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1.
BMC Immunol ; 25(1): 49, 2024 Jul 26.
Artículo en Inglés | MEDLINE | ID: mdl-39061002

RESUMEN

BACKGROUND: The systemic inflammatory syndrome called "cytokine storm" has been described in COVID-19 pathogenesis, contributing to disease severity. The analysis of cytokine and chemokine levels in the blood of 21 SARS-CoV-2 positive patients throughout the phases of the pandemic has been studied to understand immune response dysregulation and identify potential disease biomarkers for new treatments. The present work reports the cytokine and chemokine levels in sera from a small cohort of individuals primarily infected with SARS-CoV-2 during the first wave of the COVID-19 pandemic in Milan (Italy). RESULTS: Among the 27 cytokines and chemokines investigated, a significant higher expression of Interleukin-9 (IL-9), IP-10 (CXCL10), MCP-1 (CCL2) and RANTES (CCL-5) in infected patients compared to uninfected subjects was observed. When the change in cytokine/chemokine levels was monitored over time, from the hospitalization day to discharge, only IL-6 and IP-10 showed a significant decrease. Consistent with these findings, a significant negative correlation was observed between IP-10 and anti-Spike IgG antibodies in infected individuals. In contrast, IL-17 was positively correlated with the production of IgG against SARS-CoV-2. CONCLUSIONS: The cytokine storm and the modulation of cytokine levels by SARS-CoV-2 infection are hallmarks of COVID-19. The current global immunity profile largely stems from widespread vaccination campaigns and previous infection exposures. Consequently, the immunological features and dynamic cytokine profiles of non-vaccinated and primarily-infected subjects reported here provide novel insights into the inflammatory immune landscape in the context of SARS-CoV-2 infection, and offer valuable knowledge for addressing future viral infections and the development of novel treatments.


Asunto(s)
COVID-19 , Quimiocinas , Citocinas , SARS-CoV-2 , Humanos , COVID-19/inmunología , COVID-19/sangre , COVID-19/epidemiología , Italia/epidemiología , SARS-CoV-2/inmunología , Femenino , Masculino , Persona de Mediana Edad , Citocinas/sangre , Anciano , Quimiocinas/sangre , Adulto , Síndrome de Liberación de Citoquinas/sangre , Síndrome de Liberación de Citoquinas/inmunología , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Biomarcadores/sangre , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Pandemias
2.
Small ; 19(32): e2300947, 2023 08.
Artículo en Inglés | MEDLINE | ID: mdl-37060208

RESUMEN

Rapid detection of whole virus particles in biological or environmental samples represents an unmet need for the containment of infectious diseases. Here, an optical device enabling the enumeration of single virion particles binding on antibody or aptamers immobilized on a surface with anti-reflective coating is described. In this regime, nanoparticles adhering to the sensor surface provide localized contributions to the reflected field that become detectable because of their mixing with the interfering waves in the reflection direction. Thus, these settings are exploited to realize a scan-free, label-free, micro-array-type digital assay on a disposable cartridge, in which the virion counting takes place in wide field-of-view imaging. With this approach we could quantify, by enumeration, different variants of SARS-CoV-2 virions interacting with antibodies and aptamers immobilized on different spots. For all tested variants, the aptamers showed larger affinity but lower specificity relative to the antibodies. It is found that the combination of different probes on the same surface enables increasing specificity of detection and dynamic range.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , COVID-19 , Humanos , SARS-CoV-2 , Técnicas Biosensibles/métodos , Anticuerpos , Virión
3.
Mol Divers ; 2023 Jul 22.
Artículo en Inglés | MEDLINE | ID: mdl-37480422

RESUMEN

In recent years, the viral outbreak named COVID-19 showed that infectious diseases have a huge impact on both global health and the financial and economic sectors. The lack of efficacious antiviral drugs worsened the health problem. Based on our previous experience, we investigated in vitro and in silico a series of quinoline-3-carboxylate derivatives against a SARS-CoV-2 isolate. In the present study, the in-vitro antiviral activity of a series of quinoline-3-carboxylate compounds and the in silico target-based molecular dynamics (MD) and metabolic studies are reported. The compounds' activity against SARS-CoV-2 was evaluated using plaque assay and RT-qPCR. Moreover, from the docking scores, it appears that the most active compounds (1j and 1o) exhibit stronger binding affinity to the primary viral protease (NSP5) and the exoribonuclease domain of non structural protein 14 (NSP14). Additionally, the in-silico metabolic analysis of 1j and 1o defines CYP2C9 and CYP3A4 as the major P450 enzymes involved in their metabolism.

