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Article ; Online: Development and comparison of immunologic assays to detect primary RSV infections in infants.

Anderson, Larry J / Jadhao, Samadhan J / Hussaini, Laila / Ha, Binh / McCracken, Courtney E / Gibson, Theda / Yildirim, Inci / Yi, Jumi / Stephens, Kathy / Korski, Chelsea / Kao, Carol / Sun, Heying / Lee, Chun Yi / Jaunarajs, Anna / Rostad, Christina A / Anderson, Evan J

Frontiers in immunology

2024 Volume 14, Page(s) 1332772

Abstract: Effective respiratory syncytial virus (RSV) vaccines have been developed and licensed for elderly adults and pregnant women but not yet for infants and young children. The RSV immune state of the young child, i.e., previously RSV infected or not, is ... ...

Abstract	Effective respiratory syncytial virus (RSV) vaccines have been developed and licensed for elderly adults and pregnant women but not yet for infants and young children. The RSV immune state of the young child, i.e., previously RSV infected or not, is important to the conduct and interpretation of epidemiology studies and vaccine clinical trials. To address the need for sensitive assays to detect immunologic evidence of past infection, we developed, characterized, and evaluated 7 assays including 4 IgG antibody enzyme immunoassays (EIAs), two neutralizing antibody assays, and an IFN-γ EliSpot (EliSpot) assay. The four IgG EIAs used a subgroup A plus subgroup B RSV-infected Hep-2 cell lysate antigen (Lysate), an expressed RSV F protein antigen (F), an expressed subgroup A G protein antigen (Ga), or an expressed subgroup B G protein (Gb) antigen. The two neutralizing antibody assays used either a subgroup A or a subgroup B RSV strain. The EliSpot assay used a sucrose cushion purified combination of subgroup A and subgroup B infected cell lysate. All seven assays had acceptable repeatability, signal against control antigen, lower limit of detection, and, for the antibody assays, effect of red cell lysis, lipemia and anticoagulation of sample on results. In 44 sera collected from children >6 months after an RSV positive illness, the lysate, F, Ga and Gb IgG EIAs, and the subgroup A and B neutralizing antibody assays, and the EliSpot assays were positive in 100%, 100%, 86%, 95%, 43%, and 57%, respectively. The Lysate and F EIAs were most sensitive for detecting RSV antibody in young children with a documented RSV infection. Unexpectedly, the EliSpot assay was positive in 9/15 (60%) of PBMC specimens from infants not exposed to an RSV season, possibly from maternal microchimerism. The Lysate and F EIAs provide good options to reliably detect RSV antibodies in young children for epidemiologic studies and vaccine trials.
MeSH term(s)	Adult ; Infant ; Child ; Humans ; Female ; Pregnancy ; Child, Preschool ; Aged ; Respiratory Syncytial Virus Infections ; Leukocytes, Mononuclear ; Antibodies, Neutralizing ; Antibodies, Viral ; Respiratory Syncytial Virus Vaccines ; Antigens, Viral ; Immunoglobulin G ; GTP-Binding Proteins
Chemical Substances	Antibodies, Neutralizing ; Antibodies, Viral ; Respiratory Syncytial Virus Vaccines ; Antigens, Viral ; Immunoglobulin G ; GTP-Binding Proteins (EC 3.6.1.-)
Language	English
Publishing date	2024-01-12
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.2023.1332772
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Two RSV Platforms for G, F, or G+F Proteins VLPs.

Ha, Binh / Yang, Jie E / Chen, Xuemin / Jadhao, Samadhan J / Wright, Elizabeth R / Anderson, Larry J

Viruses

2020 Volume 12, Issue 9

Abstract: Respiratory syncytial virus (RSV) causes substantial lower respiratory tract disease in children and at-risk adults. Though there are no effective anti-viral drugs for acute disease or licensed vaccines for RSV, palivizumab prophylaxis is available for ... ...

