University of Nebraska at Omaha DigitalCommons@UNO Biology Faculty Publications Department of Biology 7-2019 TLR9 agonist MGN1703 enhances B cell differentiation and function in lymph nodes Mariane H. Schleimann Aarhus University Maria-Louise Kobberø Aarhus University Line K. Vibholm Aarhus University Katherine Kjær Aarhus University Leila B. Giron The Wistar Institute See next page for additional authors Follow this and additional works at: https://digitalcommons.unomaha.edu/biofacpub Part of the Biology Commons Recommended Citation Schleimann, Mariane H.; Kobberø, Maria-Louise; Vibholm, Line K.; Kjær, Katherine; Giron, Leila B.; Busman-Sahay, Kathleen; Chan, Chi Ngai; Nekorchuk, Michael; Schmidt, Manuel; Wittig, Burghardt; Damsgaard, Tine E.; Ahlburg, Peter; Hellfritzsch, Michel B.; Zuwala, Kaja; Rothemejer, Frederik H.; Olesen, Rikke; Schommers, Phillipp; Klein, Florian; Dweep, Harsh; Kossenkov, Andrew; Nyengaard, Jens R.; Estes, Jacob D.; Abdel-Mohsen, Mohamed; Østergaard, Lars; Søgaard, Ole S.; and Denton, Paul W., "TLR9 agonist MGN1703 enhances B cell differentiation and function in lymph nodes" (2019). Biology Faculty Publications. 129. https://digitalcommons.unomaha.edu/biofacpub/129 This Article is brought to you for free and open access by the Department of Biology at DigitalCommons@UNO. It has been accepted for inclusion in Biology Faculty Publications by an authorized administrator of DigitalCommons@UNO. For more information, please contact [email protected]. Authors Mariane H. Schleimann, Maria-Louise Kobberø, Line K. Vibholm, Katherine Kjær, Leila B. Giron, Kathleen Busman-Sahay, Chi Ngai Chan, Michael Nekorchuk, Manuel Schmidt, Burghardt Wittig, Tine E. Damsgaard, Peter Ahlburg, Michel B. Hellfritzsch, Kaja Zuwala, Frederik H. Rothemejer, Rikke Olesen, Phillipp Schommers, Florian Klein, Harsh Dweep, Andrew Kossenkov, Jens R. Nyengaard, Jacob D. Estes, Mohamed Abdel-Mohsen, Lars Østergaard, Ole S. Søgaard, and Paul W. Denton This article is available at DigitalCommons@UNO: https://digitalcommons.unomaha.edu/biofacpub/129 EBioMedicine 45 (2019) 328–340 Contents lists available at ScienceDirect EBioMedicine journal homepage: www.ebiomedicine.com TLR9 agonist MGN1703 enhances B cell differentiation and function in lymph nodes Mariane H. Schleimann a,b,⁎, Maria-Louise Kobberø c,LineK.Vibholma,c,KathrineKjæra,c, Leila B. Giron d, Kathleen Busman-Sahay b, Chi Ngai Chan b, Michael Nekorchuk b,ManuelSchmidte, Burghardt Wittig e,f,1, Tine E. Damsgaard c,g,PeterAhlburgh, Michel B. Hellfritzsch c,i, Kaja Zuwala a,c, Frederik H. Rothemejer c, Rikke Olesen c,PhillippSchommersj,k,l, Florian Klein j,l, Harsh Dweep m, Andrew Kossenkov m, Jens R. Nyengaard c,n, Jacob D. Estes b, Mohamed Abdel-Mohsen d,LarsØstergaarda,c, Martin Tolstrup a,c, Ole S. Søgaard a,c,PaulW.Dentona,c,⁎⁎ a Department of Infectious Diseases, Aarhus University Hospital, Denmark b Vaccine and Gene Therapy Institute, Oregon Health and Science University, Portland, OR, USA c Department of Clinical Medicine, Aarhus University, Denmark d Vaccine & Immunotherapy Center, The Wistar Institute, Philadelphia, PA, USA e Mologen AG, Berlin, Germany f MolBio2Math - Molecular Biology & Integral Biomathics, a non-profit Foundation Institute, Berlin, Germany g Department of Plastic and Breast Surgery, Plastic Surgery Research Unit, Aarhus University Hospital, Denmark h Department of Anesthesiology, Aarhus University Hospital, Denmark i Department of Radiology, Aarhus University Hospital, Denmark j Institute of Virology, Faculty of Medicine and University Hospital of Cologne, University of Cologne, 50931 Cologne, Germany k Department of Internal Medicine, Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany l German Center for Infection Research, Partner Site Bonn-Cologne, 50931 Cologne, Germany m Bioinformatics Facility, The Wistar Institute, Philadelphia, PA, USA n Core Centre for Molecular Morphology, Section for Stereology and Microscopy, Department of Clinical Medicine, Centre for Stochastic Geometry and Advanced Bioimaging, Aarhus University Hospital, Aarhus, Denmark article info abstract Article history: Background: TLR9 agonists are being developed as immunotherapy against malignancies and infections. TLR9 is Received 1 May 2019 primarily expressed in B cells and plasmacytoid dendritic cells (pDCs). TLR9 signalling may be critically impor- Received in revised form 27 June 2019 tant for B cell activity in lymph nodes but little is known about the in vivo impact of TLR9 agonism on human Accepted 2 July 2019 lymph node B cells. As a pre-defined sub-study within our clinical trial investigating TLR9 agonist MGN1703 Available online 9 