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Article ; Online: An untargeted metabolomic approach to investigate antiviral defence mechanisms in memory leukocytes secreting anti-SARS-CoV-2 IgG in vitro.

Federica, Gevi / Giuseppina, Fanelli / Veronica, Lelli / Gianpaolo, Zarletti / Massimo, Tiberi / Veronica, De Molfetta / Giuseppe, Scapigliati / Maria, Timperio Anna

Scientific reports

2023 Volume 13, Issue 1, Page(s) 629

Abstract: Evidence shows that individuals infected by SARS-CoV-2 experience an altered metabolic state in multiple organs. Metabolic activities are directly involved in modulating immune responses against infectious diseases, yet our understanding of how host ... ...

Abstract	Evidence shows that individuals infected by SARS-CoV-2 experience an altered metabolic state in multiple organs. Metabolic activities are directly involved in modulating immune responses against infectious diseases, yet our understanding of how host metabolism relates to inflammatory responses remains limited. To better elucidate the underlying biochemistry of the leukocyte response, we focused our analysis on possible relationships between SARS-CoV-2 post-infection stages and distinct metabolic pathways. Indeed, we observed a significant altered metabolism of tryptophan and urea cycle pathways in cultures of peripheral blood mononuclear cells obtained 60-90 days after infection and showing in vitro IgG antibody memory for spike-S1 antigen (n = 17). This work, for the first time, identifies metabolic routes in cell metabolism possibly related to later stages of immune defence against SARS-CoV-2 infection, namely, when circulating antibodies may be absent but an antibody memory is present. The results suggest reprogramming of leukocyte metabolism after viral pathogenesis through activation of specific amino acid pathways possibly related to protective immunity against SARS-CoV-2.
MeSH term(s)	Humans ; Leukocytes, Mononuclear ; COVID-19 ; SARS-CoV-2 ; Immunoglobulin G ; Antibodies, Viral ; Leukocytes ; Antiviral Agents
Chemical Substances	Immunoglobulin G ; Antibodies, Viral ; Antiviral Agents
Language	English
Publishing date	2023-01-12
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-022-26156-4
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: The Anti-SARS-CoV-2 Antibody Response in a Centenarian Woman: A Case of Long-Term Memory?

Toppi, Elisa / De Molfetta, Veronica / Zarletti, Gianpaolo / Tiberi, Massimo / Bossù, Paola / Scapigliati, Giuseppe

Viruses

2021 Volume 13, Issue 9

Abstract: SARS-CoV-2 is the virus responsible for the COVID-19 pandemic, causing respiratory syndrome and other manifestations. The clinical consequences of the SARS-CoV-2 infection are highly heterogeneous, ranging from asymptomatic and mild to severe and fatal ... ...

Abstract	SARS-CoV-2 is the virus responsible for the COVID-19 pandemic, causing respiratory syndrome and other manifestations. The clinical consequences of the SARS-CoV-2 infection are highly heterogeneous, ranging from asymptomatic and mild to severe and fatal conditions, with the highest mortality rate reached among elderly people. Such heterogeneity appears strongly influenced by the host immune response, which in turn is profoundly affected by aging. In fact, the occurrence of a low-grade inflammation and a decline in specific immune defense is generally reported in older people. Although the low ability of B cells to provide primary and secondary specific responses with a consequent increase in susceptibility to and severity of virus infections is generally described in elderly people, we would like to present here the particular case of a 100-year-old woman, who recovered well from COVID-19 and developed a long-term memory against SARS-CoV-2. Following the infection, the patient's blood was tested with both a classical ELISA and a specific Cell-ELISA addressed to measure the anti-spike S1 specific IgG released in plasma or produced in vitro by memory B cells, respectively. While showing negative on classical serological testing, the patient's blood was positive in Cell-ELISA up to 1 year after the infection. Our observation highlights a potential mechanism of B cell-dependent, long-term protection in response to SARS-CoV-2 infection, suggesting that in a case of successful aging, the absence of specific antibodies in serum does not necessarily mean the absence of immune memory.
MeSH term(s)	Age Factors ; Aged, 80 and over ; Antibodies, Viral/blood ; Antibodies, Viral/immunology ; Antibody Formation/immunology ; B-Lymphocytes/immunology ; B-Lymphocytes/metabolism ; COVID-19/blood ; COVID-19/diagnosis ; COVID-19/immunology ; COVID-19/virology ; Enzyme-Linked Immunosorbent Assay ; Female ; Host-Pathogen Interactions/immunology ; Humans ; Immunoglobulin G/blood ; Immunoglobulin G/immunology ; Immunologic Memory ; Radiography, Thoracic ; SARS-CoV-2/immunology
Chemical Substances	Antibodies, Viral ; Immunoglobulin G
Language	English
Publishing date	2021-08-27
Publishing country	Switzerland
Document type	Case Reports
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13091704
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: The Anti-SARS-CoV-2 Antibody Response in a Centenarian Woman: A Case of Long-Term Memory?