4.
J Neurovirol ; 28(1): 113-122, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-34997473

RESUMEN

Here we described the virological and serological assessment of 23 COVID-19 patients hospitalized and followed up in Milan, Italy, during the first wave of COVID-19 pandemic. Nasopharyngeal (NPS), anal swabs, and blood samples were collected from 23 COVID-19 patients, at hospital admission, and periodically up to discharge, for a median time of 20 days (3-83 days). RNA was isolated and tested for SARS-CoV-2 by qRT-PCR; anti-SARS-CoV-2 IgM and IgG antibody titers were evaluated in serum samples by ELISA. SARS-CoV-2 genome was detected in the NPS swabs of the 23 patients, at the admission, and 8/19 (42.1%) were still positive at the discharge. Anal swabs were positive to SARS-CoV-2 RNA detection in 20/23 (86.9%) patients; 6/19 (31.6%) were still positive at discharge. The mean time of RNA negative conversion was 17 days (4-36 days) and 33 days (4-77 days), for NPS and anal swabs, respectively. SARS-CoV-2-RNA was detected in the blood of 6/23 (26.1%) patients. Thirteen/23 (56.5%) and 17/23 (73.9%) patients were seropositive for IgM and IgG, respectively, at the admission, and the median IgM and IgG levels significantly (p < 0.05) increased after 13 days. Although the limited cohort size, our report provides evidence that SARS-CoV-2 is shed through multiple routes, with important implications in healthcare settings.


Asunto(s)
COVID-19 , Anticuerpos Antivirales , COVID-19/diagnóstico , Humanos , Inmunoglobulina G , Inmunoglobulina M , Pandemias , ARN Viral/genética , SARS-CoV-2
5.
Int J Mol Sci ; 23(7)2022 Apr 06.
Artículo en Inglés | MEDLINE | ID: mdl-35409421

RESUMEN

In the novel pandemic of Coronavirus Disease 2019, high levels of pro-inflammatory cytokines lead to endothelial activation and dysfunction, promoting a pro-coagulative state, thrombotic events, and microvasculature injuries. The aim of the present work was to investigate the effect of SARS-CoV-2 on pro-inflammatory cytokines, tissue factor, and chemokine release, with Human Microvascular Endothelial Cells (HMEC-1). ACE2 receptor expression was evaluated by western blot analysis. SARS-CoV-2 infection was assessed by one-step RT-PCR until 7 days post-infection (p.i.), and by Transmission Electron Microscopy (TEM). IL-6, TNF-α, IL-8, IFN-α, and hTF mRNA expression levels were detected by RT-PCR, while cytokine release was evaluated by ELISA. HMEC-1 expressed ACE2 receptor and SARS-CoV-2 infection showed a constant viral load. TEM analysis showed virions localized in the cytoplasm. Expression of IL-6 at 24 h and IFN-α mRNA at 24 h and 48 h p.i. was higher in infected than uninfected HMEC-1 (p < 0.05). IL-6 levels were significantly higher in supernatants from infected HMEC-1 (p < 0.001) at 24 h, 48 h, and 72 h p.i., while IL-8 levels were significantly lower at 24 h p.i. (p < 0.001). These data indicate that in vitro microvascular endothelial cells are susceptible to SARS-CoV-2 infection but slightly contribute to viral amplification. However, SARS-CoV-2 infection might trigger the increase of pro-inflammatory mediators.


Asunto(s)
COVID-19 , Enzima Convertidora de Angiotensina 2 , Quimiocinas/genética , Quimiocinas/metabolismo , Citocinas/metabolismo , Células Endoteliales/metabolismo , Humanos , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , SARS-CoV-2
6.
J Med Virol ; 93(11): 6333-6339, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-33547809

RESUMEN

Colon cancer is the third cause of cancer death in the developed countries. Some environmental factors are involved in its pathogenesis, including viral infections. The possible involvement of human polyomaviruses (HPyVs) in colon cancer pathogenesis has been previously reported, leading to inconsistent conclusions. Clinical specimens were collected from 125 colon cancer patients. Specifically, 110 tumor tissues, 55 negative surgical margins, and 39 peripheral blood samples were analyzed for the presence of six HPyVs: JC polyomavirus (JCPyV), BK polyomavirus (BKPyV), Merkel cell PyV (MCPyV), HPyV -6, -7, and -9 by means of DNA isolation and subsequent duplex Real Time quantitative polymerase chain reaction. HPyVs genome was detected in 33/204 samples (16.2%): the significant higher positivity was found in tumor tissues (26/110, 23.6%), followed by negative surgical margins (3/55, 5.5%, p < .05), and peripheral blood mononuclear cells (PBMCs) (4/39; 10.3%). HPyVs load was statistically higher only in the tumor tissues compared to negative surgical margins (p < .05). Specifically, MCPyV was detected in 19.1% (21/110) of tumor tissues, 3.6% (2/55) of negative surgical margins (p < .05), and 7.7% (3/39) of PBMCs; HPyV-6 in 2.7% (3/110) of tumor tissues, and 1.8% (1/55) of negative surgical margins; one tumor tissue (1/110, 0.9%) and one PBMCs sample (1/39, 2.6%) were positive for BKPyV; JCPyV was present in 0.9% (1/110) of tumor tissues. HPyV-7 and 9 were not detected in any sample. High prevalence and load of MCPyV genome in the tumor tissues might be indicative of a relevant rather than bystander role of the virus in the colon tumorigenesis.


