Pub Date : 2020-01-01Epub Date: 2020-09-23DOI: 10.1159/000510223
Juan Camilo Sánchez-Arcila, Jessica Badolato-Correa, Thiara Manuele Alves de Souza, Iury Amâncio Paiva, Luciana Santos Barbosa, Priscila Conrado Guerra Nunes, Monique da Rocha Queiroz Lima, Flavia Barrento Dos Santos, Paulo Vieira Damasco, Rivaldo Venancio da Cunha, Elzinandes Leal de Azeredo, Luzia Maria de Oliveira-Pinto
Background: Arboviruses co-circulating within a population that are transmitted by the same vector have the potential to cause coinfections. Coinfections with dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV) have been occurring in Brazil, but it is not well-understood how human responses vary during mono- or coinfections and whether they play different roles in pathogenesis. Methods: We investigated the clinical, virological, and immunological status during patients’ acute infections, focusing on the CCL/CXC chemokines, proinflammatory, as well as anti-inflammatory cytokines levels quantified by ELISAs. Viral load was determined by qRT-PCR in serum samples from 116 acute DENV, ZIKV, CHIKV, DENV/ZIKV, and CHIKV/ZIKV-infected adult patients from Brazil. Results: Most of the acute patients displayed fever, headache, prostration, and myalgia, regardless of the type of arbovirus infection. Zika viral load was higher in CHIKV/ZIKV coinfected patients compared with ZIKV or DENV/ZIKV infections. All infected individuals presented increased concentrations of C-X-C motif chemokine ligand 10/interferon protein-10 (CXCL10/IP-10), C-C motif chemokine ligand 2/monocyte chemoattractant protein-1 (CCL2/MCP-1), and tumor necrosis factor alpha (TNF-α) compared to healthy donors. Interestingly, the ZIKV group separated from CHIKV/ZIKV due to higher levels of interleukin-10 (IL-10) and lower levels of TNF-α. While DENV/ZIKV differentiated from CHIKV due to their higher levels of CCL2/MCP-1, in CHIKV- and CHIKV/ZIKV-infected patients, levels of CXC10/IP-10, CCL2/MCP-1, and migration inhibitory factor (MIF) were associated with CHIKV viral load. By contrast, in DENV/ZIKV- and CHIKV/ZIKV-infected patients, levels of CXCL10/IP-10, CCL2/MCP-1, and TNF-α showed a significant inverse correlation with ZIKV viral load. Conclusions: From all the circulating mediators measured, we detected differences of IL-10, TNF-α, and CCL2/MCP-1 between arbovirus groups. We hypothesize that CXC10/IP-10, CCL2/MCP-1, and MIF in the CHIKV-infected group could regulate the CHIKV viral load, while CXC10/IP-10, CCL2/MCP-1, and TNF-α in DENV/ZIKV, and CHIKV/ZIKV groups, could regulate ZIKV viral load.
{"title":"Clinical, Virological, and Immunological Profiles of DENV, ZIKV, and/or CHIKV-Infected Brazilian Patients.","authors":"Juan Camilo Sánchez-Arcila, Jessica Badolato-Correa, Thiara Manuele Alves de Souza, Iury Amâncio Paiva, Luciana Santos Barbosa, Priscila Conrado Guerra Nunes, Monique da Rocha Queiroz Lima, Flavia Barrento Dos Santos, Paulo Vieira Damasco, Rivaldo Venancio da Cunha, Elzinandes Leal de Azeredo, Luzia Maria de Oliveira-Pinto","doi":"10.1159/000510223","DOIUrl":"https://doi.org/10.1159/000510223","url":null,"abstract":"Background: Arboviruses co-circulating within a population that are transmitted by the same vector have the potential to cause coinfections. Coinfections with dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV) have been occurring in Brazil, but it is not well-understood how human responses vary during mono- or coinfections and whether they play different roles in pathogenesis. Methods: We investigated the clinical, virological, and immunological status during patients’ acute infections, focusing on the CCL/CXC chemokines, proinflammatory, as well as anti-inflammatory cytokines levels quantified by ELISAs. Viral load was determined by qRT-PCR in serum samples from 116 acute DENV, ZIKV, CHIKV, DENV/ZIKV, and CHIKV/ZIKV-infected adult patients from Brazil. Results: Most of the acute patients displayed fever, headache, prostration, and myalgia, regardless of the type of arbovirus infection. Zika viral load was higher in CHIKV/ZIKV coinfected patients compared with ZIKV or DENV/ZIKV infections. All infected individuals presented increased concentrations of C-X-C motif chemokine ligand 10/interferon protein-10 (CXCL10/IP-10), C-C motif chemokine ligand 2/monocyte chemoattractant