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Article ; Online: Comparison of multiplex cytokine assays in a pediatric cohort with epilepsy

Adam L. Numis / Christine H. Fox / Daniel J. Lowenstein / Philip J. Norris / Clara Di Germanio

Heliyon, Vol 7, Iss 3, Pp e06445- (2021)

2021

Abstract: Background: Multiplex analyses allow for detection of dozens of cytokines/chemokines in small sample volumes. Although several commercially available assay kits are available, there are no comparative data in plasma measurements among pediatric or ... ...

Abstract	Background: Multiplex analyses allow for detection of dozens of cytokines/chemokines in small sample volumes. Although several commercially available assay kits are available, there are no comparative data in plasma measurements among pediatric or epilepsy cohorts. New method: Cohort study of 38 children with epilepsy. We evaluated plasma levels of cytokines/chemokines using three different assays: Luminex® xMAP high-sensitivity (HS) and standard-sensitivity (SS) assays, and Meso-Scale Discovery (MSD). We calculated recovery rates of each analyte, correlation coefficients between assays, and level of agreement between measurements. We repeated analyses in a subset of samples after a single freeze-thaw cycle. Results: Among ten analytes common to all assays, HS had high recovery (<15% of values extrapolated or out-of- range [OOR]) for all analytes, SS for 50%, and MSD for 40%. While several analytes had a high correlation between assays, Bland-Altman plots demonstrated assays were not interchangeable. For most analytes, a single freeze-thaw cycle decreased cytokines/chemokine measurements. There was good correlation of measurements after a freeze-thaw cycle with acceptable agreement between measurements for six of 13 (46%) analytes using HS, one of 9 (11%) for SS, and none for MSD. Comparison with existing methods: HS assays may optimize yield in plasma for proteins of particular interest in epilepsy research, limit values extrapolated beyond the standard curve, and improve precision compared to other SS and MSD assays. Conclusion: Our results demonstrate assay choice may be critical to study results and support the need for a standardized approach to biomarker assessment across epilepsy research and other domains.
Keywords	Pediatric epilepsy ; Cytokines ; Chemokines ; Luminex ; Meso-scale discovery ; Neuro-inflammation ; Science (General) ; Q1-390 ; Social sciences (General) ; H1-99
Subject code	570
Language	English
Publishing date	2021-03-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Amniotic epithelial cells

Clara Di Germanio / Michel Bernier / Rafael de Cabo / Barbara Barboni

Frontiers in Cell and Developmental Biology, Vol

a new tool to combat aging and age-related diseases?

2016 Volume 4

Abstract: The number of elderly people is growing at an unprecedented rate and this increase of the aging population is expected to have a direct impact on the incidence of age-related diseases and healthcare-associated costs. Thus, it is imperative that new tools ...

Abstract	The number of elderly people is growing at an unprecedented rate and this increase of the aging population is expected to have a direct impact on the incidence of age-related diseases and healthcare-associated costs. Thus, it is imperative that new tools are developed to fight and slow age-related diseases. Regenerative medicine is a promising strategy for the maintenance of health and function late in life; however, stem cell-based therapies face several challenges including rejection and tumor transformation. As an alternative, the placenta offers an extraordinary source of fetal stem cells, including the amniotic epithelial cells (AECs), which retain some of the characteristics of embryonic stem cells, but show low immunogenicity, together with immunomodulatory and anti-inflammatory activities. Because of these characteristics, AECs have been widely utilized in regenerative medicine. This perspective highlights different mechanisms triggered by transplanted AECs that could be potentially useful for anti-aging therapies, which include: Graft and differentiation for tissue regeneration in age-related settings, anti-inflammatory behavior to combat inflammaging, anti-tumor activity, direct lifespan and healthspan extension properties, and possibly rejuvenation in a manner reminiscent of heterochronic parabiosis.Here, we critically discuss benefits and limitation of AECs-based therapies in age-related diseases.
Keywords	Aging ; Inflammation ; Regenerative Medicine ; Rejuvenation ; tumorigenesis ; Amniotic cells ; Biology (General) ; QH301-705.5
Subject code	610
Language	English
Publishing date	2016-11-01T00:00:00Z
Publisher	Frontiers Media S.A.
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Using sero-epidemiology to monitor disparities in vaccination and infection with SARS-CoV-2

Isobel Routledge / Saki Takahashi / Adrienne Epstein / Jill Hakim / Owen Janson / Keirstinne Turcios / Jo Vinden / John Tomas Risos / Margaret Rose Baniqued / Lori Pham / Clara Di Germanio / Michael Busch / Margot Kushel / Bryan Greenhouse / Isabel Rodríguez-Barraquer

Nature Communications, Vol 13, Iss 1, Pp 1-

2022 Volume 7

Abstract: Continued monitoring of SARS-CoV-2 at the population level is important for identifying at-risk groups. Here the authors analyse data from a serological surveillance platform in San Francisco and find considerable variation in infection and vaccination ... ...