Asunto(s)
Neoplasias del Colon/virología , ADN Viral/aislamiento & purificación , Genoma Viral , Infecciones por Polyomavirus/virología , Poliomavirus/genética , Poliomavirus/aislamiento & purificación , Carga Viral , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias del Colon/clasificación , ADN Viral/genética , Femenino , Humanos , Masculino , Persona de Mediana Edad , Poliomavirus/clasificación , Manejo de Especímenes , Infecciones Tumorales por Virus/virología
7.
Med Microbiol Immunol ; 209(2): 189-199, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32040616

RESUMEN

Human endogenous retroviruses (HERV) are remnants of exogenous retroviral infections, representing 8% of the human genome. Their regulation is based on the DNA methylation of promoters, the long terminal repeats (LTRs). Transcripts from HERV have been associated with cancers, but reports concerning HERV expression in colorectal cancer remain sporadic. Sixty-three patients with advanced stages of colorectal cancer were enrolled in this study. The expressions of HERV env gene, and HERV-H, -K, -R and -P LTRs and Alu, LINE-1 methylation levels, were investigated in the tumor, normal adjacent tissues, and, where possible, blood and plasmatic extracellular vesicles (EVs). Associations among HERV env expression, methylation status and clinical characteristics were evaluated. No differences were observed in HERV env gene expression levels among the clinical specimens, while Alu, LINE-1, HERV-H and -K LTRs were demethylated in the tumor compared to the normal adjacent tissues (p < 0.05).The HERV env gene was expressed in the EVs at of 54% (-H), 38% (-K), 31% (-R) patients. Association was not found between HERV env expression and LTR methylation, but significant higher expression of HERV-P and -R env was found in tumor tissues arising from the right colon. Our findings do not demonstrate significant overexpression of the studied HERV in colorectal cancer, but their association with tumor localization and specificity of the changes in DNA methylation of retroelements are shown. HERV sequences were packaged in the EVs and might be transferred from one cell to another.


Asunto(s)
Neoplasias Colorrectales/genética , Metilación de ADN , Retrovirus Endógenos/genética , Productos del Gen env/metabolismo , Secuencias Repetidas Terminales , Anciano , Anciano de 80 o más Años , Elementos Alu , Neoplasias Colorrectales/virología , Retrovirus Endógenos/metabolismo , Vesículas Extracelulares/química , Femenino , Regulación Neoplásica de la Expresión Génica , Productos del Gen env/sangre , Productos del Gen env/clasificación , Genes env , Humanos , Elementos de Nucleótido Esparcido Largo , Masculino , Regiones Promotoras Genéticas
8.
Parasitology ; 146(3): 399-406, 2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30269694

RESUMEN

Haemozoin is a by-product of haemoglobin digestion by intraerythrocytic malaria parasites, which induces immunologic responses on different tissues, including endothelial cells. In the present paper, the incubation of human microvascular endothelial cells with haemozoin significantly inhibited MTT reduction, a measure of cytotoxicity, without increasing the release of cytoplasmic lactate dehydrogenase. Moreover, haemozoin did not induce apoptosis or cell cycle arrest nor decreased the number of live cells, suggesting that cells viability itself was not affected and that the inhibition of MTT reduction was only apparent and probably due to accelerated MTT-formazan exocytosis. After 30 min of MTT addition, a significant increase in the % of cells exocytosing MTT formazan crystals was observed in haemozoin-treated cells compared with control cells. Such an effect was partially reversed by the addition of genistein, an inhibitor of MTT-formazan exocytosis. The rapid release of CXCL-8, a preformed chemokine contained in Weibel-Palade bodies, confirmed that haemozoin induces a perturbation of the intracellular endothelial trafficking, including the exocytosis of MTT-formazan containing vesicles. The haem moiety of haemozoin is responsible for the observed effect. Moreover, this work underlines that MTT assay should not be used to measure cytotoxicity induced by haemozoin and other methods should be preferred.


Asunto(s)
Células Endoteliales/fisiología , Exocitosis/fisiología , Formazáns/química , Hemoproteínas/metabolismo , Pigmentos Biológicos/metabolismo , Plasmodium falciparum/fisiología , Sales de Tetrazolio/química , Humanos
9.
J Neurovirol ; 23(1): 99-105, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-27538993

