This page has only limited features, please log in for full access.
Objectives Pertussis is a respiratory infectious disease caused by Bordetella pertussis. In the Caribbean Netherlands (CN), comprising the islands Bonaire, St. Eustatius, and Saba, registration of cases is mandatory for disease surveillance. However, insufficient laboratory facilities hamper case confirmation and circulation persists. The aim of this seroepidemiological study is to gain insight into B. pertussis circulation in CN and to investigate what factors contribute to the risk of infection. Methods Blood samples and questionnaires were collected for 1,829 participants 0-90 years old. Concentrations of B. pertussis toxin-specific IgG antibodies (anti-Pt) were determined using a bead-based immunoassay to indicate infections within the previous twelve months (based on anti-Pt ≥50IU/mL) in participants without detectable vaccine-induced humoral immunity. Risk factors for a recent infection were analyzed using logistic regression models. Results An estimated 8.2% (95% CI 6.6-10.1) of CN residents aged ≥9 years was recently infected by B. pertussis. Risk factors for a recent infection were age 12-29 years (13.8%-14.6%) and Dutch Caribbean or Surinamese origin (10.7%). Conclusions B. pertussis infections occur frequently among CN residents aged ≥9 years, although few clinical pertussis cases are reported. Transmission to vulnerable individuals seems likely and should be taken into account in optimizing the vaccination program.
Maarten M. Immink; Eric R.A. Vos; Alcira V.A. Janga-Jansen; Sharda Baboe-Kalpoe; Koen Hulshof; Jeffrey van Vliet; Jeroen Kerkhof; Gerco Den Hartog; Hester E. de Melker; Fiona R.M. van der Klis; Nicoline A.T. van der Maas. Circulation of Bordetella pertussis in the Caribbean Netherlands: a population-based seroepidemiological study. International Journal of Infectious Diseases 2021, 1 .
AMA StyleMaarten M. Immink, Eric R.A. Vos, Alcira V.A. Janga-Jansen, Sharda Baboe-Kalpoe, Koen Hulshof, Jeffrey van Vliet, Jeroen Kerkhof, Gerco Den Hartog, Hester E. de Melker, Fiona R.M. van der Klis, Nicoline A.T. van der Maas. Circulation of Bordetella pertussis in the Caribbean Netherlands: a population-based seroepidemiological study. International Journal of Infectious Diseases. 2021; ():1.
Chicago/Turabian StyleMaarten M. Immink; Eric R.A. Vos; Alcira V.A. Janga-Jansen; Sharda Baboe-Kalpoe; Koen Hulshof; Jeffrey van Vliet; Jeroen Kerkhof; Gerco Den Hartog; Hester E. de Melker; Fiona R.M. van der Klis; Nicoline A.T. van der Maas. 2021. "Circulation of Bordetella pertussis in the Caribbean Netherlands: a population-based seroepidemiological study." International Journal of Infectious Diseases , no. : 1.
Background There is an urgent need for fair and equitable access to safe and effective vaccines to end the COVID-19 pandemic. Shortages in reagents and vaccines are a major challenge, as well as limited knowledge on dose response relationship with mRNA COVID-19 vaccines. We explored intradermal fractional dose administration of a mRNA SARS-CoV-2/COVID-19 vaccine as a potential dose-sparing strategy. Methods We conducted a proof-of-concept, dose-escalation, open-label, randomised-controlled vaccine trial (IDSCOVA) in healthy adults aged 18-30 years. To test initial safety, ten participants received 10 µg mRNA-1273 vaccine through intradermal injection at day 1 and 29. Following a favourable safety review, thirty participants were 1:1 randomised to receive 20 µg mRNA-1273 either intradermally or intramuscularly. The primary endpoint was tolerability and safety. The secondary endpoint was seroconversion and specific IgG concentration against SARS-CoV-2 spike S1 and Receptor Binding Domain (RBD) after the second dose at day 43. We compared results to two historical cohorts of non-hospitalised COVID-19 patients and vaccinated individuals. Findings Thirty-eight of forty included participants (median age 25 years) completed the study. There were no serious adverse events. Self-reported local adverse reactions after intradermal delivery were mild, both in the 10 µg and the 20 µg group. In the higher dose group, systemic adverse reactions were more common, but still well tolerated. All 38 participants mounted substantially higher IgG-anti-S1 and IgG-anti-RBD concentrations at day 43 than COVID-19 controls. At day 43, anti-S1 (95% CI) was 1,696 (1,309-2,198) BAU/mL for the 10 µg intradermal group, 1,406 (953·5-2,074) BAU/mL for the 20 µg intramuscular group and 2,057 (1,421-2,975) BAU/mL for the 20 µg intradermal group. Anti-S1 was 107·2 (63-182·2) BAU/mL for the convalescent plasma control group and 1,558 (547·8-4,433) BAU/mL for the individuals vaccinated with 100 µg mRNA-1273. Interpretation Intradermal administration of 10 µg and 20 µg mRNA-1273 vaccine was well tolerated and safe, and resulted in a robust antibody response. Intradermal vaccination has the potential to be deployed for vaccine dose-sparing. Funding The trial was supported by crowdfunding (Wake Up to Corona).
Geert V.T. Roozen; Margaretha L.M. Prins; Rob van Binnendijk; Gerco Den Hartog; Vincent P. Kuiper; Corine Prins; Jacqueline J. Janse; Annelieke C. Kruithof; Mariet C.W. Feltkamp; Marjan Kuijer; Frits R. Rosendaal; Meta Roestenberg; Leo G. Visser; Anna H.E. Roukens. Tolerability, safety and immunogenicity of intradermal delivery of a fractional dose mRNA-1273 SARS-CoV-2 vaccine in healthy adults as a dose sparing strategy. 2021, 1 .
AMA StyleGeert V.T. Roozen, Margaretha L.M. Prins, Rob van Binnendijk, Gerco Den Hartog, Vincent P. Kuiper, Corine Prins, Jacqueline J. Janse, Annelieke C. Kruithof, Mariet C.W. Feltkamp, Marjan Kuijer, Frits R. Rosendaal, Meta Roestenberg, Leo G. Visser, Anna H.E. Roukens. Tolerability, safety and immunogenicity of intradermal delivery of a fractional dose mRNA-1273 SARS-CoV-2 vaccine in healthy adults as a dose sparing strategy. . 2021; ():1.
