July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
TRPV1 positive sensory neurons respond to Pseudomonas aeruginosa, increasing susceptibility to keratitis
Author Affiliations & Notes
  • Tiffany Lin
    Infectious Diseaseas, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Tiphaine Voisin
    Harvard Medical School, Massachusetts, United States
  • Pankaj Baral
    Harvard Medical School, Massachusetts, United States
  • Isaac Chiu
    Harvard Medical School, Massachusetts, United States
  • Gerald Pier
    Infectious Diseaseas, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Mihaela G Gadjeva
    Infectious Diseaseas, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Footnotes
    Commercial Relationships   Tiffany Lin, None; Tiphaine Voisin, None; Pankaj Baral, None; Isaac Chiu, None; Gerald Pier, None; Mihaela Gadjeva, None
  • Footnotes
    Support  NIH/NEI R01 EY022054
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4641. doi:
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    • Get Citation

      Tiffany Lin, Tiphaine Voisin, Pankaj Baral, Isaac Chiu, Gerald Pier, Mihaela G Gadjeva; TRPV1 positive sensory neurons respond to Pseudomonas aeruginosa, increasing susceptibility to keratitis. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4641.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Pain is a hallmark of most bacterial infections; however, the molecular mechanisms of pain are not well defined. It is unknown whether different pathogens induce distinct pain circuits and how pain-mediating nociceptor neurons affect infection outcomes. In this study, we explore the impact of nociceptors on susceptibility to keratitis in an experimental mouse model wherein treatment with Resiniferatoxin (RTX), a potent activator of Transient Receptor Potential Vanilloid 1 (TRPV1) channel, ablates corneal nociceptors. As a secondary approach, we targeted nociceptors using Nav1.8-cre driven diphtheria toxin A expression. We hypothesize that nociceptor sensory neurons regulate the magnitude and molecular characteristics of the inflammatory responses during bacterial infections

Methods : To chemically ablate TRPV1+ neurons, C57BL6 mice were treated with three consecutive doses of RTX: 30 ug/kg, 70 ug/kg, and 100 ug/kg subcutaneously. Whole mount immunohistochemical staining of mouse corneas were used to quantify sensory neuron presence. The capsaicin eye wipe test was performed to evaluate functional deficiency of nociceptors.

To determine the impact of nociceptors on keratitis susceptibility, RTX (RTX and vehicle) and Nav1.8 cohorts were infected with P. aeruginosa and bacterial burdens were quantified. Cytokine and chemokine levels in the corneal lysates were assessed using ELISA.

To monitor neuronal response to P. aeruginosa 6294, PAK and PAK mutants, trigeminal ganglia primary neuronal cultures were generated and imaged for intracellular Ca 2+ release using Fura-2 AM.

Results : Our data demonstrate that nociceptors directly sense P. aeruginosa by an increase of intracellular Ca2+, a marker for neuronal activation. The sensory neurons responded significantly less to strains of P. aeruginosa that lacked pili, flagella or a type III secretion system. Pharmacological ablation of TRPV1+ nociceptors via RTX treatments or genetic ablation of Nav1.8+ nociceptors altered susceptibility to infection. This was associated with decreased bacterial burdens and superior neutrophil bactericidal functions.

Conclusions : Our results suggest that nociceptors regulate corneal neutrophil response and bacterial burden during P. aeruginosa keratitis. Hence, therapeutic modalities that control pain sensation could favorably impact anti-infective therapy.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

Representative fields of TG neuronal response to P. aeruginosa

Representative fields of TG neuronal response to P. aeruginosa

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