RESUMEN

JC virus (JCV) is a widespread member of the Polyomaviridae family. Following primary infection, which occurs asymptomatically during childhood, JCV establishes latency in the host. JCV seroprevalence can reach 80 % in healthy adults, but the age of viral exposure has not been yet characterized. This study was conducted to define JCV seroprevalence in Italian infants and to estimate the date of primary infection. A JCV viral protein 1 (VP1)-GST fusion protein was used in conjunction with a homemade indirect enzyme-linked immunosorbent assay (ELISA) to test for the presence of IgG antibodies to JCV in 981 serum samples collected from 644 Italian infants of different ages (1 day to 3 years old) and in 102 breast milk samples. IgM antibody presence was also evaluated in longitudinally collected samples from 17 selected children. JCV antibody prevalence and normalized optical density (nOD) were calculated. For the longitudinal analysis, generalized estimating equation techniques and spline functions were used to estimate the possible non-linear effects of time on antibody production kinetics. JCV IgG was detected in 71.8 % of the sera. Prevalence increased over time from 46.1 % (1 month old) to 80.7 % (12 months old), 85.9 % (24 months old), and 85.5 % (36 months old). As determined by nOD, the longitudinal analysis of serum IgG amounts in children of this study (ages 1 day to 3 years old) illustrated IgG kinetic changes with statistically significant trends (p = 0.001). One-month-old children were largely negative for JCV IgM (82.4 %), and 58.8 % of children produced JCV IgM within the second and sixth months of life. JCV IgG was detected in 27.3 % of breast milk samples. JCV primary infection likely occurs before 6 months of age, and a sizeable percentage of Italian infants will become JCV seropositive within 2 years of age. This study can be used to determine the optimal age for potential future JCV vaccination in infants.


Asunto(s)
Anticuerpos Antivirales/sangre , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Virus JC/inmunología , Infecciones por Polyomavirus/diagnóstico , Infecciones por Polyomavirus/epidemiología , Enfermedades Asintomáticas , Preescolar , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Lactante , Recién Nacido , Italia/epidemiología , Estudios Longitudinales , Masculino , Leche Humana/virología , Infecciones por Polyomavirus/sangre , Infecciones por Polyomavirus/inmunología , Prevalencia , Estudios Seroepidemiológicos
10.
J Cell Physiol ; 231(6): 1343-9, 2016 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-26529465

RESUMEN

BK polyomavirus (BKV) has a worldwide seroprevalence of approximately 90%. After primary infection, BKV establishes a life-long latency within the urogenital tract. The severe immunological impairment occurring in renal transplant recipients leads to BKV reactivation, which may result in polyomavirus associated nephropathy (PVAN). While the transplanted kidney is transiently unperfused, Hypoxia Inducible Factors (HIFs) mediate the cellular response to hypoxia. The α-subunit of HIF isoform 1 (HIF-1α) may interact with several viruses, but until now, there has been no information regarding the interaction between BKV and HIF-1α. The aim of this study is to investigate the possible interaction between HIF-1α and BKV and its potential effect on the pathogenesis of PVAN. Screening of 17 kidney tissue samples revealed that HIF-1α expression was 13.6-fold higher in PVAN tissues compared to control tissues. A luminometric assay in co-transfected African green monkey kidney cells (VERO) demonstrated BKV promoter activation ranging from two to sixfold (P < 0.05) when HIF-1α was over-expressed. A Chromatin ImmunoPrecipitation (ChIP) assay showed structural binding between the BKV promoter and HIF-1α. The amount of BKV DNA increased by threefold in VERO infected cells that were exposed to simulated hypoxia, compared to the cells not subjected to hypoxia. Both ex vivo and in vitro interactions between HIF-1α and BKV were observed, suggesting that HIF-1α, stabilized during transplantation, may be able to bind the BKV promoter and enhance BKV replication. Thus, hypoxia should be considered a risk factor for the development of PVAN in kidney transplant recipients.


Asunto(s)
Virus BK/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Trasplante de Riñón/efectos adversos , Riñón/metabolismo , Infecciones por Polyomavirus/metabolismo , Infecciones por Polyomavirus/virología , Infecciones Tumorales por Virus/metabolismo , Infecciones Tumorales por Virus/virología , Adulto , Anciano , Animales , Virus BK/genética , Virus BK/crecimiento & desarrollo , Virus BK/aislamiento & purificación , Sitios de Unión , Hipoxia de la Célula , Chlorocebus aethiops , Replicación del ADN , ADN Viral/biosíntesis , Femenino , Regulación Viral de la Expresión Génica , Interacciones Huésped-Patógeno , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Riñón/virología , Masculino , Persona de Mediana Edad , Infecciones por Polyomavirus/genética , Regiones Promotoras Genéticas , Unión Proteica , Factores de Riesgo , Transfección , Infecciones Tumorales por Virus/genética , Regulación hacia Arriba , Células Vero , Carga Viral
11.
J Cell Physiol ; 230(7): 1457-65, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-25556900