Chicago/Turabian StyleGeert V.T. Roozen; Margaretha L.M. Prins; Rob van Binnendijk; Gerco Den Hartog; Vincent P. Kuiper; Corine Prins; Jacqueline J. Janse; Annelieke C. Kruithof; Mariet C.W. Feltkamp; Marjan Kuijer; Frits R. Rosendaal; Meta Roestenberg; Leo G. Visser; Anna H.E. Roukens. 2021. "Tolerability, safety and immunogenicity of intradermal delivery of a fractional dose mRNA-1273 SARS-CoV-2 vaccine in healthy adults as a dose sparing strategy." , no. : 1.
The emergence of SARS-CoV-2 variants harboring mutations in the spike (S) protein has raised concern about potential immune escape. Here, we studied humoral and cellular immune responses to wild type SARS-CoV-2 and the B.1.1.7 and B.1.351 variants of concern in a cohort of 121 BNT162b2 mRNA-vaccinated health care workers (HCW). Twenty-three HCW recovered from mild COVID-19 disease and exhibited a recall response with high levels of SARS-CoV-2-specific functional antibodies and virus-specific T cells after a single vaccination. Specific immune responses were also detected in seronegative HCW after one vaccination, but a second dose was required to reach high levels of functional antibodies and cellular immune responses in all individuals. Vaccination-induced antibodies cross-neutralized the variants B.1.1.7 and B.1.351, but the neutralizing capacity and Fc-mediated functionality against B.1.351 was consistently 2- to 4-fold lower than to the homologous virus. In addition, peripheral blood mononuclear cells were stimulated with peptide pools spanning the mutated S regions of B.1.1.7 and B.1.351 to detect cross-reactivity of SARS-CoV-2-specific T cells with variants. Importantly, we observed no differences in CD4+ T-cell activation in response to variant antigens, indicating that the B.1.1.7 and B.1.351 S proteins do not escape T-cell-mediated immunity elicited by the wild type S protein. In conclusion, this study shows that some variants can partially escape humoral immunity induced by SARS-CoV-2 infection or BNT162b2 vaccination, but S-specific CD4+ T-cell activation is not affected by the mutations in the B.1.1.7 and B.1.351 variants.
Daryl Geers; Marc C. Shamier; Susanne Bogers; Gerco Den Hartog; Lennert Gommers; Nella N. Nieuwkoop; Katharina S. Schmitz; Laurine C. Rijsbergen; Jolieke A.T. van Osch; Emma Dijkhuizen; Gaby Smits; Anouskha Comvalius; Djenolan van Mourik; Tom G. Caniels; Marit J. van Gils; Rogier W. Sanders; Bas B. Oude Munnink; Richard Molenkamp; Herbert J. de Jager; Bart L. Haagmans; Rik L. de Swart; Marion P.G. Koopmans; Robert S. van Binnendijk; Rory D. de Vries; Corine H. GeurtsvanKessel. SARS-CoV-2 variants of concern partially escape humoral but not T-cell responses in COVID-19 convalescent donors and vaccinees. Science Immunology 2021, 6, eabj1750 .
AMA StyleDaryl Geers, Marc C. Shamier, Susanne Bogers, Gerco Den Hartog, Lennert Gommers, Nella N. Nieuwkoop, Katharina S. Schmitz, Laurine C. Rijsbergen, Jolieke A.T. van Osch, Emma Dijkhuizen, Gaby Smits, Anouskha Comvalius, Djenolan van Mourik, Tom G. Caniels, Marit J. van Gils, Rogier W. Sanders, Bas B. Oude Munnink, Richard Molenkamp, Herbert J. de Jager, Bart L. Haagmans, Rik L. de Swart, Marion P.G. Koopmans, Robert S. van Binnendijk, Rory D. de Vries, Corine H. GeurtsvanKessel. SARS-CoV-2 variants of concern partially escape humoral but not T-cell responses in COVID-19 convalescent donors and vaccinees. Science Immunology. 2021; 6 (59):eabj1750.
Chicago/Turabian StyleDaryl Geers; Marc C. Shamier; Susanne Bogers; Gerco Den Hartog; Lennert Gommers; Nella N. Nieuwkoop; Katharina S. Schmitz; Laurine C. Rijsbergen; Jolieke A.T. van Osch; Emma Dijkhuizen; Gaby Smits; Anouskha Comvalius; Djenolan van Mourik; Tom G. Caniels; Marit J. van Gils; Rogier W. Sanders; Bas B. Oude Munnink; Richard Molenkamp; Herbert J. de Jager; Bart L. Haagmans; Rik L. de Swart; Marion P.G. Koopmans; Robert S. van Binnendijk; Rory D. de Vries; Corine H. GeurtsvanKessel. 2021. "SARS-CoV-2 variants of concern partially escape humoral but not T-cell responses in COVID-19 convalescent donors and vaccinees." Science Immunology 6, no. 59: eabj1750.
Thymic stromal lymphopoietin (TSLP) contributes to asthmatic disease. The concentrations of protective IgA may be reduced in the respiratory tract of asthma patients. We investigated how homeostatic short TSLP (shTSLP) and asthma-associated long TSLP (loTSLP) regulate IgA production. B cells from healthy donors were stimulated in the presence or absence of shTSLP or loTSLP; the concentrations of IgA, IgM, IgE, and IgG antibodies were determined in cell culture supernatants; and B cells were analyzed by flow cytometry. LoTSLP, but not shTSLP, suppressed the secretion of IgA but not of IgE. The type 2 cytokine IL-4, which in addition to loTSLP contributes to asthmatic disease, did not affect the production of IgA or the frequency of IgA+ B cells. Instead, IL-4 increased IgG production, especially of the subclasses IgG2 and IgG4. LoTSLP inhibited IgA secretion by sorted memory B cells but not by naïve B cells. Although loTSLP inhibited IgA production, the vitamin A metabolite retinoic acid promoted the secretion of IgA, also in the presence of loTSLP, suggesting that vitamin A may promote IgA production in asthma. Our data demonstrate that asthma-associated loTSLP negatively regulates the secretion of IgA, which may negatively impact the surveillance of mucosal surfaces in asthma.
Dorianne van Heerden; Robert van Binnendijk; Samantha Tromp; Huub Savelkoul; R. van Neerven; Gerco Den Hartog. Asthma-Associated Long TSLP Inhibits the Production of IgA. International Journal of Molecular Sciences 2021, 22, 3592 .
AMA StyleDorianne van Heerden, Robert van Binnendijk, Samantha Tromp, Huub Savelkoul, R. van Neerven, Gerco Den Hartog. Asthma-Associated Long TSLP Inhibits the Production of IgA. International Journal of Molecular Sciences. 2021; 22 (7):3592.