Abstract	Respiratory syncytial virus (RSV) causes substantial lower respiratory tract disease in children and at-risk adults. Though there are no effective anti-viral drugs for acute disease or licensed vaccines for RSV, palivizumab prophylaxis is available for some high risk infants. To support anti-viral and vaccine development efforts, we developed an RSV virus-like particle (VLP) platform to explore the role RSV F and G protein interactions in disease pathogenesis. Since VLPs are immunogenic and a proven platform for licensed human vaccines, we also considered these VLPs as potential vaccine candidates. We developed two RSV VLP platforms, M+P and M+M2-1 that had F and G, F and a G peptide, or a truncated F and G on their surface. Immunoblots of sucrose gradient purified particles showed co-expression of M, G, and F with both VLP platforms. Electron microscopy imaging and immunogold labeling confirmed VLP-like structures with surface exposed projections consistent with F and G proteins. In mice, the VLPs induced both anti-F and -G protein antibodies and, on challenge, reduced lung viral titer and inflammation. These data show that these RSV VLP platforms provide a tool to study the structure of F and G and their interactions and flexible platforms to develop VLP vaccines in which all components contribute to RSV-specific immune responses.
MeSH term(s)	Animals ; Antibodies, Neutralizing/immunology ; Antibodies, Viral/immunology ; Female ; Humans ; Immunization ; Lung/immunology ; Lung/virology ; Mice ; Mice, Inbred BALB C ; Respiratory Syncytial Virus Infections/immunology ; Respiratory Syncytial Virus Infections/prevention & control ; Respiratory Syncytial Virus Infections/virology ; Respiratory Syncytial Virus, Human/genetics ; Respiratory Syncytial Virus, Human/immunology ; Vaccines, Virus-Like Particle/administration & dosage ; Vaccines, Virus-Like Particle/genetics ; Vaccines, Virus-Like Particle/immunology ; Viral Proteins/administration & dosage ; Viral Proteins/genetics ; Viral Proteins/immunology
Chemical Substances	Antibodies, Neutralizing ; Antibodies, Viral ; Vaccines, Virus-Like Particle ; Viral Proteins
Language	English
Publishing date	2020-08-19
Publishing country	Switzerland
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v12090906
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Article ; Online: Characterization of Elements Regulating the Nuclear-to-Cytoplasmic Translocation of ICP0 in Late Herpes Simplex Virus 1 Infection.

Samrat, Subodh Kumar / Ha, Binh L / Zheng, Yi / Gu, Haidong

Journal of virology

2018 Volume 92, Issue 2

Abstract: Infected cell protein 0 (ICP0) of herpes simplex virus 1 (HSV-1) is an immediate early protein containing a RING-type E3 ubiquitin ligase. It targets several host factors for proteasomal degradation and subsequently activates viral expression. ICP0 has a ...

