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Article ; Online: The Natural History of Severe Acute Respiratory Syndrome Coronavirus 2 Infection: A Composite But Incomplete Picture.

Sachithanandham, Jaiprasath / Thio, Chloe L / Balagopal, Ashwin

Clinical infectious diseases : an official publication of the Infectious Diseases Society of America

2021 Volume 73, Issue 9, Page(s) e2943–e2945

MeSH term(s)	Antibodies, Viral ; COVID-19 ; Humans ; SARS Virus ; SARS-CoV-2
Chemical Substances	Antibodies, Viral
Keywords	covid19
Language	English
Publishing date	2021-09-07
Publishing country	United States
Document type	Editorial ; Comment
ZDB-ID	1099781-7
ISSN	1537-6591 ; 1058-4838
ISSN (online)	1537-6591
ISSN	1058-4838
DOI	10.1093/cid/ciaa1413
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Article: Antigen-display exosomes provide adjuvant-free protection against SARS-CoV-2 disease at nanogram levels of spike protein.

Guo, Chenxu / Sachithanandham, Jaiprasath / Zhong, William / Craney, Morgan / Villano, Jason / Pekosz, Andrew / Gould, Stephen J

bioRxiv : the preprint server for biology

2024

Abstract: As the only bionormal nanovesicle, exosomes have high potential as a nanovesicle for delivering vaccines and therapeutics. We show here that the loading of type-1 membrane proteins into the exosome membrane is induced by exosome membrane anchor domains, ... ...

Abstract	As the only bionormal nanovesicle, exosomes have high potential as a nanovesicle for delivering vaccines and therapeutics. We show here that the loading of type-1 membrane proteins into the exosome membrane is induced by exosome membrane anchor domains, EMADs, that maximize protein delivery to the plasma membrane, minimize protein sorting to other compartments, and direct proteins into exosome membranes. Using SARS-CoV-2 spike as an example and EMAD13 as our most effective exosome membrane anchor, we show that cells expressing a spike-EMAD13 fusion protein produced exosomes that carry dense arrays of spike trimers on 50% of all exosomes. Moreover, we find that immunization with spike-EMAD13 exosomes induced strong neutralizing antibody responses and protected hamsters against SARS-CoV-2 disease at doses of just 0.5-5 ng of spike protein, without adjuvant, demonstrating that antigen-display exosomes are particularly immunogenic, with important implications for both structural and expression-dependent vaccines.
Language	English
Publishing date	2024-01-05
Publishing country	United States
Document type	Preprint
DOI	10.1101/2024.01.04.574272
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Article ; Online: Antigen-display exosomes provide adjuvant-free protection against SARS-CoV-2 disease at nanogram levels of spike protein

Guo, Chenxu / Sachithanandham, Jaiprasath / Zhong, William / Craney, Morgan / Villano, Jason / Pekosz, Andrew / Gould, Stephen J.

bioRxiv

Abstract	As the only bionormal nanovesicle, exosomes have high potential as a nanovesicle for delivering vaccines and therapeutics. We show here that the loading of type-1 membrane proteins into the exosome membrane is induced by exosome membrane anchor domains, EMADs, that maximize protein delivery to the plasma membrane, minimize protein sorting to other compartments, and direct proteins into exosome membranes. Using SARS-CoV-2 spike as an example and EMAD13 as our most effective exosome membrane anchor, we show that cells expressing a spike-EMAD13 fusion protein produced exosomes that carry dense arrays of spike trimers on 50% of all exosomes. Moreover, we find that immunization with spike-EMAD13 exosomes induced strong neutralizing antibody responses and protected hamsters against SARS-CoV-2 disease at doses of just 0.5-5 ng of spike protein, without adjuvant, demonstrating that antigen-display exosomes are particularly immunogenic, with important implications for both structural and expression-dependent vaccines.
Keywords	covid19
Language	English
Publishing date	2024-01-05
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2024.01.04.574272
Database	COVID19

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Article: The Natural History of Severe Acute Respiratory Syndrome Coronavirus 2 Infection: A Composite But Incomplete Picture

Sachithanandham, Jaiprasath / Thio, Chloe L / Balagopal, Ashwin

Clin. infect. dis

Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #802767
Database	COVID19

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Article ; Online: Evaluation of Four Point of Care (POC) Antigen Assays for the Detection of the SARS-CoV-2 Variant Omicron.

