Delayed treatment with the immunotherapeutic LNFPIII ameliorates multiple neurological deficits in a pesticide-nerve agent prophylactic mouse model of Gulf War Illness

Evaluation of delayed LNFPIII treatment initiation protocol on improving long-term behavioral and neuroinflammatory pathology in a mouse model of Gulf War Illness

“…The following chemicals were used for animal treatments: pyridostigmine bromide (PB; Sigma Aldrich, St. Louis, MO), N-Diethyl-3-methylbenzamide (DEET; Sigma Aldrich), corticosterone (CORT; Steraloids, Newport, RI), and diisopropylfluorophosphate (DFP; Sigma Aldrich). Lacto-N-fucopentaose III (LNFPIII) dextran conjugate was produced as in Carpenter et al (2021) . All additional chemicals and reagents, unless otherwise noted, were of analytical or higher grade and obtained from Sigma Aldrich or Fisher Scientific (Hampton, NH).…”

“…At 7 months post GWI exposure, mice within each of the two original treatments were randomly subdivided into LNFPIII or dextran vehicle groups. Mice were treated with LNFPIII or dextran vehicle (both 35 μg/mouse diluted in sterile saline; SC) twice a week until study completion as in ( Carpenter et al, 2021 ). At this stage of the study, treatment groups were as follows: Vehicle-Vehicle (Saline/water/saline-Dextran, n = 16), Vehicle-LNFPIII (Saline/water/saline-LNFPIII, n = 12), GWI-Vehicle (PB/DEET/CORT/DFP-Dextran, n = 13), and GWI-LNFPIII (PB/DEET/CORT/DFP-LNFPIII, n = 13).…”

“…One month after LNFPIII treatment initiation, a battery of behavioral tests for assessment of cognition/memory, mood, and motor function were performed as done previously ( Carpenter et al, 2021 ). All behavioral testing, scoring, and analysis were done in a randomized, treatment-blind manner.…”

“…Considering the immune dysfunction in GWI, recent preclinical studies have explored the efficacy of several immune-targeted compounds that have shown some promising results ( Joshi et al, 2018 ; Kodali et al, 2018a ; Ribeiro et al, 2020 ; Shetty et al, 2020 ). In our studies, we have focused on one potent immunomodulatory and anti-inflammatory treatment, Lacto-N-fucopentaose III (LNFPIII), that may be an advantageous treatment option for GWI as demonstrated in two, chemically distinct GWI models ( Brown et al, 2021a , 2021b ; Carpenter et al, 2020 , 2021 ; Mote et al, 2020 ). LNFPIII is a Lewis(X)-containing immunomodulatory glycan found in human breast milk and on parasitic helminths that, to date, has had no documented adverse effects ( Atochina et al, 2008 ; Bhargava et al, 2012 ; Srivastava et al, 2014 ; Tundup et al, 2015 ; Zhu et al, 2012 ).…”

“…LNFPIII is a Lewis(X)-containing immunomodulatory glycan found in human breast milk and on parasitic helminths that, to date, has had no documented adverse effects ( Atochina et al, 2008 ; Bhargava et al, 2012 ; Srivastava et al, 2014 ; Tundup et al, 2015 ; Zhu et al, 2012 ). LNFPIII, when conjugated to a dextran carrier, activates CD14/TLR-4 signaling for extracellular signal-regulated kinase (ERK)-dependent production of anti-inflammatory mediators to skew the inflammatory balance of the innate immune system in an anti-inflammatory direction ( Atochina et al, 2008 ; Bhargava et al, 2012 ; Carpenter et al, 2020 , 2021 ; Srivastava et al, 2014 ; Tundup et al, 2015 ; Zhu et al, 2012 ). Of note, in utilizing the PB/DEET/CORT/DFP and the pyridostigmine bromide/permethrin (PB/PM) GWI models, we demonstrated that LNFPIII was beneficial in restoring acute monoaminergic disbalance and neuroinflammation (both models) as well as long-term behavioral deficits (PB/PM model), neuroinflammation, and hippocampal synaptic plasticity impairments after these chemical exposures ( Brown et al, 2021a , 2021b ; Carpenter et al, 2020 , 2021 ).…”