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Evidence of Long-range nerve pathways connecting and coordinating activity in secondary lymph organs

Victoria Cotero, Tzu-Jen Kao, John Graf, Jeffrey Ashe, Christine Morton, Sangeeta S. Chavan, Stavros Zanos, Kevin J. Tracey, Christopher M. Puleo

Bioelectronic Medicine 2020 · 10.1186/s42234-020-00056-2

rodenthealthyhistology molecular

Abstract

Background Peripheral nerve reflexes enable organ systems to maintain long-term physiological homeostasis while responding to rapidly changing environmental conditions. Electrical nerve stimulation is commonly used to activate these reflexes and modulate organ function, giving rise to an emerging class of therapeutics called bioelectronic medicines. Dogma maintains that immune cell migration to and from organs is mediated by inflammatory signals (i.e. cytokines or pathogen associated signaling molecules). However, nerve reflexes that regulate immune function have only recently been elucidated, and stimulation of these reflexes for therapeutic effect has not been fully investigated. Methods We utilized both electrical and ultrasound-based nerve stimulation to activate nerve pathways projecting to specific lymph nodes. Tissue and cell analysis of the stimulated lymph node, distal lymph nodes and immune organs is then utilized to measure the stimulation-induced changes in neurotransmitter/neuropeptide concentrations and immune cellularity in each of these sites. Results and conclusions In this report, we demonstrate that activation of nerves and stimulated release of neurotransmitters within a local lymph node results in transient retention of immune cells (e.g. lymphocytes and neutrophils) at that location. Furthermore, such stimulation results in transient changes in neurotransmitter concentrations at distal organs of the immune system, spleen and liver, and mobilization of immune cells into the circulation. This report will enable future studies in which stimulation of these long-range nerve connections between lymphatic and immune organs can be applied for clinical purpose, including therapeutic modulation of cellularity during vaccination, active allergic response, or active auto-immune disease.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjects6 animals
Sessions per subject1
Randomisednot reported
Blindingnot reported
Sham / controlundescribed
Auditory controlnot reported
Readout timingoffline
Anaesthesiaanaesthetised
Readoutshistology molecularwhite blood cell, neutrophil and lymphocyte counts (Hemavet 950 hematology analyzer) in lymph nodes, liver, spleen and lymphatic fluid
Direction of effectexcitatoryUltrasound neurostimulation directly above the popliteal lymph node produced similar increases in white blood cell counts in the targeted popliteal lymph node and liver as direct electrical sciatic nerve stimulation, though effects on the spleen and contralateral lymph node were attenuated compared with electrical stimulation.
Adverse eventsnot reported

Exposures

Exposure 1: Ultrasound neurostimulation directly above the popliteal lymph node

Target: other — “popliteal lymph node (surrounding neural tissue)
Device: Sonic Concepts · Sonic Concepts · H106

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,100✓✓
Pulse duration (ms)0.136✓✓
Pulse repetition frequency (Hz)2,000✓✓
Duty cycle (%)27pulse duration × PRF gives 27.272%✓✓
Sonication duration (s)60✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatenot reported
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Pressure, domain unspecified (kPa)830✓✓
Protocol, in the paper’s words

An ultrasound stimulus was applied directly above the popliteal lymph node using the peripheral pFUS device, with total duration of a single stimulus not surpassing a single 1-min pulse; WBC/neutrophil/lymphocyte counts were compared with and without (sham CTRL) this stimulation.

Flags from extraction

  • exposures[0].target.termsthe target is neural tissue in and around the popliteal lymph node, not a listed nervous-system structure; classified as 'other' with target.label describing the site.
  • exposures[0].timing.sonication_duration_smethods state total duration of a single stimulus 'not surpassing a single 1 min pulse', an upper bound rather than a precisely stated exposure duration; recorded as 60 s (the stated maximum) and flagged.
  • sham_typefigure legend describes a 'sham CTRL' condition (no ultrasound) but the mechanism (inactive transducer vs. simple omission) is not described; classified as 'other'.
  • unspecified_domain.pressure_kpadomain (free field vs in situ) for the 0.83 MPa peak positive pressure is not stated by the paper; recorded as unspecified_domain.