Abstract	Infected cell protein 0 (ICP0) of herpes simplex virus 1 (HSV-1) is an immediate early protein containing a RING-type E3 ubiquitin ligase. It targets several host factors for proteasomal degradation and subsequently activates viral expression. ICP0 has a nuclear localization sequence and functions in the nucleus early during infection. However, later in infection, ICP0 is found solely in the cytoplasm. The molecular mechanism and biological function of the ICP0 nuclear-to-cytoplasmic translocation are not well understood. In this study, we sought to characterize elements important for this translocation. We found that (i) in human embryonic lung fibroblast (HEL) cells, ICP0 C-terminal residues 741 to 775 were necessary but not sufficient for the nuclear-to-cytoplasmic translocation; (ii) the loss of ICP0 E3 ubiquitin ligase activity, which led to defective viral replication in nonpermissive cells, also caused mutant ICP0 to be retained in the nucleus of HEL cells; (iii) in permissive U2OS cells, however, ICP0 lacking E3 ligase activity was translocated to the cytoplasm at a pace faster than that of wild-type ICP0, suggesting that nuclear retention of ICP0 occurs in an ICP0 E3 ligase-dependent manner; and (iv) the ICP0 C terminus and late viral proteins cooperate in order to overcome nuclear retention and stimulate ICP0 cytoplasmic translocation. Taken together, less ICP0 nuclear retention may contribute to the permissiveness of U2OS cells to HSV-1 in the absence of functional ICP0.
MeSH term(s)	Amino Acid Sequence ; Cell Line ; Cell Nucleus/metabolism ; Cytoplasm/metabolism ; Gene Expression Regulation, Viral ; Herpes Simplex/virology ; Herpesvirus 1, Human/physiology ; Host-Pathogen Interactions ; Humans ; Immediate-Early Proteins/chemistry ; Immediate-Early Proteins/genetics ; Immediate-Early Proteins/metabolism ; Mutation ; Protein Binding ; Protein Interaction Domains and Motifs ; Protein Transport ; Ubiquitin-Protein Ligases/chemistry ; Ubiquitin-Protein Ligases/genetics ; Ubiquitin-Protein Ligases/metabolism
Chemical Substances	Immediate-Early Proteins ; Ubiquitin-Protein Ligases (EC 2.3.2.27) ; Vmw110 protein, Human herpesvirus 1 (EC 2.3.2.27)
Language	English
Publishing date	2018--15
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	80174-4
ISSN	1098-5514 ; 0022-538X
ISSN (online)	1098-5514
ISSN	0022-538X
DOI	10.1128/JVI.01673-17
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Article: Two RSV Platforms for G, F, or G+F Proteins VLPs

Ha, Binh / Yang, Jie E / Chen, Xuemin / Jadhao, Samadhan J / Wright, Elizabeth R / Anderson, Larry J

Viruses. 2020 Aug. 19, v. 12, no. 9

2020

Abstract	Respiratory syncytial virus (RSV) causes substantial lower respiratory tract disease in children and at-risk adults. Though there are no effective anti-viral drugs for acute disease or licensed vaccines for RSV, palivizumab prophylaxis is available for some high risk infants. To support anti-viral and vaccine development efforts, we developed an RSV virus-like particle (VLP) platform to explore the role RSV F and G protein interactions in disease pathogenesis. Since VLPs are immunogenic and a proven platform for licensed human vaccines, we also considered these VLPs as potential vaccine candidates. We developed two RSV VLP platforms, M+P and M+M2-1 that had F and G, F and a G peptide, or a truncated F and G on their surface. Immunoblots of sucrose gradient purified particles showed co-expression of M, G, and F with both VLP platforms. Electron microscopy imaging and immunogold labeling confirmed VLP-like structures with surface exposed projections consistent with F and G proteins. In mice, the VLPs induced both anti-F and -G protein antibodies and, on challenge, reduced lung viral titer and inflammation. These data show that these RSV VLP platforms provide a tool to study the structure of F and G and their interactions and flexible platforms to develop VLP vaccines in which all components contribute to RSV-specific immune responses.
Keywords	G-proteins ; Respiratory syncytial virus ; acute course ; adults at risk ; antibodies ; antiviral agents ; children ; disease prevention ; electron microscopy ; humans ; image analysis ; immune response ; infants ; inflammation ; lungs ; mice ; pathogenesis ; peptides ; respiratory tract diseases ; risk ; sucrose ; vaccine development ; viral load ; virus-like particle vaccines
Language	English
Dates of publication	2020-0819
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v12090906
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: In vitro model for the assessment of human immune responses to subunit RSV vaccines.

Chirkova, Tatiana / Ha, Binh / Rimawi, Bassam H / Oomens, Antonius G P / Hartert, Tina V / Anderson, Larry J

PloS one

2020 Volume 15, Issue 3, Page(s) e0229660

Abstract: Respiratory syncytial virus (RSV) is the single most important cause of serious lower respiratory tract disease in infants and young children worldwide and a high priority for vaccine development. Despite over 50 years of research, however, no vaccine is ...

