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Low-Dose Subcutaneous Anti-CD20 Treatment Depletes Disease Relevant B Cell Subsets and Attenuates Neuroinflammation

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Abstract

To explore the B cell depleting capacity of a low-dose (20 μg) subcutaneous mouse anti-CD20 antibody treatment on disease-relevant B cell populations within lymph nodes and the spleen. B cell depleting capacity was explored in healthy female C57BL/6 and BALB/c mice; following immune activation in two different mouse models: trinitrophenylated lipopolysaccharide model (thymus-independent response) and dinitrophenyl-keyhole limpet hemocyanin model (thymus-dependent response); and in a chronic neuroinflammation experimental autoimmune encephalomyelitis model. CD20 protein expression on B cell subpopulations was also studied. The subcutaneous anti-CD20 regimen resulted in rapid depletion of B cells in blood, lymph nodes and spleen. Low-dose subcutaneous treatment did not reduce antigen-specific immunoglobulin M and immunoglobulin G titers in all subgroups, and relatively spared splenic marginal zone (MZ) B cells in both T cell dependent and T cell independent B cell immunization models. Analysis of immune compartments during anti-CD20-modulated autoimmune neuroinflammation showed that the maximal B cell depletion was achieved within 2 days of treatment and was highest in the lymph node. Regardless of the tissues analyzed, low-dose subcutaneous treatment was characterized by rapid B cell repletion following treatment cessation. CD20 protein expression was consistent on all B cell subsets in blood, and was more pronounced in germinal center B cells of lymph nodes and MZ B-cells of the spleen. Low-dose subcutaneous anti-CD20 therapy effectively depleted B cells within lymphatic tissues and reduced the severity of neuroinflammation. These data suggest that subcutaneous anti-CD20 therapies can effectively target disease-relevant B cell populations, have shorter repletion kinetics and maintain vaccination responses, thereby achieving autoimmune amelioration without severely impacting immune surveillance functions.

*p < 0.05; **p < 0.01. CD, cluster of differentiation; DNP-KLH, dinitrophenyl-keyhole limpet hemocyanin; EC50, concentration of a drug that gives half-maximal response; Ig, immunoglobulin; MZ, marginal zone; s.c., subcutaneous; SEM, standard error of mean; TNP-LPS, trinitrophenylatedlipopolysaccharide.

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Data Availability

The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgments

This study was funded by Novartis Pharma AG. The authors would like to acknowledge the laboratory animal services technical team for in vivo support. The authors would like to thank Richa Chhabra and Sivaram Vedantam (Medical communications, Novartis Healthcare Pvt. Ltd) for medical writing assistance in developing the first draft of the manuscript, formatting, referencing and incorporating the authors’ revisions and submission, all under the direction of the authors.

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Authors and Affiliations

  1. Autoimmunity, Transplantation and Inflammation, Novartis Institutes for Biomedical Research, Basel, Switzerland

    Catherine Huck, Vanessa Wegert, Cindy Schmid, Gisbert Weckbecker & Paul A. Smith

  2. Neuroscience Development Unit, Novartis Pharma AG, Basel, Switzerland

    David Leppert

  3. Genomics Institute of the Novartis Research Foundation, San Diego, CA, USA

    Robert Dunn

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  1. Catherine Huck
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  2. David Leppert
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  5. Robert Dunn
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  6. Gisbert Weckbecker
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  7. Paul A. Smith
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Contributions

PAS and CH designed the studies, interpreted the data and drafted the manuscript. VW, CS and RD contributed experimental data. GW contributed to experimental designs, data evaluation and writing the manuscript. DL contributed to experimental designs and data evaluation. All authors have read and approved the final manuscript.

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Correspondence to Catherine Huck.

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Animal Ethics Statement

All animal work was performed according to the Swiss federal law for animal protection and approved by the Veterinary Office of the Canton Basel-Stadt.

Conflict of Interest

Catherine Huck, Cindy Schmid, Robert Dunn and Gisbert Weckbecker are employees of Novartis Pharma AG, Basel, Switzerland.

Paul Smith, David Leppert and Vanessa Wegert were employed by Novartis Pharma AG, Basel, Switzerland during the conduct of the study.

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Huck, C., Leppert, D., Wegert, V. et al. Low-Dose Subcutaneous Anti-CD20 Treatment Depletes Disease Relevant B Cell Subsets and Attenuates Neuroinflammation. J Neuroimmune Pharmacol 14, 709–719 (2019). https://doi.org/10.1007/s11481-019-09872-z

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