15 Impact Factor
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    ABSTRACT: Type 1 diabetes (T1D) is a chronic autoimmune disease characterized by T cell-mediated destruction of pancreatic β cells, resulting in insulin deficiency and hyperglycaemia. Recent studies have described that apoptosis impairment during central and peripheral tolerance is involved in T1D pathogenesis. In this study, the apoptosis-related gene expression in T1D patients was evaluated before and after treatment with high-dose immunosuppression followed by autologous haematopoietic stem cell transplantation (HDI-AHSCT). We also correlated gene expression results with clinical response to HDI-AHSCT. We observed a decreased expression of bad, bax and fasL pro-apoptotic genes and an increased expression of a1, bcl-x(L) and cIAP-2 anti-apoptotic genes in patients' peripheral blood mononuclear cells (PBMCs) compared to controls. After HDI-AHSCT, we found an up-regulation of fas and fasL and a down-regulation of anti-apoptotic bcl-x(L) genes expression in post-HDI-AHSCT periods compared to pre-transplantation. Additionally, the levels of bad, bax, bok, fasL, bcl-x(L) and cIAP-1 genes expression were found similar to controls 2 years after HDI-AHSCT. Furthermore, over-expression of pro-apoptotic noxa at 540 days post-HDI-AHSCT correlated positively with insulin-free patients and conversely with glutamic acid decarboxylase autoantibodies (GAD65) autoantibody levels. Taken together, the results suggest that apoptosis-related genes deregulation in patients' PBMCs might be involved in breakdown of immune tolerance and consequently contribute to T1D pathogenesis. Furthermore, HDI-AHSCT modulated the expression of some apoptotic genes towards the levels similar to controls. Possibly, the expression of these apoptotic molecules could be applied as biomarkers of clinical remission of T1D patients treated with HDI-AHSCT therapy.
    Clinical & Experimental Immunology 06/2012; 168(3):291-302. · 3.41 Impact Factor
  • Multiple Sclerosis 06/2012; 18(6):772-5. · 4.47 Impact Factor
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    ABSTRACT: Mesenchymal stem cells (MSCs) are known to induce the conversion of activated T cells into regulatory T cells in vitro. The marker CD69 is a target of canonical nuclear factor kappa-B (NF-κB) signalling and is transiently expressed upon activation; however, stable CD69 expression defines cells with immunoregulatory properties. Given its enormous therapeutic potential, we explored the molecular mechanisms underlying the induction of regulatory cells by MSCs. Peripheral blood CD3+ T cells were activated and cultured in the presence or absence of MSCs. CD4+ cell mRNA expression was then characterized by microarray analysis. The drug BAY11-7082 (BAY) and a siRNA against v-rel reticuloendotheliosis viral oncogene homolog B (RELB) were used to explore the differential roles of canonical and non-canonical NF-κB signalling, respectively. Flow cytometry and real-time PCR were used for analyses. Genes with immunoregulatory functions, CD69 and non-canonical NF-κB subunits (RELB and NFKB2) were all expressed at higher levels in lymphocytes co-cultured with MSCs. The frequency of CD69+ cells among lymphocytes cultured alone progressively decreased after activation. In contrast, the frequency of CD69+ cells increased significantly following activation in lymphocytes co-cultured with MSCs. Inhibition of canonical NF-κB signalling by BAY immediately following activation blocked the induction of CD69; however, inhibition of canonical NF-κB signalling on the third day further induced the expression of CD69. Furthermore, late expression of CD69 was inhibited by RELB siRNA. These results indicate that the canonical NF-κB pathway controls the early expression of CD69 after activation; however, in an immunoregulatory context, late and sustained CD69 expression is promoted by the non-canonical pathway and is inhibited by canonical NF-κB signalling.
