Summary of Study ST003425

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002117. The data can be accessed directly via it's Project DOI: 10.21228/M8ZV6G This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST003425
Study Title	Targeted free fatty acids metabolomics studies on serum, cecal content, cecum tissue, chow diet, and Tritrichomonas mu cells
Study Summary	Serum, cecal content, cecum tissue samples of murine, chow diet, and T.mu cells were collected to perform the targeted free fatty acids metabolome analysis. The aim of this study was to verify that whether T.mu could release free PUFA (especially ARA) to the intestinal tract of its host by comparing the PUFA concentration in the freshly isolated T.mu cells, chow diet, cecal content and serum of the host (Mus musculus).Based on the result of this targeted free fatty acids metabolomics studies, we found that T.mu could release ARA to the intestinal tract of its host and increase the concentration of ARA in its host's intestinal tract.
Institute	Xuzhou medical university
Last Name	Kou
First Name	Yanbo
Address	Tongshan road 209, Xuzhou, Jiangsu, 221004, China
Email	fightingkyb@163.com
Phone	+86-051683262123
Submit Date	2024-08-20
Raw Data Available	Yes
Raw Data File Type(s)	cdf
Analysis Type Detail	GC-MS
Release Date	2024-09-10
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR002117
Project DOI:	doi: 10.21228/M8ZV6G
Project Title:	A mouse protozoan boosts antigen-specific mucosal IgA responses in a specific lipid metabolism- and signaling-dependent manner
Project Summary:	IgA antibodies play an important role in mucosal immunity. However, there is still no effective way to consistently boost mucosal IgA responses, and the factors influencing these responses are not fully understood. We observed that colonization with the murine intestinal symbiotic protozoan Tritrichomonas musculis (T.mu) boosted antigen-specific mucosal IgA responses in wild-type C57BL/6 mice. This enhancement was attributed to the accumulation of free arachidonic acid (ARA) in the intestinal lumen, which served as a signal to stimulate the production of antigen-specific mucosal IgA. When ARA was prevented from undergoing its downstream metabolic transformation using the 5-lipoxygenase inhibitor zileuton or by blocking its downstream biological signaling through genetic deletion of the Leukotriene B4 receptor 1 (Blt1), the T.mu-mediated enhancement of antigen-specific mucosal IgA production was suppressed. Moreover, both T.mu transfer and dietary supplementation of ARA augmented the efficacy of an oral vaccine against Salmonella infection, with this effect being dependent on Blt1. Our findings elucidate a tripartite circuit linking nutrients from the diet or intestinal microbiota, host lipid metabolism, and the mucosal humoral immune response.
Institute:	Xuzhou medical university
Last Name:	Kou
First Name:	Yanbo
Address:	Tongshan road 209, Xuzhou, Jiangsu, 221004, China
Email:	fightingkyb@163.com
Phone:	+86-051683262123

Subject:

Subject ID:	SU003552
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Gender:	Not applicable

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id	local_sample_id	Sample source	Treatment
SA377682	T_mu_content	cecal content	colonized with Tmu
SA377683	Ctrl_content	cecal content	control
SA377684	T_mu_cecum	cecum tissue	colonized with Tmu
SA377685	Ctrl_cecum	cecum tissue	control
SA377686	Diet	chow diet	ground to power
SA377687	T_mu_serum	serum	colonized with Tmu
SA377688	Ctrl_serum	serum	control
SA377681	T_mu	T_mu cells	isolated from cecal content

Showing results 1 to 8 of 8

Showing results 1 to 8 of 8

Collection:

Collection ID:	CO003545
Collection Summary:	WT C57/BL6 mice were colonized with or without T.mu for 7 days, and the serum, cecum tissue, cecal content samples were collected on day 7. For T.mu cells sample, T.mu cells were isolated and counted from T.mu positive cecal content. Samples were washed with cold PBS and then flash-frozen in liquid nitrogen.
Sample Type:	Serum, cecum tissue, cecal content, Tritrichomonas musculis cells, chow diet

Treatment:

Treatment ID:	TR003561
Treatment Summary:	WT C57/BL6 mice were colonized with or without T.mu for 7 days, and the serum, cecum tissue, cecal content samples were collected on day 7. For T.mu cells sample, T.mu cells were isolated and counted from T.mu positive cecal content. For diet sample, the chow diet power was used.