Chicago/Turabian StyleDorianne van Heerden; Robert van Binnendijk; Samantha Tromp; Huub Savelkoul; R. van Neerven; Gerco Den Hartog. 2021. "Asthma-Associated Long TSLP Inhibits the Production of IgA." International Journal of Molecular Sciences 22, no. 7: 3592.
Background Assessing the duration of immunity following infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a first priority to gauge the degree of protection following infection. Such knowledge is lacking, especially in the general population. Here, we studied changes in immunoglobulin isotype seropositivity and immunoglobulin G (IgG) binding strength of SARS-CoV-2–specific serum antibodies up to 7 months following onset of symptoms in a nationwide sample. Methods Participants from a prospective representative serological study in the Netherlands were included based on IgG seroconversion to the spike S1 protein of SARS-CoV-2 (N = 353), with up to 3 consecutive serum samples per seroconverted participant (N = 738). Immunoglobulin M (IgM), immunoglobulin A (IgA), and IgG antibody concentrations to S1, and increase in IgG avidity in relation to time since onset of disease symptoms, were determined. Results While SARS-CoV-2–specific IgM and IgA antibodies declined rapidly after the first month after disease onset, specific IgG was still present in 92% (95% confidence interval [CI], 89%–95%) of the participants after 7 months. The estimated 2-fold decrease of IgG antibodies was 158 days (95% CI, 136–189 days). Concentrations were sustained better in persons reporting significant symptoms compared to asymptomatic persons or those with mild upper respiratory complaints only. Similarly, avidity of IgG antibodies for symptomatic persons showed a steeper increase over time compared with persons with mild or no symptoms (P = .022). Conclusions SARS-CoV-2–specific IgG antibodies persist and show increasing avidity over time, indicative of underlying immune maturation. These data support development of immune memory against SARS-CoV-2, providing insight into protection of the general unvaccinated part of the population. Clinical Trials Registration NL8473 (the Dutch trial registry).
Gerco Den Hartog; Eric R A Vos; Lotus L Van Den Hoogen; Michiel van Boven; Rutger M Schepp; Gaby Smits; Jeffrey van Vliet; Linde Woudstra; Alienke J Wijmenga-Monsuur; Cheyenne C E van Hagen; Elisabeth A M Sanders; Hester E de Melker; Fiona R M van der Klis; Robert S van Binnendijk. Persistence of Antibodies to Severe Acute Respiratory Syndrome Coronavirus 2 in Relation to Symptoms in a Nationwide Prospective Study. Clinical Infectious Diseases 2021, 1 .
AMA StyleGerco Den Hartog, Eric R A Vos, Lotus L Van Den Hoogen, Michiel van Boven, Rutger M Schepp, Gaby Smits, Jeffrey van Vliet, Linde Woudstra, Alienke J Wijmenga-Monsuur, Cheyenne C E van Hagen, Elisabeth A M Sanders, Hester E de Melker, Fiona R M van der Klis, Robert S van Binnendijk. Persistence of Antibodies to Severe Acute Respiratory Syndrome Coronavirus 2 in Relation to Symptoms in a Nationwide Prospective Study. Clinical Infectious Diseases. 2021; ():1.
Chicago/Turabian StyleGerco Den Hartog; Eric R A Vos; Lotus L Van Den Hoogen; Michiel van Boven; Rutger M Schepp; Gaby Smits; Jeffrey van Vliet; Linde Woudstra; Alienke J Wijmenga-Monsuur; Cheyenne C E van Hagen; Elisabeth A M Sanders; Hester E de Melker; Fiona R M van der Klis; Robert S van Binnendijk. 2021. "Persistence of Antibodies to Severe Acute Respiratory Syndrome Coronavirus 2 in Relation to Symptoms in a Nationwide Prospective Study." Clinical Infectious Diseases , no. : 1.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as a new human pathogen in late 2019 and it has infected over 100 million people in less than a year. There is a clear need for effective antiviral drugs to complement current preventive measures, including vaccines. In this study, we demonstrate that berberine and obatoclax, two broad-spectrum antiviral compounds, are effective against multiple isolates of SARS-CoV-2. Berberine, a plant-derived alkaloid, inhibited SARS-CoV-2 at low micromolar concentrations and obatoclax, which was originally developed as an anti-apoptotic protein antagonist, was effective at sub-micromolar concentrations. Time-of-addition studies indicated that berberine acts on the late stage of the viral life cycle. In agreement, berberine mildly affected viral RNA synthesis, but it strongly reduced infectious viral titers, leading to an increase in the particle-to-pfu ratio. In contrast, obatoclax acted at the early stage of the infection, which is in line with its activity to neutralize the acidic environment in endosomes. We assessed infection of primary human nasal epithelial cells that were cultured on an air-liquid interface and found that SARS-CoV-2 infection induced and repressed expression of specific sets of cytokines and chemokines. Moreover, both obatoclax and berberine inhibited SARS-CoV-2 replication in these primary target cells. We propose berberine and obatoclax as potential antiviral drugs against SARS-CoV-2 that could be considered for further efficacy testing.
Finny Varghese; Esther van Woudenbergh; Gijs Overheul; Marc Eleveld; Lisa Kurver; Niels van Heerbeek; Arjan van Laarhoven; Pascal Miesen; Gerco Den Hartog; Marien de Jonge; Ronald van Rij. Berberine and Obatoclax Inhibit SARS-Cov-2 Replication in Primary Human Nasal Epithelial Cells In Vitro. Viruses 2021, 13, 282 .
AMA StyleFinny Varghese, Esther van Woudenbergh, Gijs Overheul, Marc Eleveld, Lisa Kurver, Niels van Heerbeek, Arjan van Laarhoven, Pascal Miesen, Gerco Den Hartog, Marien de Jonge, Ronald van Rij. Berberine and Obatoclax Inhibit SARS-Cov-2 Replication in Primary Human Nasal Epithelial Cells In Vitro. Viruses. 2021; 13 (2):282.
