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2. Alpha-synuclein shapes monocyte and macrophage cell biology and functions by bridging alterations of autophagy and inflammatory pathways.

Author

Limanaqi, Fiona, Zecchini, Silvia, Ogno, Pasquale, Artusa, Valentina, Fenizia, Claudio, Saulle, Irma, Vanetti, Claudia, Garziano, Micaela, Strizzi, Sergio, Trabattoni, Daria, Clerici, Mario, and Biasin, Mara

Subjects
CYTOLOGY, CELL physiology, CELL migration, ALPHA-synuclein, MACROPHAGES
Abstract

Introduction: Abnormal spreading of alpha-synuclein (αS), a hallmark of Parkinson's disease, is known to promote peripheral inflammation, which occurs in part via functional alterations in monocytes/macrophages. However, underlying intracellular mechanisms remain unclear. Methods: Herein we investigate the subcellular, molecular, and functional effects of excess αS in human THP-1monocytic cell line, THP-1-derived macrophages, and at least preliminarily, in primary monocyte-derived macrophages (MDMs). In cells cultured w/wo recombinant αS (1 βM) for 4 h and 24 h, by Confocal microscopy, Western Blot, RT-qPCR, Elisa, and Flow Cytometry we assessed: i) αS internalization; ii) cytokine/chemokine expression/secretion, and C-C motif chemokine receptor 2 (CCR2) levels; iii) autophagy (LC3II/I, LAMP1/LysoTracker, p62, pS6/total S6); and iv) lipid droplets (LDs) accumulation, and cholesterol pathway gene expression. Transwell migration assay was employed to measure THP-1 cell migration/chemotaxis, while FITC-IgG-bead assay was used to analyze phagocytic capacity, and the fate of phagocytosed cargo in THP-1-derived macrophages. Results: Extracellular αSwas internalized by THP-1 cells, THP-1-derivedmacrophages, and MDMs. In THP1 cells, αS induced a general pro-inflammatory profile and conditioned media from αS-exposed THP-1 cells potently attracted unstimulated cells. However, CCL2 secretion peaked at 4 h αS, consistent with early internalization of its receptor CCR2, while this was blunted at 24 h αS exposure, when CCR2 recycled back to the plasma membrane. Again, 4 h αS-exposed THP-1 cells showed increased spontaneous migration, while 24 h αS-exposed cells showed reduced chemotaxis. This occurred in the absence of cell toxicity and was associated with upregulation of autophagy/lysosomalmarkers, suggesting a pro-survival/tolerance mechanism against stress-related inflammation. Instead, in THP-1-derived macrophages, αS time-dependently potentiated the intracellular accumulation, and release of pro-inflammatory mediators. This was accompanied by mild toxicity, reduced autophagy-lysosomal markers, defective LDs formation, as well as impaired phagocytosis, and the appearance of stagnant lysosomes engulfed with phagocytosed cargo, suggesting a status of macrophage exhaustion reminiscent of hypophagia. Discussion: In summary, despite an apparently similar pro-inflammatory phenotype, monocytes and macrophages respond differently to intracellular αS accumulation in terms of cell survival, metabolism, and functions. Our results suggest that in periphery, αS exerts cell- and context-specific biological effects bridging alterations of autophagy, lipid dynamics, and inflammatory pathways. [ABSTRACT FROM AUTHOR]

Published
2024
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3. Humoral and cell-mediated immune responses in HIV-vertically infected young patients after three doses of the BNT162b2 mRNA SARS-CoV-2 vaccine

Author

Vanetti, Claudia, primary, Stracuzzi, Marta, additional, Crivellaro, Elisa, additional, Ciciliano, Federica, additional, Garziano, Micaela, additional, Fenizia, Claudio, additional, Biasin, Mara, additional, Rubinacci, Valeria, additional, Amendola, Antonella, additional, Tanzi, Elisabetta, additional, Zuccotti, Gian Vincenzo, additional, Clerici, Mario, additional, Giacomet, Vania, additional, and Trabattoni, Daria, additional