Toppi, Elisa / De Molfetta, Veronica / Zarletti, Gianpaolo / Tiberi, Massimo / Bossù, Paola / Scapigliati, Giuseppe

Viruses. 2021 Aug. 27, v. 13, no. 9

2021

Abstract	SARS-CoV-2 is the virus responsible for the COVID-19 pandemic, causing respiratory syndrome and other manifestations. The clinical consequences of the SARS-CoV-2 infection are highly heterogeneous, ranging from asymptomatic and mild to severe and fatal conditions, with the highest mortality rate reached among elderly people. Such heterogeneity appears strongly influenced by the host immune response, which in turn is profoundly affected by aging. In fact, the occurrence of a low-grade inflammation and a decline in specific immune defense is generally reported in older people. Although the low ability of B cells to provide primary and secondary specific responses with a consequent increase in susceptibility to and severity of virus infections is generally described in elderly people, we would like to present here the particular case of a 100-year-old woman, who recovered well from COVID-19 and developed a long-term memory against SARS-CoV-2. Following the infection, the patient’s blood was tested with both a classical ELISA and a specific Cell-ELISA addressed to measure the anti-spike S1 specific IgG released in plasma or produced in vitro by memory B cells, respectively. While showing negative on classical serological testing, the patient’s blood was positive in Cell-ELISA up to 1 year after the infection. Our observation highlights a potential mechanism of B cell-dependent, long-term protection in response to SARS-CoV-2 infection, suggesting that in a case of successful aging, the absence of specific antibodies in serum does not necessarily mean the absence of immune memory.
Keywords	COVID-19 infection ; Severe acute respiratory syndrome coronavirus 2 ; antibody formation ; blood serum ; elderly ; immunologic memory ; inflammation ; memory ; mortality ; patients ; viruses ; women
Language	English
Dates of publication	2021-0827
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v13091704
Database	NAL-Catalogue (AGRICOLA)

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Article: A Cell-Based ELISA to Improve the Serological Analysis of Anti-SARS-CoV-2 IgG

Zarletti, Gianpaolo / Tiberi, Massimo / De Molfetta, Veronica / Bossù, Maurizio / Toppi, Elisa / Bossù, Paola / Scapigliati, Giuseppe

Viruses. 2020 Nov. 08, v. 12, no. 11

2020

Abstract: Knowledge of the antibody-mediated immune response to SARS-CoV-2 is crucial to understand virus immunogenicity, establish seroprevalence, and determine whether subjects or recovered patients are at risk for infection/reinfection and would therefore ... ...

Abstract	Knowledge of the antibody-mediated immune response to SARS-CoV-2 is crucial to understand virus immunogenicity, establish seroprevalence, and determine whether subjects or recovered patients are at risk for infection/reinfection and would therefore benefit from vaccination. Here, we describe a novel and simple cell-ELISA specifically designed to measure viral spike S1-specific IgG produced in vitro by B cells in peripheral blood mononuclear cell (PBMC) cultures from a cohort of 45 asymptomatic (n = 24) and symptomatic (n = 21) individuals, with age ranging from 8 to 99 years. All subjects underwent ELISA serological screening twice, at the same time as the cell-ELISA (T2) as well as 35–60 days earlier (T1). Cryopreserved PBMCs of healthy donors obtained years before the COVID-19 pandemic were also included in the analysis. The preliminary results presented here show that out of 45 tested subjects, 16 individuals (35.5%) were positive to the cell-ELISA, 11 (24.5%) were concomitantly positive in the serological screening (T1 and/or T2), and only one person was exclusively positive in ELISA (T1) and negative in cell-ELISA, though values were close to the cutoff. Of note, five individuals (11.2%) tested negative in ELISA but positive in cell-ELISA and thus, they appear to have circulating B cells that produce antibodies against SARS-CoV-2, likely at levels that are undetectable in the serum, which challenges the negative results of the serological screening. The relative level of in vitro secreted IgG was measurable in positive subjects, ranging from 7 to 50 ng/well. Accordingly, all anti-SARS-CoV-2 antibody-positive subjects previously reported moderate to severe symptoms attributable to COVID-19, even though the RT-PCR data were rarely available to confirm viral infection. Overall, the described cell-ELISA might be an effective method for detecting subjects who encountered the virus in the past, and thus helpful to improve serological ELISA tests in the case of undetectable/equivocal circulating IgG levels, and a suitable and improved tool to better evaluate SARS-CoV-2-specific humoral immunity in the COVID-19 pandemic.
Keywords	B-lymphocytes ; COVID-19 infection ; Severe acute respiratory syndrome coronavirus 2 ; antibodies ; blood serum ; cryopreservation ; enzyme-linked immunosorbent assay ; humoral immunity ; immune response ; immunogenicity ; immunoglobulin G ; patients ; reverse transcriptase polymerase chain reaction ; risk ; screening ; serodiagnosis ; seroprevalence ; vaccination ; viruses
Language	English
Dates of publication	2020-1108
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v12111274
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: A Cell-Based ELISA to Improve the Serological Analysis of Anti-SARS-CoV-2 IgG.