Chicago/Turabian StyleFinny Varghese; Esther van Woudenbergh; Gijs Overheul; Marc Eleveld; Lisa Kurver; Niels van Heerbeek; Arjan van Laarhoven; Pascal Miesen; Gerco Den Hartog; Marien de Jonge; Ronald van Rij. 2021. "Berberine and Obatoclax Inhibit SARS-Cov-2 Replication in Primary Human Nasal Epithelial Cells In Vitro." Viruses 13, no. 2: 282.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as a new human pathogen in late 2019 and has infected an estimated 10% of the global population in less than a year. There is a clear need for effective antiviral drugs to complement current preventive measures including vaccines. In this study, we demonstrate that berberine and obatoclax, two broad-spectrum antiviral compounds, are effective against multiple isolates of SARS-CoV-2. Berberine, a plant-derived alkaloid, inhibited SARS-CoV-2 at low micromolar concentrations and obatoclax, originally developed as an anti-apoptotic protein antagonist, was effective at sub-micromolar concentrations. Time-of-addition studies indicated that berberine acts on the late stage of the viral life cycle. In agreement, berberine mildly affected viral RNA synthesis, but strongly reduced infectious viral titers, leading to an increase in the particle-to-pfu ratio. In contrast, obatoclax acted at the early stage of the infection, in line with its activity to neutralize the acidic environment in endosomes. We assessed infection of primary human nasal epithelial cells cultured on an air-liquid interface and found that SARS-CoV-2 infection induced and repressed expression of a specific set of cytokines and chemokines. Moreover, both obatoclax and berberine inhibited SARS-CoV-2 replication in these primary target cells. We propose berberine and obatoclax as potential antiviral drugs against SARS-CoV-2 that could be considered for further efficacy testing.
Finny S. Varghese; Esther van Woudenbergh; Gijs J. Overheul; Marc J. Eleveld; Lisa Kurver; Niels van Heerbeek; Arjan van Laarhoven; Pascal Miesen; Gerco Den Hartog; Marien I. de Jonge; Ronald P. van Rij. Berberine and obatoclax inhibit SARS-CoV-2 replication in primary human nasal epithelial cells in vitro. 2020, 1 .
AMA StyleFinny S. Varghese, Esther van Woudenbergh, Gijs J. Overheul, Marc J. Eleveld, Lisa Kurver, Niels van Heerbeek, Arjan van Laarhoven, Pascal Miesen, Gerco Den Hartog, Marien I. de Jonge, Ronald P. van Rij. Berberine and obatoclax inhibit SARS-CoV-2 replication in primary human nasal epithelial cells in vitro. . 2020; ():1.
Chicago/Turabian StyleFinny S. Varghese; Esther van Woudenbergh; Gijs J. Overheul; Marc J. Eleveld; Lisa Kurver; Niels van Heerbeek; Arjan van Laarhoven; Pascal Miesen; Gerco Den Hartog; Marien I. de Jonge; Ronald P. van Rij. 2020. "Berberine and obatoclax inhibit SARS-CoV-2 replication in primary human nasal epithelial cells in vitro." , no. : 1.
Background We aimed to detect SARS-CoV-2 serum antibodies in the general population of the Netherlands and identify risk factors for seropositivity amidst the first COVID-19 epidemic wave. Methods Participants (n=3207, aged 2–90 years), enrolled from a previously established nationwide serosurveillance study, provided a self-collected fingerstick blood sample and completed a questionnaire (median inclusion date 3 April 2020). IgG antibodies targeted against the spike S1-protein of SARS-CoV-2 were quantified using a validated multiplex-immunoassay. Seroprevalence was estimated controlling for survey design, individual pre-pandemic concentration, and test performance. Random-effects logistic regression identified risk factors for seropositivity. Results Overall seroprevalence in the Netherlands was 2.8% (95% CI 2.1 to 3.7), with no differences between sexes or ethnic background, and regionally ranging between 1.3 and 4.0%. Estimates were highest among 18–39 year-olds (4.9%), and lowest in children 2–17 years (1.7%). Multivariable analysis revealed that persons taking immunosuppressants and those from the Orthodox-Reformed Protestant community had over four times higher odds of being seropositive compared to others. Anosmia/ageusia was the most discriminative symptom between seropositive (53%) and seronegative persons (4%, p<0.0001). Antibody concentrations in seropositive persons were significantly higher in those with fever or dyspnoea in contrast to those without (p=0.01 and p=0.04, respectively). Conclusions In the midst of the first epidemic wave, 2.8% of the Dutch population was estimated to be infected with SARS-CoV-2, that is, 30 times higher than reported. This study identified independent groups with increased odds for seropositivity that may require specific surveillance measures to guide future protective interventions internationally, including vaccination once available.
Eric R A Vos; Gerco Den Hartog; Rutger M Schepp; Patricia Kaaijk; Jeffrey van Vliet; Kina Helm; Gaby Smits; Alienke Wijmenga-Monsuur; Janneke D M Verberk; Michiel van Boven; Rob S van Binnendijk; Hester E de Melker; Liesbeth Mollema; Fiona R M van der Klis. Nationwide seroprevalence of SARS-CoV-2 and identification of risk factors in the general population of the Netherlands during the first epidemic wave. Journal of Epidemiology & Community Health 2020, 75, 489 -495.
AMA StyleEric R A Vos, Gerco Den Hartog, Rutger M Schepp, Patricia Kaaijk, Jeffrey van Vliet, Kina Helm, Gaby Smits, Alienke Wijmenga-Monsuur, Janneke D M Verberk, Michiel van Boven, Rob S van Binnendijk, Hester E de Melker, Liesbeth Mollema, Fiona R M van der Klis. Nationwide seroprevalence of SARS-CoV-2 and identification of risk factors in the general population of the Netherlands during the first epidemic wave. Journal of Epidemiology & Community Health. 2020; 75 (6):489-495.
Chicago/Turabian StyleEric R A Vos; Gerco Den Hartog; Rutger M Schepp; Patricia Kaaijk; Jeffrey van Vliet; Kina Helm; Gaby Smits; Alienke Wijmenga-Monsuur; Janneke D M Verberk; Michiel van Boven; Rob S van Binnendijk; Hester E de Melker; Liesbeth Mollema; Fiona R M van der Klis. 2020. "Nationwide seroprevalence of SARS-CoV-2 and identification of risk factors in the general population of the Netherlands during the first epidemic wave." Journal of Epidemiology & Community Health 75, no. 6: 489-495.