Abstract	Respiratory syncytial virus (RSV) is the single most important cause of serious lower respiratory tract disease in infants and young children worldwide and a high priority for vaccine development. Despite over 50 years of research, however, no vaccine is yet available. One block to vaccine development is an incomplete understanding of the aberrant memory response to the formalin-inactivated RSV vaccine (FI-RSV) given to children in the 1960s. This vaccine caused enhanced respiratory disease (ERD) with later natural RSV infection. Concern that any non-live virus vaccine may also cause ERD has blocked development of subunit vaccines for young children. A number of animal FI-RSV studies suggest various immune mechanisms behind ERD. However, other than limited data from the original FI-RSV trial, there is no information on the human ERD-associated responses. An in vitro model with human blood specimens may shed light on the immune memory responses likely responsible for ERD. Memory T cell responses to an antigen are guided by the innate responses, particularly dendritic cells that present an antigen in conjunction with co-stimulatory molecules and cytokine signaling. Our in vitro model involves human monocyte derived dendritic cells (moDC) and allogenic T cell cultures to assess innate responses that direct T cell responses. Using this model, we evaluated human responses to live RSV, FI-RSV, and subunit RSV G vaccines (G-containing virus-like particles, G-VLP). Similar to findings in animal studies, FI-RSV induced prominent Th2/Th17-biased responses with deficient type-1 responses compared to live virus. Responses to G-VLPs were similar to live virus, i.e. biased towards a Th1 and not a Th2/Th17. Also mutating CX3C motif in G gave a more pronounced moDC responses associated with type-1 T cell responses. This in vitro model identifies human immune responses likely associated with ERD and provides another pre-clinical tool to assess the safety of RSV vaccines.
MeSH term(s)	Animals ; Antigen Presentation ; Antigens, Viral/immunology ; Child, Preschool ; Dendritic Cells/immunology ; Dendritic Cells/virology ; Humans ; Immunity, Innate ; Immunologic Memory ; In Vitro Techniques ; Infant ; Models, Immunological ; Respiratory Syncytial Virus Infections/etiology ; Respiratory Syncytial Virus Infections/immunology ; Respiratory Syncytial Virus Infections/prevention & control ; Respiratory Syncytial Virus Vaccines/adverse effects ; Respiratory Syncytial Virus Vaccines/immunology ; Respiratory Syncytial Virus, Human/immunology ; T-Lymphocyte Subsets/immunology ; T-Lymphocyte Subsets/virology ; Vaccines, Subunit/adverse effects ; Vaccines, Subunit/immunology
Chemical Substances	Antigens, Viral ; Respiratory Syncytial Virus Vaccines ; Vaccines, Subunit
Language	English
Publishing date	2020-03-19
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0229660
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Article: Mutation of Respiratory Syncytial Virus G Protein's CX3C Motif Attenuates Infection in Cotton Rats and Primary Human Airway Epithelial Cells.

Ha, Binh / Chirkova, Tatiana / Boukhvalova, Marina S / Sun, He Ying / Walsh, Edward E / Anderson, Christopher S / Mariani, Thomas J / Anderson, Larry J

Vaccines

2019 Volume 7, Issue 3

Abstract: Despite being a high priority for vaccine development, no vaccine is yet available for respiratory syncytial virus (RSV). A live virus vaccine is the primary type of vaccine being developed for young children. In this report, we describe our studies of ... ...

Abstract	Despite being a high priority for vaccine development, no vaccine is yet available for respiratory syncytial virus (RSV). A live virus vaccine is the primary type of vaccine being developed for young children. In this report, we describe our studies of infected cotton rats and primary human airway epithelial cells (pHAECs) using an RSV r19F with a mutation in the CX3C chemokine motif in the RSV G protein (CX4C). Through this CX3C motif, RSV binds to the corresponding chemokine receptor, CX3CR1, and this binding contributes to RSV infection of pHAECs and virus induced host responses that contribute to disease. In both the cotton rat and pHAECs, the CX4C mutation decreased virus replication and disease and/or host responses to infection. Thus, this mutation, or other mutations that block binding to CX3CR1, has the potential to improve a live attenuated RSV vaccine by attenuating both infection and disease pathogenesis.
Language	English
Publishing date	2019-07-19
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines7030069
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Article ; Online: Evaluation of a SARS-CoV-2 Capture IgM Antibody Assay in Convalescent Sera.