protein-1 (CCL2/MCP-1), and tumor necrosis factor alpha (TNF-α) compared to healthy donors. Interestingly, the ZIKV group separated from CHIKV/ZIKV due to higher levels of interleukin-10 (IL-10) and lower levels of TNF-α. While DENV/ZIKV differentiated from CHIKV due to their higher levels of CCL2/MCP-1, in CHIKV- and CHIKV/ZIKV-infected patients, levels of CXC10/IP-10, CCL2/MCP-1, and migration inhibitory factor (MIF) were associated with CHIKV viral load. By contrast, in DENV/ZIKV- and CHIKV/ZIKV-infected patients, levels of CXCL10/IP-10, CCL2/MCP-1, and TNF-α showed a significant inverse correlation with ZIKV viral load. Conclusions: From all the circulating mediators measured, we detected differences of IL-10, TNF-α, and CCL2/MCP-1 between arbovirus groups. We hypothesize that CXC10/IP-10, CCL2/MCP-1, and MIF in the CHIKV-infected group could regulate the CHIKV viral load, while CXC10/IP-10, CCL2/MCP-1, and TNF-α in DENV/ZIKV, and CHIKV/ZIKV groups, could regulate ZIKV viral load.","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"63 1-6","pages":"33-45"},"PeriodicalIF":4.6,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000510223","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38413981","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
J. Soares, A. Ferreira, A. Silva-Pinto, F. Almeida, C. Piñeiro, R. Serrão, A. Sarmento
Background: The role of antiretroviral therapy (ART) for Hepatitis C viral load (HCV-VL) and liver fibrosis is poorly understood. This study aimed at evaluating the influence of ART on HCV-VL and liver fibrosis in human immunodeficiency virus (HIV)/HCV-coinfected patients. Methods: We conducted a retrospective cohort study of HIV/HCV-coinfected patients followed at a tertiary university hospital. Results: In total, 143 patients were included. In 61 patients, ART initiation was accompanied by an increase in HCV-VL and a decrease in HIV viral load (HIV-VL), whereas ART suspension led to a decrease in HCV-VL and an increase in HIV-VL. Among the 55 HIV-suppressed patients who switched to a raltegravir (RAL)-containing regimen, median HCV-VL levels decreased significantly, while switching to a rilpivirine-containing regimen did not yield a significant reduction. Discussion: If the treatment of chronic hepatitis starts before ART, ART initiation should be delayed as much as possible. If ART has been started, it is advisable to wait 1 year before initiating chronic hepatitis treatment. RAL as the third agent in an ART regimen could be beneficial in HIV/HCV-coinfected patients, in comparison to other antiretroviral drugs. Conclusion: The start and the suspension of ART significantly interferes with HCV-VL in HIV/HCV-coinfected patients.
{"title":"The Influence of Antiretroviral Therapy on Hepatitis C Virus Viral Load and Liver Fibrosis in Human Immunodeficiency Virus-Coinfected Patients: An Observational Study","authors":"J. Soares, A. Ferreira, A. Silva-Pinto, F. Almeida, C. Piñeiro, R. Serrão, A. Sarmento","doi":"10.1159/000503631","DOIUrl":"https://doi.org/10.1159/000503631","url":null,"abstract":"Background: The role of antiretroviral therapy (ART) for Hepatitis C viral load (HCV-VL) and liver fibrosis is poorly understood. This study aimed at evaluating the influence of ART on HCV-VL and liver fibrosis in human immunodeficiency virus (HIV)/HCV-coinfected patients. Methods: We conducted a retrospective cohort study of HIV/HCV-coinfected patients followed at a tertiary university hospital. Results: In total, 143 patients were included. In 61 patients, ART initiation was accompanied by an increase in HCV-VL and a decrease in HIV viral load (HIV-VL), whereas ART suspension led to a decrease in HCV-VL and an increase in HIV-VL. Among the 55 HIV-suppressed patients who switched to a raltegravir (RAL)-containing regimen, median HCV-VL levels decreased significantly, while switching to a rilpivirine-containing regimen did not yield a significant reduction. Discussion: If the treatment of chronic hepatitis starts before ART, ART initiation should be delayed as much as possible. If ART has been started, it is advisable to wait 1 year before initiating chronic hepatitis treatment. RAL as the third agent in an ART regimen could be beneficial in HIV/HCV-coinfected patients, in comparison to other antiretroviral drugs. Conclusion: The start and the suspension of ART significantly interferes with HCV-VL in HIV/HCV-coinfected patients.","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"62 1","pages":"182 - 190"},"PeriodicalIF":4.6,"publicationDate":"2019-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000503631","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42655247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