Hardick, Justin / Gallagher, Nicholas / Sachithanandham, Jaiprasath / Fall, Amary / Siddiqui, Zishan / Pekosz, Andrew / Manabe, Yukari C / Mostafa, Heba H

Microbiology spectrum

2022 Volume 10, Issue 3, Page(s) e0102522

Abstract: Ensuring SARS-CoV-2 diagnostics that can reliably detect emerging variants has been an ongoing challenge. Due to the rapid spread of the Omicron variant, point-of-care (POC) antigen tests have become more widely used. This study aimed at (i) comparing ... ...

Abstract	Ensuring SARS-CoV-2 diagnostics that can reliably detect emerging variants has been an ongoing challenge. Due to the rapid spread of the Omicron variant, point-of-care (POC) antigen tests have become more widely used. This study aimed at (i) comparing the analytical sensitivity (LOD) of 4 POC antigen assays, BD Veritor, Abbott BinaxNow, Orasure InteliSwab and Quidel QuickVue, for the Omicron versus the Delta variant and (ii) verifying the reproducible detection of Omicron by the 4 antigen assays. The LOD for all four assays were evaluated using Omicron and Delta virus stocks quantified for infectivity and genome copies. The four assays detected all replicates of Omicron and Delta dilutions at 10
MeSH term(s)	COVID-19/diagnosis ; Humans ; Point-of-Care Systems ; SARS-CoV-2/genetics ; Sensitivity and Specificity
Language	English
Publishing date	2022-05-26
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2807133-5
ISSN	2165-0497 ; 2165-0497
ISSN (online)	2165-0497
ISSN	2165-0497
DOI	10.1128/spectrum.01025-22
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Article ; Online: A SARS-CoV-2 RBD vaccine fused to the chemokine MIP-3α elicits sustained murine antibody responses over 12 months and enhanced lung T-cell responses.

Gordy, James Tristan / Hui, Yinan / Schill, Courtney / Wang, Tianyin / Chen, Fengyixin / Fessler, Kaitlyn / Meza, Jacob / Li, Yangchen / Taylor, Alannah D / Bates, Rowan E / Karakousis, Petros C / Pekosz, Andrew / Sachithanandham, Jaiprasath / Li, Maggie / Karanika, Styliani / Markham, Richard B

Frontiers in immunology

2024 Volume 15, Page(s) 1292059

Abstract: Background: Previous studies have demonstrated enhanced efficacy of vaccine formulations that incorporate the chemokine macrophage inflammatory protein 3α (MIP-3α) to direct vaccine antigens to immature dendritic cells. To address the reduction in ... ...

