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Frontiers in immunology2022; 13; 805026; doi: 10.3389/fimmu.2022.805026

CD154 Expression Indicates T Cell Activation Following Tetanus Toxoid Vaccination of Horses.

Abstract: Despite the relevance of adaptive immunity against equine pathogens antigen-specific T cell responses of horses are not well characterized and the lack of insight into T cell responses hampers the understanding of the pathogeneses of important diseases. In this study we used tetanus toxoid (TT) as a well-defined antigen to characterize antigen-reactive T cells. Six healthy adult horses received a routine booster against tetanus with an immune stimulating complex (ISCOM)-based vaccine and were followed for 28 days. TT-specific serum antibodies were quantified by ELISA and increased in all horses by day 7 after vaccination. CD154 is an established indicator of antigen-reactive T helper cells in other species, but has not been characterized in horses. CD154 detection in equine PBMC by an anti-human CD154 antibody (clone 5C8) was confirmed by Western blots and then applied for flow cytometry. As a common indicator of equine T cell activation, cytokine induction was studied in parallel. T cells were analyzed by multicolor flow cytometry of PBMC after re-stimulation with TT . Reactive T helper (Th) cells were characterized by increased frequencies of CD4CD154 lymphocytes in TT-re-stimulated PBMC on day 14 after vaccination of the horses compared to pre-vaccination. The majority of all CD154 cells after TT re-stimulation were CD4 Th cells, but CD154 was also induced on CD4 cells albeit in lower frequencies. CD154CD4 Th cells were enriched in cytokine-expressing cells compared to CD154CD4 Th cells. Similar to the CD4CD154 frequencies, CD4IL-4, CD4IFN-γ and CD4TNF-α were increased after vaccination, but IL-4 increased later than IFN-γ and CD4TNF-α, which already exceeded pre-vaccination frequencies on day 7. CD4CD154 frequencies correlated positively with those of CD4IL-4 (Th2) on day 14, and negatively with CD4IFN-γ induction on day 7, but did not correlate with CD4TNF-α frequencies or TT-specific antibody concentrations. CD154 appears to be a useful marker of antigen-reactive equine Th cells in combination with cytokine expression. The T cell analyses established here with TT can be applied to other antigens relevant for infections or allergies of horses and in horse models for translational research.
Copyright © 2022 Schnabel, Fletemeyer, Lübke, Marti, Wagner and Alber.
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Publication Date: 2022-04-13 PubMed ID: 35493462PubMed Central: PMC9043809DOI: 10.3389/fimmu.2022.805026Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates how the expression of CD154, a protein marker, relates to the activation of T cells in horses after tetanus toxoid vaccination. This study provides an important step in understanding the immune response processes of horses against pathogens, and provides a basis for studying diseases in this particular animal species.

Methodology

  • The study involved six healthy adult horses which were given a regular booster dose of a tetanus vaccine that uses an Immune Stimulating Complex (ISCOM).
  • These horses were observed over a 28-day period. Tetanus-specific antibodies in their serum were measured using an ELISA assay, which confirmed increasing antibody levels by the seventh day after vaccination.
  • The researchers primarily focused on the detection of CD154 in horses’ Peripheral Blood Mononuclear Cells (PBMC), a protein that is known to indicate the activation of T-helper cells in other species.
  • The detection was carried out with an anti-human CD154 antibody and confirmed with Western blots, a technique commonly used to detect specific proteins.
  • Simultaneously, the scientists studied the induction of cytokines, substances secreted by immune cells that influence other cells.
  • The researchers examined T cells using multicolor flow cytometry, a method for analyzing cell populations based on their properties.

Results

  • The study discovered that reactive T-helper cells in horses could be described by an increased amount of CD4CD154 lymphocytes in tetanus toxoid-restimulated PMBC, particularly by the fourteenth day after vaccination.
  • Most CD154 cells after TT restimulation were CD4 T-helper cells. CD154 was also found on CD4 cells, but less frequently.
  • Findings revealed that CD154CD4 T-helper cells had a higher percentage of cytokine-expressing cells compared to CD154CD4 T-helper cells.
  • Similar to the frequencies of CD4CD154, the levels of the cytokines IL-4, IFN-γ, and TNF-α cytokines also increased after vaccination.
  • However, IL-4 levels rose later than IFN-γ and CD4TNF-α, which already exceeded pre-vaccination frequencies by the seventh day.
  • CD4CD154 frequencies correlated positively with CD4IL-4 on the fourteenth day, negatively with CD4IFN-γ induction on the seventh day, but showed no correlation with CD4TNF- α frequencies or TT-specific antibody concentrations.

Conclusion

  • The study draws the conclusion that CD154 can serve as a useful marker for identifying antigen-reactive T-helper cells in horses, especially when used in combination with cytokine expression study.
  • This research establishes a foundation for analyzing T cells in horses in response to other antigens, creating a crucial framework for understanding infectious diseases or allergies in horses, and aids in translational research involving horse models.

Cite This Article

APA
Schnabel CL, Fletemeyer B, Lübke S, Marti E, Wagner B, Alber G. (2022). CD154 Expression Indicates T Cell Activation Following Tetanus Toxoid Vaccination of Horses. Front Immunol, 13, 805026. https://doi.org/10.3389/fimmu.2022.805026

Publication

Frontiers in immunology
ISSN: 1664-3224
NlmUniqueID: 101560960
Country: Switzerland
Language: English
Volume: 13
Pages: 805026
PII: 805026

Researcher Affiliations

Schnabel, Christiane L
  • Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
Fletemeyer, Babette
  • Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
Lübke, Sabrina
  • Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
Marti, Eliane
  • Clinical Immunology Group, Department for Clinical Research and Veterinary Public Health (VPH), Vetsuisse Faculty of the University of Bern, Bern, Switzerland.
Wagner, Bettina
  • Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
Alber, Gottfried
  • Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.

MeSH Terms

  • Animals
  • Antibodies, Bacterial
  • CD40 Ligand
  • Cytokines
  • Horses
  • Interleukin-4
  • Leukocytes, Mononuclear
  • Tetanus
  • Tetanus Toxoid
  • Toxoids
  • Tumor Necrosis Factor-alpha
  • Vaccination

Conflict of Interest Statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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