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3. Automated, Point-of-Care mobile flow cytometry: Bringing the laboratory to the sample

Author

CTI, Experimentele Afdeling Longziekten, HAG Hart- Vaatziekten, HAG Infectieziekten, Infection & Immunity, HAG Onderzoek, Child Health, Cancer, Jukema, B. N., Pelgrim, T. C., Spoelder, M., Bongers, C. C.W.G., Hopman, M. T.E., Smit, K., Rijk, M. H., Venekamp, R. P., Vrisekoop, N., Koenderman, L., CTI, Experimentele Afdeling Longziekten, HAG Hart- Vaatziekten, HAG Infectieziekten, Infection & Immunity, HAG Onderzoek, Child Health, Cancer, Jukema, B. N., Pelgrim, T. C., Spoelder, M., Bongers, C. C.W.G., Hopman, M. T.E., Smit, K., Rijk, M. H., Venekamp, R. P., Vrisekoop, N., and Koenderman, L.

Published
2024

4. Nuclear segmentation facilitates neutrophil migration.

Author

Shen, C., Mulder, E. de, Buitenwerf, W., Postat, J., Jansen, A., Kox, M., Mandl, J.N., Vrisekoop, N., Shen, C., Mulder, E. de, Buitenwerf, W., Postat, J., Jansen, A., Kox, M., Mandl, J.N., and Vrisekoop, N.

Abstract

Item does not contain fulltext, Neutrophils are among the fastest-moving immune cells. Their speed is critical to their function as 'first responder' cells at sites of damage or infection, and it has been postulated that the unique segmented nucleus of neutrophils functions to assist their rapid migration. Here, we tested this hypothesis by imaging primary human neutrophils traversing narrow channels in custom-designed microfluidic devices. Individuals were given an intravenous low dose of endotoxin to elicit recruitment of neutrophils into the blood with a high diversity of nuclear phenotypes, ranging from hypo- to hyper-segmented. Both by cell sorting of neutrophils from the blood using markers that correlate with lobularity and by directly quantifying the migration of neutrophils with distinct lobe numbers, we found that neutrophils with one or two nuclear lobes were significantly slower to traverse narrower channels, compared to neutrophils with more than two nuclear lobes. Thus, our data show that nuclear segmentation in primary human neutrophils provides a speed advantage during migration through confined spaces.

Published
2023

7. Longitudinal assessment of the inflammatory response: The next step in personalized medicine after severe trauma

Author

Zorgeenheid Traumatologie, Experimentele Afdeling Longziekten, Infection & Immunity, Cancer, de Fraiture, E J, Vrisekoop, N, Leenen, L P H, van Wessem, K J P, Koenderman, L, Hietbrink, F, Zorgeenheid Traumatologie, Experimentele Afdeling Longziekten, Infection & Immunity, Cancer, de Fraiture, E J, Vrisekoop, N, Leenen, L P H, van Wessem, K J P, Koenderman, L, and Hietbrink, F

Published
2022

9. Analysis of human neutrophil phenotypes as biomarker to monitor exercise-induced immune changes

Author

Spijkerman, R., Hesselink, L., Bertinetto, C.G., Bongers, C.C.W.G., Hietbrink, F., Vrisekoop, N., Leenen, L.P.H., Hopman, M.T.E., Jansen, J.J., Koenderman, L., Spijkerman, R., Hesselink, L., Bertinetto, C.G., Bongers, C.C.W.G., Hietbrink, F., Vrisekoop, N., Leenen, L.P.H., Hopman, M.T.E., Jansen, J.J., and Koenderman, L.

Abstract

Contains fulltext : 231632.pdf (Publisher’s version ) (Open Access)

Published
2021

10. Hoe schadelijk is plastic voor onze gezondheid?

Author

Vrisekoop, N. and Vrisekoop, N.

Abstract

Microplastics, je ademt ze in en eet ze op. En helaas, je kunt er niet aan ontkomen, want ze zitten tegenwoordig overal. Wetenschappers maken zich zorgen om al die plasticdeeltjes in ons lichaam en doen er daarom veel onderzoek naar. Immunoloog Nienke Vrisekoop (UMC Utrecht) praat je bij over de meest recente kennis en onderzoeken over die piepkleine plastics. Om nu eindelijk antwoord te krijgen op die ene vraag: hoe #schadelijk zijn die microplastics nou echt?

Published
2021

11. An increase in CD62Ldim neutrophils precedes the development of pulmonary embolisms in COVID-19 patients

Author

Experimentele Afdeling Longziekten, Cancer, Trialbureau Beeld, Zorgeenheid Traumatologie, MS Orthopaedie Algemeen, Infection & Immunity, Acute geneeskunde, MS Interne Geneeskunde, Circulatory Health, Medische Staf Intensive Care, Spijkerman, Roy, Jorritsma, Nikita Kn, Bongers, Suzanne H, Bindels, Bas Jj, Jukema, Bernard N, Hesselink, Lillian, Hietbrink, Falco, Leenen, Luke Ph, van Goor, Harriët Mr, Vrisekoop, N, Kaasjager, Karin Ah, Koenderman, Leo, COVPACH study group, Experimentele Afdeling Longziekten, Cancer, Trialbureau Beeld, Zorgeenheid Traumatologie, MS Orthopaedie Algemeen, Infection & Immunity, Acute geneeskunde, MS Interne Geneeskunde, Circulatory Health, Medische Staf Intensive Care, Spijkerman, Roy, Jorritsma, Nikita Kn, Bongers, Suzanne H, Bindels, Bas Jj, Jukema, Bernard N, Hesselink, Lillian, Hietbrink, Falco, Leenen, Luke Ph, van Goor, Harriët Mr, Vrisekoop, N, Kaasjager, Karin Ah, Koenderman, Leo, and COVPACH study group

Published
2021

12. Differentiation and activation of eosinophils in the human bone marrow during experimental human endotoxemia

Author

Hassani, M., Leijte, G.P., Bruse, N., Kox, M., Pickkers, P., Vrisekoop, N., Koenderman, L., Hassani, M., Leijte, G.P., Bruse, N., Kox, M., Pickkers, P., Vrisekoop, N., and Koenderman, L.