July 2019 (lefitolimod) treatment in the context of developing HIV cure strategies (NCT02443935), we assessed TLR9 agonist-mediated effects in lymph nodes. Keywords: Methods: Participants received MGN1703 for 24 weeks concurrent with antiretroviral therapy. Seven participants HIV cure TLR9 agonist completed the sub-study including lymph node resection at baseline and after 24 weeks of treatment. A variety of B cell differentiation tissue-based immunologic and virologic parameters were assessed. B cell follicle Findings: MGN1703 dosing increased B cell differentiation; activated pDCs, NK cells, and T cells; and induced a Antibody glycosylation robust interferon response in lymph nodes. Expression of Activation-Induced cytidine Deaminase, an essential regulator of B cell diversification and somatic hypermutation, was highly elevated. During MGN1703 treatment IgG production increased and antibody glycosylation patterns were changed. Interpretation: Our data present novel evidence that the TLR9 agonist MGN1703 modulates human lymph node B cells in vivo. These findings warrant further considerations in the development of TLR9 agonists as immunother- apy against cancers and infectious diseases. ⁎ Correspondence to: M. H. Schleimann, Postdoc at Research Department of Infectious Diseases, Aarhus University Hospital, Skejby, Palle Juul Jensens Boulevard 99, 8200 Aarhus, Denmark. ⁎⁎ Correspondence to: P. W. Denton University of Nebraska at Omaha, Allwine Hall 109B, 6001 Dodge Street, Omaha, NE 68182-0040, USA. E-mail addresses: [email protected] (M.H. Schleimann), [email protected] (P.W. Denton). 1 Senior advisor to Mologen AG, Berlin, Germany. https://doi.org/10.1016/j.ebiom.2019.07.005 2352-3964/© 2019 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). M.H. Schleimann et al. / EBioMedicine 45 (2019) 328–340 329 Fund: This work was supported by Aarhus University Research Foundation, the Danish Council for Independent Research and the NovoNordisk Foundation. Mologen AG provided study drug free of charge. © 2019 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/). 1. Introduction stimulation in human pDCs are well described ex vivo and in vivo [1–8]. In short, TLR9 stimulated pDCs produce interferon α (IFN-α). Pattern recognition receptors are innate molecules that sense This type I interferon production leads to a stimulatory environment pathogen-associated or danger-associated molecular patterns [1]. One that results in T, NKT, and NK cells becoming activated with improved such receptor, the toll-like receptor 9 (TLR9), is found primarily in capacity for cytotoxic effector functions. Since engaging TLR9 leads to endosomes of plasmacytoid dendritic cells (pDCs) and B cells where it heightened activity of immune effector cells, the development of TLR9 detects non-methylated cytosine-guanine (CpG) DNA motifs [2,3]. agonists as immunotherapy for cancers and infectious diseases is a These dinucleotide motifs are frequently found in bacterial DNA as field of intense research. well as in transformed cells [1–4]. The downstream effects of TLR9 TLR9 agonists include multiple classes of CpG DNA oligonucleo- tides (CpG ODNs) as well as members of the double stem loop immu- nomodulator (dSLIM) class of molecules [2,4]. CpG ODNs are linear DNA fragments that harbor CpG motifs and these fragments are typ- fi Research in context ically protected from nuclease degradation by a modi ed phospho- rothioate backbone. To avoid chemical modification leading to unwanted side effects, the dSLIM molecules utilize a covalently- Evidence before this study closed dumbbell-shaped configuration offering protection from nucleases. Because the major effects of TLR9 stimulation occur in Synthetic TLR9 agonists are being developed as immunotherapy lymphoid tissue, we need to study this specific anatomical compart- for the treatment of cancers and chronic infections as well as vac- ment to fully understand how TLR9 agonists engage the immune sys- cine adjuvants. Few human PBMC in vitro or murine in vivo studies tem. Such investigations provide profound insights into how have examined the direct effect of TLR9 agonists on B cells. Al- responses to TLR9 agonism differ between anatomical compart- though the analyses utilized in these studies were not designed ments. For example, we previously reported compartmentalized for in depth interrogation of B cells, they indicated that B cell pro- human intestinal responses to MGN1703 (lefitolimod; a member of liferation and differentiation are stimulated by TLR9 agonism. the dSLIM class of TLR9 agonists [4,9–11]) dosing that were quite nu- However, no previous studies have investigated the in