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Home / Equine Research Bank / Vet Ophthalmol / Effect of gentamicin on CD3+ T-lymphocyte proliferation for ...
Veterinary ophthalmology2023; 26(4); 347-354; doi: 10.1111/vop.13098

Effect of gentamicin on CD3+ T-lymphocyte proliferation for treatment of equine recurrent uveitis: An in vitro study.

Abstract: The objective of the study was to determine the effect of gentamicin on CD3+ T-lymphocyte proliferation and cell viability using an in vitro cell culture model as a means of investigating the mechanism of action of low-dose intravitreal gentamicin injection. Methods: Three adult horses with no evidence of ophthalmic or systemic disease. Methods: Peripheral blood lymphocytes were treated with gentamicin at concentrations 37.5 μg/mL, 112.5 μg/mL, 187 μg/mL, 375 μg/mL, or 750 μg/mL then stimulated to proliferate with concanavalin A (ConA). 4',6-diamidino-2-phenylindole (DAPI) and carboxyfluoroscein succinimidyl ester (CSFE) were used as markers of cell viability and cell proliferation, respectively. Following 5-day culture, live cell counts and CSFE fluorescent intensity data were collected via automated cell count and flow cytometry. The experimental design was duplicated using preservative-free gentamicin and a proprietary brand formulation. Statistical analysis was performed using two-way ANOVA with Tukey's multiple comparison test. Results: No statistically significant comparisons in CD3+ T-lymphocyte live cell counts and geometric mean fluorescent intensity of CSFE were identified between gentamicin concentrations or formulations. Conclusions: Gentamicin had no effect on equine peripheral blood CD3+ T-lymphocyte cell viability and proliferation in concentrations ranging from "safe" to "retinotoxic" in relation to intravitreal injection volumes. Low-dose intravitreal gentamicin may not suppress the Th1- and Th17-mediated immune response.
© 2023 The Authors. Veterinary Ophthalmology published by Wiley Periodicals LLC on behalf of American College of Veterinary Ophthalmologists.
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Publication Date: 2023-04-28 PubMed ID: 37116984DOI: 10.1111/vop.13098Google Scholar: Lookup
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  • Journal Article

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 investigated the effects of the antibiotic gentamicin on certain immune cells (CD3+ T-lymphocytes) in horses, to understand if the drug could be used to treat a common eye disease (uveitis). The study found no significant effects on these immune cells, even at varying concentrations of gentamicin, suggesting it may not be an effective treatment.

Objective of the Research

  • The research aimed to explore the effect of the antibiotic gentamicin on the proliferation and vitality of CD3+ T-lymphocytes (a type of white blood cell vital to the immune system) in horses.
  • This was performed as an in-vitro study, using cultured cells, to further understand the potential usage of gentamicin for treatment of equine recurrent uveitis, a commonly occurring eye disease in horses.

Methodology

  • The study employed peripheral blood lymphocytes from three healthy adult horses. These cells were exposed to varying concentrations of gentamicin and then stimulated to proliferate with a substance called Concanavalin A (ConA).
  • Two markers were used to measure cell viability and proliferation, DAPI and CSFE, providing data on living cells and their rate of multiplication respectively.
  • This experimental design was also tested using two different types of gentamicin: a preservative-free form and a proprietary brand formulation.
  • Statistical analysis was performed using a two-way ANOVA with Tukey’s multiple comparison test to identify any significant differences in results.

Results

  • The study found no significant differences in the live cell counts and rate of proliferation of the CD3+ T-lymphocytes across the different concentrations and formulations of gentamicin.
  • This suggests that gentamicin, in the concentrations tested, does not affect the viability or proliferation of these immune cells in horses.

Conclusions

  • The findings indicate that gentamicin, even at concentrations deemed “retinotoxic,” does not influence the behavior of equine CD3+ T-lymphocytes.
  • Therefore, the study concludes that low-dose intravitreal (injected into the eye) gentamicin may not be effective in suppressing the Th1- and Th17- mediated immune response, which are thought to be involved in the development of equine recurrent uveitis.

Cite This Article

APA
Smith HL, Berglund AK, Robertson JB, Schnabel LV, McMullen RJ, Gilger BC, Oh A. (2023). Effect of gentamicin on CD3+ T-lymphocyte proliferation for treatment of equine recurrent uveitis: An in vitro study. Vet Ophthalmol, 26(4), 347-354. https://doi.org/10.1111/vop.13098

Publication

Veterinary ophthalmology
ISSN: 1463-5224
NlmUniqueID: 100887377
Country: England
Language: English
Volume: 26
Issue: 4
Pages: 347-354

Researcher Affiliations

Smith, Hannah L
  • Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, North Carolina, Raleigh, USA.
Berglund, Alix K
  • Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, North Carolina, Raleigh, USA.
  • Comparative Medicine Institute, North Carolina State University, North Carolina, Raleigh, USA.
Robertson, James B
  • Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, North Carolina, Raleigh, USA.
  • Office of Research, College of Veterinary Medicine, North Carolina State University, North Carolina, Raleigh, USA.
Schnabel, Lauren V
  • Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, North Carolina, Raleigh, USA.
  • Comparative Medicine Institute, North Carolina State University, North Carolina, Raleigh, USA.
McMullen, Richard J
  • JT Vaughan Large Animal Teaching Hospital, College of Veterinary Medicine, Auburn University, Alabama, Auburn, USA.
Gilger, Brian C
  • Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, North Carolina, Raleigh, USA.
Oh, Annie
  • Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, North Carolina, Raleigh, USA.

MeSH Terms

  • Animals
  • Horses
  • Gentamicins / pharmacology
  • Gentamicins / therapeutic use
  • Research Design
  • Uveitis / drug therapy
  • Uveitis / veterinary
  • T-Lymphocytes
  • Cell Proliferation
  • Horse Diseases / drug therapy

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