Dataset Information

Mouse spleen LC-MSMS from C57BL/6J WT, CD38-deficient and B6.C-H2bm12/KHEg (bm12) mice in cGVHD lupus-like model

ABSTRACT: Proteins from post-nuclear cell lysates from C57BL/6J WT, CD38-deficient and B6.C-H2bm12/KHEg (bm12) recipient mice at 2 weeks after the adoptive transfer of co-isogenic spleen cells were identified using a proteomic approach. The relative abundances of the proteins identified were calculated using emPAIs scores.

INSTRUMENT(S): amaZon Speed ETD

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Spleen

DISEASE(S): Systemic Lupus Erythematosus

SUBMITTER: Jaime Sancho

LAB HEAD: Jaime Sancho

PROVIDER: PXD026947 | Pride | 2021-10-11

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	1_BM12__NT__210119_2926.dat	Other
	1_BM12__NT__210119_2926.mzid.gz	Mzid
	1_BM12__NT__210119_2926.yep	Other
	1_BM12__NT__210119_2928.dat	Other
	1_BM12__NT__210119_2928.mzid.gz	Mzid

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Publications

CD38 Deficiency Ameliorates Chronic Graft-Versus-Host Disease Murine Lupus via a B-Cell-Dependent Mechanism.

Martínez-Blanco África Á Domínguez-Pantoja Marilú M Botía-Sánchez María M Pérez-Cabrera Sonia S Bello-Iglesias Nerea N Carrillo-Rodríguez Paula P Martin-Morales Natividad N Lario-Simón Antonio A Pérez-Sánchez-Cañete María M MM Montosa-Hidalgo Laura L Guerrero-Fernández Salvador S Longobardo-Polanco Victoria M VM Redondo-Sánchez Sandra S Cornet-Gomez Alberto A Torres-Sáez María M Fernández-Ibáñez Ana A Terrón-Camero Laura L Andrés-León Eduardo E O'Valle Francisco F Merino Ramón R Zubiaur Mercedes M Sancho Jaime J

Frontiers in immunology 20210824

The absence of the mouse cell surface receptor CD38 in Cd38-/- mice suggests that this receptor acts as a positive regulator of inflammatory and autoimmune responses. Here, we report that, in the context of the chronic graft-versus-host disease (cGVHD) lupus inducible model, the transfer of B6.C-H2bm12/KhEg(bm12) spleen cells into co-isogenic Cd38-/- B6 mice causes milder lupus-like autoimmunity with lower levels of anti-ssDNA autoantibodies than the tr ...[more]

PMID: 34504495

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Project description:During reproductive life, the human endometrium undergoes around 480 cycles of growth, breakdown and regeneration should pregnancy not be achieved. This outstanding regenerative capacity is the basis for women’s cycling and its dysfunction may be involved in the etiology of pathological disorders. Therefore, the human endometrial tissue must rely on a remarkable endometrial somatic stem cells (SSC) population. Here we explore the hypothesis that human endometrial side population (SP) cells correspond to somatic stem cells. We isolated, identified and characterized the SP corresponding to the stromal and epithelial compartments using endometrial SP genes signature, immunophenotyping and characteristic telomerase pattern. We analyzed the clonogenic activity of SP cells under hypoxic conditions and the differentiation capacity in vitro to adipogenic and osteogenic lineages. Finally, we demonstrated the functional capability of endometrial SP to develop human endometrium after subcutaneous injection in NOD-SCID mice. Briefly, SP cells of human endometrium from epithelial and stromal compartments display genotypic, phenotypic and functional features of SSC. Human tissue collection: Human endometrial biopsies were taken using a Pipelle catheter under sterile conditions (from 18 to 48 years) throughout the menstrual cycle. Epithelial and stromal separation: Epithelial and stromal fractions were isolated using an established protocol with modifications. Briefly, samples were carefully dissected and minced into 1-2mm fragments, and enzymatically digested in DMEM containing 10 mg/ml collagenase type IA. Stromal cells (single cells or small aggregates) and epithelial glands were separated on a size basis using gravity sedimentation and membrane filtration. Cell suspensions were treated with erythrocyte lysis solution and evaluation of cell viability was performed with Propidium Iodide (PI; 5 μg/ml). Hoechst 33342 labeling: Cells isolated from the endometrial epithelial and stromal fractions were resuspended in DMEM prewarmed at 37ºC and supplemented with 2% FBS and 10mM HEPES. The cell suspension was labeled in the same media with 5 μg/ml of Hoechst 33342 dye (Ho-33342), either alone or in combination with 100 μM verapamil (Vp), in a water bath at 37ºC for 90-120 minutes. Then, cells were centrifuged for 6 minutes at 4ºC, and were resuspended in cold HBSS supplemented with 2%FBS and 10mM HEPES. PI permits to exclude dead cells prior to the flow cytometric analysis and sorting. Isolation of human endometrial SP cells: Cells were analyzed and sorted by a MoFlo® jet-in-air high speed sorter. Excitation was performed with a water cooled Enterprise II ion laser which operated at the 351 nm and 488 nm wavelengths, and worked at 30 mW. Hoechst 33342 blue and red fluorescences were detected through 405/30 and 670/20 nm band-pass filters respectively, by measuring the signals on a linear scale. PI fluorescence was detected through a band-pass filter of 613/20 on a logarithmic scale. The gates for cell sorting were defined to collect live cells with a low Hoechst 33342 fluorescence (SP fraction), as well as live cells with a high Hoechst 33342 fluorescence (NSP fraction). Microarrays experiments: Endometrial tissue consisted in a total of 8 endometrial biopsies (epithelial (n=8) and stromal (n=8) endometrial cell suspensions, epithelial SP fraction (n=8) and stromal SP fractions (n=8) separately) which were pooled in pairs at the RNA levels. Four microarrays were analyzed per group.

Dataset Information

Mouse spleen LC-MSMS from C57BL/6J WT, CD38-deficient and B6.C-H2bm12/KHEg (bm12) mice in cGVHD lupus-like model

Dataset's files

Publications

CD38 Deficiency Ameliorates Chronic Graft-<i>Versus</i>-Host Disease Murine Lupus <i>via</i> a B-Cell-Dependent Mechanism.

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