Abstract	Continued monitoring of SARS-CoV-2 at the population level is important for identifying at-risk groups. Here the authors analyse data from a serological surveillance platform in San Francisco and find considerable variation in infection and vaccination history by race/ethnicity and socioeconomic status.
Keywords	Science ; Q
Language	English
Publishing date	2022-05-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Distinct SARS-CoV-2 antibody reactivity patterns in coronavirus convalescent plasma revealed by a coronavirus antigen microarray

Rafael Assis / Aarti Jain / Rie Nakajima / Algis Jasinskas / Saahir Khan / Huw Davies / Laurence Corash / Larry J. Dumont / Kathleen Kelly / Graham Simmons / Mars Stone / Clara Di Germanio / Michael Busch / Philip L. Felgner

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 12

Abstract: Abstract A coronavirus antigen microarray (COVAM) was constructed containing 11 SARS-CoV-2, 5 SARS-1, 5 MERS, and 12 seasonal coronavirus recombinant proteins. The array is designed to measure immunoglobulin isotype and subtype levels in serum or plasma ... ...

Abstract	Abstract A coronavirus antigen microarray (COVAM) was constructed containing 11 SARS-CoV-2, 5 SARS-1, 5 MERS, and 12 seasonal coronavirus recombinant proteins. The array is designed to measure immunoglobulin isotype and subtype levels in serum or plasma samples against each of the individual antigens printed on the array. We probed the COVAM with COVID-19 convalescent plasma (CCP) collected from 99 donors who recovered from a PCR+ confirmed SARS-CoV-2 infection. The results were analyzed using two computational approaches, a generalized linear model (glm) and random forest (RF) prediction model, to classify individual specimens as either Reactive or non-reactive against the SARS-CoV-2 antigens. A training set of 88 pre-COVID-19 specimens (PreCoV) collected in August 2019 and102 positive specimens from SARS-CoV-2 PCR+ confirmed COVID-19 cases was used for these analyses. Results compared with an FDA emergency use authorized (EUA) SARS-CoV2 S1-based total Ig chemiluminescence immunoassay (Ortho Clinical Diagnostics VITROS Anti-SARS-CoV-2 Total, CoV2T) and with a SARS-CoV-2 S1-S2 spike-based pseudovirus micro neutralization assay (SARS-CoV-2 reporter viral particle neutralization titration (RVPNT) showed high concordance between the three assays. Three CCP specimens that were negative by the VITROS CoV2T immunoassay were also negative by both COVAM and the RVPNT assay. Concordance between VITROS CoV2T and COVAM was 96%, VITROS CoV2T and RVPNT 93%, and RVPNT and COVAM 91%. The discordances were all weakly reactive samples near the cutoff threshold of the VITROS CoV2T immunoassay. The multiplex COVAM allows CCP to be grouped according to antibody reactivity patterns against 11 SARS-CoV-2 antigens. Unsupervised K-means analysis, via the gap statistics, as well as hierarchical clustering analysis revealed three main clusters with distinct reactivity intensities and patterns. These patterns were not recapitulated by adjusting the VITROS CoV2T or RVPNT assay thresholds. Plasma classified by COVAM reactivity patterns ...
Keywords	Medicine ; R ; Science ; Q
Subject code	630
Language	English
Publishing date	2021-04-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Characterization of Endocannabinoid System and Interleukin Profiles in Ovine AEC

Luana Greco / Valentina Russo / Cinzia Rapino / Clara Di Germanio / Filomena Fezza / Nicola Bernabò / Paolo Berardinelli / Alessia Peserico / Domenico Fazio / Mauro Maccarrone / Mauro Mattioli / Barbara Barboni

Cells, Vol 9, Iss 1008, p

Cannabinoid Receptors Type-1 and Type-2 as Key Effectors of Pro-Inflammatory Response

2020 Volume 1008

Abstract: Amniotic epithelial cells (AEC) have been proposed as promising clinical candidates for regenerative medicine therapies due to their immunomodulatory capacity. In this context, the endocannabinoid system (ECS) has been identified as mediating the immune- ... ...