Abstract

Contains fulltext : 229398.pdf (Publisher’s version ) (Open Access), Acute infection is characterized by eosinopenia. However, the underlying mechanism(s) are poorly understood and it is unclear whether decreased mobilization/production of eosinophils in the bone marrow (BM) and/or increased homing to the tissues play a role. The objective of this study was to investigate the differentiation and activation status of eosinophils in the human BM and blood upon experimental human endotoxemia, a standardized, controlled, and reproducible model of acute systemic inflammation. A BM aspirate and venous blood was obtained from seven healthy volunteers before, 4 h after, and 1 week after intravenous challenge with 2 ng/kg endotoxin. Early progenitors (CD34+/IL-5Rα+), eosinophil promyelocytes, myelocytes, metamyelocytes, and mature eosinophils were identified and quantified in the bone marrow and blood samples using flowcytometry based on specific eosinophil markers (CD193 and IL-5Rα). Activation status was assessed using antibodies against known markers on eosinophils: Alpha-4 (CD49d), CCR3 (CD193), CR1 (CD35), CEACAM-8 (CD66b), CBRM 1/5 (activation epitope of MAC-1), and by plasma cytokine analysis. Four hours after endotoxin administration, numbers of mature eosinophils in the blood and in the BM markedly declined compared with baseline, whereas numbers of all eosinophil progenitors did not change. The remaining eosinophils did not show signs of activation or degranulation despite significantly increased circulating levels of eotaxin-1. Furthermore, the expression of CD49d and CD193 on eosinophils was lower compared to baseline, but normalized after 7 days. Together these data imply that circulatory eosinopenia after an innate immune challenge is mediated by CD49d-mediated homing of eosinophils to the tissues.

Published
2020

13. Characterization of the phenotype of human eosinophils and their progenitors in the bone marrow of healthy individuals

Author

Hassani, M., Staveren, S. van, Grinsven, E. van, Bartels, M., Tesselaar, K., Leijte, G.P., Kox, M., Pickkers, P., Vrisekoop, N., Koenderman, L., Hassani, M., Staveren, S. van, Grinsven, E. van, Bartels, M., Tesselaar, K., Leijte, G.P., Kox, M., Pickkers, P., Vrisekoop, N., and Koenderman, L.

Abstract

Contains fulltext : 218545.pdf (publisher's version ) (Open Access)

Published
2020

14. Plasticity of Lgr5-Negative Cancer Cells Drives Metastasis in Colorectal Cancer

Author

Fumagalli, A. (Arianna), Oost, K.C. (Koen C.), Kester, L. (Lennart), Morgner, J. (Jessica), Bornes, L. (Laura), Bruens, L. (Lotte), Spaargaren, L. (Lisa), Azkanaz, M. (Maria), Schelfhorst, T. (Tim), Beerling, E. (Evelyne), Heinz, M.C. (Maria C.), Postrach, D. (Daniel), Seinstra, D. (Danielle), Sieuwerts, A.M. (Anieta), Martens, J.W.M. (John), van der Elst, S. (Stefan), van Baalen, M. (Martijn), Bhowmick, D. (Debajit), Vrisekoop, N. (Nienke), Ellenbroek, S.I.J. (Saskia I J), Suijkerbuijk, S.J.E. (Saskia J E), Snippert, H.J. (Hugo J.), van Rheenen, J. (Jacco), Fumagalli, A. (Arianna), Oost, K.C. (Koen C.), Kester, L. (Lennart), Morgner, J. (Jessica), Bornes, L. (Laura), Bruens, L. (Lotte), Spaargaren, L. (Lisa), Azkanaz, M. (Maria), Schelfhorst, T. (Tim), Beerling, E. (Evelyne), Heinz, M.C. (Maria C.), Postrach, D. (Daniel), Seinstra, D. (Danielle), Sieuwerts, A.M. (Anieta), Martens, J.W.M. (John), van der Elst, S. (Stefan), van Baalen, M. (Martijn), Bhowmick, D. (Debajit), Vrisekoop, N. (Nienke), Ellenbroek, S.I.J. (Saskia I J), Suijkerbuijk, S.J.E. (Saskia J E), Snippert, H.J. (Hugo J.), and van Rheenen, J. (Jacco)

Abstract

Colorectal cancer stem cells (CSCs) express Lgr5 and display extensive stem cell-like multipotency and self-renewal and are thought to seed metastatic disease. Here, we used a mouse model of colorectal cancer (CRC) and human tumor xenografts to investigate the cell of origin of metastases. We found that most disseminated CRC cells in circulation were Lgr5- and formed distant metastases in which Lgr5+ CSCs appeared. This pl

Published
2020
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15. Plasticity of Lgr5-Negative Cancer Cells Drives Metastasis in Colorectal Cancer