R. Čivljak, T. Košutić-Gulija, A. Slović, Eva Huljev, N. Turčić, T. Meštrović, J. Vraneš, S. Ljubin-Sternak
Introduction: Although highly pertinent for children, outbreaks of human parainfluenza virus (HPIV) may cause up to 15% of all respiratory illnesses in adults and predispose them to serious adverse outcomes, with HPIV serotype 3 (HPIV3) being the most common. This study represents the first report of an HPIV3 outbreak among adults at a long-term health-care facility in Croatia. Methods: A retrospective study was conducted to investigate an outbreak of acute respiratory infection (ARI) at a single residential care facility for the disabled in Croatia. Demographic, epidemiological, and clinical data were collected for all residents, while hospitalized patients were appraised in detail by laboratory/radiological methods. Multiplex PCR for respiratory viruses and sequencing was performed. Partial HPIV3 HN 581 nt sequences were aligned with HPIV3 sequences from the GenBank database to conduct a phylogenetic analysis, where different bioinformatic approaches were employed. Results: In late June 2018, 5 of the 10 units at the facility were affected by the outbreak. Among the 106 residents, 23 (21.7%) developed ARI, and 6 (26.1%) of them were hospitalized. HPIV3 was identified in 18 (73%) of the residents and 5 (83%) of the hospitalized individuals. Isolated HPIV3 strains were classified within the phylogenetic subcluster C5 but grouped on 2 separate branches of the phylogenetic tree. During the entire outbreak period, none of the institution’s employees reported symptoms of ARI. Conclusions: Our study has shown that this health care-associated outbreak of HPIV3 infection could have been linked to multiple importation events. Preventive measures in curbing such incidents should be enforced vigorously.
{"title":"An Outbreak of Human Parainfluenza Virus 3 (Phylogenetic Subcluster C5) Infection among Adults at a Residential Care Facility for the Disabled in Croatia, 2018","authors":"R. Čivljak, T. Košutić-Gulija, A. Slović, Eva Huljev, N. Turčić, T. Meštrović, J. Vraneš, S. Ljubin-Sternak","doi":"10.1159/000503630","DOIUrl":"https://doi.org/10.1159/000503630","url":null,"abstract":"Introduction: Although highly pertinent for children, outbreaks of human parainfluenza virus (HPIV) may cause up to 15% of all respiratory illnesses in adults and predispose them to serious adverse outcomes, with HPIV serotype 3 (HPIV3) being the most common. This study represents the first report of an HPIV3 outbreak among adults at a long-term health-care facility in Croatia. Methods: A retrospective study was conducted to investigate an outbreak of acute respiratory infection (ARI) at a single residential care facility for the disabled in Croatia. Demographic, epidemiological, and clinical data were collected for all residents, while hospitalized patients were appraised in detail by laboratory/radiological methods. Multiplex PCR for respiratory viruses and sequencing was performed. Partial HPIV3 HN 581 nt sequences were aligned with HPIV3 sequences from the GenBank database to conduct a phylogenetic analysis, where different bioinformatic approaches were employed. Results: In late June 2018, 5 of the 10 units at the facility were affected by the outbreak. Among the 106 residents, 23 (21.7%) developed ARI, and 6 (26.1%) of them were hospitalized. HPIV3 was identified in 18 (73%) of the residents and 5 (83%) of the hospitalized individuals. Isolated HPIV3 strains were classified within the phylogenetic subcluster C5 but grouped on 2 separate branches of the phylogenetic tree. During the entire outbreak period, none of the institution’s employees reported symptoms of ARI. Conclusions: Our study has shown that this health care-associated outbreak of HPIV3 infection could have been linked to multiple importation events. Preventive measures in curbing such incidents should be enforced vigorously.","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"62 1","pages":"174 - 181"},"PeriodicalIF":4.6,"publicationDate":"2019-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000503630","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41414695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-30Print Date: 2019-10-15DOI: 10.1128/JVI.00719-19