Sample Preparation:

Sampleprep ID:	SP003559
Sampleprep Summary:	The weighed cecal content, cecum tissue, serum, animal chow diet powder, or enumerated T.mu cells were transferred into new 2 mL EP tubes followed extraction with 500 μL extracting solution [Isopropanol : n-Hexane = 2:3 (V:V)], including 0.2 mg/L internal standard). After homogenized in a ball mill for 4 min at 40 Hz and a 5 min ultrasound treatment, the samples were centrifuged 16200 × g for 15 min at 4 °C. Then the supernatants were transferred into new 2 mL EP tubes followed with nitrogen blow dry. The dried samples were resuspended in 500 μL of methanol: trimethylsilyl diazomethane solution (1:2), after standing 30 min at room temperature, another nitrogen blow dry was performed. Then, the samples were resuspended in 160 μL of n-hexane followed with a centrifugation of 16200 × g for 1 min. Finally, the supernatants were transferred into new vials for GC-MS analysis.

Sampleprep ID:

SP003559

Sampleprep Summary:

The weighed cecal content, cecum tissue, serum, animal chow diet powder, or enumerated T.mu cells were transferred into new 2 mL EP tubes followed extraction with 500 μL extracting solution [Isopropanol : n-Hexane = 2:3 (V:V)], including 0.2 mg/L internal standard). After homogenized in a ball mill for 4 min at 40 Hz and a 5 min ultrasound treatment, the samples were centrifuged 16200 × g for 15 min at 4 °C. Then the supernatants were transferred into new 2 mL EP tubes followed with nitrogen blow dry. The dried samples were resuspended in 500 μL of methanol: trimethylsilyl diazomethane solution (1:2), after standing 30 min at room temperature, another nitrogen blow dry was performed. Then, the samples were resuspended in 160 μL of n-hexane followed with a centrifugation of 16200 × g for 1 min. Finally, the supernatants were transferred into new vials for GC-MS analysis.

Combined analysis:

Analysis ID	AN005624
Analysis type	MS
Chromatography type	GC
Chromatography system	Agilent 7890B
Column	Agilent DB-FastFAME capillary column (90 m × 0.25 mm × 0.25 um)
MS Type	EI
MS instrument type	Single quadrupole
MS instrument name	Agilent 5977B
Ion Mode	POSITIVE
Units	μg/ml for serum, μg/g for cecal content, μg/g for diet, μg/8*10^7 T.mu cells

Chromatography:

Chromatography ID:	CH004274
Chromatography Summary:	DB-FastFAME capillary column (90 m × 0.25 mm × 0.25 µm, Agilent Technologies)
Instrument Name:	Agilent 7890B
Column Name:	Agilent DB-FastFAME capillary column (90 m × 0.25 mm × 0.25 um)
Column Temperature:	230
Flow Gradient:	-
Flow Rate:	-
Solvent A:	-
Solvent B:	-
Chromatography Type:	GC

MS:

MS ID:	MS005348
Analysis ID:	AN005624
Instrument Name:	Agilent 5977B
Instrument Type:	Single quadrupole
MS Type:	EI
MS Comments:	Mass spectrometry data were acquired in Scan/SIM mode over an m/z range of 33-400 after a solvent delay of 7 minutes. Fatty acid methyl esters (FAMEs) were identified by comparing them to commercial FAME standards (ANPEL, Shanghai, China, Cat# CDAA-252). The absolute concentrations of FAMEs were calculated using calibration curves based on the internal standard method.
Ion Mode:	POSITIVE