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  1. Article: The nearest neighbor nuclei method to objectify analysis of pertussis toxin-induced clustering.

    Hoonakker, Marieke E / Remarque, Ed / Veth, Jennifer / Sloots, Arjen / Bajramovic, Jeffrey J

    ALTEX

    2021  Volume 39, Issue 1, Page(s) 140–148

    Abstract: The in vivo histamine sensitization test (HIST) has historically been performed to guarantee the safety of acellular per­tussis vaccine batches. Non-compliance of batches is primarily associated with the presence of low levels of pertussis toxin (PTx). ... ...

    Abstract The in vivo histamine sensitization test (HIST) has historically been performed to guarantee the safety of acellular per­tussis vaccine batches. Non-compliance of batches is primarily associated with the presence of low levels of pertussis toxin (PTx). Because of ethical, standardization and scientific reasons, a variety of alternative in vitro approaches have been studied to replace the lethal HIST. A broadly applied and partially accepted method is the CHO cell clustering test, which is based on the clustered growth pattern of CHO cells when exposed to minute amounts of PTx. One of the major hurdles for global application of the CHO clustering test is the manual assessment of the clusters, which is associated with suboptimal reproducibility of test outcomes and is time-consuming. Here, various parameters of CHO cell nuclei were evaluated in search for a reliable, objective read-out parameter. We demonstrate that the distance between each nucleus and its nearest neighbor (3N method) is the most suitable parameter to assess clustered cell growth. This method detects 2.8 mIU PTx/mL and thereby complies with the requirement set for the sensitivity of the CHO clustering test based on visual reading. In commercial acellular pertussis vaccines spiked with PTx, the method detects 45 mIU/mL PTx, which is substantially lower than the 181-725 mIU/mL PTx detected by visual interpretation. The 3N method thus allows objective and sensitive assessment of CHO clustering and thereby encourages broad and global implementation of the in vitro test as an alternative to the HIST.
    MeSH term(s) Animal Testing Alternatives ; Animals ; Cell Nucleus ; Cluster Analysis ; Cricetinae ; Cricetulus ; Pertussis Toxin ; Reproducibility of Results ; Vaccines, Acellular
    Chemical Substances Vaccines, Acellular ; Pertussis Toxin (EC 2.4.2.31)
    Language English
    Publishing date 2021-10-15
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 165707-0
    ISSN 1868-596X ; 1018-4562 ; 0946-7785
    ISSN 1868-596X ; 1018-4562 ; 0946-7785
    DOI 10.14573/altex.2012171
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Reporter cell lines for detection of pertussis toxin in acellular pertussis vaccines as a functional animal-free alternative to the in vivo histamine sensitization test.

    Hoonakker, Marieke E / Verhagen, Lisa M / van der Maas, Larissa / Sloots, Arjen / Hendriksen, Coenraad F M

    Vaccine

    2017  Volume 35, Issue 8, Page(s) 1152–1160

    Abstract: Detoxified pertussis toxin (pertussis toxoid) is a major antigen in acellular pertussis vaccines. Testing these vaccines on the presence of residual pertussis toxin (PTx) and reversion to toxicity is performed by the regulatory required in vivo Histamine ...

    Abstract Detoxified pertussis toxin (pertussis toxoid) is a major antigen in acellular pertussis vaccines. Testing these vaccines on the presence of residual pertussis toxin (PTx) and reversion to toxicity is performed by the regulatory required in vivo Histamine Sensitization test (HIST). Lack of mechanistic understanding of the HIST, technical handicaps and animal welfare concerns, have promoted the development of alternative methods. As the majority of the cellular effects of PTx depend on its ability to activate intracellular pathways involving cAMP, the in vitro cAMP-PTx assay was developed. Although this assay could be used to detect PTx activity, it lacked sensitivity and robustness for use in a quality control setting. In the present study, novel reporter cell lines (CHO-CRE and A10-CRE) were generated that stably express a reporter construct responsive to changes in intracellular cAMP levels. These reporter cell lines were able to detect PTx in a concentration-dependent manner when combined with fixed amounts of forskolin. The CHO-CRE cell line enabled detection of PTx in the context of a multivalent vaccine containing aP, with a sensitivity equal to the HIST. However, the sensitivity of the A10-CRE cells was insufficient for this purpose. The experiments also suggest that the CHO-CRE reporter cell line might be suitable for assessment of cellular effects of PTd reverted to PTx. The CHO-CRE reporter cell line provides a platform that meets the criteria for specificity and sensitivity and is a promising in vitro model with potential to replace the HIST.
    Language English
    Publishing date 2017-02-22
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 605674-x
    ISSN 1873-2518 ; 0264-410X
    ISSN (online) 1873-2518
    ISSN 0264-410X
    DOI 10.1016/j.vaccine.2017.01.011
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1930: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
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  3. Article: Reporter cell lines for detection of pertussis toxin in acellular pertussis vaccines as a functional animal-free alternative to the in vivo histamine sensitization test