RESUMEN

This study designs a strategy for an adoptive cellular therapy (ACT) protocol based on the ex-vivo selection of autologous peripheral blood-derived CD8-enriched T-cells, stimulated with dendritic cells (DCs) that had been pulsed with apoptotic tumor cells to generate cytotoxic T lymphocytes (CTLs) with anti-tumor activity. Seventy-eight colorectal cancer (CRC) patients were enrolled in this study. Tumor tissues and peripheral blood (PB) were obtained at surgery. Tissues were mechanically dissociated and cultured to obtain a primary tumor cell line from each patient. DCs were derived from peripheral blood mononuclear cells (PBMCs) using magnetic positive selection of CD14+ monocytes. Anti-tumor CTLs were elicited in co-/micro-cultures using DCs as antigen-presenting cells, autologous apoptotic tumor cells as a source of antigens, and CD8+ T lymphocytes as effectors. Interferon-γ (IFN-γ) secretion was assessed by ELISpot assays to evaluate the activation of the CTLs against the autologous tumor cells. Primary tumor cell lines were obtained from 20 of 78 patients (25.6%). DCs were generated from 26 patients, and of them, corresponding tumor cell lines were derived from six patients. ELISpot results showed that significant IFN-γ secretion was detected after different numbers of stimulations for two patients, whereas weak secretion was observed for three patients. Despite difficulties due to contamination of several primary tumor cell lines with gut intestinal flora, the results suggest that the generation of tumor-specific CTLs is feasible from patients with CRC, and could be useful for supporting an ACT approach in CRC.


Asunto(s)
Traslado Adoptivo , Neoplasias Colorrectales/terapia , Linfocitos T Citotóxicos/fisiología , Anciano , Anciano de 80 o más Años , Línea Celular Tumoral , Femenino , Humanos , Masculino , Persona de Mediana Edad , Linfocitos T Citotóxicos/citología
12.
J Neurovirol ; 21(6): 645-52, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-25052191

RESUMEN

The risk of developing progressive multifocal leukoencephalopathy (PML), as a consequence of infection/reactivation with JC virus (JCV), is consistent in natalizumab-treated multiple sclerosis (MS) patients, with 430 cases of PML reported so far. The risk of PML is higher in JCV seropositive patients, and it is recommended that only MS patients without JCV antibodies should be enrolled in the treatment postulating that they do not have JCV infection.We have studied forty-two natalizumab-treated MS patients, and urine and blood were collected monthly for up to 60 months. JCV and BK virus (BKV) DNA presence was verified using quantitative real-time PCR assays, and serum anti-JCV antibodies were measured with the Stratify and/or Stratify DxSelect tests.JCV and BKV DNA were not found in the blood samples, whereas they were found at least once in the urine of 21 of 42 (50 %) and of 25/42 (59.5 %) patients, respectively. JCV DNA urinary shedding increased up to month 24 of natalizumab treatment (45.2 %), and the effect of time was significant for JCV (p = 0.04), but not for BKV (p = 0.39). JCV viruria and seropositivity did not completely correlate, since three patients shedding JCV DNA in the urine were seronegative according to the serological tests.The results indicated that natalizumab therapy may increase the rate of JCV urinary shedding. Additionally, we confirmed that the identification of JCV carriers cannot solely rely on serological tests, but sensitive methods for viral DNA detection should be adopted to more precisely identify the truly JCV uninfected cases.


Asunto(s)
Anticuerpos Antivirales/orina , ADN Viral/orina , Factores Inmunológicos/uso terapéutico , Esclerosis Múltiple Recurrente-Remitente/tratamiento farmacológico , Esclerosis Múltiple Recurrente-Remitente/virología , Natalizumab/uso terapéutico , Adulto , Anticuerpos Antivirales/sangre , ADN Viral/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Virus JC/aislamiento & purificación , Masculino , Persona de Mediana Edad , Infecciones por Polyomavirus/virología , Reacción en Cadena en Tiempo Real de la Polimerasa , Estudios Seroepidemiológicos , Esparcimiento de Virus/fisiología , Adulto Joven
13.
Clin Exp Med ; 24(1): 3, 2024 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-38231278

RESUMEN

INTRODUCTION: Immunosuppression after kidney transplantation (KTx) exposes recipients to Human Polyomaviruses (HPyVs) infections, whose natural history is still misunderstood. METHODS: Allograft biopsies, and urine from 58 donor-recipient pairs were collected before KTx (T0) and 1 (T1), 15 (T2), 30 (T3), 60 (T4), 90 (T5), 180 (T6), 270 (T7), 360 (T8), and 540 (T9) days after transplant. Specimens were tested for JC (JCPyV) and BK (BKPyV), by quantitative Real-Time PCR. The course of post-KTx HPyVs viruria, and the association between JCPyV viruria in recipients and donors, were evaluated. RESULTS: HPyVs were detected in 3/58 (5.2%) allograft biopsies. HPyVs viruria was present in 29/58 (50%) donors and 41/58 (70.7%) recipients. JCPyV DNA was detected in 26/58 (44.8%) donors and 25/58 recipients (43.1%), 19 of whom received kidney from JCPyV positive donor, whereas BKPyV genome was detected in 3 (5.2%) donors and 22 (37.9%) recipients. The median time of JCPyV, and BKPyV first episode of replication was 1, and 171 days post KTx, respectively. At T0, JCPyV viruria of donors was associated with increased risk of JCPyV replication post-KTx; recipients with JCPyV positive donors showed lower risk of BKPyV replication post-KTx. CONCLUSIONS: The results suggested that JCPyV may be transmitted by allograft, and that its replication post KTx might prevent BKPyV reactivation. Future investigation regarding correlation between chronic exposure to immunosuppressive agents and HPyVs urinary replication are warranted.