Abstract	Amniotic epithelial cells (AEC) have been proposed as promising clinical candidates for regenerative medicine therapies due to their immunomodulatory capacity. In this context, the endocannabinoid system (ECS) has been identified as mediating the immune-stem cell dialogue, even if no information on AEC is available to date. Therefore, this study was designed to assess whether ECS is involved in tuning the constitutive and lipopolysaccharide (LPS)-induced ovine AEC anti-inflammatory and pro-inflammatory interleukin (IL-10, IL-4, and IL-12) profiles. Firstly, interleukins and ECS expressions were studied at different stages of gestation. Then, the role of cannabinoid receptors 1 and 2 (CB1 and CB2) on interleukin expression and release was investigated in middle stage AEC using selective agonists and antagonists. AEC displayed a degradative more than a synthetic endocannabinoid metabolism during the early and middle stages of gestation. At the middle stage, cannabinoid receptors mediated the balance between pro-inflammatory (IL-12) and anti-inflammatory (IL-4 and IL-10) interleukins. The activation of both receptors mediated an overall pro-inflammatory shift—CB1 reduced the anti-inflammatory and CB2 increased the pro-inflammatory interleukin release, particularly after LPS stimulation. Altogether, these data pave the way for the comprehension of AEC mechanisms tuning immune-modulation, crucial for the development of new AEC-based therapy protocols.
Keywords	amniotic epithelial cells ; cannabinoid receptors ; immunomodulation ; Biology (General) ; QH301-705.5
Subject code	616
Language	English
Publishing date	2020-04-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Compartmentalization of cerebrospinal fluid inflammation across the spectrum of untreated HIV-1 infection, central nervous system injury and viral suppression.

Magnus Gisslen / Sheila M Keating / Serena Spudich / Victor Arechiga / Sophie Stephenson / Henrik Zetterberg / Clara Di Germanio / Kaj Blennow / Dietmar Fuchs / Lars Hagberg / Philip J Norris / Julia Peterson / Barbara L Shacklett / Constantin T Yiannoutsos / Richard W Price

PLoS ONE, Vol 16, Iss 5, p e

2021 Volume 0250987

Abstract: Objective To characterize the evolution of central nervous system (CNS) inflammation in HIV-1 infection applying a panel of cerebrospinal fluid (CSF) inflammatory biomarkers to grouped subjects representing a broad spectrum of systemic HIV-1 immune ... ...

Abstract	Objective To characterize the evolution of central nervous system (CNS) inflammation in HIV-1 infection applying a panel of cerebrospinal fluid (CSF) inflammatory biomarkers to grouped subjects representing a broad spectrum of systemic HIV-1 immune suppression, CNS injury and viral control. Methods This is a cross-sectional analysis of archived CSF and blood samples, assessing concentrations of 10 functionally diverse soluble inflammatory biomarkers by immunoassays in 143 HIV-1-infected subjects divided into 8 groups: untreated primary HIV-1 infection (PHI); four untreated groups defined by their blood CD4+ T lymphocyte counts; untreated patients presenting with subacute HIV-associated dementia (HAD); antiretroviral-treated subjects with ≥1 years of plasma viral suppression; and untreated elite controllers. Twenty HIV-1-uninfected controls were included for comparison. Background biomarkers included blood CD4+ and CD8+ T lymphocytes, CSF and blood HIV-1 RNA, CSF white blood cell (WBC) count, CSF/blood albumin ratio, CSF neurofilament light chain (NfL), and CSF t-tau. Findings HIV-1 infection was associated with a broad compartmentalized CSF inflammatory response that developed early in its course and changed with systemic disease progression, development of neurological injury, and viral suppression. CSF inflammation in untreated individuals without overt HAD exhibited at least two overall patterns of inflammation as blood CD4+ T lymphocytes decreased: one that peaked at 200-350 blood CD4+ T cells/μL and associated with lymphocytic CSF inflammation and HIV-1 RNA concentrations; and a second that steadily increased through the full range of CD4+ T cell decline and associated with macrophage responses and increasing CNS injury. Subacute HAD was distinguished by a third inflammatory profile with increased blood-brain barrier permeability and robust combined lymphocytic and macrophage CSF inflammation. Suppression of CSF and blood HIV-1 infections by antiretroviral treatment and elite viral control were associated with reduced CSF inflammation, though not fully to levels found in HIV-1 seronegative controls.
Keywords	Medicine ; R ; Science ; Q
Subject code	610
Language	English
Publishing date	2021-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: SARS-CoV-2 Antibody persistence in COVID-19 convalescent plasma donors

Clara Di Germanio, Clara Di / Simmons, Graham / Kelly, Kathleen / Martinelli, Rachel / Darst, Orsolya / Azimpouran, Mahzad / Stone, Mars / Hazegh, Kelsey / Grebe, Eduard / Zhang, Shuting / Ma, Peijun / Orzechowski, Marek / Livny, Jonathan / Hung, Deborah T / Vassallo, Ralph R / Busch, Michael P / Dumont, Larry J

medRxiv

Abstract: Background Antibody response duration following SARS-CoV-2 infection tends to be variable and depends on severity of disease and method of detection. Study design and methods COVID-19 convalescent plasma (CCP) from 18 donors was collected longitudinally ... ...