The spike protein of the pandemic human Corona virus is essential for virus entry into human cells. In fact, most neutralizing antibodies against Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) are directed against the Virus-surface exposed spike protein, making it the antigen of choice for use in vaccines and diagnostic tests. In the current pandemic context, global demand for spike proteins has rapidly increased and could exceed hundreds of grams to kilograms annually. Coronavirus spikes are large heavily glycosylated homo-trimeric complexes, with inherent instability. The poor manufacturability now threatens the availability of these proteins for vaccines and diagnostic tests. Here, we outline scalable, Good Manufacturing Practice (GMP) compliant, and chemically defined processes for the production of two cell-secreted stabilized forms of the trimeric spike proteins (Wuhan and D614G variant). The processes are chemically defined and based on clonal suspension-CHO cell populations and on protein purification via a two-step scalable downstream process. The trimeric conformation was confirmed using electron microscopy and HPLC analysis. Binding to susceptible cells was shown using a virus-inhibition assay. The diagnostic sensitivity and specificity for detection of serum SARS-CoV-2-specific-immunoglobulin molecules was found to exceed that of spike fragments (Spike fragment-1, S1 and Receptor Binding Domain, RBD). The process described here will enable production of sufficient high-quality trimeric spike protein to meet the global demand for SARS-CoV-2 diagnostic tests and potentially vaccines.
Paco Pino; Joeri Kint; Divor Kiseljak; Valentina Agnolon; Giampietro Corradin; Andrey Kajava; Paolo Rovero; Ronald Dijkman; Gerco Den Hartog; Jason McLellan; Patrick Byrne; Maria Wurm; Florian Wurm. Trimeric SARS-CoV-2 Spike Proteins Produced from CHO Cells in Bioreactors are High-Quality Antigens. Processes 2020, 8, 1539 .
AMA StylePaco Pino, Joeri Kint, Divor Kiseljak, Valentina Agnolon, Giampietro Corradin, Andrey Kajava, Paolo Rovero, Ronald Dijkman, Gerco Den Hartog, Jason McLellan, Patrick Byrne, Maria Wurm, Florian Wurm. Trimeric SARS-CoV-2 Spike Proteins Produced from CHO Cells in Bioreactors are High-Quality Antigens. Processes. 2020; 8 (12):1539.
Chicago/Turabian StylePaco Pino; Joeri Kint; Divor Kiseljak; Valentina Agnolon; Giampietro Corradin; Andrey Kajava; Paolo Rovero; Ronald Dijkman; Gerco Den Hartog; Jason McLellan; Patrick Byrne; Maria Wurm; Florian Wurm. 2020. "Trimeric SARS-CoV-2 Spike Proteins Produced from CHO Cells in Bioreactors are High-Quality Antigens." Processes 8, no. 12: 1539.
Background Respiratory syncytial virus (RSV) generally causes mild disease but can cause severe infections in (premature) infants and elderly adults. Here, we studied RSV-specific antibody concentrations throughout life with emphasis on infants and chronic obstructive pulmonary disease (COPD) patients. Methods Sera (N = 2655) from 2 nationwide cross-sectional studies in the Netherlands including individuals aged 0–90 years were analyzed for IgG and IgA antibodies to RSV prefusion F, postfusion F, N, Ga, and Gb proteins and for antibody avidity in 42 COPD patients. Results Maternal IgG concentrations declined to age 10–12 months. After the first year of life, approximately 40% of children lacked infection-induced IgA antibodies and may therefore be uninfected. All Dutch children showed serological evidence of RSV infection by age 3 years. Antibody concentrations reached a plateau by age 5–9 years and remains constant throughout life. COPD patients had similar levels and avidity of RSV-specific IgG antibodies compared with age-matched healthy controls. Conclusions RSV-IgG antibody patterns throughout life can be used to estimate the degree of immunity acquisition to RSV and to identify groups at increased risk of infection. Seroprevalence of IgA could be a proxy to determine RSV infection in children younger than 1 year.
Guy Berbers; Liesbeth Mollema; Fiona Van Der Klis; Gerco Den Hartog; Rutger Schepp. Antibody Responses to Respiratory Syncytial Virus: A Cross-Sectional Serosurveillance Study in the Dutch Population Focusing on Infants Younger Than 2 Years. The Journal of Infectious Diseases 2020, 224, 269 -278.
AMA StyleGuy Berbers, Liesbeth Mollema, Fiona Van Der Klis, Gerco Den Hartog, Rutger Schepp. Antibody Responses to Respiratory Syncytial Virus: A Cross-Sectional Serosurveillance Study in the Dutch Population Focusing on Infants Younger Than 2 Years. The Journal of Infectious Diseases. 2020; 224 (2):269-278.
Chicago/Turabian StyleGuy Berbers; Liesbeth Mollema; Fiona Van Der Klis; Gerco Den Hartog; Rutger Schepp. 2020. "Antibody Responses to Respiratory Syncytial Virus: A Cross-Sectional Serosurveillance Study in the Dutch Population Focusing on Infants Younger Than 2 Years." The Journal of Infectious Diseases 224, no. 2: 269-278.
Background The COVID-19 pandemic necessitates better understanding of the kinetics of antibody production induced by infection with SARS-CoV-2. We aimed to develop a high-throughput multiplex assay to detect antibodies to SARS-CoV-2 to assess immunity to the virus in the general population. Methods Spike protein subunits S1 and receptor binding domain, and nucleoprotein were coupled to microspheres. Sera collected before emergence of SARS-CoV-2 (n = 224) and of non-SARS-CoV-2 influenza-like illness (n = 184), and laboratory-confirmed cases of SARS-CoV-2 infection (n = 115) with various severities of COVID-19 were tested for SARS-CoV-2–specific IgG concentrations. Results Our assay discriminated SARS-CoV-2–induced antibodies and those induced by other viruses. The assay specificity was 95.1%–99.0% with sensitivity 83.6%–95.7%. By merging the test results for all 3 antigens a specificity of 100% was achieved with a sensitivity of at least 90%. Hospitalized COVID-19 patients developed higher IgG concentrations and the rate of IgG production increased faster compared to nonhospitalized cases. Conclusions The bead-based serological assay for quantitation of SARS-CoV-2–specific antibodies proved to be robust and can be conducted in many laboratories. We demonstrated that testing of antibodies against multiple antigens increases sensitivity and specificity compared to single-antigen–specific IgG determination.
Gerco Den Hartog; Rutger M Schepp; Marjan Kuijer; Corine GeurtsvanKessel; Josine van Beek; Nynke Rots; Marion P G Koopmans; Fiona R M van der Klis; Robert S van Binnendijk. SARS-CoV-2–Specific Antibody Detection for Seroepidemiology: A Multiplex Analysis Approach Accounting for Accurate Seroprevalence. Journal of Infectious Diseases 2020, 222, 1452 -1461.