Thomas van Stigt Thans, Janet I Akko, Annika Niehrs, Wilfredo F Garcia-Beltran, Laura Richert, Christina M Stürzel, Christopher T Ford, Hui Li, Christina Ochsenbauer, John C Kappes, Beatrice H Hahn, Frank Kirchhoff, Glòria Martrus, Daniel Sauter, Marcus Altfeld, Angelique Hölzemer
Human immunodeficiency virus type 1 (HIV-1) has evolved elaborate ways to evade immune cell recognition, including downregulation of classical HLA class I (HLA-I) from the surfaces of infected cells. Recent evidence identified HLA-E, a nonclassical HLA-I, as an important part of the antiviral immune response to HIV-1. Changes in HLA-E surface levels and peptide presentation can prompt both CD8+ T-cell and natural killer (NK) cell responses to viral infections. Previous studies reported unchanged or increased HLA-E levels on HIV-1-infected cells. Here, we examined HLA-E surface levels following infection of CD4+ T cells with primary HIV-1 strains and observed that a subset downregulated HLA-E. Two primary strains of HIV-1 that induced the strongest reduction in surface HLA-E expression were chosen for further testing. Expression of single Nef or Vpu proteins in a T-cell line, as well as tail swap experiments exchanging the cytoplasmic tail of HLA-A2 with that of HLA-E, demonstrated that Nef modulated HLA-E surface levels and targeted the cytoplasmic tail of HLA-E. Furthermore, infection of primary CD4+ T cells with HIV-1 mutants showed that a lack of functional Nef (and Vpu to some extent) impaired HLA-E downmodulation. Taken together, the results of this study demonstrate for the first time that HIV-1 can downregulate HLA-E surface levels on infected primary CD4+ T cells, potentially rendering them less vulnerable to CD8+ T-cell recognition but at increased risk of NKG2A+ NK cell killing.IMPORTANCE For almost two decades, it was thought that HIV-1 selectively downregulated the highly expressed HLA-I molecules HLA-A and HLA-B from the cell surface in order to evade cytotoxic-T-cell recognition, while leaving HLA-C and HLA-E molecules unaltered. It was stipulated that HIV-1 infection thereby maintained inhibition of NK cells via inhibitory receptors that bind HLA-C and HLA-E. This concept was recently revised when a study showed that primary HIV-1 strains reduce HLA-C surface levels, whereas the cell line-adapted HIV-1 strain NL4-3 lacks this ability. Here, we demonstrate that infection with distinct primary HIV-1 strains results in significant downregulation of surface HLA-E levels. Given the increasing evidence for HLA-E as an important modulator of CD8+ T-cell and NKG2A+ NK cell functions, this finding has substantial implications for future immunomodulatory approaches aimed at harnessing cytotoxic cellular immunity against HIV.
人类免疫缺陷病毒 1 型（HIV-1）已经进化出了躲避免疫细胞识别的复杂方法，包括下调受感染细胞表面的经典 HLA I 类（HLA-I）。最近的证据表明，HLA-E（一种非经典的 HLA-I）是 HIV-1 抗病毒免疫反应的重要组成部分。HLA-E 表面水平和肽呈现的变化可促使 CD8+ T 细胞和自然杀伤（NK）细胞对病毒感染做出反应。以前的研究报告称，HIV-1 感染细胞的 HLA-E 水平没有变化或有所增加。在这里，我们检测了 CD4+ T 细胞感染原代 HIV-1 株系后的 HLA-E 表面水平，并观察到一个亚群下调了 HLA-E。我们选择了两种导致 HLA-E 表面表达下降最强的 HIV-1 原始菌株进行进一步检测。在T细胞系中表达单个Nef或Vpu蛋白，以及将HLA-A2的胞浆尾部与HLA-E的胞浆尾部交换的尾部交换实验表明，Nef可调节HLA-E的表面水平，并以HLA-E的胞浆尾部为靶标。此外，用HIV-1突变体感染原代CD4+T细胞表明，缺乏功能性Nef（在一定程度上也缺乏Vpu）会影响HLA-E的下调。综上所述，本研究结果首次证明了 HIV-1 能下调受感染的原代 CD4+ T 细胞的 HLA-E 表面水平，从而降低它们被 CD8+ T 细胞识别的可能性，但增加它们被 NKG2A+ NK 细胞杀伤的风险。重要意义 近二十年来，人们一直认为 HIV-1 选择性地下调细胞表面高表达的 HLA-I 分子 HLA-A 和 HLA-B，以逃避细胞毒性 T 细胞的识别，而 HLA-C 和 HLA-E 分子则保持不变。据此推断，HIV-1 感染通过与 HLA-C 和 HLA-E 结合的抑制性受体来维持对 NK 细胞的抑制。最近的一项研究表明，原生 HIV-1 株系能降低 HLA-C 表面水平，而细胞系适应 HIV-1 株系 NL4-3 则缺乏这种能力，从而修正了这一概念。在这里，我们证明了感染不同的原代 HIV-1 株系会导致表面 HLA-E 水平显著下调。鉴于越来越多的证据表明 HLA-E 是 CD8+ T 细胞和 NKG2A+ NK 细胞功能的重要调节因子，这一发现对未来旨在利用细胞毒性细胞免疫对抗 HIV 的免疫调节方法具有重大意义。
