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Article ; Online: Inflammatory and Prothrombotic Biomarkers Contribute to the Persistence of Sequelae in Recovered COVID-19 Patients.

Garcia-Larragoiti, Nallely / Cano-Mendez, Alan / Jimenez-Vega, Yeny / Trujillo, Mercedes / Guzman-Cancino, Patricia / Ambriz-Murillo, Yesenia / Viveros-Sandoval, Martha Eva

International journal of molecular sciences

2023 Volume 24, Issue 24

Abstract: The presence of long COVID (LC) following SARS-CoV-2 infection is a common condition that affects the quality of life of patients and represents a diagnostic challenge due to the diversity of symptoms that may coexist. We still do not have accurate ... ...

Abstract	The presence of long COVID (LC) following SARS-CoV-2 infection is a common condition that affects the quality of life of patients and represents a diagnostic challenge due to the diversity of symptoms that may coexist. We still do not have accurate information regarding the pathophysiological pathways that generate the presence of LC, and so it is important to know the inflammatory and immunothrombotic biomarker profiles and their implications in order to characterize risk subgroups and establish early therapeutic strategies. We performed the determination of inflammatory and immunothrombotic biomarkers in volunteers with previous diagnoses of SARS-CoV-2. The inflammatory biomarkers were analyzed in plasma by flow cytometry, and we analyzed the von Willebrand factor (vWF) in the plasma samples using ELISA. The clinical variables and the presence or absence of long COVID symptoms were then analyzed. IL-6, sCD40L, p-Selectin, PSGL-1, PAI-1, tPA, D-Dimer, TF, and Factor IX levels were elevated in the groups with LC, especially in the subgroup of patients with metabolic syndrome (MetS). VWF levels were found to be increased in patients with sequelae and MetS. Our results confirmed the persistence of an active immunothrombotic state, and so it is important to identify the population at risk in order to provide adequate clinical follow-up.
MeSH term(s)	Humans ; von Willebrand Factor/metabolism ; COVID-19/complications ; Post-Acute COVID-19 Syndrome ; Quality of Life ; SARS-CoV-2/metabolism ; Biomarkers ; Disease Progression ; Metabolic Syndrome
Chemical Substances	von Willebrand Factor ; Biomarkers
Language	English
Publishing date	2023-12-14
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms242417468
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Article ; Online: Platelet Reactivity and Inflammatory Phenotype Induced by Full-Length Spike SARS-CoV-2 Protein and Its RBD Domain.

Cano-Mendez, Alan / García-Larragoiti, Nallely / Damian-Vazquez, Maria / Guzman-Cancino, Patricia / Lopez-Castaneda, Sandra / Ochoa-Zarzosa, Alejandra / Viveros-Sandoval, Martha Eva

International journal of molecular sciences

2022 Volume 23, Issue 23

Abstract: A state of immunothrombosis has been reported in COVID-19. Platelets actively participate in this process. However, little is known about the ability of SARS-CoV-2 virus proteins to induce platelet activity. Platelet-rich plasma (PRP) was incubated with ... ...

Abstract	A state of immunothrombosis has been reported in COVID-19. Platelets actively participate in this process. However, little is known about the ability of SARS-CoV-2 virus proteins to induce platelet activity. Platelet-rich plasma (PRP) was incubated with spike full-length protein and the RBD domain in independent assays. We evaluated platelet activation through the expression of P-selectin and activation of glicoprotein IIbIIIa (GP IIbIIIa), determined by flow cytometry and the ability of the proteins to induce platelet aggregation. We determined concentrations of immunothrombotic biomarkers in PRP supernatant treated with the proteins. We determined that the spike full-length proteins and the RBD domain induced an increase in P-selectin expression and GP IIbIIIa activation (p < 0.0001). We observed that the proteins did not induce platelet aggregation, but favored a pro-aggregating state that, in response to minimal doses of collagen, could re-establish the process (p < 0.0001). On the other hand, the viral proteins stimulated the release of interleukin 6, interleukin 8, P-selectin and the soluble fraction of CD40 ligand (sCD40L), molecules that favor an inflammatory state p < 0.05. These results indicate that the spike full-length protein and its RBD domain can induce platelet activation favoring an inflammatory phenotype that might contribute to the development of an immunothrombotic state.
MeSH term(s)	Humans ; Blood Platelets/metabolism ; COVID-19/metabolism ; Platelet Activation ; SARS-CoV-2/metabolism ; Spike Glycoprotein, Coronavirus/metabolism ; Protein Domains
Chemical Substances	Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2
Language	English
Publishing date	2022-12-02
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms232315191
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Article ; Online: Production and Purification of Zika Virus NS1 Glycoprotein in HEK293 Cells.