AMA StyleGerco Den Hartog, Rutger M Schepp, Marjan Kuijer, Corine GeurtsvanKessel, Josine van Beek, Nynke Rots, Marion P G Koopmans, Fiona R M van der Klis, Robert S van Binnendijk. SARS-CoV-2–Specific Antibody Detection for Seroepidemiology: A Multiplex Analysis Approach Accounting for Accurate Seroprevalence. Journal of Infectious Diseases. 2020; 222 (9):1452-1461.
Chicago/Turabian StyleGerco Den Hartog; Rutger M Schepp; Marjan Kuijer; Corine GeurtsvanKessel; Josine van Beek; Nynke Rots; Marion P G Koopmans; Fiona R M van der Klis; Robert S van Binnendijk. 2020. "SARS-CoV-2–Specific Antibody Detection for Seroepidemiology: A Multiplex Analysis Approach Accounting for Accurate Seroprevalence." Journal of Infectious Diseases 222, no. 9: 1452-1461.
BackgroundThe COVID-19 pandemic demands detailed understanding of the kinetics of antibody production induced by infection with SARS-CoV-2. We aimed to develop a high throughput multiplex assay to detect antibodies to SARS-CoV-2 to assess immunity to the virus in the general population.MethodsSpike protein subunits S1 and RBD, and Nucleoprotein were coupled to distinct microspheres. Sera collected before the emergence of SARS-CoV-2 (N=224), and of non-SARS-CoV-2 influenza-like illness (N=184), and laboratory-confirmed cases of SARS-CoV-2 infection (N=115) with various severity of COVID-19 were tested for SARS-CoV-2-specific concentrations of IgG.ResultsOur assay discriminated SARS-CoV-2-induced antibodies and those induced by other viruses. The assay obtained a specificity between 95.1 and 99.0% with a sensitivity ranging from 83.6-95.7%. By merging the test results for all 3 antigens a specificity of 100% was achieved with a sensitivity of at least 90%. Hospitalized COVID-19 patients developed higher IgG concentrations and the rate of IgG production increased faster compared to non-hospitalized cases.ConclusionsThe bead-based serological assay for quantitation of SARS-CoV-2-specific antibodies proved to be robust and can be conducted in many laboratories. Finally, we demonstrated that testing of antibodies against different antigens increases sensitivity and specificity compared to single antigen-specific IgG determination.
Gerco Den Hartog; Rutger M. Schepp; Marjan Kuijer; Corine GeurtsvanKessel; Josine Van Beek; Nynke Rots; Marion P.G. Koopmans; Fiona R.M. Van Der Klis; Robert S. Van Binnendijk. SARS-CoV-2-specific antibody detection for sero-epidemiology: a multiplex analysis approach accounting for accurate seroprevalence. 2020, 1 .
AMA StyleGerco Den Hartog, Rutger M. Schepp, Marjan Kuijer, Corine GeurtsvanKessel, Josine Van Beek, Nynke Rots, Marion P.G. Koopmans, Fiona R.M. Van Der Klis, Robert S. Van Binnendijk. SARS-CoV-2-specific antibody detection for sero-epidemiology: a multiplex analysis approach accounting for accurate seroprevalence. . 2020; ():1.
Chicago/Turabian StyleGerco Den Hartog; Rutger M. Schepp; Marjan Kuijer; Corine GeurtsvanKessel; Josine Van Beek; Nynke Rots; Marion P.G. Koopmans; Fiona R.M. Van Der Klis; Robert S. Van Binnendijk. 2020. "SARS-CoV-2-specific antibody detection for sero-epidemiology: a multiplex analysis approach accounting for accurate seroprevalence." , no. : 1.
Introduction: Immunesurveillance is an important tool to monitor the protection of the population against vaccine-preventable diseases, which is currently mostly based on the detection of specific serum antibodies. However, the landscape of immune surveillance is changing, driven by emerging and evolving pathogens, changes in the age distribution of the population and scientific understanding of protective immunity, necessitating a comprehensive review. Areas covered: To anticipate these changes, reliable and high-throughput detection of antibody levels is desired to enable screening in larger population settings. Antibody levels alone do not always equate with protection and may require additional functional testing of the antibodies or immune cell-based assays. In addition, the location (systemic or locally mucosal) of the infection and whether the antibodies are induced through infection or vaccination have implications for both immune protection and assessing immune status. Expert commentary: In order to perform multicenter studies on many samples for multiple antigens, more validated reference materials and wider adoption of high-throughput techniques are needed. The field of serosurveillance will also benefit from better correlates of protection and understanding of (local) mechanisms of protection. Here we give an overview of the current state-of-the-art of serosurveillance and how the field could move forward.
Gerco Den Hartog; Rob van Binnendijk; Anne-Marie Buisman; Guy A. M. Berbers; Fiona R. M. van der Klis. Immune surveillance for vaccine-preventable diseases. Expert Review of Vaccines 2020, 19, 327 -339.
AMA StyleGerco Den Hartog, Rob van Binnendijk, Anne-Marie Buisman, Guy A. M. Berbers, Fiona R. M. van der Klis. Immune surveillance for vaccine-preventable diseases. Expert Review of Vaccines. 2020; 19 (4):327-339.
Chicago/Turabian StyleGerco Den Hartog; Rob van Binnendijk; Anne-Marie Buisman; Guy A. M. Berbers; Fiona R. M. van der Klis. 2020. "Immune surveillance for vaccine-preventable diseases." Expert Review of Vaccines 19, no. 4: 327-339.
Background Whooping cough is caused by infection of the airways with Bordetella pertussis (Bp). As interferon gamma (IFN-γ) is essential for protective immunity against Bp, we investigated how IFN-γ is induced by Bp or the virulence antigens filamentous hemagglutinin adhesin, pertactin, or pertussis toxin, and how IFN-γ contributes to local immune responses in humans. Methods Peripheral blood mononuclear cells (PBMCs) from healthy donors and/or respiratory epithelial cells were stimulated with soluble antigens or inactivated intact Bp and the presence or absence of blocking antibodies or chemokines. Supernatants and cells were analyzed for IFN-γ and chemokine production, and lymphocyte migration was tested using epithelial supernatants. Results The soluble antigens failed to induce IFN-γ production, whereas inactivated Bp induced IFN-γ production. Natural killer (NK) cells were the main source of IFN-γ production, which was enhanced by interleukin 15. Epithelial–PBMC co-cultures showed robust IFN-γ–dependent CXCL9 and CXCL10 production by the epithelial cells following stimulation with IFN-γ and Bp. The epithelial-derived chemokines resulted in CXCR3-dependent recruitment of NK and T cells. Conclusions Inactivated Bp, but not antigens, induced potent IFN-γ production by NK cells, resulting in chemoattraction of lymphocytes toward the respiratory epithelium. These data provide insight into the requirements for IFN-γ production and how IFN-γ enhances local immune responses to prevent Bp-mediated disease.