Abstract	Background Antibody response duration following SARS-CoV-2 infection tends to be variable and depends on severity of disease and method of detection. Study design and methods COVID-19 convalescent plasma (CCP) from 18 donors was collected longitudinally for a maximum of 63 - 129 days following resolution of symptoms. All the samples were initially screened by the Ortho Total Ig test to confirm positivity and subsequently tested with 7 additional direct sandwich or indirect binding assays (Ortho, Roche, Abbott, Broad Institute) directed against a variety of antigen targets (S1, RBD, and NC), along with 2 neutralization assays (Broad Institute live virus PRNT and Vitalant Research Institute Pseudovirus RVPN). Results The direct detection assays (Ortho Total Ig total and Roche Total Ig) showed increasing levels of antibodies over the time period, in contrast to the indirect IgG assays that showed a decline. Neutralization assays also demonstrated declining responses; the VRI RVPN pseudovirus had a greater rate of decline than the Broad PRNT live virus assay. Discussion These data show that in addition to variable individual responses and associations with disease severity, the detection assay chosen contributes to the heterogeneous results in antibody stability over time. Depending on the scope of the research, one assay may be preferable over another. For serosurveillance studies, direct, double Ag-sandwich assays appear to be the best choice due to their stability; in particular, algorithms that include both S1 and NC based assays can help reduce the rate of false-positivity and discriminate between natural infection and vaccine-derived seroreactivity.
Keywords	covid19
Language	English
Publishing date	2021-03-26
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2021.03.24.21254260
Database	COVID19

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Article ; Online: Evaluation of Commercially Available High-Throughput SARS-CoV-2 Serologic Assays for Serosurveillance and Related Applications

Emerging Infectious Diseases, Vol 28, Iss 3, Pp 672-

2022 Volume 683

Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serosurveys can estimate cumulative incidence for monitoring epidemics, requiring assessment of serologic assays to inform testing algorithm development and interpretation of results. We ... ...

Abstract	Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serosurveys can estimate cumulative incidence for monitoring epidemics, requiring assessment of serologic assays to inform testing algorithm development and interpretation of results. We conducted a multilaboratory evaluation of 21 commercial high-throughput SARS-CoV-2 serologic assays using blinded panels of 1,000 highly characterized specimens. Assays demonstrated a range of sensitivities (96%–63%), specificities (99%–96%), and precision (intraclass correlation coefficient 0.55–0.99). Durability of antibody detection was dependent on antigen and immunoglobulin targets; antispike and total Ig assays demonstrated more stable longitudinal reactivity than antinucleocapsid and IgG assays. Assays with high sensitivity, specificity, and durable antibody detection are ideal for serosurveillance, but assays demonstrating waning reactivity are appropriate for other applications, including correlation with neutralizing activity and detection of anamnestic boosting by reinfections. Assay performance must be evaluated in context of intended use, particularly in the context of widespread vaccination and circulation of SARS-CoV-2 variants.
Keywords	COVID-19 ; coronavirus disease ; SARS-CoV-2 ; severe acute respiratory syndrome coronavirus 2 ; viruses ; respiratory infections ; Medicine ; R ; Infectious and parasitic diseases ; RC109-216
Language	English
Publishing date	2022-03-01T00:00:00Z
Publisher	Centers for Disease Control and Prevention
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Cerebrospinal fluid soluble CD30 elevation despite suppressive antiretroviral therapy in individuals living with HIV-1

Michael J. Peluso / Cassandra Thanh / Cecilia A. Prator / Louise E. Hogan / Victor M. Arechiga / Sophie Stephenson / Philip J. Norris / Clara Di Germanio / Dietmar Fuchs / Henrik Zetterberg / Steven G. Deeks / Magnus Gisslén / Richard W. Price / Timothy J. Henrich

Journal of Virus Eradication, Vol 6, Iss 1, Pp 19-

2020 Volume 26

Abstract: Objectives: The aim of this study was to assess soluble CD30 (sCD30), a protein that colocalises with HIV-1 RNA and DNA in lymphoid cells and tissues, in cerebrospinal fluid (CSF) as a marker of HIV-1 infection in the central nervous system (CNS). ... ...