Zarletti, Gianpaolo / Tiberi, Massimo / De Molfetta, Veronica / Bossù, Maurizio / Toppi, Elisa / Bossù, Paola / Scapigliati, Giuseppe

Viruses

2020 Volume 12, Issue 11

Abstract	Knowledge of the antibody-mediated immune response to SARS-CoV-2 is crucial to understand virus immunogenicity, establish seroprevalence, and determine whether subjects or recovered patients are at risk for infection/reinfection and would therefore benefit from vaccination. Here, we describe a novel and simple cell-ELISA specifically designed to measure viral spike S1-specific IgG produced in vitro by B cells in peripheral blood mononuclear cell (PBMC) cultures from a cohort of 45 asymptomatic (
MeSH term(s)	Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Viral/blood ; Betacoronavirus/immunology ; COVID-19 ; COVID-19 Testing ; Child ; Clinical Laboratory Techniques/methods ; Coronavirus Infections/diagnosis ; Enzyme-Linked Immunosorbent Assay/methods ; Female ; Humans ; Immunoglobulin G/blood ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral/diagnosis ; SARS-CoV-2 ; Serologic Tests ; Spike Glycoprotein, Coronavirus/immunology ; Young Adult
Chemical Substances	Antibodies, Viral ; Immunoglobulin G ; Spike Glycoprotein, Coronavirus
Keywords	covid19
Language	English
Publishing date	2020-11-08
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v12111274
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Oral High-Dose Thiamine Improves the Symptoms of Chronic Cluster Headache.

Antonio, Costantini / Massimo, Tiberi / Gianpaolo, Zarletti / Immacolata, Pala Maria / Erika, Trevi

Case reports in neurological medicine

2018 Volume 2018, Page(s) 3901619

Abstract: Cluster headache is a rare painful primary disorder occurring in either episodic or chronic patterns. Several authors found that the hypothalamus, the brain region regulating endocrine function and autonomic system, is involved in the pathophysiology of ... ...

Abstract	Cluster headache is a rare painful primary disorder occurring in either episodic or chronic patterns. Several authors found that the hypothalamus, the brain region regulating endocrine function and autonomic system, is involved in the pathophysiology of cluster headache. Some authors have found in patients affected by this disease abnormality in glucose metabolism. Considering the role of thiamine in brain function, in energetic metabolism, and in pain modulation, we treated a patient affected by cluster headache with oral high-dose thiamine. We report a 41-year-old man suffering from primary chronic cluster headache since the age of 15 years. The patient began oral therapy with high-dose thiamine in December 2016. Oral thiamine supplementation led to a dramatic improvement of the symptoms. The therapy was effective in reversing all the symptoms of the disease. Our observation suggests that a thiamine deficiency due to enzymatic abnormalities or to dysfunction of the circulation of thiamine in the intracellular space could cause a neuronal selective impairment in the centers that are involved in this disease and could have an important role in the pathogenesis of the symptoms of cluster headache.
Language	English
Publishing date	2018-04-18
Publishing country	United States
Document type	Case Reports
ZDB-ID	2629909-4
ISSN	2090-6676 ; 2090-6668
ISSN (online)	2090-6676
ISSN	2090-6668
DOI	10.1155/2018/3901619
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: A Cell-Based ELISA to Improve the Serological Analysis of Anti-SARS-CoV-2 IgG

Gianpaolo Zarletti / Massimo Tiberi / Veronica De Molfetta / Maurizio Bossù / Elisa Toppi / Paola Bossù / Giuseppe Scapigliati