Gerco Den Hartog; Marcel A Schijf; Guy A M Berbers; Fiona R M Van Der Klis; Anne-Marie Buisman. Bordetella pertussis Induces Interferon Gamma Production by Natural Killer Cells, Resulting in Chemoattraction by Respiratory Epithelial Cells. Journal of Infectious Diseases 2020, 1 .
AMA StyleGerco Den Hartog, Marcel A Schijf, Guy A M Berbers, Fiona R M Van Der Klis, Anne-Marie Buisman. Bordetella pertussis Induces Interferon Gamma Production by Natural Killer Cells, Resulting in Chemoattraction by Respiratory Epithelial Cells. Journal of Infectious Diseases. 2020; ():1.
Chicago/Turabian StyleGerco Den Hartog; Marcel A Schijf; Guy A M Berbers; Fiona R M Van Der Klis; Anne-Marie Buisman. 2020. "Bordetella pertussis Induces Interferon Gamma Production by Natural Killer Cells, Resulting in Chemoattraction by Respiratory Epithelial Cells." Journal of Infectious Diseases , no. : 1.
Latent infection with Cytomegalovirus (CMV) or Epstein-Bar virus (EBV) is associated with compromised immune responses. The commercially available ELISA-kits provide a semi-quantitative or qualitative detection of IgG antibodies against either CMV or EBV. To reduce the amount of sample needed and improve throughput and range of quantitation compared to ELISAs, a multiplex immunoassay (MIA) for the simultaneous quantitative detection of antibodies against CMV and two antigens of EBV (EBV capsid antigen (EBV-VCA) and EBV nuclear antigen 1 (EBNA-1)) was developed and standardized. Our assay shows a good correlation with the separate ELISAs, has a high specificity and is much more sensitive than the ELISAs. The MIA allows faster analysis of samples and can be combined with the MIA for detection of antibodies to measles, mumps, rubella and varicella zoster (MMRV) as published previously. In conclusion, the CMV-EBV MIA is a reliable assay that allows for more efficient detection of specific antibodies to CMV and EBV. This MIA therefore is especially useful for large-scale surveillance studies.
Irina Tcherniaeva; Gerco Den Hartog; Guy Berbers; Fiona van der Klis. The development of a bead-based multiplex immunoassay for the detection of IgG antibodies to CMV and EBV. Journal of Immunological Methods 2018, 462, 1 -8.
AMA StyleIrina Tcherniaeva, Gerco Den Hartog, Guy Berbers, Fiona van der Klis. The development of a bead-based multiplex immunoassay for the detection of IgG antibodies to CMV and EBV. Journal of Immunological Methods. 2018; 462 ():1-8.
Chicago/Turabian StyleIrina Tcherniaeva; Gerco Den Hartog; Guy Berbers; Fiona van der Klis. 2018. "The development of a bead-based multiplex immunoassay for the detection of IgG antibodies to CMV and EBV." Journal of Immunological Methods 462, no. : 1-8.
Lindsay D. Butcher; Gerco Den Hartog; Amber Ablack; Mason Matsubara; Jailal Ablack; Taren Thron; Sarkis Mazmanian; Lars Eckmann; Peter Ernst; Tadahide Izumi; Sheila E. Crowe. 149 - Gastrointestinal Microbes Enhance the Accumulation of Reactive Oxygen Species (ROS) and Dna Damage that are Regulated by Apurinic/Apyrimidinic Endonuclease 1 (APE1). Gastroenterology 2018, 154, S-41 -41.
AMA StyleLindsay D. Butcher, Gerco Den Hartog, Amber Ablack, Mason Matsubara, Jailal Ablack, Taren Thron, Sarkis Mazmanian, Lars Eckmann, Peter Ernst, Tadahide Izumi, Sheila E. Crowe. 149 - Gastrointestinal Microbes Enhance the Accumulation of Reactive Oxygen Species (ROS) and Dna Damage that are Regulated by Apurinic/Apyrimidinic Endonuclease 1 (APE1). Gastroenterology. 2018; 154 (6):S-41-41.
Chicago/Turabian StyleLindsay D. Butcher; Gerco Den Hartog; Amber Ablack; Mason Matsubara; Jailal Ablack; Taren Thron; Sarkis Mazmanian; Lars Eckmann; Peter Ernst; Tadahide Izumi; Sheila E. Crowe. 2018. "149 - Gastrointestinal Microbes Enhance the Accumulation of Reactive Oxygen Species (ROS) and Dna Damage that are Regulated by Apurinic/Apyrimidinic Endonuclease 1 (APE1)." Gastroenterology 154, no. 6: S-41-41.
The gastrointestinal barrier is more than a tissue with physiological function. As reviewed by others, it is now recognized as a complex ecosystem that requires a delicate homeostatic balance in order for the tissue to remain healthy and functional. The niche involves a multifaceted and dynamic mixture of host cells and molecules; microbes and their metabolites; as well as environmental factors that may reflect diet or contamination with toxins. Furthermore, gene expression in the host can modulate transcription in microbes resulting in changes in the metabolic profile and local immune responses that resonate throughout the body. Thus, these biological spheres are highly interconnected. A more recent and important concept is that the microbial-host interactions that begin in the digestive tract not only impact gastrointestinal inflammation and risks of local cancer, but they also affect the hosťs susceptibility to conditions ranging from diabetes, obesity to various neurological conditions. This chapter summarizes the key elements whereby the gut barrier contributes to innate immunity in the digestive tract.
Gerco Den Hartog; Sheila E. Crowe; Soumita Das; Peter B. Ernst. Gut Barrier: Innate Immunity. Physiology of the Gastrointestinal Tract 2018, 663 -670.
AMA StyleGerco Den Hartog, Sheila E. Crowe, Soumita Das, Peter B. Ernst. Gut Barrier: Innate Immunity. Physiology of the Gastrointestinal Tract. 2018; ():663-670.
Chicago/Turabian StyleGerco Den Hartog; Sheila E. Crowe; Soumita Das; Peter B. Ernst. 2018. "Gut Barrier: Innate Immunity." Physiology of the Gastrointestinal Tract , no. : 663-670.