Author

Fumagalli, A, Oost, KC, Kester, L, Morgner, J, Bornes, L, Bruens, L, Spaargaren, L, Azkanaz, M, Schelfhorst, T, Beerling, E, Heinz, MC, Postrach, D, Seinstra, D, Sieuwerts, Anieta, Martens, John, van der Elst, S, van Baalen, M, Bhowmick, D, Vrisekoop, N, Ellenbroek, SIJ, Suijkerbuijk, SJE, Snippert, HJ, van Rheenen, J, Fumagalli, A, Oost, KC, Kester, L, Morgner, J, Bornes, L, Bruens, L, Spaargaren, L, Azkanaz, M, Schelfhorst, T, Beerling, E, Heinz, MC, Postrach, D, Seinstra, D, Sieuwerts, Anieta, Martens, John, van der Elst, S, van Baalen, M, Bhowmick, D, Vrisekoop, N, Ellenbroek, SIJ, Suijkerbuijk, SJE, Snippert, HJ, and van Rheenen, J

Published
2020

17. Human CD62Ldim neutrophils identified as a separate subset by proteome profiling and in vivo pulse-chase labeling

Author

Tak, T., Wijten, P., Heeres, M., Pickkers, P., Scholten, A., Heck, A.J.R. van, Vrisekoop, N., Leenen, L.P.H., Borghans, J.A.M., Tesselaar, K., Koenderman, L., Tak, T., Wijten, P., Heeres, M., Pickkers, P., Scholten, A., Heck, A.J.R. van, Vrisekoop, N., Leenen, L.P.H., Borghans, J.A.M., Tesselaar, K., and Koenderman, L.

Abstract

Contains fulltext : 177600.pdf (publisher's version ) (Closed access), During acute inflammation, 3 neutrophil subsets are found in the blood: neutrophils with a conventional segmented nucleus, neutrophils with a banded nucleus, and T-cell-suppressing CD62Ldim neutrophils with a high number of nuclear lobes. In this study, we compared the in vivo kinetics and proteomes of banded, mature, and hypersegmented neutrophils to determine whether these cell types represent truly different neutrophil subsets or reflect changes induced by lipopolysaccharide (LPS) activation. Using in vivo pulse-chase labeling of neutrophil DNA with 6,6-2H2-glucose, we found that 2H-labeled banded neutrophils appeared much earlier in blood than labeled CD62Ldim and segmented neutrophils, which shared similar label kinetics. Comparison of the proteomes by cluster analysis revealed that CD62Ldim neutrophils were clearly separate from conventional segmented neutrophils despite having similar kinetics in peripheral blood. Interestingly, the conventional segmented cells were more related at a proteome level to banded cells despite a 2-day difference in maturation time. The differences between CD62Ldim and mature neutrophils are unlikely to have been a direct result of LPS-induced activation, because of the extremely low transcriptional capacity of CD62Ldim neutrophils and the fact that neutrophils do not directly respond to the low dose of LPS used in the study (2 ng/kg body weight). Therefore, we propose CD62Ldim neutrophils are a truly separate neutrophil subset that is recruited to the bloodstream in response to acute inflammation. This trial was registered at www.clinicaltrials.gov as #NCT01766414.

Published
2017

18. Quantification of naive and memory T-cell turnover during HIV-1 infection

Author

Vrisekoop, N, Drylewicz, J, van Gent, R., Mugwagwa, T., van Lelyveld, SFL, Veel, E.M., Otto, S.A., Ackermans, M.T., Vermeulen, J.N., Huidekoper, H.H., Prins, J.M., Miedema, F, de Boer, Rob J., Tesselaar, NA, Borghans, JAM, Vrisekoop, N, Drylewicz, J, van Gent, R., Mugwagwa, T., van Lelyveld, SFL, Veel, E.M., Otto, S.A., Ackermans, M.T., Vermeulen, J.N., Huidekoper, H.H., Prins, J.M., Miedema, F, de Boer, Rob J., Tesselaar, NA, and Borghans, JAM

Published
2015

19. Quantification of naive and memory T-cell turnover during HIV-1 infection

Author

Experimentele Afdeling Longziekten, CTI Borghans, Infection & Immunity, MS Infectieziekten, CTI, Directie Raad van Bestuur, Vrisekoop, N, Drylewicz, J, van Gent, R., Mugwagwa, T., van Lelyveld, SFL, Veel, E.M., Otto, S.A., Ackermans, M.T., Vermeulen, J.N., Huidekoper, H.H., Prins, J.M., Miedema, F, de Boer, Rob J., Tesselaar, NA, Borghans, JAM, Experimentele Afdeling Longziekten, CTI Borghans, Infection & Immunity, MS Infectieziekten, CTI, Directie Raad van Bestuur, Vrisekoop, N, Drylewicz, J, van Gent, R., Mugwagwa, T., van Lelyveld, SFL, Veel, E.M., Otto, S.A., Ackermans, M.T., Vermeulen, J.N., Huidekoper, H.H., Prins, J.M., Miedema, F, de Boer, Rob J., Tesselaar, NA, and Borghans, JAM

Published
2015

20. Intravital imaging of cancer stem cell plasticity in mammary tumors

Author

Zomer, A., Ellenbroek, S.I., Ritsma, L., Beerling, E., Vrisekoop, N., van Rheenen, J., and Hubrecht Institute for Developmental Biology and Stem Cell Research

Abstract

It is widely debated whether all tumor cells in mammary tumors have the same potential to propagate and maintain tumor growth or whether there is a hierarchical organization. Evidence for the latter theory is mainly based on the ability or failure of transplanted tumor cells to produce detectable tumors in mice with compromised immune systems; however, this assay has lately been disputed to accurately reflect cell behavior in unperturbed tumors. Lineage tracing experiments have recently shown the existence of a small population of cells, referred to as cancer stem cells (CSCs), that maintains and provides growth of squamous skin tumors and intestinal adenomas. However, the lineage tracing techniques used in these studies provide static images and lack the ability to study whether stem cell properties can be obtained or lost, a process referred to as stem cell plasticity. Here, by intravital lineage tracing, we report for the first time the existence of CSCs in unperturbed mammary tumors and demonstrate CSC plasticity. Our data indicate that existing CSCs disappear and new CSCs form during mammary tumor growth, illustrating the dynamic nature of these cells.