Viruses, Vol 12, Iss 1274, p

2020 Volume 1274

Abstract	Knowledge of the antibody-mediated immune response to SARS-CoV-2 is crucial to understand virus immunogenicity, establish seroprevalence, and determine whether subjects or recovered patients are at risk for infection/reinfection and would therefore benefit from vaccination. Here, we describe a novel and simple cell-ELISA specifically designed to measure viral spike S1-specific IgG produced in vitro by B cells in peripheral blood mononuclear cell (PBMC) cultures from a cohort of 45 asymptomatic ( n = 24) and symptomatic ( n = 21) individuals, with age ranging from 8 to 99 years. All subjects underwent ELISA serological screening twice, at the same time as the cell-ELISA (T2) as well as 35–60 days earlier (T1). Cryopreserved PBMCs of healthy donors obtained years before the COVID-19 pandemic were also included in the analysis. The preliminary results presented here show that out of 45 tested subjects, 16 individuals (35.5%) were positive to the cell-ELISA, 11 (24.5%) were concomitantly positive in the serological screening (T1 and/or T2), and only one person was exclusively positive in ELISA (T1) and negative in cell-ELISA, though values were close to the cutoff. Of note, five individuals (11.2%) tested negative in ELISA but positive in cell-ELISA and thus, they appear to have circulating B cells that produce antibodies against SARS-CoV-2, likely at levels that are undetectable in the serum, which challenges the negative results of the serological screening. The relative level of in vitro secreted IgG was measurable in positive subjects, ranging from 7 to 50 ng/well. Accordingly, all anti-SARS-CoV-2 antibody-positive subjects previously reported moderate to severe symptoms attributable to COVID-19, even though the RT-PCR data were rarely available to confirm viral infection. Overall, the described cell-ELISA might be an effective method for detecting subjects who encountered the virus in the past, and thus helpful to improve serological ELISA tests in the case of undetectable/equivocal circulating IgG levels, and a suitable and improved tool to better evaluate SARS-CoV-2-specific humoral immunity in the COVID-19 pandemic.
Keywords	SARS-CoV-2 ; COVID-19 ; in vitro IgG ; B cell memory ; cell-ELISA ; spike S1 protein ; Microbiology ; QR1-502 ; covid19
Subject code	571
Language	English
Publishing date	2020-11-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: An untargeted metabolomic approach to identify antiviral defense mechanisms in memory leukocytes secreting in vitro IgG anti-SARS-Cov-2

Timperio, Anna Maria / Gevi, Federica / Fanelli, Giuseppina / Lelli, Veronica / Zarletti, Gianpaolo / Tiberi, Massimo / De Molfetta, Veronica / Scapigliati, Giuseppe

bioRxiv

Abstract: Available knowledge shows that individuals infected by SARS-CoV-2 undergo an altered metabolic state in multiple organs. Metabolic activities are directly involved in modulating the immune responses against infectious diseases, yet our understanding ... ...

Abstract	Available knowledge shows that individuals infected by SARS-CoV-2 undergo an altered metabolic state in multiple organs. Metabolic activities are directly involved in modulating the immune responses against infectious diseases, yet our understanding remains limited on how host metabolism relates with inflammatory responses. To better elucidate the underlying biochemistry of leukocytes response, we focused our analysis on the possible relationships between SARS-CoV-2 post-infection stages and distinct metabolic pathways. Indeed, in cultures of peripheral blood mononuclear cells (PBMC, n=48) obtained 60-90 days after infection and showing in vitro IgG antibody memory for spike-S1 antigen (n=19), we observed a significant altered metabolism of tryptophan and urea cycle pathways. This work for the first time identifies metabolic routes in cell metabolism possibly related to later stages of immune defense against SARS-Cov-2 infection, namely when circulating antibodies may be absent, but an antibody memory is present. The results suggest a reprogramming of leukocyte metabolism after viral pathogenesis through activation of specific amino acid pathways possibly related to protective immunity against SARS-CoV-2.
Keywords	covid19
Language	English
Publishing date	2021-07-21
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2021.07.20.453042
Database	COVID19

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Article: A Cell-Based ELISA to Improve the Serological Analysis of Anti-SARS-CoV-2 IgG

Zarletti, Gianpaolo / Tiberi, Massimo / De Molfetta, Veronica / Bossù, Maurizio / Toppi, Elisa / Bossù, Paola / Scapigliati, Giuseppe