    Hoonakker, Marieke E / Arjen Sloots / Coenraad F.M. Hendriksen / Larissa van der Maas / Lisa M. Verhagen

    Vaccine. 2017 Feb. 22, v. 35, no. 8

    2017  

    Abstract: Detoxified pertussis toxin (pertussis toxoid) is a major antigen in acellular pertussis vaccines. Testing these vaccines on the presence of residual pertussis toxin (PTx) and reversion to toxicity is performed by the regulatory required in vivo Histamine ...

    Abstract Detoxified pertussis toxin (pertussis toxoid) is a major antigen in acellular pertussis vaccines. Testing these vaccines on the presence of residual pertussis toxin (PTx) and reversion to toxicity is performed by the regulatory required in vivo Histamine Sensitization test (HIST). Lack of mechanistic understanding of the HIST, technical handicaps and animal welfare concerns, have promoted the development of alternative methods. As the majority of the cellular effects of PTx depend on its ability to activate intracellular pathways involving cAMP, the in vitro cAMP-PTx assay was developed. Although this assay could be used to detect PTx activity, it lacked sensitivity and robustness for use in a quality control setting. In the present study, novel reporter cell lines (CHO-CRE and A10-CRE) were generated that stably express a reporter construct responsive to changes in intracellular cAMP levels. These reporter cell lines were able to detect PTx in a concentration-dependent manner when combined with fixed amounts of forskolin. The CHO-CRE cell line enabled detection of PTx in the context of a multivalent vaccine containing aP, with a sensitivity equal to the HIST. However, the sensitivity of the A10-CRE cells was insufficient for this purpose. The experiments also suggest that the CHO-CRE reporter cell line might be suitable for assessment of cellular effects of PTd reverted to PTx. The CHO-CRE reporter cell line provides a platform that meets the criteria for specificity and sensitivity and is a promising in vitro model with potential to replace the HIST.
    Keywords animal welfare ; antigens ; cyclic AMP ; forskolin ; histamine ; models ; pertussis toxin ; quality control ; toxicity ; vaccines
    Language English
    Dates of publication 2017-0222
    Size p. 1152-1160.
    Publishing place Elsevier Ltd
    Document type Article
    ZDB-ID 605674-x
    ISSN 1873-2518 ; 0264-410X
    ISSN (online) 1873-2518
    ISSN 0264-410X
    DOI 10.1016/j.vaccine.2017.01.011
    Database NAL-Catalogue (AGRICOLA)

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    In stock of ZB MED Cologne/Königswinter

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  4. Article ; Online: The cAMP assay: a functional in vitro alternative to the in vivo Histamine Sensitization test.

    Hoonakker, Marieke E / Ruiterkamp, Nicole / Hendriksen, Coenraad F M

    Vaccine

    2010  Volume 28, Issue 5, Page(s) 1347–1352

    Abstract: Safety requirements stipulate the performance of the in vivo Histamine Sensitization (HS) test for quality control of acellular pertussis (aP) vaccines. For reasons of reproducibility and animal welfare concern, an in vitro assay was developed. The assay ...

