Preprint
Generation and characterization of a novel MHC-II tetramer for tracking and characterization of toxin B-specific CD4 + T cell responses
bioRxiv
19 Feb 2026
PMID: 41757099
Abstract
The gastrointestinal pathogen
, is a major burden for health systems due to high rates of recurrence.
pathogenesis is mediated by two virulence factors, toxin A (TcdA) and Toxin B (TcdB). Antibodies specific for TcdA and TcdB are correlated with protection from symptomatic recurrence, however, the role for CD4
T cells is poorly understood in part due to the lack of tools to study the toxin-specific CD4
T cell response. Our group recently demonstrated the antibody and CD4
T cell response to
toxins is impaired via the glucosyltransferase activity of the toxins; however, tools do not exist to study the protective capacity and the phenotype of toxin-specific CD4
T cells. Therefore, we developed an MHC-II tetramer to identify TcdB-specific CD4
T cells via flow cytometry. Herein, we identified an immunodominant epitope (TcdB
) in the CROPs region of TcdB and optimized an MHC-II tetramer for use in tracking and phenotyping TcdB-specific CD4
T cell responses following multiple different immunization strategies in mice. Utilizing the tetramer, TcdB-specific T follicular helper (Tfh) cells were detected following TcdB-CROPs mRNA-LNP vaccination validating the advantage of the tetramer. Furthermore, using a modular mRNA vector expressing the TcdB
peptide covalently bound to the beta chain of MHC-II (MHC-IIβ) we were able to generate a robust population of TcdB-specific CD4
T cells. These data outline the generation of new tools for the
field and lay the groundwork for future studies of toxin-specific CD4
T cell responses.
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Details
- Title
- Generation and characterization of a novel MHC-II tetramer for tracking and characterization of toxin B-specific CD4 + T cell responses
- Creators
- Jeffrey R Maslanka - University of PennsylvaniaQianxuan She - Children's Hospital of PhiladelphiaKathleen S Krauss - Children's Hospital of PhiladelphiaEmily N Konopka - Drexel UniversityNile U Bayard - Children's Hospital of PhiladelphiaJennifer Londregan - University of PennsylvaniaMohamad-Gabriel AlamehLaurence C Eisenlohr - Children's Hospital of PhiladelphiaMichele A KutzlerJoseph P Zackular - Children's Hospital of PhiladelphiaMichael C Abt - University of Pennsylvania
- Publication Details
- bioRxiv
- Publisher
- United States
- Resource Type
- Preprint
- Language
- English
- Academic Unit
- Microbiology and Immunology; Infectious Diseases (and HIV Medicine)
- Other Identifier
- 991022164018704721