{"title":"Primary HIV-1 Strains Use Nef To Downmodulate HLA-E Surface Expression.","authors":"Thomas van Stigt Thans, Janet I Akko, Annika Niehrs, Wilfredo F Garcia-Beltran, Laura Richert, Christina M Stürzel, Christopher T Ford, Hui Li, Christina Ochsenbauer, John C Kappes, Beatrice H Hahn, Frank Kirchhoff, Glòria Martrus, Daniel Sauter, Marcus Altfeld, Angelique Hölzemer","doi":"10.1128/JVI.00719-19","DOIUrl":"10.1128/JVI.00719-19","url":null,"abstract":"<p><p>Human immunodeficiency virus type 1 (HIV-1) has evolved elaborate ways to evade immune cell recognition, including downregulation of classical HLA class I (HLA-I) from the surfaces of infected cells. Recent evidence identified HLA-E, a nonclassical HLA-I, as an important part of the antiviral immune response to HIV-1. Changes in HLA-E surface levels and peptide presentation can prompt both CD8<sup>+</sup> T-cell and natural killer (NK) cell responses to viral infections. Previous studies reported unchanged or increased HLA-E levels on HIV-1-infected cells. Here, we examined HLA-E surface levels following infection of CD4<sup>+</sup> T cells with primary HIV-1 strains and observed that a subset downregulated HLA-E. Two primary strains of HIV-1 that induced the strongest reduction in surface HLA-E expression were chosen for further testing. Expression of single Nef or Vpu proteins in a T-cell line, as well as tail swap experiments exchanging the cytoplasmic tail of HLA-A2 with that of HLA-E, demonstrated that Nef modulated HLA-E surface levels and targeted the cytoplasmic tail of HLA-E. Furthermore, infection of primary CD4<sup>+</sup> T cells with HIV-1 mutants showed that a lack of functional Nef (and Vpu to some extent) impaired HLA-E downmodulation. Taken together, the results of this study demonstrate for the first time that HIV-1 can downregulate HLA-E surface levels on infected primary CD4<sup>+</sup> T cells, potentially rendering them less vulnerable to CD8<sup>+</sup> T-cell recognition but at increased risk of NKG2A<sup>+</sup> NK cell killing.<b>IMPORTANCE</b> For almost two decades, it was thought that HIV-1 selectively downregulated the highly expressed HLA-I molecules HLA-A and HLA-B from the cell surface in order to evade cytotoxic-T-cell recognition, while leaving HLA-C and HLA-E molecules unaltered. It was stipulated that HIV-1 infection thereby maintained inhibition of NK cells via inhibitory receptors that bind HLA-C and HLA-E. This concept was recently revised when a study showed that primary HIV-1 strains reduce HLA-C surface levels, whereas the cell line-adapted HIV-1 strain NL4-3 lacks this ability. Here, we demonstrate that infection with distinct primary HIV-1 strains results in significant downregulation of surface HLA-E levels. Given the increasing evidence for HLA-E as an important modulator of CD8<sup>+</sup> T-cell and NKG2A<sup>+</sup> NK cell functions, this finding has substantial implications for future immunomodulatory approaches aimed at harnessing cytotoxic cellular immunity against HIV.</p>","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"59 1","pages":""},"PeriodicalIF":5.4,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/JVI.00719-19","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"63837986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective: Respiratory syncytial virus (RSV) infection causes lower respiratory tract infection primarily in infants and toddlers. RSV reinfection also occurs throughout life and can be a significant cause of pneumonia and mortality in the elderly. Surges in physician offices, emergency department visits, and hospitalization often result from RSV illness. Point-of-care (POC) testing reduces healthcare costs and permits informed decisions on treatment, further testing, or hospitalization to occur during the physician-patient encounter. Optimal POC assays must be sensitive, easy to perform, and provide rapid results. Methods: In this study, 2 POC assays (Alere i; Abbot Rapid Diagnostics and cobas Liat, Roche Molecular, Inc.) and a laboratory-based assay (Solana; Quidel, Inc.) were evaluated using 133 patient nasopharyngeal specimens. Results: Sensitivity/specificity values (%) of 94.7/96.1, 98.2/96.1, and 96.5/94.7 were obtained for the Alere i, Liat, and Solana assays, respectfully. These values approximated those stated in each assay’s package insert. Conclusion: Rapid molecular assays for RSV are sensitive and accurate. The choice of assay should reflect each healthcare institution’s specific testing needs with respect to the benefits and drawbacks of each product.