Published
2024
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4. Human motor neurons derived from induced pluripotent stem cells are susceptible to SARS-CoV-2 infection

Author

Cappelletti, Gioia, primary, Colombrita, Claudia, additional, Limanaqi, Fiona, additional, Invernizzi, Sabrina, additional, Garziano, Micaela, additional, Vanetti, Claudia, additional, Moscheni, Claudia, additional, Santangelo, Serena, additional, Zecchini, Silvia, additional, Trabattoni, Daria, additional, Silani, Vincenzo, additional, Clerici, Mario, additional, Ratti, Antonia, additional, and Biasin, Mara, additional

Published
2023
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6. Saliva and Plasma Neutralizing Activity Induced by the Administration of a Third bnt162b2 Vaccine Dose

Author

Garziano, Micaela, primary, Utyro, Olga, additional, Strizzi, Sergio, additional, Vanetti, Claudia, additional, Saulle, Irma, additional, Conforti, Chiara, additional, Cicilano, Federica, additional, Ardizzone, Francesco, additional, Cappelletti, Gioia, additional, Clerici, Mario, additional, Limanaqi, Fiona, additional, and Biasin, Mara, additional

Published
2022
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7. Leishmania tarentolae as an Antigen Delivery Platform: Dendritic Cell Maturation after Infection with a Clone Engineered to Express the SARS-CoV-2 Spike Protein

Author

Varotto-Boccazzi, Ilaria, primary, Garziano, Micaela, additional, Cattaneo, Giulia Maria, additional, Bisaglia, Beatrice, additional, Gabrieli, Paolo, additional, Biasin, Mara, additional, Manenti, Alessandro, additional, Rubolini, Diego, additional, Clerici, Mario, additional, Montomoli, Emanuele, additional, Zuccotti, Gian Vincenzo, additional, Trabattoni, Daria, additional, Epis, Sara, additional, and Bandi, Claudio, additional

Published
2022
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11. Severity of COVID-19 Patients Predicted by Serum Sphingolipids Signature

Author

Torretta, Enrica, Garziano, Micaela, Poliseno, Mariacristina, Capitanio, Daniele, Biasin, Mara, Santantonio, Teresa Antonia, Clerici, Mario, Lo Caputo, Sergio, Trabattoni, Daria, and Gelfi, Cecilia

Subjects
Adult, Male, QH301-705.5, frailty, Severity of Illness Index, Article, serine palmitoyltransferase, Young Adult, caspase 3, Humans, acid sphingomyelinase, Biology (General), QD1-999, Aged, mass spectrometry, Aged, 80 and over, sphingolipids, SARS-CoV-2, COVID-19, Middle Aged, COVID-19 severity, Prognosis, Sphingomyelins, Chemistry, Lipidomics, Female, lipids (amino acids, peptides, and proteins)
Abstract

The reason behind the high inter-individual variability in response to SARS-CoV-2 infection and patient’s outcome is poorly understood. The present study targets the sphingolipid profile of twenty-four healthy controls and fifty-nine COVID-19 patients with different disease severity. Sera were analyzed by untargeted and targeted mass spectrometry and ELISA. Results indicated a progressive increase in dihydrosphingosine, dihydroceramides, ceramides, sphingosine, and a decrease in sphingosine-1-phosphate. These changes are associated with a serine palmitoyltransferase long chain base subunit 1 (SPTLC1) increase in relation to COVID-19 severity. Severe patients showed a decrease in sphingomyelins and a high level of acid sphingomyelinase (aSMase) that influences monosialodihexosyl ganglioside (GM3) C16:0 levels. Critical patients are characterized by high levels of dihydrosphingosine and dihydroceramide but not of glycosphingolipids. In severe and critical patients, unbalanced lipid metabolism induces lipid raft remodeling, leads to cell apoptosis and immunoescape, suggesting active sphingolipid participation in viral infection. Furthermore, results indicated that the sphingolipid and glycosphingolipid metabolic rewiring promoted by aSMase and GM3 is age-dependent but also characteristic of severe and critical patients influencing prognosis and increasing viral load. AUCs calculated from ROC curves indicated ceramides C16:0, C18:0, C24:1, sphingosine and SPTLC1 as putative biomarkers of disease evolution