Kim, Young Chan / Garcia-Larragoiti, Nallely / Lopez-Camacho, Cesar / Viveros-Sandoval, Martha Eva / Reyes-Sandoval, Arturo

Methods in molecular biology (Clifton, N.J.)

2020 Volume 2142, Page(s) 93–102

Abstract: In this chapter, we describe production and purification of the Zika virus NS1 glycoprotein in human embryonic kidney (HEK293T) cells at small, research laboratory scale. The expression of secreted NS1 (sNS1) and the C-terminal β-ladder domain in HEK293T ...

Abstract	In this chapter, we describe production and purification of the Zika virus NS1 glycoprotein in human embryonic kidney (HEK293T) cells at small, research laboratory scale. The expression of secreted NS1 (sNS1) and the C-terminal β-ladder domain in HEK293T cells were tested in a small-scale transfection before scaling up to a larger-scale transfection using roller bottles. Two different purification approaches have been applied to obtain purified NS1 (sNS1) and the C-terminal β-ladder domain ready for clinical applications.
MeSH term(s)	Cell Culture Techniques/methods ; Cloning, Molecular/methods ; HEK293 Cells ; Histidine/chemistry ; Humans ; Oligopeptides/chemistry ; Proteomics/methods ; Recombinant Proteins/genetics ; Recombinant Proteins/isolation & purification ; Recombinant Proteins/metabolism ; Secretory Pathway ; Serologic Tests/methods ; Transfection/methods ; Viral Nonstructural Proteins/genetics ; Viral Nonstructural Proteins/isolation & purification ; Viral Nonstructural Proteins/metabolism ; Zika Virus/genetics
Chemical Substances	His-His-His-His-His-His ; NS1 protein, zika virus ; Oligopeptides ; Recombinant Proteins ; Viral Nonstructural Proteins ; Histidine (4QD397987E)
Language	English
Publishing date	2020-05-04
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-0581-3_8
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Article: Temperature-dependent secretion of Zika virus envelope and non-structural protein 1 in mammalian cells for clinical applications

Kim, Young Chan / Nettleship, Joanne E / García-Larragoiti, Nallely / Mar, Maria Antonieta / Mondragón-García, Ariadna Lorena / Ríos-Valencia, Javier / Figueroa-Aguilar, Gloria / Viveros-Sandoval, Martha Eva / Owens, Raymond J / Reyes-Sandoval, Arturo

Journal of virological methods. 2021 Aug., v. 294

2021

Abstract: Zika virus (ZIKV) is an emerging mosquito-borne flavivirus associated with congenital Zika syndrome and Guillain-Barré syndrome in adults. The recombinant ZIKV envelope (E) antigen can be useful for serodiagnosis of ZIKV infection and for monitoring ... ...

Abstract	Zika virus (ZIKV) is an emerging mosquito-borne flavivirus associated with congenital Zika syndrome and Guillain-Barré syndrome in adults. The recombinant ZIKV envelope (E) antigen can be useful for serodiagnosis of ZIKV infection and for monitoring immune responses during preclinical and clinical ZIKV vaccine development. In this study, we describe production of ZIKV E using the modified polyethyleneimine (PEI) transfection in HEK293 cells to improve cost-effective large-scale production. We show that the secretion of ZIKV E in HEK293 cells is dependent on cell culture incubation temperatures where incubation at a low temperature of 28 °C improved protein secretion of both, E-CD4 and E, whereas a substantial decrease in secretion was observed at 37 °C. The resulting E-CD4 produced at low temperature yielded similar binding profiles in ELISAs in comparison with a commercially available E protein using human seropositive sera to ZIKV. We also show that ZIKV NS1 and NS1 β-ladder antigens produced in HEK293 cells, have similar binding profiles in ELISA which suggests that both NS1 or NS1 β-ladder can be used for serodiagnosis of ZIKV. In conclusion, we propose a cost-effective production of the ZIKV E and NS1, suitable for both, clinical and research applications in endemic countries.
Keywords	Zika virus ; antigens ; cell culture ; cost effectiveness ; humans ; polyethyleneimine ; protein secretion ; serodiagnosis ; seroprevalence ; temperature ; transfection ; vaccine development ; viral nonstructural proteins
Language	English
Dates of publication	2021-08
Publishing place	Elsevier B.V.
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	8013-5
ISSN	1879-0984 ; 0166-0934
ISSN (online)	1879-0984
ISSN	0166-0934
DOI	10.1016/j.jviromet.2021.114175
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Article ; Online: Platelet activation and aggregation response to dengue virus nonstructural protein 1 and domains.