Asunto(s)
Trasplante de Riñón , Poliomavirus , Humanos , Poliomavirus/genética , Trasplante de Riñón/efectos adversos , Estudios Longitudinales , Riñón , Receptores de Trasplantes
14.
Sci Rep ; 14(1): 13134, 2024 06 07.
Artículo en Inglés | MEDLINE | ID: mdl-38849411

RESUMEN

The pandemic of coronavirus disease 19 (COVID-19), caused by severe respiratory syndrome coronavirus 2 (SARS-CoV-2), had severe repercussions for breast cancer patients. Increasing evidence indicates that SARS-CoV-2 infection may directly impact breast cancer biology, but the effects of SARS-CoV-2 on breast tumor cells are still unknown. Here, we analyzed the molecular events occurring in the MCF7, MDA-MB-231 and HCC1937 breast cancer cell lines, representative of the luminal A, basal B/claudin-low and basal A subtypes, respectively, upon SARS-CoV-2 infection. Viral replication was monitored over time, and gene expression profiling was conducted. We found that MCF7 cells were the most permissive to viral replication. Treatment of MCF7 cells with Tamoxifen reduced the SARS-CoV-2 replication rate, suggesting an involvement of the estrogen receptor in sustaining virus replication in malignant cells. Interestingly, a metagene signature based on genes upregulated by SARS-CoV-2 infection in all three cell lines distinguished a subgroup of premenopausal luminal A breast cancer patients with a poor prognosis. As SARS-CoV-2 still spreads among the population, it is essential to understand the impact of SARS-CoV-2 infection on breast cancer, particularly in premenopausal patients diagnosed with the luminal A subtype, and to assess the long-term impact of COVID-19 on breast cancer outcomes.


Asunto(s)
Neoplasias de la Mama , COVID-19 , SARS-CoV-2 , Tamoxifeno , Replicación Viral , Humanos , Neoplasias de la Mama/virología , Neoplasias de la Mama/patología , COVID-19/virología , Femenino , SARS-CoV-2/fisiología , Línea Celular Tumoral , Tamoxifeno/farmacología , Células MCF-7 , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica
15.
Photochem Photobiol ; 99(1): 101-105, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-35578782

RESUMEN

The ongoing emergency provoked by the SARS-CoV-2 pandemic demands the development of technologies to mitigate the spread of infection, and UV irradiation is a technique that can efficiently address this issue. However, proper use of UV equipment for disinfection requires an understanding of how the effects on SARS-CoV-2 are dependent on certain parameters. In this work, we determined the UV-C inactivation constant k for SARS-CoV-2 using an LED source at λ = 280 nm. Specifically, a Log3 reduction was measured after irradiation for 24 min with a delivered UV-C dose of 23 J m-2 . By multitarget model fitting, n = 2 and k = 0.32 ± 0.02 m2 J-1 were obtained. A lag time for the inactivation effect was also observed, which was attributed to the low irradiation levels used to perform the study. The combination of k and delay time allows for reliable estimation of disinfection times in small, closed environments.


Asunto(s)
COVID-19 , SARS-CoV-2 , Humanos , COVID-19/prevención & control , Rayos Ultravioleta , Desinfección/métodos , Pandemias/prevención & control , Inactivación de Virus/efectos de la radiación
16.
Sci Rep ; 13(1): 15521, 2023 09 19.
Artículo en Inglés | MEDLINE | ID: mdl-37726309

RESUMEN

Breastmilk protects newborns from infections through specific and nonspecific compounds. This study investigated the neutralizing activity against SARS-CoV-2 of breastmilk from SARS-CoV-2 negative, unvaccinated mothers, and compared it to that from infected nursing mothers. We enrolled women after COVID-19 swab testing results upon maternity admission, and divided them into two groups: group A, COVID-19-positive mothers, and group B, negative mothers. Breastmilk was randomly sampled at 2, 7, and 20 days postpartum. We collected 19 samples for Group A and 41 for Group B. A microneutralization assay was used to determine the 50% neutralization (NT50) titre. The presence of neutralizing antibodies was also determined. Group A had 100% neutralizing samples at 2 days postpartum (T0), declining 7 days postpartum (T1) and 20 days postpartum (T2). Group B samples exhibited neutralizing activity mostly at 7 days postpartum (T1) (90%). Negative mothers' samples showed no correlation between NT50 titres and antibodies' presence, suggesting that non-specific breastmilk components may exert antiviral action against SARS-CoV-2.