Published
2021

12. MiRNA Profiling in Plasma and Placenta of SARS-CoV-2-Infected Pregnant Women

Author

Saulle, Irma, Garziano, Micaela, Fenizia, Claudio, Cappelletti, Gioia, Parisi, Francesca, Clerici, Mario, Cetin, Irene, Savasi, Valeria, and Biasin, Mara

Subjects
Adult, SARS-CoV-2, QH301-705.5, viruses, Placenta, COVID-19, MicroRNA, Article, immune response, MicroRNAs, Young Adult, Pregnancy, Humans, Female, Pregnancy Complications, Infectious, Biology (General), Transcriptome, pregnant women
Abstract

MicroRNAs are gene expression regulators associated with several human pathologies, including those generated by viral infections. Their role in SARS-CoV-2 infection and COVID-19 has been investigated and reviewed in many informative studies, however, a thorough miRNA outline in SARS-CoV-2-infected pregnant women (SIPW), at both systemic and placental levels, is missing. To fill this gap, blood and placenta biopsies collected at delivery from 15 asymptomatic SIPW were immediately analysed for: miRNA expression (n = 84) (QPCR array), antiviral/immune mRNA target expression (n = 74) (QGene) and cytokine/chemokines production (n = 27) (Multiplex ELISA). By comparing these results with those obtained from six uninfected pregnant women (UPW), we observed that, following SARS-CoV-2 infection, the transcriptomic profile of pregnant women is significantly altered in different anatomical districts, even in the absence of clinical symptoms and vertical transmission. This characteristic combination of miRNA and antiviral/immune factors seems to control both the infection and the dysfunctional immune reaction, thus representing a positive correlate of protection and a potential therapeutic target against SARS-CoV-2.

Published
2021

16. A 6-amino acid insertion/deletion polymorphism in the mucin domain of TIM-1 confers protections against HIV-1 infection

Author

Biasin, Mara, Sironi, Manuela, Saulle, Irma, Pontremoli, Chiara, Garziano, Micaela, Cagliani, Rachele, Trabattoni, Daria, Lo Caputo, Sergio, Vichi, Francesca, Mazzotta, Francesco, Forni, Diego, Riva, Stefania, Aguilar-Jimenez, Wbeimar, Cedeño, Samandhy, Sanchez, Jorge, Brander, Christian, Zapata, Wildeman, Rugeles, Maria Teresa, and Clerici, Mario

Published
2017
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17. IMMUNOMODULATORY EFFECTS OF PIDOTIMOD IN PATIENTS HOSPITALIZED FOR COMMUNITY-ACQUIRED PNEUMONIA

