Combined Glycoprotein Mutations in Rabies Virus Promote Astrocyte Tropism and Protective CNS Immunity in Mice

Authors

Mirjam Anna Rita Bertoune
Corinna Kolbe
Ann-Cathrin Werner
Maren Steinmetz
Bernhard Dietzschold, Thomas Jefferson UniversityFollow
Eberhard Weihe

Document Type

Article

Publication Date

1-29-2026

Comments

This article is the author's final published version in Viruses, Volume 18, Issue 2, February 2026, Article number 181.

The published version is available at https://doi.org/10.3390/v18020181. Copyright © 2026 by the authors.

Abstract

Rabies virus (RABV) causes fatal encephalitis once it invades the central nervous system (CNS), and treatment options are extremely limited at this stage. We investigated the recombinant RABV variants SPBN, SPBNGA (glycoprotein substitution R333E), SPBNGAK (R333E plus N194K), SPBNGAS (R333E plus N194S), and TriGAS (three copies of the R333E/N194S glycoprotein). We evaluated their cellular tropism and immune activation in an intracerebral mouse infection model using immunohistochemistry and confocal immunofluorescence. SPBNGAK (R333E/N194K) resulted in mixed neuronal and astrocytic infection and lethal disease. In contrast, the R333E/N194S mutations in the GAS variants were associated with reduced neuronal infection and apparent astrocyte-restricted infection patterns. This tropism shift coincided with microglial activation (allograft inflammatory factor 1, amoeboid transformation) and astrocytic activation (nestin), along with T-cell infiltration and endothelial activation that persisted beyond viral clearance. SPBNGAK-infected astrocytes expressed nestin, while GAS variant-infected astrocytes remained nestin-negative and were rapidly cleared. Intracerebral co-inoculation of astrocytotropic TriGAS with the lethal neurotropic DOG4 strain was associated with survival and a marked reduction in detectable DOG4 neuronal infection. These findings suggest that glycoprotein-mediated astrocyte tropism may be associated with altered immune responses after rabies CNS invasion. While mechanistic causality cannot be inferred, these observations may inform the design of future studies exploring astrocyte-restricted RABV infection in therapeutic-related contexts.

Recommended Citation

Bertoune, Mirjam Anna Rita; Kolbe, Corinna; Werner, Ann-Cathrin; Steinmetz, Maren; Dietzschold, Bernhard; and Weihe, Eberhard, "Combined Glycoprotein Mutations in Rabies Virus Promote Astrocyte Tropism and Protective CNS Immunity in Mice" (2026). Department of Microbiology and Immunology Faculty Papers. Paper 201.
https://jdc.jefferson.edu/mifp/201

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 License.

PubMed ID

41754524

Language

English