Ha, Binh / Jadhao, Samadhan / Hussaini, Laila / Gibson, Theda / Stephens, Kathy / Salazar, Luis / Ciric, Caroline / Taylor, Meg / Rouphael, Nadine / Edupuganti, Srilatha / Rostad, Christina A / Tompkins, S Mark / Anderson, Evan J / Anderson, Larry J

Microbiology spectrum

2021 Volume 9, Issue 2, Page(s) e0045821

Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for a global pandemic with over 152 million cases and 3.19 million deaths reported by early May 2021. Understanding the serological response to SARS-CoV-2 is critical to ... ...

Abstract	Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for a global pandemic with over 152 million cases and 3.19 million deaths reported by early May 2021. Understanding the serological response to SARS-CoV-2 is critical to determining the burden of infection and disease (coronavirus disease 2019 [COVID-19]) and transmission dynamics. We developed a capture IgM assay because it should have better sensitivity and specificity than the commonly used indirect assay. Here, we report the development and performance of a capture IgM enzyme-linked immunosorbent assay (ELISA) and a companion indirect IgG ELISA for the spike (S) and nucleocapsid (N) proteins and the receptor-binding domain (RBD) of S. We found that among the IgM ELISAs, the S ELISA was positive in 76% of 55 serum samples from SARS-CoV-2 PCR-positive patients, the RBD ELISA was positive in 55% of samples, and the N ELISA was positive in 15% of samples. The companion indirect IgG ELISAs were positive for S in 89% of the 55 serum samples, RBD in 78%, and N in 85%. While the specificities for IgM RBD, S, and N ELISAs and IgG S and RBD ELISAs were 97% to 100%, the specificity of the N IgG ELISA was lower (89%). RBD-specific IgM antibodies became undetectable by 3 to 6 months, and S IgM reached low levels at 6 months. The corresponding IgG S, RBD, and N antibodies persisted with some decreases in levels over this time period. These capture IgM ELISAs and the companion indirect IgG ELISAs should enhance serologic studies of SARS-CoV-2 infections.
MeSH term(s)	Antibodies, Viral/immunology ; COVID-19/diagnosis ; COVID-19/therapy ; COVID-19 Serological Testing/methods ; Coronavirus Nucleocapsid Proteins/immunology ; Enzyme-Linked Immunosorbent Assay/methods ; Humans ; Immunization, Passive ; Immunoglobulin G/blood ; Immunoglobulin M/blood ; Phosphoproteins/immunology ; SARS-CoV-2/immunology ; Sensitivity and Specificity ; Serologic Tests/methods ; Spike Glycoprotein, Coronavirus/immunology ; COVID-19 Serotherapy
Chemical Substances	Antibodies, Viral ; Coronavirus Nucleocapsid Proteins ; Immunoglobulin G ; Immunoglobulin M ; Phosphoproteins ; Spike Glycoprotein, Coronavirus ; nucleocapsid phosphoprotein, SARS-CoV-2 ; spike protein, SARS-CoV-2
Language	English
Publishing date	2021-09-08
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.00458-21
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Article: Dietary exposure and health risk characterization of aflatoxin B1, ochratoxin A, fumonisin B1, and zearalenone in food from different provinces in Northern Vietnam

Do, Tuan Huu / Tran, Son Cao / Le, Chi Dinh / Nguyen, Ha-Binh Thi / Le, Phuong-Thao Thi / Le, Hong-Hao Thi / Le, Tuyen Danh / Thai-Nguyen, Hung-Thu

Food control. 2020 June, v. 112

2020

Abstract: A dietary exposure and health risk assessment of mycotoxins including aflatoxin B1, fumonisin B1, ochratoxin A, and zearalenone was conducted in 3 provinces in Northern Vietnam namely Hanoi, Thanh Hoa, and Ha Giang. Results of the analysis of samples of ... ...