Author

GARZIANO, MICAELA

Abstract

Introduzione: Il pidotimod (PDT) � una molecola dipeptidica di sintesi ad azione immunomodulante. Tale composto � studiato gi� da diversi anni con incoraggianti risultati a carico sia del sistema immunitario innato che adattativo. Studi in vitro ed in vivo ne hanno gi� dimostrato i benefici effetti nel trattamento di molte patologie. Col nostro studio abbiamo voluto porre l?accento sugli effetti del PDT associato alla terapia antibiotica standard in pazienti ospedalizzati per polmonite contratta in comunit� (CAP). Metodi: 36 pazienti ospedalizzati per CAP, di cui 20 bambini e 16 adulti, sono stati arruolati nello studio secondo specifici criteri di inclusione. I pazienti sono stati suddivisi in due gruppi di trattamento, al primo � stato somministrato PDT associato alla terapia antibiotica standard, al secondo la sola terapia antibiotica. I campioni di sangue sono stati prelevati a diverse tempistiche, i bambini sono stati prelevati al momento dell?arruolamento (T0), a 3 e 5 giorni dall? inizio della terapia (T3 e T5), e a 7 giorni dalla fine della somministrazione farmacologica (T21), Gli adulti sono stati invece prelevati al momento dell?arruolamento (T0), e rispettivamente a 1, 3, e 5 giorni dall?inizio della terapia (T1, T3, T5). Le PBMC sono state isolate da sangue periferico e stimolate in presenza di pneumococco. Dopo 3 ore di stimolazione una parte delle cellule � stata utilizzata per l?estrazione dell?RNA per la valutazione dell?espressione di specifici geni coinvolti nella risposta antibatterica; mentre dopo 18 ore di stimolazione le restanti cellule sono state marcate per le analisi citofluorimetriche. Risultati: I risultati ottenuti nei bambini hanno evidenziato un?azione del PDT su cellule denritiche, dove modula l?espressione di marker di attivazione e molecole costimolatorie quali HLA DRII, CD80 e CD86; e monociti, sui quali incrementa l?espressione del TLR2. Inoltre il gruppo trattato con PDT manifesta anche un?incremento nella secrezione di citochine proinfiammatorie TNF? e IL-12. Negli adulti � stato anche osservato un aumento nell? espressione del TLR4 a livello dei monociti, mentre nessuna differenza � stata osservata nella secrezione delle citochine proinfiammatorie. In aggiunta, in entrambi i gruppi si � osservata una modulazione nell? espressione di geni coinvolti nella risposta infiammatoria e antimicrobica. Conclusioni: Questi risultati confermano gli effetti immunomodulanti del PDT gi� descritti in letteratura, nonch� i benefici della molecola associata alla terapia antibiotica standard nei pazienti affetti da CAP, grazie alla sua azione a livello dell?immunit� innata.

Published
2017
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19. Precursor Forms of Vitamin D Reduce HIV-1 Infection In Vitro

Author

Aguilar-Jimenez, Wbeimar, primary, Villegas-Ospina, Simon, additional, Gonzalez, Sandra, additional, Zapata, Wildeman, additional, Saulle, Irma, additional, Garziano, Micaela, additional, Biasin, Mara, additional, Clerici, Mario, additional, and Rugeles, Maria T., additional

Published
2016
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21. Plasma and PBMC miRNA Profile in Sexually HIV-1 Exposed Seronegative Individuals

Author

Biasin, Mara, primary, Yahyaei, Sara, additional, De Luca, Mariacristina, additional, Saulle, Irma, additional, Gnudi, Federica, additional, Ibba, Salomè, additional, Garziano, Micaela, additional, Berzi, Angela, additional, Rainone, Veronica, additional, Trabattoni, Daria, additional, Caputo, Sergio Lo, additional, Mazzotta, Francesco, additional, and Clerici, Mario, additional

Published
2014
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22. Evolutionary Analysis Identifies an MX2 Haplotype Associated with Natural Resistance to HIV-1 Infection

Author

Saulle, Irma, primary, Biasin, Mara, additional, Gnudi, Federica, additional, Ibba, Salomè, additional, Garziano, Micaela, additional, Sironi, Manuela, additional, Trabattoni, Daria, additional, Caputo, Sergio Lo, additional, Mazzotta, Francesco, additional, Caruz, Antonio, additional, De Gioia, Luca, additional, and Clerici, Mario Salvatore, additional

Published
2014
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23. A Regulatory Polymorphism in HAVCR2 Modulates Susceptibility to HIV-1 Infection

Author

Sironi, Manuela, primary, Biasin, Mara, additional, Gnudi, Federica, additional, Cagliani, Rachele, additional, Saulle, Irma, additional, Forni, Diego, additional, Rainone, Veronica, additional, Trabattoni, Daria, additional, Garziano, Micaela, additional, Mazzotta, Francesco, additional, Real, Luis Miguel, additional, Rivero-Juarez, Antonio, additional, Caruz, Antonio, additional, Caputo, Sergio Lo, additional, and Clerici, Mario, additional

Published
2014
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25. Evolutionary Analysis Identifies an MX2 Haplotype Associated with Natural Resistance to HIV-1 Infection.