García-Larragoiti, Nallely / Kim, Young Chan / López-Camacho, César / Cano-Méndez, Alan / López-Castaneda, Sandra / Hernández-Hernández, Darinel / Vargas-Ruiz, Ángel G / Vázquez-Garcidueñas, Ma Soledad / Reyes-Sandoval, Arturo / Viveros-Sandoval, Martha E

Journal of thrombosis and haemostasis : JTH

2021 Volume 19, Issue 10, Page(s) 2572–2582

Abstract: Background: Platelets are now recognized as immunological sentries in the first line of defense that participate in the detection and response to pathogens. This frequently results in a decrease in the number of circulating platelets. Different ... ...

Abstract	Background: Platelets are now recognized as immunological sentries in the first line of defense that participate in the detection and response to pathogens. This frequently results in a decrease in the number of circulating platelets. Different mechanisms have been hypothesized to explain the thrombocytopenia in patients with severe dengue, one of them is the participation of the non-structural protein 1 (NS1) of dengue virus (DENV), which can be secreted into circulation during DENV infection and promotes a more efficient infection. Objective: The present study aimed to investigate the ability of platelet response to stimulation with full-length DENV NS1 protein and its domains. Methods: DENV NS1 plasmid was transfected into HEK-293T. Proteins were purified by Niquel Sepharose affinity chromatography. Secreted proteins were assessed by sodium dodecylsulfate polyacrylamide gel electrophoresis, Coomassie staining and western blot. Platelet-rich plasma was directly incubated with DENV NS1 proteins. Platelet activation was confirmed by expression of αIIbβIII and P-selectin by flow cytometry. Platelet aggregation was also assessed using DENV NS1 protein and its individual domains as agonists. Results: DENV NS1 protein and its domains induce P-selectin and αIIbβ3 complex expression on platelet surfaces. DENV NS1 induce a stable platelet aggregation after the addition of a minimal dose of adenosine diphosphate (ADP), epinephrine (EPI), or collagen. Interestingly, only EPI could induce the formation of platelet aggregates after incubation with the protein domains of NS1. Conclusion: Our results suggest that the full DENV NS1 protein and also its domains promote platelet recognition, activation, and aggregation.
MeSH term(s)	Blood Platelets ; Dengue ; Dengue Virus ; Humans ; Platelet Aggregation ; Viral Nonstructural Proteins
Chemical Substances	Viral Nonstructural Proteins
Language	English
Publishing date	2021-07-20
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2112661-6
ISSN	1538-7836 ; 1538-7933
ISSN (online)	1538-7836
ISSN	1538-7933
DOI	10.1111/jth.15431
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Zs.A 5805: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Inflammatory and Prothrombotic Biomarkers Associated With the Severity of COVID-19 Infection.

Lopez-Castaneda, Sandra / García-Larragoiti, Nallely / Cano-Mendez, Alan / Blancas-Ayala, Kenia / Damian-Vázquez, Guadalupe / Perez-Medina, Ana Itzel / Chora-Hernández, Luis David / Arean-Martínez, Carlos / Viveros-Sandoval, Martha Eva

Clinical and applied thrombosis/hemostasis : official journal of the International Academy of Clinical and Applied Thrombosis/Hemostasis

2021 Volume 27, Page(s) 1076029621999099

Abstract: Among COVID-19 hospitalized patients, high incidence of alterations in inflammatory and coagulation biomarkers correlates with a poor prognosis. Comorbidities such as chronic degenerative diseases are frequently associated with complications in COVID-19 ... ...