{"title":"Evaluation of Rapid, Molecular-Based Assays for the Detection of Respiratory Syncytial Virus","authors":"G. P. Leonardi","doi":"10.1159/000502995","DOIUrl":"https://doi.org/10.1159/000502995","url":null,"abstract":"Objective: Respiratory syncytial virus (RSV) infection causes lower respiratory tract infection primarily in infants and toddlers. RSV reinfection also occurs throughout life and can be a significant cause of pneumonia and mortality in the elderly. Surges in physician offices, emergency department visits, and hospitalization often result from RSV illness. Point-of-care (POC) testing reduces healthcare costs and permits informed decisions on treatment, further testing, or hospitalization to occur during the physician-patient encounter. Optimal POC assays must be sensitive, easy to perform, and provide rapid results. Methods: In this study, 2 POC assays (Alere i; Abbot Rapid Diagnostics and cobas Liat, Roche Molecular, Inc.) and a laboratory-based assay (Solana; Quidel, Inc.) were evaluated using 133 patient nasopharyngeal specimens. Results: Sensitivity/specificity values (%) of 94.7/96.1, 98.2/96.1, and 96.5/94.7 were obtained for the Alere i, Liat, and Solana assays, respectfully. These values approximated those stated in each assay’s package insert. Conclusion: Rapid molecular assays for RSV are sensitive and accurate. The choice of assay should reflect each healthcare institution’s specific testing needs with respect to the benefits and drawbacks of each product.","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"62 1","pages":"112 - 115"},"PeriodicalIF":4.6,"publicationDate":"2019-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000502995","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46303581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Huu Chien Nguyen, H. Park, Ho-Joon Shin, N. Nguyen, Anh Thi Viet Nguyen, T. Trinh, Thi Hai Yen Duong, H. Tuong, Vui Thi Hoang, G. Seo, H. Sohn, Seon-Ju Yeo
Background: When infected with the chikungunya virus (CHIKV), 3% to 28% of CHIKV-infected individuals remain asymptomatic, necessitating the development of improved high-throughput screening methods to overcome the limitations of molecular diagnostics or enzyme-linked immunosorbent assays (ELISAs). Objective: In this study, two novel monoclonal antibodies (mAbs) targeting envelope 1 (E1) of CHIKV were developed and applied in a fluorescence-linked immunosorbent assay (FLISA) using coumarin-derived dendrimer as the fluorophore. Methods: The performance of the FLISA was compared with that of ELISA. Results: Using the two novel mAbs (2B5 and 2C8), FLISA could detect 1 × 105 PFU/mL of CHIKV, exhibiting a 2-fold lower limit of detection (LOD) compared to ELISA. The LOD of FICT corresponded to a comparative threshold value of 23.95 and 4 × 106 of RNA copy number/µL. In the presence of human sera and blood, virus detection by FLISA was 3-fold better than ELISA, with an LOD of 2 × 105 PFU/mL. Sera and blood interfered with the ELISA, resulting in 6 × 105 PFU/mL as the LOD. Conclusions: FLISA using two novel mAbs and coumarin-derived dendrimer is a superior diagnostic assay for detecting CHIKV in human sera and blood, compared to conventional ELISA.