Published
2013

21. Peering Into Cancer

Author

Ritsma, L., Steller, E.J.A., Beerling, E., Loomans, C.J.M., Zomer, A., Gerlach, C., Vrisekoop, N., Seinstra, D., Gurp, L. van, Schafer, R., Raats, D.A., Graaff, A. de, Schumacher, T.N., Koning, E.J.P. de, Rinkes, I.H.B., Kranenburg, O., and Rheenen, J. van

Published
2012

22. Response: The in vivo half-life of human neutrophils

Author

Pillay, J., den Braber, A.J., Vrisekoop, N., Kwast, L.M., de Boer, R.J., Borghans, J.A.M., Tesselaar, N.A., Theoretical Biology and Bioinformatics, and Sub Theoretical Biology & Bioinformatics

Published
2011

23. T-cell dynamics in healthy and HIV-infected individuals

Author

Vrisekoop, N. and University Utrecht

Subjects
Geneeskunde, HAART, T-cell production, chronic immune activation, T-cell reconstitution, T-cell proliferation, T-cell activation, HIV, T-cell life-span, thymus production, recent thymic emigrant
Abstract

This thesis focuses on T-cell dynamics in healthy and both treated and untreated HIV-infected individuals. Although the progressive decline in CD4+ T-cell numbers is the hallmark of HIV infection, the mechanisms behind this depletion remain controversial. Currently the most prevailing ideas include direct HIV-induced cytopathicity, thymic impairment and chronic immune activation. We show that direct virus kill is insufficient to fully explain CD4+ T-cell depletion and that chronic immune activation plays a predominant role in CD4+ T-cell loss. To investigate the effects of non-HIV related chronic immune activation, we studied healthy Ethiopians who are known to have increased exposure to environmental pathogens. While immune characteristics of Ethiopian and Dutch neonates are similar, and therefore differences are not genetic, the CD4+ TREC content and fraction naive CD4+ T cells decrease at very early childhood in Ethiopia. We surprisingly found that when Dutch and Ethiopian HIV-infected patients were matched for CD4+ T-cell count, the percentages proliferating CD4+ and CD8+ T cells were lower in Ethiopians. This is in line with the slower CD4+ T-cell decline found in HIV+ Ethiopians compared to Dutch HIV-infected individuals. In chickens and mice it has been shown that recent thymic emigrants (RTE) form a substantial pool of short-lived naive T cells. In humans however, the lifespan and number of RTE are unknown. Currently there is no unambiguous marker for RTE and therefore the contribution of the thymus to the maintenance of the T-cell pool during aging, the role of thymic impairment in HIV-related CD4+ T-cell depletion and the role of thymic output during reconstitution following HAART are difficult to assess. By using in vivo heavy water labeling of T cells from young healthy adults, we determined the production of naive and memory CD4+ and CD8+ T cells, and followed the survival of these recently produced T cells. The average life-span of naive CD4+ en CD8+ T cells is 4.2 and 6.6 years and of memory CD4+ en CD8+ T cells 0.4 en 0.7 years. Furthermore, we show that in humans, recently produced naive T cells are long-lived and even preferentially incorporated into the naive T-cell pool. Our data thereby provide the first conclusive experimental evidence that there is no substantial population of short-lived RTE in healthy humans. Although children could recover to normal levels within 1 year after treatment even when HAART was initiated at an extremely lymphopenic stage, adults with a CD4+ nadir below 200 cells/l persistently sustained lower CD4+ T-cell counts than adults with high pre-therapy CD4+ T-cell counts throughout follow-up, but nevertheless normalized CD4+ T-cell counts after 7 years of HAART. Absolute numbers of naive CD4+ T cells normalized in all children and in adults with high baseline CD4+ T-cell counts, whereas naive CD4+ T-cell counts in adults with low CD4+ nadirs lagged behind. The reconstitution of the peripheral T-cell pool of adults and children was attributed to both thymic output and peripheral T-cell proliferation and did not seem to cause more 'proliferative history' of the T-cell pool as compared to age-matched controls.

Published
2007

24. In vivo imaging and histochemistry are combined in the cryosection labelling and intravital microscopy technique

Author

Ritsma, L., Vrisekoop, N., van Rheenen, J., Ritsma, L., Vrisekoop, N., and van Rheenen, J.

Abstract

Intravital microscopy has been used extensively to study dynamic processes in the context of a living animal; however, only a limited number of fluorescent probes and mouse models are available. By contrast, many dyes and antibodies exist for the immuno-labelling of fixed tissue. Here we report a method that combines the advantages of histochemistry and in vivo imaging by correlating cryosection labelling with corresponding intravital microscopy images (CLIM). Using CLIM, we find that the presence of CD3(+) T cells correlates with mammary tumour cell migration. When CD4(+) and CD8(+) T-cell subsets are depleted, reduced tumour cell migration is observed. From these data we conclude that CLIM is a powerful tool to correlate intravital microscopy data with cryosection labelling data., Intravital microscopy has been used extensively to study dynamic processes in the context of a living animal; however, only a limited number of fluorescent probes and mouse models are available. By contrast, many dyes and antibodies exist for the immuno-labelling of fixed tissue. Here we report a method that combines the advantages of histochemistry and in vivo imaging by correlating cryosection labelling with corresponding intravital microscopy images (CLIM). Using CLIM, we find that the presence of CD3(+) T cells correlates with mammary tumour cell migration. When CD4(+) and CD8(+) T-cell subsets are depleted, reduced tumour cell migration is observed. From these data we conclude that CLIM is a powerful tool to correlate intravital microscopy data with cryosection labelling data.