Abstract	Background: Previous studies have demonstrated enhanced efficacy of vaccine formulations that incorporate the chemokine macrophage inflammatory protein 3α (MIP-3α) to direct vaccine antigens to immature dendritic cells. To address the reduction in vaccine efficacy associated with a mutation in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mutants, we have examined the ability of receptor-binding domain vaccines incorporating MIP-3α to sustain higher concentrations of antibody when administered intramuscularly (IM) and to more effectively elicit lung T-cell responses when administered intranasally (IN). Methods: BALB/c mice aged 6-8 weeks were immunized intramuscularly or intranasally with DNA vaccine constructs consisting of the SARS-CoV-2 receptor-binding domain alone or fused to the chemokine MIP-3α. In a small-scale ( Results: At 12 months postprimary vaccination, recipients of the IM vaccine incorporating MIP-3α had significantly, approximately threefold, higher serum antibody concentrations than recipients of the vaccine not incorporating MIP-3α. The area-under-the-curve analyses of the 12-month observation interval demonstrated significantly greater antibody concentrations over time in recipients of the MIP-3α vaccine formulation. At 12 months postprimary immunization, only recipients of the fusion vaccine had concentrations of serum-neutralizing activity deemed to be effective. After intranasal immunization, only recipients of the MIP-3α vaccine formulations developed T-cell responses in the lungs significantly above those of PBS controls. Low levels of serum antibody responses were obtained following IN immunization. Conclusion: Although requiring separate IM and IN immunizations for optimal immunization, incorporating MIP-3α in a SARS-CoV-2 vaccine construct demonstrated the potential of a stable and easily produced vaccine formulation to provide the extended antibody and T-cell responses that may be required for protection in the setting of emerging SARS-CoV-2 variants. Without electroporation, simple, uncoated plasmid DNA incorporating MIP-3α administered intranasally elicited lung T-cell responses.
MeSH term(s)	Animals ; Mice ; Antibody Formation ; Chemokines ; COVID-19/prevention & control ; COVID-19 Vaccines/immunology ; DNA ; Lung ; SARS-CoV-2 ; T-Lymphocytes
Chemical Substances	Chemokines ; COVID-19 Vaccines ; DNA (9007-49-2)
Language	English
Publishing date	2024-02-02
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2024.1292059
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Article ; Online: Second-Phase Hepatitis C Plasma Viral Kinetics Directly Reflects Reduced Intrahepatic Burden of Hepatitis C Virus.

Sachithanandham, Jaiprasath / Balagopal, Ashwin / Leep-Lazar, Julia / Quinn, Jeffrey / Bowden, Kenneth / Ward, Kathleen / Ribeiro, Ruy M / Sulkowski, Mark S

The Journal of infectious diseases

2022 Volume 228, Issue 3, Page(s) 311–320

Abstract: Background: Mathematical models explain how antivirals control viral infections. Hepatitis C virus (HCV) treatment results in at least 2 phases of decline in viremia. The first phase reflects clearance of rapidly produced virions. The second phase is ... ...

Abstract	Background: Mathematical models explain how antivirals control viral infections. Hepatitis C virus (HCV) treatment results in at least 2 phases of decline in viremia. The first phase reflects clearance of rapidly produced virions. The second phase is hypothesized to derive from loss of infected cells but has been challenging to prove. Methods: Using single-cell methods, we quantified the number of hepatitis C virus (HCV)-infected hepatocytes in liver biopsies taken before and within 7 days of initiating direct-acting antivirals (DAAs) in a double-blinded randomized controlled trial testing 2 (sofosbuvir-velpatasvir) versus 3 (sofosbuvir-velpatasvir-voxilaprevir) DAAs. Results: We employed thousands of intrahepatic measurements in 10 persons with chronic genotype 1a HCV infection: median proportion of infected hepatocytes declined from 11.3% (range, 1.3%-59%) to 0.6% (range, <0.3%-5.8%), a loss of 75%-95% infected hepatocytes. Plasma viremia correlated with numbers of HCV-infected hepatocytes (r = 0.77; P < .0001). Second-phase plasma dynamics and changes in infected hepatocytes were indistinct (P = .16), demonstrating that second-phase viral dynamics derive from loss of infected cells. DAAs led to a decline in intracellular HCV RNA and interferon-stimulated gene expression (P < .05 for both). Conclusions: We proved that second-phase viral dynamics reflect decay of intrahepatic burden of HCV, partly due to clearance of HCV RNA from hepatocytes. Clinical trials registration: NCT02938013.
MeSH term(s)	Humans ; Sofosbuvir/therapeutic use ; Antiviral Agents/therapeutic use ; Hepacivirus/genetics ; Hepatitis C, Chronic/drug therapy ; Viremia/drug therapy ; Kinetics ; Lactams, Macrocyclic/therapeutic use ; Hepatitis C/drug therapy ; RNA, Viral ; Genotype
Chemical Substances	Sofosbuvir (WJ6CA3ZU8B) ; Antiviral Agents ; velpatasvir (KCU0C7RS7Z) ; Lactams, Macrocyclic ; RNA, Viral
Language	English
Publishing date	2022-04-21
Publishing country	United States
Document type	Randomized Controlled Trial ; Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	3019-3
ISSN	1537-6613 ; 0022-1899
ISSN (online)	1537-6613
ISSN	0022-1899
DOI	10.1093/infdis/jiad025
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Article ; Online: Convergent Evolution of A-Lineage (Clade 19B) SARS-CoV-2 Spike Sequences with B-Lineage Variants of Concern Affects Virus Replication in a Temperature-Dependent Manner on Human Nasal Epithelial Cell Cultures