{"title":"Fluorescent Immunosorbent Assay for Chikungunya Virus Detection","authors":"Huu Chien Nguyen, H. Park, Ho-Joon Shin, N. Nguyen, Anh Thi Viet Nguyen, T. Trinh, Thi Hai Yen Duong, H. Tuong, Vui Thi Hoang, G. Seo, H. Sohn, Seon-Ju Yeo","doi":"10.1159/000502823","DOIUrl":"https://doi.org/10.1159/000502823","url":null,"abstract":"Background: When infected with the chikungunya virus (CHIKV), 3% to 28% of CHIKV-infected individuals remain asymptomatic, necessitating the development of improved high-throughput screening methods to overcome the limitations of molecular diagnostics or enzyme-linked immunosorbent assays (ELISAs). Objective: In this study, two novel monoclonal antibodies (mAbs) targeting envelope 1 (E1) of CHIKV were developed and applied in a fluorescence-linked immunosorbent assay (FLISA) using coumarin-derived dendrimer as the fluorophore. Methods: The performance of the FLISA was compared with that of ELISA. Results: Using the two novel mAbs (2B5 and 2C8), FLISA could detect 1 × 105 PFU/mL of CHIKV, exhibiting a 2-fold lower limit of detection (LOD) compared to ELISA. The LOD of FICT corresponded to a comparative threshold value of 23.95 and 4 × 106 of RNA copy number/µL. In the presence of human sera and blood, virus detection by FLISA was 3-fold better than ELISA, with an LOD of 2 × 105 PFU/mL. Sera and blood interfered with the ELISA, resulting in 6 × 105 PFU/mL as the LOD. Conclusions: FLISA using two novel mAbs and coumarin-derived dendrimer is a superior diagnostic assay for detecting CHIKV in human sera and blood, compared to conventional ELISA.","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"62 1","pages":"145 - 155"},"PeriodicalIF":4.6,"publicationDate":"2019-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000502823","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43370273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Annick Heykers, A. Leemans, W. Van der Gucht, M. De Schryver, P. Cos, P. Delputte
Objectives: Differences have been observed in the susceptibility of macrophage cell lines to respiratory syncytial virus (RSV) infection. In this study, we evaluated whether the type of macrophage cell line and RSV strain used have an influence on the infectivity and production of progeny virus. Methods: Both human and murine macrophage-like cell lines were infected with different RSV strains, both lab strains as well as clinical isolates. The infection was evaluated after 24 and 72 h by immunofluorescence staining and microscopic analysis, and the production of new virus particles was determined by plaque assay. Results: Susceptibility of macrophages to RSV was influenced by the RSV strain used but was mostly dependent on the macrophage cell line. Numbers of infected cells and virus production were generally very low or absent in murine cell lines. In human cell lines, clear infection was observed associated with production of new virus particles. Conclusion: Differences in susceptibility of macrophage cell lines to RSV infection are primarily related to the species of origin of the cell line but are also influenced by the RSV strain.
{"title":"Differences in Susceptibility of Human and Mouse Macrophage Cell Lines to Respiratory Syncytial Virus Infection","authors":"Annick Heykers, A. Leemans, W. Van der Gucht, M. De Schryver, P. Cos, P. Delputte","doi":"10.1159/000502674","DOIUrl":"https://doi.org/10.1159/000502674","url":null,"abstract":"Objectives: Differences have been observed in the susceptibility of macrophage cell lines to respiratory syncytial virus (RSV) infection. In this study, we evaluated whether the type of macrophage cell line and RSV strain used have an influence on the infectivity and production of progeny virus. Methods: Both human and murine macrophage-like cell lines were infected with different RSV strains, both lab strains as well as clinical isolates. The infection was evaluated after 24 and 72 h by immunofluorescence staining and microscopic analysis, and the production of new virus particles was determined by plaque assay. Results: Susceptibility of macrophages to RSV was influenced by the RSV strain used but was mostly dependent on the macrophage cell line. Numbers of infected cells and virus production were generally very low or absent in murine cell lines. In human cell lines, clear infection was observed associated with production of new virus particles. Conclusion: Differences in susceptibility of macrophage cell lines to RSV infection are primarily related to the species of origin of the cell line but are also influenced by the RSV strain.","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"62 1","pages":"134 - 144"},"PeriodicalIF":4.6,"publicationDate":"2019-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000502674","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47383894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zabihollah Shoja, M. Farahmand, N. Hosseini, S. Jalilvand
Introduction: To date, the human papillomavirus (HPV) vaccine has not been integrated into the national vaccination program of most countries of the WHO Eastern Mediterranean Region (EMRO), except for the United Arab Emirates and Libya. The knowledge of HPV genotype distribution in cervical neoplasia is valuable to predict the impact of current HPV vaccines on cancer prevention and can help the health policymakers to select the most appropriate vaccine types in their countries. Methods: Hence, this meta-analysis recapitulates all available data on HPV prevalence and genotypes in women with atypical squamous cells of undetermined significance (ASCUS), cervical intraepithelial neoplasia (CIN) I–III or low- and high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively), and invasive cervical cancer (ICC) in EMRO countries. Results: The meta-analysis included 5,990 cases of cervical precancer and cancer. The overall HPV prevalence was 85.4, 71.3, 59.2, and 34.8% in women with ICC, CIN II–III or HSIL, CIN I or LSIL, and ASCUS, respectively. HPV 16 was the most common genotype followed by HPV 18, representing 58 and 16.5% in ICC cases, respectively. Conclusion: This meta-analysis showed that the introduction of current HPV vaccines into national vaccination programs and the establishment of comprehensive screening programs in EMRO countries is beneficial by preventing 74.5% of cervical neoplasia.