Abstract	Among COVID-19 hospitalized patients, high incidence of alterations in inflammatory and coagulation biomarkers correlates with a poor prognosis. Comorbidities such as chronic degenerative diseases are frequently associated with complications in COVID-19 patients. The aim of this study was to evaluate inflammatory and procoagulant biomarkers in COVID-19 patients from a public hospital in Mexico. Blood was sampled within the first 48 h after admission in 119 confirmed COVID-19 patients that were classified in 3 groups according to oxygen demand, evolution and the severity of the disease as follows: 1) Non severe: nasal cannula or oxygen mask; 2) Severe: high flow nasal cannula and 3) Death: mechanical ventilation eventually leading to fatal outcome. Blood samples from 20 healthy donors were included as a Control Group. Analysis of inflammatory and coagulation biomarkers including D-dimer, interleukin 6, interleukin 8, PAI-1, P-selectin and VWF was performed in plasma. Routine laboratory and clinical biomarkers were also included and compared among groups. Concentrations of D-dimer (14.5 ± 13.8 µg/ml) and PAI-1 (1223 ± 889.6 ng/ml) were significantly elevated in severe COVID-19 patients (P < 0.0001). A significant difference was found in interleukin-6, PAI-1 and P-selectin in non-severe and healthy donors when compared to Severe COVID-19 and deceased patients (P < 0.001). VWF levels were also significantly different between severe patients (153.5 ± 24.3 UI/dl) and non-severe ones (133.9 ± 20.2 UI/dl) (P < 0.0001). WBC and glucose levels were also significantly elevated in patients with Severe COVID-19. Plasma concentrations of all prothrombotic biomarkers were significantly higher in patients with a fatal outcome.
MeSH term(s)	Adult ; Aged ; Biomarkers/blood ; COVID-19/blood ; COVID-19/complications ; COVID-19/epidemiology ; Case-Control Studies ; Female ; Fibrin Fibrinogen Degradation Products/metabolism ; Hospitalization ; Humans ; Inflammation Mediators/blood ; Interleukin-6/blood ; Male ; Mexico/epidemiology ; Middle Aged ; P-Selectin/blood ; Pandemics ; Plasminogen Activator Inhibitor 1/blood ; Prognosis ; SARS-CoV-2 ; Severity of Illness Index ; Thrombosis/blood ; Thrombosis/etiology ; von Willebrand Factor/metabolism
Chemical Substances	Biomarkers ; Fibrin Fibrinogen Degradation Products ; IL6 protein, human ; Inflammation Mediators ; Interleukin-6 ; P-Selectin ; Plasminogen Activator Inhibitor 1 ; SELP protein, human ; SERPINE1 protein, human ; fibrin fragment D ; von Willebrand Factor
Language	English
Publishing date	2021-04-09
Publishing country	United States
Document type	Journal Article
ZDB-ID	1237357-6
ISSN	1938-2723 ; 1076-0296
ISSN (online)	1938-2723
ISSN	1076-0296
DOI	10.1177/1076029621999099
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Article ; Online: Temperature-dependent secretion of Zika virus envelope and non-structural protein 1 in mammalian cells for clinical applications.

Journal of virological methods

2021 Volume 294, Page(s) 114175

Abstract	Zika virus (ZIKV) is an emerging mosquito-borne flavivirus associated with congenital Zika syndrome and Guillain-Barré syndrome in adults. The recombinant ZIKV envelope (E) antigen can be useful for serodiagnosis of ZIKV infection and for monitoring immune responses during preclinical and clinical ZIKV vaccine development. In this study, we describe production of ZIKV E using the modified polyethyleneimine (PEI) transfection in HEK293 cells to improve cost-effective large-scale production. We show that the secretion of ZIKV E in HEK293 cells is dependent on cell culture incubation temperatures where incubation at a low temperature of 28 °C improved protein secretion of both, E-CD4 and E, whereas a substantial decrease in secretion was observed at 37 °C. The resulting E-CD4 produced at low temperature yielded similar binding profiles in ELISAs in comparison with a commercially available E protein using human seropositive sera to ZIKV. We also show that ZIKV NS1 and NS1 β-ladder antigens produced in HEK293 cells, have similar binding profiles in ELISA which suggests that both NS1 or NS1 β-ladder can be used for serodiagnosis of ZIKV. In conclusion, we propose a cost-effective production of the ZIKV E and NS1, suitable for both, clinical and research applications in endemic countries.
MeSH term(s)	Animals ; Antibodies, Viral ; HEK293 Cells ; Humans ; Temperature ; Viral Envelope ; Viral Nonstructural Proteins/genetics ; Zika Virus ; Zika Virus Infection/diagnosis
Chemical Substances	Antibodies, Viral ; Viral Nonstructural Proteins
Language	English
Publishing date	2021-05-19
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	8013-5
ISSN	1879-0984 ; 0166-0934
ISSN (online)	1879-0984
ISSN	0166-0934
DOI	10.1016/j.jviromet.2021.114175
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Zs.A 1565: Show issues

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Article: Development of an E2 ELISA Methodology to Assess Chikungunya Seroprevalence in Patients from an Endemic Region of Mexico

Kim, Young Chan / López-Camacho, César / Garcia-Larragoiti, Nallely / Cano-Mendez, Alan / Hernandez-Flores, Karina Guadalupe / Domínguez-Alemán, Carlos Alonso / Antonieta Mar, Maria / Vivanco-Cid, Héctor / Viveros-Sandoval, Martha Eva / Reyes-Sandoval, Arturo

Viruses. 2019 May 01, v. 11, no. 5

2019

Abstract: Chikungunya fever is a debilitating disease caused by Chikungunya virus (CHIKV) that can result in long-lasting arthralgias. The early diagnosis of CHIKV relies on PCR during the acute infection phase to allow differential diagnosis with other co- ... ...