    Abstract Safety requirements stipulate the performance of the in vivo Histamine Sensitization (HS) test for quality control of acellular pertussis (aP) vaccines. For reasons of reproducibility and animal welfare concern, an in vitro assay was developed. The assay reflects the mechanism of histamine sensitization and is based on cAMP production in A10 cells to residual pertussis toxin (PT). We showed that PT induces cAMP levels in a dose-dependent manner while the sensitivity of the assay equals the sensitivity of the HS test. Neither the individual components nor the combination vaccine DTaP-IP did affect the assay. The cAMP assay meets the criteria for specificity and sensitivity and therefore might be a promising candidate to replace the HS test.
    MeSH term(s) Animals ; Cyclic AMP/analysis ; Dose-Response Relationship, Drug ; Histamine/analysis ; Humans ; Pertussis Toxin/analysis ; Pertussis Toxin/pharmacology ; Pertussis Vaccine/analysis ; Pertussis Vaccine/pharmacology ; Quality Control ; Rats ; Sensitivity and Specificity
    Chemical Substances Pertussis Vaccine ; Histamine (820484N8I3) ; Cyclic AMP (E0399OZS9N) ; Pertussis Toxin (EC 2.4.2.31)
    Language English
    Publishing date 2010-02-03
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 605674-x
    ISSN 1873-2518 ; 0264-410X
    ISSN (online) 1873-2518
    ISSN 0264-410X
    DOI 10.1016/j.vaccine.2009.11.009
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1930: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)
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  5. Article ; Online: Vaccine-Mediated Activation of Human TLR4 Is Affected by Modulation of Culture Conditions during Whole-Cell Pertussis Vaccine Preparation.

    Hoonakker, Marieke E / Verhagen, Lisa M / Pupo, Elder / de Haan, Alex / Metz, Bernard / Hendriksen, Coenraad F M / Han, Wanda G H / Sloots, Arjen

    PloS one

    2016  Volume 11, Issue 8, Page(s) e0161428

    Abstract: The potency of whole-cell pertussis (wP) vaccines is still determined by an intracerebral mouse protection test. To allow development of suitable in vitro alternatives to this test, insight into relevant parameters to monitor the consistency of vaccine ... ...

    Abstract The potency of whole-cell pertussis (wP) vaccines is still determined by an intracerebral mouse protection test. To allow development of suitable in vitro alternatives to this test, insight into relevant parameters to monitor the consistency of vaccine quality is essential. To this end, a panel of experimental wP vaccines of varying quality was prepared by sulfate-mediated suppression of the BvgASR master virulence regulatory system of Bordetella pertussis during cultivation. This system regulates the transcription of a range of virulence proteins, many of which are considered important for the induction of effective host immunity. The protein compositions and in vivo potencies of the vaccines were BvgASR dependent, with the vaccine containing the highest amount of virulence proteins having the highest in vivo potency. Here, the capacities of these vaccines to stimulate human Toll-like receptors (hTLR) 2 and 4 and the role these receptors play in wP vaccine-mediated activation of antigen-presenting cells in vitro were studied. Prolonged BvgASR suppression was associated with a decreased capacity of vaccines to activate hTLR4. In contrast, no significant differences in hTLR2 activation were observed. Similarly, vaccine-induced activation of MonoMac-6 and monocyte-derived dendritic cells was strongest with the highest potency vaccine. Blocking of TLR2 and TLR4 showed that differences in antigen-presenting cell activation could be largely attributed to vaccine-dependent variation in hTLR4 signalling. Interestingly, this BvgASR-dependent decrease in hTLR4 activation coincided with a reduction in GlcN-modified lipopolysaccharides in these vaccines. Accordingly, expression of the lgmA-C genes, required for this glucosamine modification, was significantly reduced in bacteria exposed to sulfate. Together, these findings demonstrate that the BvgASR status of bacteria during wP vaccine preparation is critical for their hTLR4 activation capacity and suggest that including such parameters to assess consistency of newly produced vaccines could bring in vitro testing of vaccine quality a step closer.
    MeSH term(s) Antigen Presentation ; Bacterial Proteins/genetics ; Bacterial Proteins/immunology ; Biological Assay ; Bordetella pertussis/drug effects ; Bordetella pertussis/genetics ; Bordetella pertussis/immunology ; Bordetella pertussis/pathogenicity ; Carbohydrate Sequence ; Dendritic Cells/drug effects ; Dendritic Cells/immunology ; Dendritic Cells/microbiology ; Gene Expression ; HEK293 Cells ; Host-Pathogen Interactions ; Humans ; Lipopolysaccharides/pharmacology ; Magnesium Sulfate/pharmacology ; Monocytes/drug effects ; Monocytes/immunology ; Monocytes/microbiology ; Pertussis Vaccine/pharmacology ; Plasmids/chemistry ; Plasmids/metabolism ; Primary Cell Culture ; Toll-Like Receptor 2/genetics ; Toll-Like Receptor 2/immunology ; Toll-Like Receptor 4/genetics ; Toll-Like Receptor 4/immunology ; Trans-Activators/genetics ; Trans-Activators/immunology ; Transfection ; Transgenes ; Vaccines, Attenuated ; Virulence Factors/genetics ; Virulence Factors/immunology ; Whooping Cough/prevention & control
    Chemical Substances Bacterial Proteins ; Bvg accessory factor protein, Bordetella pertussis ; Lipopolysaccharides ; Pertussis Vaccine ; TLR2 protein, human ; TLR4 protein, human ; Toll-Like Receptor 2 ; Toll-Like Receptor 4 ; Trans-Activators ; Vaccines, Attenuated ; Virulence Factors ; Magnesium Sulfate (7487-88-9)
    Language English
    Publishing date 2016
    Publishing country United States
    Document type Journal Article
    ISSN 1932-6203
    ISSN (online) 1932-6203
    DOI 10.1371/journal.pone.0161428
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: The crystal structure of titanium dioxide nanoparticles influences immune activity in vitro and in vivo.