Published
2013

25. Maintenance of peripheral naive T cells is sustained by thymus output in mice but not humans.

Author

den Braber, A.J., Mugwagwa, T., Vrisekoop, N., Westera, L., Mögling, R, de Boer, A.B., Willems, N., Schrijver, E.H.R., Spierenburg, G.Th., Gaiser, J.F., Mul, E., Otto, S.A., Ruiter, A.F., Ackermans, M.T., Miedema, F., Borghans, J.A.M., de Boer, R.J., Tesselaar, Kiki, den Braber, A.J., Mugwagwa, T., Vrisekoop, N., Westera, L., Mögling, R, de Boer, A.B., Willems, N., Schrijver, E.H.R., Spierenburg, G.Th., Gaiser, J.F., Mul, E., Otto, S.A., Ruiter, A.F., Ackermans, M.T., Miedema, F., Borghans, J.A.M., de Boer, R.J., and Tesselaar, Kiki

Published
2012

26. Intravital microscopy through an abdominal imaging window reveals a pre-micrometastasis stage during liver metastasis

Author

Ritsma, L., Steller, E.J., Beerling, E., Loomans, C.J., Zomer, A., Gerlach, C., Vrisekoop, N., Seinstra, D., van Gurp, L., Schafer, R., Raats, D.A., de Graaff, A., Schumacher, T.N., de Koning, E., Rinkes, I.H., Kranenburg, O., van Rheenen, J., Ritsma, L., Steller, E.J., Beerling, E., Loomans, C.J., Zomer, A., Gerlach, C., Vrisekoop, N., Seinstra, D., van Gurp, L., Schafer, R., Raats, D.A., de Graaff, A., Schumacher, T.N., de Koning, E., Rinkes, I.H., Kranenburg, O., and van Rheenen, J.

Abstract

Cell dynamics in subcutaneous and breast tumors can be studied through conventional imaging windows with intravital microscopy. By contrast, visualization of the formation of metastasis has been hampered by the lack of long-term imaging windows for metastasis-prone organs, such as the liver. We developed an abdominal imaging window (AIW) to visualize distinct biological processes in the spleen, kidney, small intestine, pancreas, and liver. The AIW can be used to visualize processes for up to 1 month, as we demonstrate with islet cell transplantation. Furthermore, we have used the AIW to image the single steps of metastasis formation in the liver over the course of 14 days. We observed that single extravasated tumor cells proliferated to form "pre-micrometastases," in which cells lacked contact with neighboring tumor cells and were active and motile within the confined region of the growing clone. The clones then condensed into micrometastases where cell migration was strongly diminished but proliferation continued. Moreover, the metastatic load was reduced by suppressing tumor cell migration in the pre-micrometastases. We suggest that tumor cell migration within pre-micrometastases is a contributing step that can be targeted therapeutically during liver metastasis formation., Cell dynamics in subcutaneous and breast tumors can be studied through conventional imaging windows with intravital microscopy. By contrast, visualization of the formation of metastasis has been hampered by the lack of long-term imaging windows for metastasis-prone organs, such as the liver. We developed an abdominal imaging window (AIW) to visualize distinct biological processes in the spleen, kidney, small intestine, pancreas, and liver. The AIW can be used to visualize processes for up to 1 month, as we demonstrate with islet cell transplantation. Furthermore, we have used the AIW to image the single steps of metastasis formation in the liver over the course of 14 days. We observed that single extravasated tumor cells proliferated to form "pre-micrometastases," in which cells lacked contact with neighboring tumor cells and were active and motile within the confined region of the growing clone. The clones then condensed into micrometastases where cell migration was strongly diminished but proliferation continued. Moreover, the metastatic load was reduced by suppressing tumor cell migration in the pre-micrometastases. We suggest that tumor cell migration within pre-micrometastases is a contributing step that can be targeted therapeutically during liver metastasis formation.

Published
2012

27. Maintenance of peripheral naive T cells is sustained by thymus output in mice but not humans

Author

Infection & Immunity, CTI, Directie Raad van Bestuur, den Braber, A.J., Mugwagwa, T., Vrisekoop, N., Westera, L., Mögling, R, de Boer, A.B., Willems, N., Schrijver, E.H.R., Spierenburg, G.Th., Gaiser, J.F., Mul, E., Otto, S.A., Ruiter, A.F., Ackermans, M.T., Miedema, F., Borghans, J.A.M., de Boer, R.J., Tesselaar, Kiki, Infection & Immunity, CTI, Directie Raad van Bestuur, den Braber, A.J., Mugwagwa, T., Vrisekoop, N., Westera, L., Mögling, R, de Boer, A.B., Willems, N., Schrijver, E.H.R., Spierenburg, G.Th., Gaiser, J.F., Mul, E., Otto, S.A., Ruiter, A.F., Ackermans, M.T., Miedema, F., Borghans, J.A.M., de Boer, R.J., and Tesselaar, Kiki

Published
2012

28. Response: The in vivo half-life of human neutrophils

Author

Theoretical Biology and Bioinformatics, Sub Theoretical Biology & Bioinformatics, Pillay, J., den Braber, A.J., Vrisekoop, N., Kwast, L.M., de Boer, R.J., Borghans, J.A.M., Tesselaar, N.A., Theoretical Biology and Bioinformatics, Sub Theoretical Biology & Bioinformatics, Pillay, J., den Braber, A.J., Vrisekoop, N., Kwast, L.M., de Boer, R.J., Borghans, J.A.M., and Tesselaar, N.A.