Abstract	Chikungunya fever is a debilitating disease caused by Chikungunya virus (CHIKV) that can result in long-lasting arthralgias. The early diagnosis of CHIKV relies on PCR during the acute infection phase to allow differential diagnosis with other co-circulating arboviruses such as dengue and Zika. Alternatively, serology can support diagnosis and provide epidemiological information on current and past outbreaks. Many commercial serological ELISA assays are based on the inactivated whole CHIKV, but their sensitivity and specificity show great variability. We produced recombinant CHIKV E2 that is suitable for ELISA assays, which was used for the serodiagnosis of CHIKV infections occurring in an arbovirus endemic Mexican region within Michoacán state. A cross-sectional study was conducted in 2016–2017; sera was obtained from 15 healthy donors and 68 patients presenting undifferentiated febrile illness. Serum samples were screened by RT-PCR and by our in-house ELISA assay. Our results indicate that IgM and IgG anti-CHIKV E2 antibodies were detected with our ELISA assay with higher sensitivity than a commercially available CHIKV ELISA kit. Our simple and sensitive ELISA assay for the serodiagnosis of CHIKV infections can be applied to population-based seroprevalence surveys and has potential for monitoring vaccine immunogenicity in CHIKV vaccine clinical trials.
Keywords	Chikungunya fever ; Chikungunya virus ; acute course ; antibodies ; arboviruses ; blood serum ; clinical trials ; cross-sectional studies ; dengue ; early diagnosis ; enzyme-linked immunosorbent assay ; immunogenicity ; immunoglobulin G ; immunoglobulin M ; monitoring ; patients ; reverse transcriptase polymerase chain reaction ; serodiagnosis ; serology ; seroprevalence ; surveys ; vaccines ; Mexico
Language	English
Dates of publication	2019-0501
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v11050407
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Article ; Online: Development of an E2 ELISA Methodology to Assess Chikungunya Seroprevalence in Patients from an Endemic Region of Mexico.

Kim, Young Chan / López-Camacho, César / Garcia-Larragoiti, Nallely / Cano-Mendez, Alan / Hernandez-Flores, Karina Guadalupe / Domínguez-Alemán, Carlos Alonso / Mar, Maria Antonieta / Vivanco-Cid, Héctor / Viveros-Sandoval, Martha Eva / Reyes-Sandoval, Arturo

Viruses

2019 Volume 11, Issue 5

Abstract	Chikungunya fever is a debilitating disease caused by Chikungunya virus (CHIKV) that can result in long-lasting arthralgias. The early diagnosis of CHIKV relies on PCR during the acute infection phase to allow differential diagnosis with other co-circulating arboviruses such as dengue and Zika. Alternatively, serology can support diagnosis and provide epidemiological information on current and past outbreaks. Many commercial serological ELISA assays are based on the inactivated whole CHIKV, but their sensitivity and specificity show great variability. We produced recombinant CHIKV E2 that is suitable for ELISA assays, which was used for the serodiagnosis of CHIKV infections occurring in an arbovirus endemic Mexican region within Michoacán state. A cross-sectional study was conducted in 2016-2017; sera was obtained from 15 healthy donors and 68 patients presenting undifferentiated febrile illness. Serum samples were screened by RT-PCR and by our in-house ELISA assay. Our results indicate that IgM and IgG anti-CHIKV E2 antibodies were detected with our ELISA assay with higher sensitivity than a commercially available CHIKV ELISA kit. Our simple and sensitive ELISA assay for the serodiagnosis of CHIKV infections can be applied to population-based seroprevalence surveys and has potential for monitoring vaccine immunogenicity in CHIKV vaccine clinical trials.
MeSH term(s)	Chikungunya Fever/epidemiology ; Chikungunya Fever/immunology ; Chikungunya Fever/virology ; Chikungunya virus/genetics ; Chikungunya virus/immunology ; Cross-Sectional Studies ; Enzyme-Linked Immunosorbent Assay/methods ; Humans ; Immunoglobulin G/blood ; Immunoglobulin G/immunology ; Immunoglobulin M/blood ; Immunoglobulin M/immunology ; Mexico/epidemiology ; Public Health Surveillance ; Real-Time Polymerase Chain Reaction ; Sensitivity and Specificity ; Seroepidemiologic Studies ; Viral Proteins/immunology
Chemical Substances	Immunoglobulin G ; Immunoglobulin M ; Viral Proteins
Language	English
Publishing date	2019-05-01
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v11050407
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