    Vandebriel, Rob J / Vermeulen, Jolanda P / van Engelen, Laurens B / de Jong, Britt / Verhagen, Lisa M / de la Fonteyne-Blankestijn, Liset J / Hoonakker, Marieke E / de Jong, Wim H

    Particle and fibre toxicology

    2018  Volume 15, Issue 1, Page(s) 9

    Abstract: Background: The use of engineered nanoparticles (NP) is widespread and still increasing. There is a great need to assess their safety. Newly engineered NP enter the market in a large variety; therefore safety evaluation should preferably be in a high- ... ...

    Abstract Background: The use of engineered nanoparticles (NP) is widespread and still increasing. There is a great need to assess their safety. Newly engineered NP enter the market in a large variety; therefore safety evaluation should preferably be in a high-throughput fashion. In vitro screening is suitable for this purpose. TiO
    Methods: Immature DC were differentiated in vitro from human peripheral blood monocytes. Exposure effects of a series of fourteen TiO
    Results: All NP dispersions contained NP aggregates. The anatase NP and anatase/rutile mixture NP induced a higher CD83 and CD86 expression and a higher IL-12p40 production in vitro than the rutile NP (including coated rutile NP and a rutile NP of a 10-fold larger primary diameter). OVA-specific serum IgE and IgG1 were increased by anatase NP, rutile NP, and CB, in the order rutile<anatase<CB. The three particles similarly increased IL-4 and IL-5 production by bronchial LN cells and eosinophils and lymphocytes in the BALF. Neutrophils were induced by rutile NP and CB but not by anatase NP.<br />Conclusions: Our data show that measuring CD83 and CD86 expression and IL-12p40 and TNF-α production in DC in vitro may provide an efficient way to screen NP for potential adjuvant activity; future studies should establish whether this also holds for other NP. Based on antigen-specific IgE and IgG1, anatase NP have higher adjuvant activity than rutile NP, confirming our in vitro data. Other parameters of the allergic response showed a similar response for the two NP crystal structures. From the viewpoint of safe(r) by design products, rutile NP may be preferred over anatase NP, especially when inhalation exposure can be expected during production or application of the product.
    MeSH term(s) Animals ; Bronchoalveolar Lavage Fluid/cytology ; Bronchoalveolar Lavage Fluid/immunology ; Crystallization ; Dendritic Cells/drug effects ; Dendritic Cells/immunology ; Humans ; Immunity, Cellular/drug effects ; Immunoglobulin E/blood ; Immunoglobulin G/blood ; Interleukin-4/immunology ; Interleukin-5/immunology ; Lung/drug effects ; Lung/immunology ; Mice, Inbred BALB C ; Nanoparticles/chemistry ; Nanoparticles/toxicity ; Particle Size ; Surface Properties ; Titanium/chemistry ; Titanium/toxicity
    Chemical Substances Immunoglobulin G ; Interleukin-5 ; titanium dioxide (15FIX9V2JP) ; Interleukin-4 (207137-56-2) ; Immunoglobulin E (37341-29-0) ; Titanium (D1JT611TNE)
    Language English
    Publishing date 2018-01-30
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2170936-1
    ISSN 1743-8977 ; 1743-8977
    ISSN (online) 1743-8977
    ISSN 1743-8977
    DOI 10.1186/s12989-018-0245-5
    Database MEDical Literature Analysis and Retrieval System OnLINE

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