Published
2011

29. Intravital microscopy: new insights into metastasis of tumors

Author

Beerling, E., Ritsma, L., Vrisekoop, N., Derksen, P.W., van Rheenen, J., Beerling, E., Ritsma, L., Vrisekoop, N., Derksen, P.W., and van Rheenen, J.

Abstract

Metastasis, the process by which cells spread from the primary tumor to a distant site to form secondary tumors, is still not fully understood. Although histological techniques have provided important information, they give only a static image and thus compromise interpretation of this dynamic process. New advances in intravital microscopy (IVM), such as two-photon microscopy, imaging chambers, and multicolor and fluorescent resonance energy transfer imaging, have recently been used to visualize the behavior of single metastasizing cells at subcellular resolution over several days, yielding new and unexpected insights into this process. For example, IVM studies showed that tumor cells can switch between multiple invasion strategies in response to various densities of extracellular matrix. Moreover, other IVM studies showed that tumor cell migration and blood entry take place not only at the invasive front, but also within the tumor mass at tumor-associated vessels that lack an intact basement membrane. In this Commentary, we will give an overview of the recent advances in high-resolution IVM techniques and discuss some of the latest insights in the metastasis field obtained with IVM., Metastasis, the process by which cells spread from the primary tumor to a distant site to form secondary tumors, is still not fully understood. Although histological techniques have provided important information, they give only a static image and thus compromise interpretation of this dynamic process. New advances in intravital microscopy (IVM), such as two-photon microscopy, imaging chambers, and multicolor and fluorescent resonance energy transfer imaging, have recently been used to visualize the behavior of single metastasizing cells at subcellular resolution over several days, yielding new and unexpected insights into this process. For example, IVM studies showed that tumor cells can switch between multiple invasion strategies in response to various densities of extracellular matrix. Moreover, other IVM studies showed that tumor cell migration and blood entry take place not only at the invasive front, but also within the tumor mass at tumor-associated vessels that lack an intact basement membrane. In this Commentary, we will give an overview of the recent advances in high-resolution IVM techniques and discuss some of the latest insights in the metastasis field obtained with IVM.

Published
2011

30. Sparse production but preferential incorporation of recently produced naive T cells in the human peripheral pool.

Author

Vrisekoop, N., den Braber, I., de Boer, A.B., Ruiter, A.F.C., Ackermans, M.T., van der Crabben, S.N., Schrijver, E.H.R., Spierenburg, G.Th., Sauerwein, H.P., Hazenberg, M.D., de Boer, R.J., Miedema, F., Borghans, J.A.M., Tesselaar, K., Vrisekoop, N., den Braber, I., de Boer, A.B., Ruiter, A.F.C., Ackermans, M.T., van der Crabben, S.N., Schrijver, E.H.R., Spierenburg, G.Th., Sauerwein, H.P., Hazenberg, M.D., de Boer, R.J., Miedema, F., Borghans, J.A.M., and Tesselaar, K.

Published
2008

31. In vivo dynamics of stable chronic lymphocytic leukemia inversely correlate with somatic hypermutation levels and suggest no major leukemic turnover in bone marrow.

Author

van Gent, R., Kater, A.P., Otto, S.A., Jaspers, A., Borghans, J.A.M., Vrisekoop, N., Ackermans, M.A., Ruiter, A.F.C., Wittebol, S., Eldering, E., van Oers, M.H.J., Tesselaar, N.A., Kersten, M.J., Miedema, F., van Gent, R., Kater, A.P., Otto, S.A., Jaspers, A., Borghans, J.A.M., Vrisekoop, N., Ackermans, M.A., Ruiter, A.F.C., Wittebol, S., Eldering, E., van Oers, M.H.J., Tesselaar, N.A., Kersten, M.J., and Miedema, F.

Published
2008

32. Restoration of the CD4 T cell compartment after long-term highly active antiretroviral therapy without phenotypical signs of accelerated immunological aging

Author

Vrisekoop, N., van Gent, R., de Boer, A.B., Otto, S.A., Borleffs, J.C.C., Steingrover, R., Prins, J.M., Kuijpers, T.W., Wolfs, T.F.W., Geelen, S.P.M., Vulto, I., Lansdorp, P., Tesselaar, N.A., Borghans, J.A.M., Miedema, F., Vrisekoop, N., van Gent, R., de Boer, A.B., Otto, S.A., Borleffs, J.C.C., Steingrover, R., Prins, J.M., Kuijpers, T.W., Wolfs, T.F.W., Geelen, S.P.M., Vulto, I., Lansdorp, P., Tesselaar, N.A., Borghans, J.A.M., and Miedema, F.

