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Application of light sheet microscopy for qualitative and quantitative analysis of bronchus-associated lymphoid tissue in mice

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Cellular & Molecular Immunology 2018年第10期15卷 875-887页

作者： David Twapokera Mzinza Henrike Fleige Kristin Laarmann stefanie willenzon Jasmin Ristenpart Julia Spanier Gerd Sutter Ulrich Kalinke Peter Valentin-Weigand Reinhold Förster Institute of Immunology Hannover Medical SchoolHannoverGermany Institute for Microbiology University of Veterinary Medicine HannoverHannoverGermany Institute for Experimental Infection Research TWINCORECentre for Experimental and Clinical Infection Researcha joint venture between the Helmholtz-Centre for Infection Research and the Hannover Medical SchoolHannoverGermany Institute for Infectious Diseases and Zoonoses University of Munich LMUMunichGermany

Bronchus-associated lymphoid tissue(BALT)develops at unpredictable locations around lung bronchi following pulmonary inflammation.The formation and composition of BALT have primarily been investigated by immunohistology that,due to the size of the invested organ,is usually restricted to a limited number of histological sections.To assess the entire BALT of the lung,other approaches are urgently needed.Here,we introduce a novel light sheet microscopy-based approach for assessing lymphoid tissue in the lung.Using antibody staining of whole lung lobes and optical clearing by organic solvents,we present a method that allows in-depth visualization of the entire bronchial tree,the lymphatic vasculature and the immune cell composition of the induced BALT.Furthermore,three-dimensional analysis of the entire lung allows the qualitative and quantitative enumeration of the induced BALT.Using this approach,we show that a single intranasal application of the replication-deficient poxvirus MVA induces BALT that constitutes up to 8%of the entire lung volume in mice deficient in CCR7,in contrast to wild type mice(WT).Furthermore,BALT induced by heat-inactivated E.coli is dominated by a pronounced T cell infiltration in Cxcr5-deficient mice,in contrast to WT mice.

关键词： Lung BALT light sheet microscopy E.coli MVA

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Low serum neutralizing anti-SARS-CoV-2 S antibody levels in mildly affected COVID-19 convalescent patients revealed by two different detection methods

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Cellular & Molecular Immunology 2021年第4期18卷 936-944页

作者： Berislav Bosnjak Saskia Catherina Stein stefanie willenzon Anne Katrin Cordes Wolfram Puppe Gunter Bernhardt Inga Ravens Christiane Ritter Christian R.Schultze-Florey Nina Godecke Jorg Martens Hannah Kleine-Weber Markus Hoffmann Anne Cossmann Mustafa Yilmaz Isabelle Pink Marius M.Hoeper Georg M.N.Behrens Stefan Pohlm ann Rainer Blasczyk Thomas F.Schulz Reinhold Forster Institute of Immunology Hannover Medical SchoolHannoverGermany lnstitute of Virology Hannover Medical SchoolHannoverGermany Department of Hematology HemostasisOncology and Stem Cell TransplantationHannover Medical SchoolHannoverGermany Institute of Transfusion Medicine and Transplant Engineering Hannover Medical SchoolHannoverGermany German Primate Center-Leibniz Institute for Primate Research GottingenGermany Faculty of Biology and Psychology University GottingenGottingenGermany Department of Rheum atology and Immunology Hannover Medical SchoolHannoverGermany Public Health Department HannoverGermany Department of Pneumology and German Centre of Lung Research(DZL) Hannover Medical SchoolHannoverGermany German Center for Infection Research(DZIF) partner site Hannover-BraunschweigHannoverGermany Cluster of Excellence RESIST(EXC 2155) Hannover Medical SchoolHannoverGermany

Neutralizing antibodies targeting the receptor-binding domain(RBD)of the SARS-CoV-2 spike(S)block severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)entry into cells via surface-expressed angiotensin-converting enzyme 2(ACE2).We used a surrogate virus neutralization test(sVNT)and SARS-CoV-2 S protein-pseudotyped vesicular stomatitis virus(VSV)vector-based neutralization assay(pVNT)to assess the degree to which serum antibodies from coronavirus disease 2019(COVID-19)convalescent patients interfere with the binding of SARS-CoV-2 S to ACE2.Both tests revealed neutralizing anti-SARS-CoV-2 S antibodies in the sera of ~90% of mildly and 100% of severely affected COVID-19 convalescent patients.Importantly,sVNT and pVNT results correlated strongly with each other and to the levels of anti-SARS-CoV-2 S1 IgG and IgA antibodies.Moreover,levels of neutralizing antibodies correlated with the duration and severity of clinical symptoms but not with patient age.Compared to pVNT,sVNT is less sophisticated and does not require any biosafety labs.Since this assay is also much faster and cheaper,sVNT will not only be important for evaluating the prevalence of neutralizing antibodies in a population but also for identifying promising plasma donors for successful passive antibody therapy.

关键词： COVID-19 SARS-CoV-2 Neutralizing antibody Serum ELISA

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