Yoon, Steve / Anaya, Eduardo U. / Sachithanandham, Jaiprasath / Pinsky, Benjamin A. / Sullivan, David J / Mostafa, Heba H / Pekosz, Andrew

bioRxiv

Abstract: The first three months of the COVID-19 pandemic was dominated by two SARS-CoV-2 lineages: A-lineages (Clade 19B) and B-lineages (Clade 19A). However, with the emergence of the Spike D614G substitution in B.1 lineages (Clade 20A), both early lineages were ...

Abstract	The first three months of the COVID-19 pandemic was dominated by two SARS-CoV-2 lineages: A-lineages (Clade 19B) and B-lineages (Clade 19A). However, with the emergence of the Spike D614G substitution in B.1 lineages (Clade 20A), both early lineages were outcompeted and remained near-extinction from mid-2020 onwards. In early-2021, there was a re-emergence and persistence of novel A-lineage variants with substitutions in the Spike gene resembling those found in Variants of Concern (VOCs). An early A.3 variant (MD-HP00076/2020) and three A.2.5 variants (MD-HP02153/2021, MD-HP05922/2021 and CA-VRLC091/2021) were isolated and characterized for their genomic sequences, antibody neutralization, and in vitro replication. All A.2.5 isolates had five Spike mutations relative to the A.3 variant sequence: D614G, L452R, Δ141-143, D215A, and ins215AGY. Plaque reduction neutralization assays demonstrated that A.2.5 isolates had a 2.5 to 5-fold reduction in neutralization using contemporaneous COVID-19 convalescent plasma when compared to A.3. In vitro viral characterization in VeroE6 cell lines revealed that the A.3 isolate grew faster and spread more than A.2.5. On VeroE6-TMPRSS2 cells, significant syncytia formation was also observed with the A.2.5 isolates, however Spike cleavage efficiency did not explain these differences. In human nasal epithelial cell (hNEC) cultures, the A.2.5 isolates grew significantly faster and to higher total infectious virus titers than A.3. All A.2.5 lineage isolates grew significantly faster at 37°C than at 33°C irrespective of cell type, and to higher peak titers except compared to A.3. This suggests A.2.59s adapted to improve replication using similar mutations found in the B-lineage SARS-CoV-2 variants.
Keywords	covid19
Language	English
Publishing date	2023-03-06
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2023.03.03.531067
Database	COVID19

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Article ; Online: Reduced control of SARS-CoV-2 infection associates with lower mucosal antibody responses in pregnancy.

mSphere

2024 Volume 9, Issue 3, Page(s) e0081223

Abstract: Pregnant patients are at greater risk of hospitalization with severe COVID-19 than non-pregnant people. This was a retrospective observational cohort study of remnant clinical specimens from patients who visited acute care hospitals within the Johns ... ...