Asunto(s)
COVID-19 , Leche Humana , Recién Nacido , Embarazo , Femenino , Humanos , SARS-CoV-2 , Lactancia , Madres , Anticuerpos Neutralizantes
17.
J Cell Physiol ; 227(10): 3511-7, 2012 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22253012

RESUMEN

Progressive multifocal leukoencephalopathy (PML) is a severe disease of the central nervous system (CNS), caused by infection with the Polyomavirus JC virus (JCV). Because there are no known treatments or prognostic factors, we performed a long-term study focusing mainly on cerebrospinal fluid (CSF) samples from PML patients to describe the virological features akin to the different forms of the disease. Twenty-eight PML patients were enrolled: 10 HIV-1+ patients with classical PML (CPML), 9 HIV-1+ patients with slowly progressing or stable neurological symptoms (benign PML), 3 HIV-1+ asymptomatic patients, and 6 HIV-1-negative patients. CSF, urine, and blood samples were collected at the enrollment (baseline) and every 6 months afterwards when possible. The JCV DNA and HIV-1 RNA loads were determined, and the JCV strains were characterized. At baseline, the mean CSF JCV load was log 6.0 ± 1.2 copies/ml for CPML patients, log 4.0 ± 1.0 copies/ml for benign PML patients, log 4.2 ± 0.5 copies/ml for asymptomatic PML patients, and log 5.8 ± 1.3 copies/ml for HIV-1-negative PML patients (CPML vs. benign: P < 0.01; CPML vs. asymptomatic: P < 0.05; HIV-1 negative vs. benign: P < 0.01). Organization of the JCV transcriptional control region (TCR) showed unusual archetype structures in two long-term survival patients; the NF1 sequence was found most commonly, whereas the Sp1 binding site was the most common for both CPML patients and HIV-1 negative patients. Our results suggest that the JCV load in the CSF and the organization of the TCR should be considered as indicators of PML clinical outcome.


Asunto(s)
Líquido Cefalorraquídeo/virología , Regulación Viral de la Expresión Génica , Virus JC/fisiología , Leucoencefalopatía Multifocal Progresiva/virología , Adulto , ADN Viral/líquido cefalorraquídeo , ADN Viral/genética , Femenino , Estudios de Seguimiento , Infecciones por VIH/líquido cefalorraquídeo , Infecciones por VIH/virología , Duplicado del Terminal Largo de VIH/genética , VIH-1/genética , Humanos , Virus JC/genética , Leucoencefalopatía Multifocal Progresiva/líquido cefalorraquídeo , Masculino , Neurofibromina 1/genética , ARN Viral/genética , Transcripción Genética , Carga Viral
18.
J Med Virol ; 84(9): 1464-70, 2012 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-22825826

RESUMEN

Rituximab is a chimeric monoclonal antibody reacting with the CD20 antigen on B cells. It has been proposed as treatment for the idiopathic nephrotic syndrome, recurrent idiopathic nephropathy, and focal segmental glomerulosclerosis refractory to steroids. Rituximab influences T-cell immunity and may predispose the patients to opportunistic infections, such as progressive multifocal leukoencephalopathy caused by the polyomavirus JC (JCV). The risk of latent viruses infections/reactivations in pediatric patients receiving monoclonal antibodies is not well known yet. In this longitudinal 6-month study, the effects of rituximab on JCV and BK virus (BKV) replication have been investigated. Blood, serum, and urine samples have been collected monthly from 11 pediatric patients (mean age: 11 years) with the idiopathic nephrotic syndrome and recurrent idiopathic nephropathy, under rituximab therapy. JCV and BKV real-time PCRs and sequencing of the viral protein 1 and the non-coding control region have been conducted. The same investigations have been undertaken on samples collected from eight pediatric patients (controls, mean age: 6 years), with idiopathic nephrotic syndrome or focal segmental glomerulosclerosis, treated with conventional chemotherapy. JCV was detected in the urine of one patient (9%), and one control (12.5%); BKV was found in the urine of 7/11 patients (63.6%) and 2/8 controls (25%) and in blood samples from four patients. No significant difference was found in the mean viral loads and in the viral molecular characterizations between the two groups. The polyomaviruses replication was not associated with rituximab therapy in children.


Asunto(s)
Anticuerpos Monoclonales de Origen Murino/farmacología , Virus BK/fisiología , Factores Inmunológicos/farmacología , Virus JC/fisiología , Síndrome Nefrótico/sangre , Replicación Viral , Adolescente , Anticuerpos Monoclonales de Origen Murino/uso terapéutico , Virus BK/genética , Niño , Preescolar , ADN Viral/sangre , ADN Viral/orina , Femenino , Genotipo , Humanos , Factores Inmunológicos/uso terapéutico , Virus JC/genética , Trasplante de Riñón/efectos adversos , Estudios Longitudinales , Masculino , Tipificación Molecular , Síndrome Nefrótico/orina , Síndrome Nefrótico/virología , Infecciones por Polyomavirus/sangre , Infecciones por Polyomavirus/orina , Infecciones por Polyomavirus/virología , Secuencias Reguladoras de Ácidos Nucleicos , Rituximab , Análisis de Secuencia de ADN , Infecciones Tumorales por Virus/sangre , Infecciones Tumorales por Virus/orina , Infecciones Tumorales por Virus/virología , Proteínas Virales/genética , Activación Viral
19.
Influenza Other Respir Viruses ; 16(3): 481-491, 2022 05.
Artículo en Inglés | MEDLINE | ID: mdl-34921508