Abstract	A dietary exposure and health risk assessment of mycotoxins including aflatoxin B1, fumonisin B1, ochratoxin A, and zearalenone was conducted in 3 provinces in Northern Vietnam namely Hanoi, Thanh Hoa, and Ha Giang. Results of the analysis of samples of maize, rice, peanut, and sesame revealed the presence of these mycotoxins in all samples and sampling locations. Aflatoxin B1 was the most frequently detected (19.1%) and widely distributed among different types of samples, whereas the percentage occurrence of fumonisin B1, ochratoxin A, and zearalenone were 11.2, 5.9 and 6.3, respectively. The later three mycotoxins were detected mostly in maize. The exposure to aflatoxin B1 at detected levels could lead to 0.23, 0.65 and 21.0 cases of liver cancer per 100,000 adult people per year in Hanoi, Thanh Hoa and Ha Giang, respectively. The risk assessment also showed the unsafe exposure to ochratoxin A and fumonisin B1 in the highland region where the people consume a large amount of foods derived from maize. In Ha Giang, the mean exposures to fumonisin B1 were lower than its PMTDI (Provisional Maximum Tolerable Daily Intake), however, the 95th percentile values were 1.1–1.9 times of the PMTDI. The mean exposures to ochratoxin A in Ha Giang were about 2.4–3.6 times higher than its PMTWI (Provisional Maximum Tolerable Weekly Intake). There was no risk of fumonisin B1 and ochratoxin A in Hanoi and Thanh Hoa. The dietary exposure to zearalenone was within its PMTDI in all locations. The results pointed out the need for further improvement of the control of these mycotoxins in Vietnam, especially in some highland provinces.
Keywords	acceptable daily intake ; adults ; aflatoxin B1 ; corn ; dietary exposure ; foods ; fumonisin B1 ; health effects assessments ; liver neoplasms ; ochratoxin A ; peanuts ; rice ; risk characterization ; zearalenone ; Vietnam
Language	English
Dates of publication	2020-06
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	1027805-9
ISSN	0956-7135
ISSN	0956-7135
DOI	10.1016/j.foodcont.2020.107108
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Performance evaluation of antibody tests for detecting infant respiratory syncytial virus infection.

Jadhao, Samadhan J / Ha, Binh / McCracken, Courtney / Gebretsadik, Tebeb / Rosas-Salazar, Christian / Chappell, James / Das, Suman / Hartert, Tina / Anderson, Larry J

Journal of medical virology

2020 Volume 93, Issue 6, Page(s) 3439–3445

Abstract: Respiratory syncytial virus (RSV) infection is a major cause of respiratory tract disease in young children and throughout life. Infant infection is also associated with later respiratory morbidity including asthma. With a prospective birth cohort study ... ...