Class‐switching of B cells to IgA can be induced via both T‐cell‐dependent and T‐cell‐independent mechanisms. IgA is most predominantly produced mucosally and is important for combating infections and allergies. In contrast to mice, humans have two forms of IgA; IgA1 and IgA2 with diverse tissue distribution. In early life, IgA levels might be sub‐optimal especially during the fall season when bacterial and viral infections are more common. Therefore, we investigated using human B cells whether T‐cell‐independent factors ‐promoting cell survival, class switching and immunoglobulin secretion‐ BAFF, APRIL, IL‐10 and retinoic acid can boost IgA production in the context of viral or bacterial infection. To this end total and naive peripheral blood B cells were stimulated with these factors for 6 days in the presence or absence of TLR7/8 agonist R848 (mimicking viral infection) or TLR9 agonist CpG‐ODN (mimicking bacterial infection). We show that BAFF significantly augments IgA2 production in TLR7/8 stimulated mature, but not naïve B cells. In addition, BAFF augments IL‐10 production and viability in TLR7/8 and TLR9 stimulated mature B cells. These data warrant further investigation of its role in immune regulation both in the periphery and mucosal tissues in early life or during disease.
Gerco Den Hartog; Thijs L.J. Van Osch; Martijn Vos; Ben Meijer; Huub F.J. Savelkoul; R.J. Joost Van Neerven; Sylvia Brugman. BAFF augments IgA2 and IL‐10 production by TLR7/8 stimulated total peripheral blood B cells. European Journal of Immunology 2017, 48, 283 -292.
AMA StyleGerco Den Hartog, Thijs L.J. Van Osch, Martijn Vos, Ben Meijer, Huub F.J. Savelkoul, R.J. Joost Van Neerven, Sylvia Brugman. BAFF augments IgA2 and IL‐10 production by TLR7/8 stimulated total peripheral blood B cells. European Journal of Immunology. 2017; 48 (2):283-292.
Chicago/Turabian StyleGerco Den Hartog; Thijs L.J. Van Osch; Martijn Vos; Ben Meijer; Huub F.J. Savelkoul; R.J. Joost Van Neerven; Sylvia Brugman. 2017. "BAFF augments IgA2 and IL‐10 production by TLR7/8 stimulated total peripheral blood B cells." European Journal of Immunology 48, no. 2: 283-292.
Helicobacter pylori is a gram-negative, microaerophilic bacterium that infects the stomach and can lead to, among other disorders, the development of gastric cancer. The inability of the host to clear the infection results in a chronic inflammatory state with continued oxidative stress within the tissue. Reactive oxygen species and reactive nitrogen species produced by the immune and epithelial cells damage the host cells and can result in DNA damage. H pylori has evolved to evoke this damaging response while blunting the host's efforts to kill the bacteria. This long-lasting state with inflammation and oxidative stress can result in gastric carcinogenesis. Continued efforts to better understand the bacterium and the host response will serve to prevent or provide improved early diagnosis and treatment of gastric cancer.
Lindsay D. Butcher; Gerco Den Hartog; Peter Ernst; Sheila E. Crowe. Oxidative Stress Resulting From Helicobacter pylori Infection Contributes to Gastric Carcinogenesis. Cellular and Molecular Gastroenterology and Hepatology 2017, 3, 316 -322.
AMA StyleLindsay D. Butcher, Gerco Den Hartog, Peter Ernst, Sheila E. Crowe. Oxidative Stress Resulting From Helicobacter pylori Infection Contributes to Gastric Carcinogenesis. Cellular and Molecular Gastroenterology and Hepatology. 2017; 3 (3):316-322.
Chicago/Turabian StyleLindsay D. Butcher; Gerco Den Hartog; Peter Ernst; Sheila E. Crowe. 2017. "Oxidative Stress Resulting From Helicobacter pylori Infection Contributes to Gastric Carcinogenesis." Cellular and Molecular Gastroenterology and Hepatology 3, no. 3: 316-322.
Background:Inhibition assays are an useful tool to identify the allergen of primary sensitization of cross-reactive allergens. Classical ELISA-based inhibition assays are limited by both the availability of commercial standardized allergen extracts and the experience and knowledge needed for making home-made extracts. Moreover the direct comparison of the inhibition ELISAs outcomes between different laboratories is difficult because of different sources of used allergen extracts and a number of methodological variations. Therefore, we propose a novel ImmunoCap (Phadia, Thermofisher Scientific) based immunoinhibition method with the use of commercially available Caps as the allergen source.Methods:The novel ImmunoCap based immunoinhibition method was developed and tested with sera from patients with a well-known cross-reactive sensitization for fig (Results:The amount of allergens (fig and ficus extracts) needed to reach a similar degree of inhibition was comparable for both inhibition methods.Conclusions:The ImmunoCap based inhibition assay, in addition to classical inhibition methods, is a valuable tool as the ImmunoCap analyzer and commercial allergens (Caps) are more widely available which makes the outcomes of inhibition tests comparable between different laboratories. Furthermore, in the ImmunoCap inhibition method the same protein source is used for both the inhibition of sIgE and sIgE measurement, which might be even more relevant when multiple cross-reactive allergens are tested.
Yvonne Schmidt-Hieltjes; Malgorzata Teodorowicz; Ad Jansen; Gerco Den Hartog; Lisette Elfvering-Berendsen; Nicolette W. De Jong; Huub F.J. Savelkoul; Janneke Ruinemans-Koerts. An alternative inhibition method for determining cross-reactive allergens. Clinical Chemistry and Laboratory Medicine (CCLM) 2017, 55, 248 -253.
AMA StyleYvonne Schmidt-Hieltjes, Malgorzata Teodorowicz, Ad Jansen, Gerco Den Hartog, Lisette Elfvering-Berendsen, Nicolette W. De Jong, Huub F.J. Savelkoul, Janneke Ruinemans-Koerts. An alternative inhibition method for determining cross-reactive allergens. Clinical Chemistry and Laboratory Medicine (CCLM). 2017; 55 (2):248-253.
Chicago/Turabian StyleYvonne Schmidt-Hieltjes; Malgorzata Teodorowicz; Ad Jansen; Gerco Den Hartog; Lisette Elfvering-Berendsen; Nicolette W. De Jong; Huub F.J. Savelkoul; Janneke Ruinemans-Koerts. 2017. "An alternative inhibition method for determining cross-reactive allergens." Clinical Chemistry and Laboratory Medicine (CCLM) 55, no. 2: 248-253.