Published
2008

33. Sparse production but preferential incorporation of recently produced naive T cells in the human peripheral pool.

Author

Evolution of major histocompatibility complex (MHC) polymorphisms, Theoretical Biology and Bioinformatics, Dep Biologie, Vrisekoop, N., den Braber, I., de Boer, A.B., Ruiter, A.F.C., Ackermans, M.T., van der Crabben, S.N., Schrijver, E.H.R., Spierenburg, G.Th., Sauerwein, H.P., Hazenberg, M.D., de Boer, R.J., Miedema, F., Borghans, J.A.M., Tesselaar, K., Evolution of major histocompatibility complex (MHC) polymorphisms, Theoretical Biology and Bioinformatics, Dep Biologie, Vrisekoop, N., den Braber, I., de Boer, A.B., Ruiter, A.F.C., Ackermans, M.T., van der Crabben, S.N., Schrijver, E.H.R., Spierenburg, G.Th., Sauerwein, H.P., Hazenberg, M.D., de Boer, R.J., Miedema, F., Borghans, J.A.M., and Tesselaar, K.

Published
2008

34. Sparse production but preferential incorporation of recently produced naive T cells in the human peripheral pool.

Author

Infection & Immunity, CTI, Genetica, Vrisekoop, N., den Braber, I., de Boer, A.B., Ruiter, A.F.C., Ackermans, M.T., van der Crabben, S.N., Schrijver, E.H.R., Spierenburg, G.Th., Sauerwein, H.P., Hazenberg, M.D., de Boer, R.J., Miedema, F., Borghans, J.A.M., Tesselaar, K., Infection & Immunity, CTI, Genetica, Vrisekoop, N., den Braber, I., de Boer, A.B., Ruiter, A.F.C., Ackermans, M.T., van der Crabben, S.N., Schrijver, E.H.R., Spierenburg, G.Th., Sauerwein, H.P., Hazenberg, M.D., de Boer, R.J., Miedema, F., Borghans, J.A.M., and Tesselaar, K.

Published
2008

35. Restoration of the CD4 T cell compartment after long-term highly active antiretroviral therapy without phenotypical signs of accelerated immunological aging

Author

Infection & Immunity, CTI, MS Infectieziekten, Infectiezieken, Vrisekoop, N., van Gent, R., de Boer, A.B., Otto, S.A., Borleffs, J.C.C., Steingrover, R., Prins, J.M., Kuijpers, T.W., Wolfs, T.F.W., Geelen, S.P.M., Vulto, I., Lansdorp, P., Tesselaar, N.A., Borghans, J.A.M., Miedema, F., Infection & Immunity, CTI, MS Infectieziekten, Infectiezieken, Vrisekoop, N., van Gent, R., de Boer, A.B., Otto, S.A., Borleffs, J.C.C., Steingrover, R., Prins, J.M., Kuijpers, T.W., Wolfs, T.F.W., Geelen, S.P.M., Vulto, I., Lansdorp, P., Tesselaar, N.A., Borghans, J.A.M., and Miedema, F.

Published
2008

36. In vivo dynamics of stable chronic lymphocytic leukemia inversely correlate with somatic hypermutation levels and suggest no major leukemic turnover in bone marrow

Author

Infection & Immunity, CTI, van Gent, R., Kater, A.P., Otto, S.A., Jaspers, A., Borghans, J.A.M., Vrisekoop, N., Ackermans, M.A., Ruiter, A.F.C., Wittebol, S., Eldering, E., van Oers, M.H.J., Tesselaar, N.A., Kersten, M.J., Miedema, F., Infection & Immunity, CTI, van Gent, R., Kater, A.P., Otto, S.A., Jaspers, A., Borghans, J.A.M., Vrisekoop, N., Ackermans, M.A., Ruiter, A.F.C., Wittebol, S., Eldering, E., van Oers, M.H.J., Tesselaar, N.A., Kersten, M.J., and Miedema, F.

Published
2008

43. Neutrophils in cancer: from biology to therapy.

Author

Koenderman L and Vrisekoop N

Abstract

The view of neutrophils has shifted from simple phagocytic cells, whose main function is to kill pathogens, to very complex cells that are also involved in immune regulation and tissue repair. These cells are essential for maintaining and regaining tissue homeostasis. Neutrophils can be viewed as double-edged swords in a range of situations. The potent killing machinery necessary for immune responses to pathogens can easily lead to collateral damage to host tissues when inappropriately controlled. Furthermore, some subtypes of neutrophils are potent pathogen killers, whereas others are immunosuppressive or can aid in tissue healing. Finally, in tumor immunology, many examples of both protumorigenic and antitumorigenic properties of neutrophils have been described. This has important consequences for cancer therapy, as targeting neutrophils can lead to either suppressed or stimulated antitumor responses. This review will discuss the current knowledge regarding the pro- and antitumorigenic roles of neutrophils, leading to the concept of a confused state of neutrophil-driven pro-/antitumor responses., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Published
2024
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44. Eosinophils are sparse in homeostatic rectal tissue which impedes studying resident eosinophils.

Author

Pelgrim TC, Jukema BN, Vrisekoop N, and Koenderman L

Abstract

Competing Interests: Declaration of competing interest The study was financially supported by GlaxoSmithKline. This company had no role in the study's design, the data analysis, the article's preparation, or the decision to submit the article for publication. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Published
2024
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47. Nuclear segmentation facilitates neutrophil migration.