Abstract	Objectives: The aim of this study was to assess soluble CD30 (sCD30), a protein that colocalises with HIV-1 RNA and DNA in lymphoid cells and tissues, in cerebrospinal fluid (CSF) as a marker of HIV-1 infection in the central nervous system (CNS). Methods: This was a cross-sectional study using archived samples from two clinical cohorts. Soluble CD30 concentrations were measured in paired CSF and plasma from untreated viraemic individuals (n=52), individuals on suppressive antiretroviral therapy (ART) (n=33), HIV-1 controllers (n=10), participants with CSF HIV-1 ‘escape’ (n=11) and controls without HIV-1 infection (n=16). Nonparametric tests were used to compare levels across groups and evaluate correlations with HIV-1 RNA, CSF neurofilament light chain protein (NFL) and neopterin. Results: Compared with controls (median 30 ng/mL, interquartile range [IRQ] 23–50), plasma sCD30 levels were elevated in viraemic participants (75 ng/mL, 52–116; P<0.001), but not in those on suppressive ART (38 ng/mL, 32–62). In contrast, CSF sCD30 levels were elevated in ART-suppressed individuals (34 ng/mL, 19–46; P=0.001) and in those with CSF ‘escape’ (33 ng/mL, 27–40; P=0.004) compared with controls (18 ng/mL, 11–23), but not in untreated viraemic individuals. No association was observed between CSF sCD30 and plasma HIV-1 RNA, concurrent or nadir CD4+ T cell count, duration of infection or plasma sCD30. CSF sCD30 correlated with CSF NFL (r=0.34, P=0.001). Conclusions: In contrast to plasma, sCD30 levels are elevated in the CSF of individuals with HIV-1 infection who are on suppressive ART. Elevated levels of sCD30 in the CSF may be an indicator of persistent CNS HIV-1 infection, although the mechanism underlying this elevation warrants further investigation.
Keywords	HIV-1 ; reservoir ; central nervous system (CNS) ; cerebrospinal fluid (CSF) ; CD30 ; Microbiology ; QR1-502 ; Public aspects of medicine ; RA1-1270
Subject code	610
Language	English
Publishing date	2020-01-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Frequent detection but lack of infectivity of SARS-CoV-2 RNA in presymptomatic, infected blood donor plasma

Paula Saá / Rebecca V. Fink / Sonia Bakkour / Jing Jin / Graham Simmons / Marcus O. Muench / Hina Dawar / Clara Di Germanio / Alvin J. Hui / David J. Wright / David E. Krysztof / Steven H. Kleinman / Angela Cheung / Theresa Nester / Debra A. Kessler / Rebecca L. Townsend / Bryan R. Spencer / Hany Kamel / Jacquelyn M. Vannoy /

Honey Dave / Michael P. Busch / Susan L. Stramer / Mars Stone / Rachael P. Jackman / Philip J. Norris

The Journal of Clinical Investigation, Vol 132, Iss

2022 Volume 17

Abstract: Respiratory viruses such as influenza do not typically cause viremia; however, SARS-CoV-2 has been detected in the blood of COVID-19 patients with mild and severe symptoms. Detection of SARS-CoV-2 in blood raises questions about its role in pathogenesis ... ...

Abstract	Respiratory viruses such as influenza do not typically cause viremia; however, SARS-CoV-2 has been detected in the blood of COVID-19 patients with mild and severe symptoms. Detection of SARS-CoV-2 in blood raises questions about its role in pathogenesis as well as transfusion safety concerns. Blood donor reports of symptoms or a diagnosis of COVID-19 after donation (post-donation information, PDI) preceded or coincided with increased general population COVID-19 mortality. Plasma samples from 2,250 blood donors who reported possible COVID-19–related PDI were tested for the presence of SARS-CoV-2 RNA. Detection of RNAemia peaked at 9%–15% of PDI donors in late 2020 to early 2021 and fell to approximately 4% after implementation of widespread vaccination in the population. RNAemic donors were 1.2- to 1.4-fold more likely to report cough or shortness of breath and 1.8-fold more likely to report change in taste or smell compared with infected donors without detectable RNAemia. No infectious virus was detected in plasma from RNAemic donors; inoculation of permissive cell lines produced less than 0.7–7 plaque-forming units (PFU)/mL and in susceptible mice less than 100 PFU/mL in RNA-positive plasma based on limits of detection in these models. These findings suggest that blood transfusions are highly unlikely to transmit SARS-CoV-2 infection.
Keywords	COVID-19 ; Medicine ; R
Language	English
Publishing date	2022-09-01T00:00:00Z
Publisher	American Society for Clinical Investigation
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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