    Journal of Cellular and Molecular Medicine 05/2012; 16(6):1232 - 1244. · 4.75 Impact Factor
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    ABSTRACT: A balance between proinflammatory (Th17 and Tc17) and anti-inflammatory (regulatory T cells) subsets of T cells is essential to maintain immunological tolerance and prevent the onset of several autoimmune diseases, including type 1 diabetes. However, the kinetics of these subsets and disease severity during the streptozotocin (STZ)-induced diabetes course has not been determined. Thus, susceptible C57BL/6 mice were administrated with multiple low doses of STZ and we evaluated the frequency/absolute number of these T cell subsets in the pancreatic lymph nodes (PLNs) and spleen and Th1, Th17, Treg cytokine production in the pancreatic tissue. At different time points of the disease progression (6, 11, 18 and 25 days after the last STZ administration), the histopathological alterations were also evaluated by H&E and immunohistochemistry staining. During the initial phase of diabetes development (day 6), we noted increased numbers of CD4(+) and CD8(+) T cells in spleen and PLNs. At the same time, the frequencies of Th17 and Tc17 cells in PLNs were also enhanced. In addition, the early augment of interferon gamma (IFN-γ), tumoral necrosis factor (TNF-α), IL-6 and IL-17 levels in pancreatic tissue correlated with pancreatic islet inflammation and mild β-cell damage. Notably, the absolute number of Treg cells increased in PLNs during over time when compared to control group. Interestingly, increased IL-10 levels were associated with control of the inflammatory process during the late phase of the type 1 diabetes (day 25). In agreement, mice lacking the expression of IL-17 receptor (Il17r) showed impairment in STZ-induced diabetes progression, reduced peri-insulitis and beta cells preservation when compared with wild-type mice. Our findings suggest that dynamic changes of pathogenic Th17/Tc17 and regulatory T cell subsets numbers is associated with early strong inflammation in the pancreatic islets followed by late regulatory profile during the experimental STZ-induced diabetes course.
    Immunobiology 05/2012; · 2.81 Impact Factor
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    ABSTRACT: Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative disease with a complicated and poorly understood pathogenesis. Strong evidence indicates impairment of all neurovascular unit components including the blood-brain and blood-spinal cord barriers (BBB/BSCB) in both patients and animal models. The present review provides an updated analysis of the microvascular pathology and impaired BBB/BSCB in ALS. Based on experimental and clinical ALS studies, the roles of cellular components, cell interactions, tight junctions, transport systems, cytokines, matrix metalloproteinases, and free radicals in the BBB/BSCB disruption are discussed. The impact of BBB/BSCB damage in ALS pathogenesis is a novel research topic, and this review will reveal some aspects of microvascular pathology involved in the disease and hopefully engender new therapeutic approaches.
    International Review of Neurobiology 01/2012; 102:91-106. · 2.46 Impact Factor
  • Immunology 01/2012; 137:683-684. · 3.71 Impact Factor
  • Immunology 01/2012; 137:354-354. · 3.71 Impact Factor
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    ABSTRACT: Peripheral nerve injuries are a frequent and disabling condition, which affects 13 to 23 per 100.000 persons each year. Severe cases, with structural disruption of the nerve, are associated with poor functional recovery. The experimental treatment using nerve grafts to replace damaged or shortened axons is limited by technical difficulties, invasiveness, and mediocre results. Other therapeutic choices include the adjunctive application of cultured Schwann cells and nerve conduits to guide axonal growth. The bone marrow is a rich source of mesenchymal cells, which can be differentiated in vitro into Schwann cells and subsequently engrafted into the damaged nerve. Alternatively, undifferentiated bone marrow mesenchymal cells can be associated with nerve conduits and afterward transplanted. Experimental studies provide evidence of functional, histological, and electromyographical improvement following transplantation of bone-marrow-derived cells in animal models of peripheral nerve injury. This paper focuses on this new therapeutic approach highlighting its direct translational and clinical utility in promoting regeneration of not only acute but perhaps also chronic cases of peripheral nerve damage.