Abstract	Respiratory syncytial virus (RSV) infection is a major cause of respiratory tract disease in young children and throughout life. Infant infection is also associated with later respiratory morbidity including asthma. With a prospective birth cohort study of RSV and asthma, we evaluated the performance of an RSV antibody enzyme-linked immunoassay (EIA) for detecting prior infant RSV infection. Infant RSV infection was determined by biweekly respiratory illness surveillance plus RSV polymerase chain reaction (PCR) testing in their first RSV season and serum RSV antibodies after the season at approximately 1 year of age. RSV antibodies were detected by RSV A and B lysate EIA. Antibody and PCR results on 1707 children included 327 RSV PCR positive (PCR+) and 1380 not RSV+. Of 327 PCR+ children, 314 (96%) were lysate EIA positive and 583 out of 1380 (42%) children not PCR+ were positive. We compared the lysate EIA to RSV F, group A G (Ga), and group B G (Gb) protein antibody EIAs in a subset of 226 sera, 118 PCR+ children (97 group A and 21 group B) and 108 not PCR+. In this subset, 117 out of 118 (99%) RSV PCR+ children were positive by both the F and lysate EIAs and 103 out of 118 (87%) were positive by the Ga and/or Gb EIAs. Comparison of the two G EIAs indicated the infecting group correctly in 100 out of 118 (86%) and incorrectly in 1 out of 118 (1%). The lysate and F EIAs are sensitive for detecting infant infection and the two G EIAs can indicate the group of an earlier primary infection.
MeSH term(s)	Antibodies, Viral/blood ; Asthma/diagnosis ; Asthma/immunology ; Female ; Humans ; Immunoenzyme Techniques/methods ; Immunoenzyme Techniques/standards ; Infant ; Male ; Prospective Studies ; Respiratory Syncytial Virus Infections/diagnosis ; Respiratory Syncytial Virus Infections/immunology ; Respiratory Syncytial Virus, Human/genetics ; Respiratory Syncytial Virus, Human/immunology ; Respiratory Tract Infections/diagnosis ; Respiratory Tract Infections/immunology ; Respiratory Tract Infections/virology
Chemical Substances	Antibodies, Viral
Language	English
Publishing date	2020-12-29
Publishing country	United States
Document type	Evaluation Study ; Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	752392-0
ISSN	1096-9071 ; 0146-6615
ISSN (online)	1096-9071
ISSN	0146-6615
DOI	10.1002/jmv.26736
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Article ; Online: 'Clustering' SIRPα into the plasma membrane lipid microdomains is required for activated monocytes and macrophages to mediate effective cell surface interactions with CD47.

Ha, Binh / Lv, Zhiyuan / Bian, Zhen / Zhang, Xiugen / Mishra, Aarti / Liu, Yuan

PloS one

2013 Volume 8, Issue 10, Page(s) e77615

Abstract: SIRPα, an ITIMs-containing signaling receptor, negatively regulates leukocyte responses through extracellular interactions with CD47. However, the dynamics of SIRPα-CD47 interactions on the cell surface and the governing mechanisms remain unclear. Here ... ...

Abstract	SIRPα, an ITIMs-containing signaling receptor, negatively regulates leukocyte responses through extracellular interactions with CD47. However, the dynamics of SIRPα-CD47 interactions on the cell surface and the governing mechanisms remain unclear. Here we report that while the purified SIRPα binds to CD47 and that SIRPα is expressed on monocytes and monocytic THP-1 or U937, these SIRPα are ineffective to mediate cell binding to immobilized CD47. However, cell binding to CD47 is significantly enhanced when monocytes transmigrating across endothelia, or being differentiated into macrophages. Cell surface labeling reveals SIRPα to be diffused on naïve monocytes but highly clustered on transmigrated monocytes and macrophages. Protein crosslink and equilibrium centrifugation confirm that SIRPα in the latter cells forms oligomerized complexes resulting in increased avidity for CD47 binding. Furthermore, formation of SIRPα complexes/clusters requires the plasma membrane 'lipid rafts' and the activity of Src family kinase during macrophage differentiation. These results together suggest that 'clustering' SIRPα into plasma membrane microdomains is essential for activated monocytes and macrophages to effectively interact with CD47 and initiate intracellular signaling.
MeSH term(s)	Actin Cytoskeleton/metabolism ; Animals ; Antigens, Differentiation/metabolism ; CD47 Antigen/metabolism ; Cell Differentiation ; Cell Line ; Cholesterol/metabolism ; Macrophages/cytology ; Macrophages/metabolism ; Membrane Microdomains/metabolism ; Mice ; Monocytes/cytology ; Monocytes/metabolism ; Protein Binding ; Receptors, Immunologic/metabolism ; src-Family Kinases/metabolism
Chemical Substances	Antigens, Differentiation ; CD47 Antigen ; Ptpns1 protein, mouse ; Receptors, Immunologic ; SIRPA protein, human ; Cholesterol (97C5T2UQ7J) ; src-Family Kinases (EC 2.7.10.2)
Language	English
Publishing date	2013-10-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0077615
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