{"title":"A Meta-Analysis on Human Papillomavirus Type Distribution among Women with Cervical Neoplasia in the WHO Eastern Mediterranean Region","authors":"Zabihollah Shoja, M. Farahmand, N. Hosseini, S. Jalilvand","doi":"10.1159/000502824","DOIUrl":"https://doi.org/10.1159/000502824","url":null,"abstract":"Introduction: To date, the human papillomavirus (HPV) vaccine has not been integrated into the national vaccination program of most countries of the WHO Eastern Mediterranean Region (EMRO), except for the United Arab Emirates and Libya. The knowledge of HPV genotype distribution in cervical neoplasia is valuable to predict the impact of current HPV vaccines on cancer prevention and can help the health policymakers to select the most appropriate vaccine types in their countries. Methods: Hence, this meta-analysis recapitulates all available data on HPV prevalence and genotypes in women with atypical squamous cells of undetermined significance (ASCUS), cervical intraepithelial neoplasia (CIN) I–III or low- and high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively), and invasive cervical cancer (ICC) in EMRO countries. Results: The meta-analysis included 5,990 cases of cervical precancer and cancer. The overall HPV prevalence was 85.4, 71.3, 59.2, and 34.8% in women with ICC, CIN II–III or HSIL, CIN I or LSIL, and ASCUS, respectively. HPV 16 was the most common genotype followed by HPV 18, representing 58 and 16.5% in ICC cases, respectively. Conclusion: This meta-analysis showed that the introduction of current HPV vaccines into national vaccination programs and the establishment of comprehensive screening programs in EMRO countries is beneficial by preventing 74.5% of cervical neoplasia.","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"62 1","pages":"101 - 111"},"PeriodicalIF":4.6,"publicationDate":"2019-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000502824","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48374112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Vontas, C. Hadjicristodoulou, Vasilis Krikelis, E. Petinaki
Despite the significant medical advances which have taken place in the last decades, acute diarrhoea cases remain a public health issue of major significance, with gastroenteritis agents being associated with severe symptoms in adults and high morbidity in infants and children. Regarding rotaviruses, while children are the predominant victims of rotavirus infection, adults (often caretakers or parents of these children) may experience the same symptoms of fever, vomiting, and non-bloody diarrhoea. Three different routine schemes for the detection of rotaviruses in archived stool samples were evaluated in terms of diagnostic performance. A total of 640 archived stool samples were included in the study. The samples were screened with three different techniques: a commercial rapid immunochromatographic test, a modified in-house conventional one-step reverse-transcription polymerase chain reaction (PCR) screen protocol, and a commercial one-step real-time PCR kit. Technical aspects and considerations are discussed.
{"title":"The Use of Immunochromatographic Technique for Rotavirus Detection: Experience from a Tertiary Care Hospital in Central Greece","authors":"A. Vontas, C. Hadjicristodoulou, Vasilis Krikelis, E. Petinaki","doi":"10.1159/000502007","DOIUrl":"https://doi.org/10.1159/000502007","url":null,"abstract":"Despite the significant medical advances which have taken place in the last decades, acute diarrhoea cases remain a public health issue of major significance, with gastroenteritis agents being associated with severe symptoms in adults and high morbidity in infants and children. Regarding rotaviruses, while children are the predominant victims of rotavirus infection, adults (often caretakers or parents of these children) may experience the same symptoms of fever, vomiting, and non-bloody diarrhoea. Three different routine schemes for the detection of rotaviruses in archived stool samples were evaluated in terms of diagnostic performance. A total of 640 archived stool samples were included in the study. The samples were screened with three different techniques: a commercial rapid immunochromatographic test, a modified in-house conventional one-step reverse-transcription polymerase chain reaction (PCR) screen protocol, and a commercial one-step real-time PCR kit. Technical aspects and considerations are discussed.","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"62 1","pages":"164 - 168"},"PeriodicalIF":4.6,"publicationDate":"2019-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000502007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41508141","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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