Abstract	Pregnant patients are at greater risk of hospitalization with severe COVID-19 than non-pregnant people. This was a retrospective observational cohort study of remnant clinical specimens from patients who visited acute care hospitals within the Johns Hopkins Health System in the Baltimore, MD-Washington DC, area between October 2020 and May 2022. Participants included confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected pregnant people and matched non-pregnant people (the matching criteria included age, race/ethnicity, area deprivation index, insurance status, and vaccination status to ensure matched demographics). The primary dependent measures were clinical COVID-19 outcomes, infectious virus recovery, viral RNA levels, and mucosal anti-spike (S) IgG titers from upper respiratory tract samples. A total of 452 individuals (117 pregnant and 335 non-pregnant) were included in the study, with both vaccinated and unvaccinated individuals represented. Pregnant patients were at increased risk of hospitalization (odds ratio [OR] = 4.2; confidence interval [CI] = 2.0-8.6), intensive care unit admittance (OR = 4.5; CI = 1.2-14.2), and being placed on supplemental oxygen therapy (OR = 3.1; CI = 1.3-6.9). Individuals infected during their third trimester had higher mucosal anti-S IgG titers and lower viral RNA levels (
MeSH term(s)	Pregnancy ; Humans ; Female ; COVID-19 ; SARS-CoV-2 ; Antibody Formation ; Breakthrough Infections ; Cohort Studies ; Retrospective Studies ; RNA, Viral ; Immunoglobulin G ; Pregnancy Complications, Infectious
Chemical Substances	RNA, Viral ; Immunoglobulin G
Language	English
Publishing date	2024-03-01
Publishing country	United States
Document type	Observational Study ; Journal Article
ISSN	2379-5042
ISSN (online)	2379-5042
DOI	10.1128/msphere.00812-23
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Article ; Online: Spike-protein proteolytic antibodies in COVID-19 convalescent plasma contribute to SARS-CoV-2 neutralization.

McConnell, Scott A / Sachithanandham, Jaiprasath / Mudrak, Nathan J / Zhu, Xianming / Farhang, Parsa Alba / Cordero, Radames J B / Wear, Maggie P / Shapiro, Janna R / Park, Han-Sol / Klein, Sabra L / Tobian, Aaron A R / Bloch, Evan M / Sullivan, David J / Pekosz, Andrew / Casadevall, Arturo

Cell chemical biology

2023 Volume 30, Issue 7, Page(s) 726–738.e4

Abstract: Understanding the mechanisms of antibody-mediated neutralization of SARS-CoV-2 is critical in combating the COVID-19 pandemic. Based on previous reports of antibody catalysis, we investigated the proteolysis of spike (S) by antibodies in COVID-19 ... ...

Abstract	Understanding the mechanisms of antibody-mediated neutralization of SARS-CoV-2 is critical in combating the COVID-19 pandemic. Based on previous reports of antibody catalysis, we investigated the proteolysis of spike (S) by antibodies in COVID-19 convalescent plasma (CCP) and its contribution to viral neutralization. Quenched fluorescent peptides were designed based on S epitopes to sensitively detect antibody-mediated proteolysis. We observed epitope cleavage by CCP from different donors which persisted when plasma was heat-treated or when IgG was isolated from plasma. Further, purified CCP antibodies proteolyzed recombinant S domains, as well as authentic viral S. Cleavage of S variants suggests CCP antibody-mediated proteolysis is a durable phenomenon despite antigenic drift. We differentiated viral neutralization occurring via direct interference with receptor binding from that occurring by antibody-mediated proteolysis, demonstrating that antibody catalysis enhanced neutralization. These results suggest that antibody-catalyzed damage of S is an immunologically relevant function of neutralizing antibodies against SARS-CoV-2.
MeSH term(s)	Humans ; Proteolysis ; SARS-CoV-2 ; Pandemics ; COVID-19/therapy ; COVID-19 Serotherapy ; Spike Glycoprotein, Coronavirus ; Peptide Hydrolases ; Antibodies, Neutralizing ; Epitopes ; Antibodies, Viral
Chemical Substances	spike protein, SARS-CoV-2 ; Spike Glycoprotein, Coronavirus ; Peptide Hydrolases (EC 3.4.-) ; Antibodies, Neutralizing ; Epitopes ; Antibodies, Viral
Language	English
Publishing date	2023-06-23
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	2451-9448
ISSN (online)	2451-9448
DOI	10.1016/j.chembiol.2023.05.011
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