RESUMEN

INTRODUCTION: Respiratory syncytial virus (RSV) is the major cause of lower respiratory tract illness in young children and can also cause influenza-like illness (ILI). Here we investigated the epidemiological features of RSV infection in pediatric ILI cases in Lombardy (a region in Northern Italy accounting nearly 10 million inhabitants) from 2014-2015 to 2020-2021 winter seasons. MATERIAL AND METHODS: Data for this study were retrieved and statistically analyzed from the database of virological influenza surveillance of the regional reference laboratory for Lombardy within the Italian influenza surveillance network (InfluNet). RESULTS: RSV accounted for nearly 19% of pediatric ILI with a risk of infection nearly two-fold greater than that of individuals ≥15 years. RSV positivity rate increased to 28% considering 0-5 years old children. Although in children ≤5 years the risk of infection from influenza viruses resulted nearly two-fold higher than the risk of RSV infection, the age group 4-6 months and 7-12 months showed a five-fold greater risk of infection from RSV than from influenza. Children ≤5 years of age with pre-existing underlying health conditions had a nearly five-fold greater risk of getting RSV infection than otherwise healthy 0-5 years old children. RSV was identified in ILI cases <15 years of age in all considered winter seasons except in the 2020-2021 season. DISCUSSION: Sentinel surveillance of ILI allowed us to identify groups at higher risk of RSV and influenza infection and to define the start, duration, timing, and intensity of the RSV and influenza community circulation. This surveillance approach can be implemented to assess the RSV circulation and impact in a real-time manner.


Asunto(s)
Gripe Humana , Orthomyxoviridae , Infecciones por Virus Sincitial Respiratorio , Virus Sincitial Respiratorio Humano , Niño , Preescolar , Humanos , Lactante , Recién Nacido , Gripe Humana/epidemiología , Infecciones por Virus Sincitial Respiratorio/epidemiología , Estaciones del Año
20.
Transplant Proc ; 54(7): 2035-2041, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35659782

RESUMEN

BACKGROUND: Human polyomavirus BK (BKPyV) is the etiologic agent of polyomavirus-associated nephropathy, a leading cause of kidney transplant dysfunction. Because of the lack of antiviral therapies, immunosuppression minimization is the recommended treatment. This strategy offers suboptimal outcomes and entails a significant risk of rejection. Our aim was to evaluate the effect of different immunosuppressive drugs (leflunomide, tacrolimus, mycophenolic acid, sirolimus, and everolimus) and their combinations in an in vitro model of BKPyV infection. METHODS: Human renal tubular epithelial cells were infected with BKPyV and treated with leflunomide, tacrolimus, mycophenolic acid, sirolimus, and everolimus, administered alone or in some combination thereof. Viral replication was assessed every 24 hours (up to 72 hours) by BKPyV-specific quantitative real-time polymerized chain reaction for the VIRAL PROTEIN 1 sequence in cell supernatants and by western blot analysis targeting the viral protein 1 and the glyceraldehyde 3-phosphate dehydrogenase on total protein lysates. Results were described as viral copies/mL and compared between treatments at any prespecified time point of the study. RESULTS: The highest inhibitory effects were observed using leflunomide or everolimus plus mycophenolic acid (mean BKPyV replication log reduction 0.28). The antiviral effect of everolimus persisted when it was used in combination with tacrolimus (mean BKPyV replication log reduction 0.27). CONCLUSIONS: Our experience confirms that everolimus has anti-BKPyV properties and prompts future research to investigate possible mechanisms of action. It also provides a rational basis for targeted clinical trials evaluating alternative immunosuppressive modification strategies.


Asunto(s)
Virus BK , Trasplante de Riñón , Infecciones por Polyomavirus , Infecciones Tumorales por Virus , Humanos , Inmunosupresores/farmacología , Inmunosupresores/uso terapéutico , Tacrolimus/farmacología , Tacrolimus/uso terapéutico , Infecciones por Polyomavirus/tratamiento farmacológico , Leflunamida/farmacología , Leflunamida/uso terapéutico , Everolimus/farmacología , Everolimus/uso terapéutico , Ácido Micofenólico/farmacología , Ácido Micofenólico/uso terapéutico , Trasplante de Riñón/efectos adversos , Sirolimus/farmacología , Sirolimus/uso terapéutico , Antivirales/farmacología , Antivirales/uso terapéutico , Proteínas Virales
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