Viruses

Abstract	Knowledge of the antibody-mediated immune response to SARS-CoV-2 is crucial to understand virus immunogenicity, establish seroprevalence, and determine whether subjects or recovered patients are at risk for infection/reinfection and would therefore benefit from vaccination. Here, we describe a novel and simple cell-ELISA specifically designed to measure viral spike S1-specific IgG produced in vitro by B cells in peripheral blood mononuclear cell (PBMC) cultures from a cohort of 45 asymptomatic (n = 24) and symptomatic (n = 21) individuals, with age ranging from 8 to 99 years. All subjects underwent ELISA serological screening twice, at the same time as the cell-ELISA (T2) as well as 35-60 days earlier (T1). Cryopreserved PBMCs of healthy donors obtained years before the COVID-19 pandemic were also included in the analysis. The preliminary results presented here show that out of 45 tested subjects, 16 individuals (35.5%) were positive to the cell-ELISA, 11 (24.5%) were concomitantly positive in the serological screening (T1 and/or T2), and only one person was exclusively positive in ELISA (T1) and negative in cell-ELISA, though values were close to the cutoff. Of note, five individuals (11.2%) tested negative in ELISA but positive in cell-ELISA and thus, they appear to have circulating B cells that produce antibodies against SARS-CoV-2, likely at levels that are undetectable in the serum, which challenges the negative results of the serological screening. The relative level of in vitro secreted IgG was measurable in positive subjects, ranging from 7 to 50 ng/well. Accordingly, all anti-SARS-CoV-2 antibody-positive subjects previously reported moderate to severe symptoms attributable to COVID-19, even though the RT-PCR data were rarely available to confirm viral infection. Overall, the described cell-ELISA might be an effective method for detecting subjects who encountered the virus in the past, and thus helpful to improve serological ELISA tests in the case of undetectable/equivocal circulating IgG levels, and a suitable and improved tool to better evaluate SARS-CoV-2-specific humoral immunity in the COVID-19 pandemic.
Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #918921
Database	COVID19

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Article: Cellular activities during a mixed leucocyte reaction in the teleost sea bass Dicentrarchus labrax.

Meloni, Sabrina / Zarletti, Gianpaolo / Benedetti, Stefania / Randelli, Elisa / Buonocore, Francesco / Scapigliati, Giuseppe

Fish & shellfish immunology

2006 Volume 20, Issue 5, Page(s) 739–749

Abstract: In this investigation a number of "in vitro" activities of sea bass peripheral blood leucocytes (PBL) against allogeneic PBL inactivated by irradiation were studied. Stimulator PBL were cultured with inactivated allogeneic PBL, and direct counting of ... ...

Abstract	In this investigation a number of "in vitro" activities of sea bass peripheral blood leucocytes (PBL) against allogeneic PBL inactivated by irradiation were studied. Stimulator PBL were cultured with inactivated allogeneic PBL, and direct counting of lymphocytes was done after 2 weeks by immunofluorescence and flow cytometry using mAbs DLT15 and DLIg3 specific for T-cells and B-cells, respectively. In a one-way mixed leucocyte reaction (MLR), results showed an increase of T lymphocytes, whereas B lymphocytes had values similar to those in control PBL. The increase of T-cells in MLR cultures was also confirmed using RT-PCR by analyzing the expression of the T-cell receptor (beta-subunit) mRNA. The addition of 5 microg/ml of cyclosporin A (CsA) to the MLR caused a significant decrease in T-cell proliferation. Leucocytes from MLR cultures displayed an enhanced cytotoxic activity against xenogeneic target cells with respect to control PBL, raising the possibility of the presence of cytotoxic-like T lymphocytes. Cellular activation of PBL was confirmed in 2 weeks MLR by measuring antibody-induced intracellular Ca(++) mobilization with Fura-2 AM. This work represents the first direct quantitative determination of an "in vitro" T-cell activity in a teleost species.
MeSH term(s)	Adenosine Triphosphate/analysis ; Analysis of Variance ; Animals ; B-Lymphocytes/drug effects ; B-Lymphocytes/immunology ; Bass/immunology ; Cell Line ; Cyclosporine/pharmacology ; Cytotoxicity Tests, Immunologic ; Flow Cytometry/methods ; Fluorescent Antibody Technique, Indirect/methods ; Immunosuppressive Agents/pharmacology ; Leukocytes/drug effects ; Leukocytes/immunology ; Leukocytes/radiation effects ; Lymphocyte Culture Test, Mixed/methods ; Reverse Transcriptase Polymerase Chain Reaction/methods ; T-Lymphocytes/drug effects ; T-Lymphocytes/immunology ; T-Lymphocytes/physiology
Chemical Substances	Immunosuppressive Agents ; Cyclosporine (83HN0GTJ6D) ; Adenosine Triphosphate (8L70Q75FXE)
Language	English
Publishing date	2006-05
Publishing country	England
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1067738-0
ISSN	1095-9947 ; 1050-4648
ISSN (online)	1095-9947
ISSN	1050-4648
DOI	10.1016/j.fsi.2005.10.001
Database	MEDical Literature Analysis and Retrieval System OnLINE

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