Author

Shen C, Mulder E, Buitenwerf W, Postat J, Jansen A, Kox M, Mandl JN, and Vrisekoop N

Subjects
Humans, Cell Movement physiology, Neutrophils physiology, Cell Nucleus
Abstract

Neutrophils are among the fastest-moving immune cells. Their speed is critical to their function as 'first responder' cells at sites of damage or infection, and it has been postulated that the unique segmented nucleus of neutrophils functions to assist their rapid migration. Here, we tested this hypothesis by imaging primary human neutrophils traversing narrow channels in custom-designed microfluidic devices. Individuals were given an intravenous low dose of endotoxin to elicit recruitment of neutrophils into the blood with a high diversity of nuclear phenotypes, ranging from hypo- to hyper-segmented. Both by cell sorting of neutrophils from the blood using markers that correlate with lobularity and by directly quantifying the migration of neutrophils with distinct lobe numbers, we found that neutrophils with one or two nuclear lobes were significantly slower to traverse narrower channels, compared to neutrophils with more than two nuclear lobes. Thus, our data show that nuclear segmentation in primary human neutrophils provides a speed advantage during migration through confined spaces., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)

Published
2023
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48. Visualization of the inflammatory response to injury by neutrophil phenotype categories : Neutrophil phenotypes after trauma.

Author

de Fraiture EJ, Bongers SH, Jukema BN, Koenderman L, Vrisekoop N, van Wessem KJP, Leenen LPH, and Hietbrink F

Subjects
Humans, Cohort Studies, Phenotype, Neutrophils
Abstract

Purpose: The risk of infectious complications after trauma is determined by the amount of injury-related tissue damage and the resulting inflammatory response. Recently, it became possible to measure the neutrophil phenotype in a point-of-care setting. The primary goal of this study was to investigate if immunophenotype categories based on visual recognition of neutrophil subsets are applicable to interpret the inflammatory response to trauma. The secondary goal was to correlate these immunophenotype categories with patient characteristics, injury severity and risk of complications., Methods: A cohort study was conducted with patients presented at a level 1 trauma center with injuries of any severity, who routinely underwent neutrophil phenotyping. Data generated by automated point-of-care flow cytometry were prospectively gathered. Neutrophil phenotypes categories were defined by visual assessment of two-dimensional CD16/CD62L dot plots. All patients were categorized in one of the immunophenotype categories. Thereafter, the categories were validated by multidimensional analysis of neutrophil populations, using FlowSOM. All clinical parameters and endpoints were extracted from the trauma registry., Results: The study population consisted of 380 patients. Seven distinct immunophenotype Categories (0-6) were defined, that consisted of different neutrophil populations as validated by FlowSOM. Injury severity scores and risk of infectious complications increased with ascending immunophenotype Categories 3-6. Injury severity was similarly low in Categories 0-2., Conclusion: The distribution of neutrophil subsets that were described in phenotype categories is easily recognizable for clinicians at the bedside. Even more, multidimensional analysis demonstrated these categories to be distinct subsets of neutrophils. Identification of trauma patients at risk for infectious complications by monitoring the immunophenotype category is a further improvement of personalized and point-of-care decision-making in trauma care., (© 2022. The Author(s).)

Published
2023
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49. Human neutrophil kinetics: a call to revisit old evidence.

Author

Koenderman L, Tesselaar K, and Vrisekoop N

Subjects
Humans, Kinetics, Bone Marrow, Adaptive Immunity, Neutrophils, Bone Marrow Cells
Abstract

The half-life of human neutrophils is still controversial, with estimates ranging from 7-9 h to 3.75 days. This debate should be settled to understand neutrophil production in the bone marrow (BM) and the potential and limitations of emergency neutropoiesis following infection or trauma. Furthermore, cellular lifespan greatly influences the potential effect(s) neutrophils have on the adaptive immune response. We posit that blood neutrophils are in exchange with different tissues, but particularly the BM, as it contains the largest pool of mature neutrophils. Furthermore, we propose that the oldest neutrophils are the first to die following a so-called conveyor belt model. These guiding principles shed new light on our interpretation of existing neutrophil lifespan data and offer recommendations for future research., Competing Interests: Declaration of interests No interests are declared., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Published
2022
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50. Neutrophils Promote Glioblastoma Tumor Cell Migration after Biopsy.

Author

Chen N, Alieva M, van der Most T, Klazen JAZ, Vollmann-Zwerenz A, Hau P, and Vrisekoop N

Subjects
Animals, Biopsy, Cell Movement, Humans, Mice, Neutrophils metabolism, Brain Neoplasms metabolism, Glioblastoma metabolism, Glioma
Abstract

Glioblastoma is diagnosed by biopsy or, if clinically feasible, tumor resection. However, emerging evidence suggests that this surgical intervention may increase the risk of tumor cell spread. It has been hypothesized that the damage to the tumor leads to infiltration of immune cells that consequently form an environment that favors tumor cell motility. In mouse glioma models, it was previously found that biopsy induced migration of tumor cells in vivo and that recruitment of monocytes from the blood was involved in this effect. However, the role of neutrophils in this process is still unclear. Here, we study the contribution of neutrophils on the pro-migratory effect of surgical interventions in glioma. Using repetitive intravital microscopy, in vivo migration of glioma tumor cells before and after biopsy was compared in mice systemically depleted of neutrophils. Interestingly, macrophages/microglia were almost completely absent from neutrophil-depleted tumors, indicating that neutrophils may be indirectly involved in biopsy-induced migration of glioma tumor cells through the recruitment of macrophages to the tumor. To further investigate whether neutrophils have the potential to also directly promote glioblastoma tumor cell migration, we performed in vitro migration assays using human neutrophils. Indeed, wound-healing of human primary glioblastoma tumor cell lines was promoted by human neutrophils. The pro-migratory effects of human neutrophils on glioblastoma tumor cells could also be recapitulated in transwell migration assays, indicating that soluble factor(s) are involved. We therefore provide evidence for both an indirect and direct involvement of neutrophils in tumor spread following biopsy of glioblastoma tumors.

Published
2022
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