    The Scientific World Journal 01/2012; 2012:413091. · 1.73 Impact Factor
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    ABSTRACT: The main current therapies for inflammatory bowel diseases (IBD) are aimed at controlling the exacerbated inflammation in the gut. Although these therapies have been successful, they are not curative and it is not possible to predict whether a beneficial response will occur or which patients will be refractory to the treatment. Total body irradiation (TBI) associated with chemotherapy is the first choice in the treatment of some hematological disorders and is an applicable option in the preparation of patients with hematologic diseases for hematopoietic stem cell transplantation. Then, in this study we investigated the association of TBI as immunosuppressive therapy and bone marrow cell (BMC) transplantation as a strategy to induce colitis recovery and immune reconstitution in the TNBS model of intestinal inflammation. TNBS mice treated with TBI associated with BMC transplantation presented elevated gain of weight and an overall better outcome of the disease when compared to those treated only with TBI. In addition, TBI associated or not with BMC reduced the frequency of inflammatory cells in the gut and restored the goblet cell counts. These results were accompanied by a down regulation in the production of inflammatory cytokines in the colon of mice treated with TBI alone or in association with BMC transplantation. The BMC infused were able to repopulate the ablated immune system and accumulate in the site of inflammation. However, although both treatments (TBI or TBI+BMC) were able to reduce gut inflammation, TBI alone was not enough to fully restore mice weight and these animals presented an extremely reduced survival rate when their immune system was not promptly reconstituted with BMC transplantation. Finally, these evidences suggest that the BMC transplantation is an efficient strategy to reduce the harmful effects of TBI in the colitis treatment, suggesting that radiotherapy may be an important immunosuppressive therapy in patients with IBD, by modulating the local inflammatory response.
    Immunobiology 01/2012; · 2.81 Impact Factor
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    ABSTRACT: Cerebrovascular diseases are the third leading cause of death and the primary cause of long-term disability in the United States. Most patients, excluded from the available treatment with plasminogen activator (tPA), present permanent neurological impairment and may benefit from restorative treatments with stem cells. Inflammation is a key feature in stroke and it plays a dual role, either increasing injury in early phases or impairing neural survival at later stages. Stem cells can be opportunely used to modulate inflammation, abrogate cell death and, therefore, preserve neural function. To date, there is no consensus about the most adequate cell type, route of delivery or timing for transplantation, as experimental and clinical studies are still inconclusive. Menstrual blood stem cells have been recently studied for their availability, proliferative capacity, pluripotentiality and angiogenic features, which make them a relevant resource for the treatment of stroke. KeywordsCerebrovascular diseases-Plasminogen activator-Neutrophils-Central nervous system (CNS)-Stem cell-Endometrium
    12/2011: pages 191-197;

Publication Stats

2k Citations
417.13 Total Impact Points

Institutions

  • 2010–2013
    • Northwestern University
      • • Division of Immunotherapy
      • • Department of Medicine
      Evanston, IL, United States
    • Universidade Estadual de Londrina
      • Department of Pathological Sciences
      Londrina, Estado do Parana, Brazil
  • 2004–2013
    • Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo
      San Paulo, São Paulo, Brazil
  • 2011
    • Universidade Federal de São Carlos
      • Departamento de Medicina (DMed)
      São Carlos, Estado de Sao Paulo, Brazil
  • 1990–2010
    • Universidade de Ribeirão Preto
      Entre Rios, São Paulo, Brazil
  • 1983–2010
    • University of São Paulo
      • • Faculdade de Medicina de Ribeirão Preto (FMRP)
      • • Departamento de Clínica Médica (VCM)
      • • Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP)
      • • Faculty of Medicine (FM)
      São Paulo, Estado de Sao Paulo, Brazil
  • 2009
    • Hospital Israelita Albert Einstein
      San Paulo, São Paulo, Brazil
    • Pontifícia Universidade Católica de Goiás (PUC Goiás)
      Goyaz, Goiás, Brazil