Author

Biasin, Mara, Gnudi, Federica, Ibba, Salomè, Garziano, Micaela, Sironi, Manuela, Trabattoni, Daria, Caputo, Sergio Lo, Mazzotta, Francesco, Caruz, Antonio, De Gioia, Luca, and Clerici, Mario Salvatore

Abstract

An abstract of the article "Evolutionary Analysis Identifies an MX2 Haplotype Associated with Natural Resistance to HIV-1 Infection" by Irma Saulle is presented.

Published
2014
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26. Plasma and PBMC miRNA Profile in Sexually HIV-1 Exposed Seronegative Individuals.

Author

Yahyaei, Sara, De Luca, Mariacristina, Saulle, Irma, Gnudi, Federica, Ibba, Salomè, Garziano, Micaela, Berzi, Angela, Rainone, Veronica, Trabattoni, Daria, Caputo, Sergio Lo, Mazzotta, Francesco, and Clerici, Mario

Abstract

An abstract of the article "Plasma and PBMC miRNA Profile in Sexually HIV-1 Exposed Seronegative Individuals" by Sara Yahyaei, Mariacristina De Luca and colleagues is presented.

Published
2014
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27. Precursor Forms of Vitamin D Reduce HIV-1 Infection In Vitro.

Author

Aguilar-Jimenez W, Villegas-Ospina S, Gonzalez S, Zapata W, Saulle I, Garziano M, Biasin M, Clerici M, and Rugeles MT

Subjects
ADP-ribosyl Cyclase 1 analysis, Cells, Cultured, Colombia, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Gene Expression Profiling, HIV Core Protein p24 analysis, HLA-DR Antigens analysis, Humans, Immunity, Innate drug effects, Italy, Leukocytes, Mononuclear chemistry, Leukocytes, Mononuclear virology, Membrane Glycoproteins analysis, Real-Time Polymerase Chain Reaction, Anti-HIV Agents pharmacology, Calcifediol pharmacology, Cholecalciferol pharmacology, HIV-1 drug effects, HIV-1 growth & development, Immunologic Factors pharmacology
Abstract

Background: Although the anti-HIV-1 effects of vitamin D (VitD) have been reported, mechanisms behind such protection remain largely unexplored., Methods: The effects of two precursor forms (cholecalciferol/calciol at 0.01, 1 and 100 nM and calcidiol at 100 and 250 nM) on HIV-1 infection, immune activation, and gene expression were analyzed in vitro in cells of Colombian and Italian healthy donors. We quantified levels of released p24 by enzyme-linked immunosorbent assay, of intracellular p24 and cell-surface expression of CD38 and HLA-DR by flow cytometry, and mRNA expression of antiviral and immunoregulatory genes by real-time reverse transcription-polymerase chain reaction., Results: Cholecalciferol decreased the frequency of HIV-1-infected p24CD4 T cells and levels of p24 in supernatants in a dose-dependent manner. Moreover, the CD4CD38HLA-DR and CD4CD38HLA-DR subpopulations were more susceptible to infection but displayed the greatest cholecalciferol-induced decreases in infection rate by an X4-tropic strain. Likewise, cholecalciferol at its highest concentration decreased the frequency of CD38HLA-DR but not of CD38HLA-DR T-cell subsets. Analyzing the effects of calcidiol, the main VitD source for immune cells and an R5-tropic strain as the most frequently transmitted virus, a reduction in HIV-1 productive infection was also observed. In addition, an increase in mRNA expression of APOBEC3G and PI3 and a reduction of TRIM22 and CCR5 expression, this latter positively correlated with p24 levels, was noted., Conclusions: VitD reduces HIV-1 infection in T cells possibly by inducing antiviral gene expression, reducing the viral co-receptor CCR5 and, at least at the highest cholecalciferol concentration, by promoting an HIV-1-restrictive CD38HLA-DR immunophenotype.

Published
2016
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