Beneficial reconstitution of gut microbiota and control of alpha-synuclein and curli-amyloids-producing enterobacteria, by beta 1,3-1,6 glucans in a clinical pilot study of autism and potentials in neurodegenerative diseases

AbstractBackground/objectiveGut dysbiosis is one of the major pathologies in children with autism spectrum disorder (ASD). In previous studies, Aureobasidium pullulans (i.e., black yeast AFO-202-produced beta glucan found in Nichi Glucan) yielded beneficial clinical outcomes related to sleep and behaviour. Evaluation of gut microbiota of the subjects in the present randomized pilot clinical study was undertaken and compared with an aim of gaining a mechanistic insight.MethodsThe study involved 18 subjects with ASD who were randomly allocated: six subjects in the control group (Group 1) underwent conventional treatment comprising remedial behavioural therapies and L-carnosine 500 mg per day, and 12 subjects (Group 2) underwent supplementation with Nichi Glucan 0.5 g twice daily along with the conventional treatment for 90 days. The subjects’ stool samples were collected at baseline and after the intervention. Whole genome metagenome (WGM) sequencing was performed.ResultsWGM sequencing followed by bioinformatic analysis in 13 subjects who completed the study showed that among genera of relevance, the abundance of Enterobacteria was decreased almost to zero in Group 2 after intervention, whereas it increased from 0.36% to 0.85% in Group 1. The abundance of Bacteroides increased from 16.84% to 19.09% in Group 1, whereas it decreased from 11.60% to 11.43% in Group 2. The abundance of Prevotella increased in both Group 1 and Group 2. The decrease in abundance of lactobacillus was significant in Group 2 compared to Group 1. Among species, a decrease was seen in Escherichia coli, Akkermansia muciniphila CAG:154, Blautia spp., Coprobacillus sp., and Clostridium bolteae CAG:59, with an increase of Faecalibacterium prausnitzii and Prevotella copri, which are both beneficial.ConclusionAFO-202 beta 1,3-1,6 glucan was able to balance the gut microbiome, which is considered beneficial in children with ASD. Effective control of curli-producing enterobacteria that leads to α-synuclein (αSyn) misfolding and accumulation, which apart from being advantageous in alleviating ASD symptoms, may have a prophylactic role in Parkinson’s and Alzheimer’s diseases where the αSyn misfolding and amyloid deposition are central to their pathogenesis. Additionally, stimulation of natural killer cells to help clear accumulated αSyn amyloids, beneficial microbiome reconstitution, and microglial rejuvenation lead us to recommend larger clinical studies in neurodevelopmental and neurodegenerative diseases of this safety-proven food supplement.Graphical AbstractThe above illustration explains, stepwise, the pathogenesis as well as the way beta glucan tackles each stage of the disease process: (A) & (B) Enterobacteria secretion of curli that causes misfolding of α-synuclein (αSyn); its aggregation in enteric neuronal cells is tackled by (1) control of enterobacteria, (2) scavenging of the accumulated amyloids by activated natural killer cells, and (3) reconstitution of beneficial microbiome. (C) The prion like propagation may not occur because the accumulation of curli proteins and amyloids is controlled at the level of production and aggregation (1) as well as clearing of already accumulated deposits (3). (D) Deposition of Lewy bodies, amyloid fibrils, and misfolded αSyn are tackled by (4) microglial-based scavenging.

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Circulating CD56 Bright Natural Killer Cells after Allogeneic Stem Cell Trasnplantation Express Functional TRAIL; Implication for a Novel Target of Immunotherapy.

Blood ◽

10.1182/blood.v106.11.5403.5403 ◽

2005 ◽

Vol 106 (11) ◽

pp. 5403-5403

Author(s):

Sumiko Takao ◽

Takayuki Ishikawa ◽

Katsuyuki Ohmori ◽

Atsumi Ishida ◽

Takashi Uchiyama

Keyword(s):

Stem Cell ◽

Natural Killer Cells ◽

Healthy Volunteers ◽

Nk Cells ◽

Natural Killer ◽

Nk Cell ◽

Killer Cells ◽

Concanamycin A ◽

Group 2 ◽

Group 1

Abstract Relapse of malignancies remains to be one of the major problems after allogeneic stem cell transplantation (allo-SCT). It is well-recognized that natural killer cells (NK-cells) are predominated in early phase of immune reconstitution after allo-SCT, and several studies demonstrated that CD56 bright CD16 negative (CD56++CD16−) NK-cells, which account for only a few percentage of peripheral NK-cells in healthy individuals, constitute a large subset of NK-cells at this phase. Although CD56++CD16− NK-cells possess unique ability to proliferate and produce proinflammatory cytokines in response to monokines or IL-2, they have been regarded to be less cytotoxic and unfavorable for graft-versus leukemia effects. To verify this issue, we compared the frequency of peripheral CD56++ NK-cells among total NK-cells with subsequent relapse in 25 allo-SCT recipients. Although the ratio of CD56++ NK-cells was gradually decreased as the increased duration between phlebotomy and allo-SCT, we could divide these patients into two groups. Group 1 was consisted of patients who showed consistently elevated ratio of CD56++ NK-cells, and the remainder was categorized into group 2. The relapse after allo-SCT was seen in 1 out of 8 patients in group 1, whereas it was documented in 5 out of 17 patients in group 2. This finding suggested that CD56++ NK-cells might also have a role in preventing relapse. We have found that peripheral CD56++CD16− NK-cells from patients after allo-SCT consistently expressed TNF-related apoptosis-inducing ligand (TRAIL), although its expression was faintly detectable on circulating NK-cells from healthy volunteers. As reported, stimulation with IL-2 or IL-15 resulted in the increased expression of TRAIL on NK-cells from healthy volunteers as well as the recipients of allo-SCT. However, its expression was always stronger in the CD16- subset than CD16+ in both groups. Cultivation of purified NK-cells from healthy volunteers with 0.5 nM of IL-2 for more than 2 weeks resulted in the expansion of both NK-cell subsets, and after sorting into CD16− and CD16+ NK-cells, cytotoxic assays against Jurkat were performed in the presence or absence of concanamycin A, neutralizing anti-Fas antibody, and neutralizing anti-TRAIL antibody. Cytotoxicity was more prominent in the CD16− subset than CD16+, and blocking study revealed that TRAIL expressed on CD16− NK-cells was strongly involved in the killing of Jurkat. We could not detect TRAIL-mediated cytotoxicity in the CD16+ subset, because the expression of TRAIL was much lower in the CD16+ subset than CD16−. Next, NK-cells purified from allo-SCT recipients and healthy volunteers were overnight cultured with 0.5 nM of IL-2 and their cytotoxicity against Jurkat was examined. NK-cells from patients who received allo-SCT within 3 months and those from healthy volunteers showed equivalent cytotoxicity. In patients who showed increased ratio of CD56++CD16− NK-cells, TRAIL was strongly expressed on overnight cultured CD56++CD16− NK cells, and TRAIL-mediated cytotoxicity was also detected. In murine models, TRAIL has been reported to exert strong graft-versus-tumor effects without causing GVHD. As CD56++CD16− NK cells readily express functional TRAIL on cytokine stimulation, and they usually reconstituted shortly early after allo-SCT, they may become promising targets for immunological intervention.

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