Journal: Advanced Science

Article Title: Islet‐Resident Memory T Cells Orchestrate the Immunopathogenesis of Type 1 Diabetes through the FABP4‐CXCL10 Axis

doi: 10.1002/advs.202308461

Figure Lengend Snippet: T RM cells enhance the immunopathologic development of T1D through the FABP4‐CXCL10 axis. A) Representative immunoblots of CXCL10 and HSP90 in the pancreas of FABP4 +/+ NOD and FABP4 −/− NOD mice. The right panel is the quantification of the band intensity of CXCL10 relative to HSP90 ( n = 4). B) The mRNA abundance of FABP4, IFNγ, and CXCL10 in FABP4 +/+ T RM cells and FABP4 −/− T RM cells isolated from NOD mice ( n = 6). C) Representative FACS plots showing the abundance of IFNγ and CXCL10 in FABP4 +/+ T RM cells or FABP4 −/− T RM cells. The right panel is the quantification of positive cell percentage ( n = 5). D) Concentration of IFNγ and CXCL10 in the medium of FABP4 +/+ T RM cells or FABP4 −/− T RM cells stimulated with CD3 + CD28 + beads or vehicle ( n = 6). E,F) Representative images (E) and quantification (F) of displacement length (left) and average speed (right) analyzed by Imaris showing the motility of GFP‐labeled CD8 + T cells cultured in the conditioned media of FABP4 +/+ T RM or FABP4 −/− T RM cells in the presence or absence of recombinant CXCL10 protein (2 µg mL −1 ) or IgG control (2 µg mL −1 ) for 24 h (scale bar 50 µm). G) The number of residual (upper) and migrated (lower) CD8 + T cells cultured in the trans‐well plate with a conditioned medium of FABP4 +/+ T RM cells or FABP4 −/− T RM cells in the presence or absence of recombinant CXCL10 protein (2 µg mL −1 ) or IgG (2 µg mL −1 ) for 24 h ( n = 3). H,I) Representative pictures (H) and quantification (I) of displacement length (left) and average speed (right) showing the motility of GFP‐labeled CD8 + T cells cultured in the conditioned medium of FABP4 +/+ T RM or FABP4 −/− T RM cells treated with siCXCL10 (5 nmol) or siControl (siCtrl, 5 nmol) for 24 h (scale bar 50 µm). J) The number of residual (upper) and migrated (lower) CD8 + T cells cultured in the trans‐well plate with the conditioned medium from FABP4 +/+ T RM cells or FABP4 −/− T RM cells treated with siCXCL10 or siCtrl (5 nmol) for 24 h ( n = 5). Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. Unpaired Student's t test or Mann‐Whitney U test was used in (A), (B), and (C). ANOVA followed by Sidak's test was used in (D), (F), (G), (I), and (J).

Article Snippet: To assess the alarming function of FABP4 +/+ T RM and FABP4 −/− T RM cells and the involvement of CXCL10, the conditioned medium of respective cells were added to the lower wells. and supplemented with recombinant murine CXCL10 (2 g mL −1 , PeproTech) or IgG as control.

Techniques: Western Blot, Isolation, Concentration Assay, Labeling, Cell Culture, Recombinant, Control, MANN-WHITNEY

Journal: Advanced Science

Article Title: Islet‐Resident Memory T Cells Orchestrate the Immunopathogenesis of Type 1 Diabetes through the FABP4‐CXCL10 Axis

doi: 10.1002/advs.202308461

Figure Lengend Snippet: Targeting FABP4 resembles T RM cell depletion in alleviating T1D development. FABP4 +/+ NOD and FABP4 −/− NOD mice were subjected to the treatment of anti‐CD69 neutralizing antibody to deplete T RM cells. A) Schematic diagram showing the protocol of T RM cell depletion in NOD mice. Six‐week‐old female FABP4 +/+ NOD and FABP4 −/− NOD mice were injected with anti‐CD69 antibody (40 µg/mouse) or IgG every week (i.v.) for 8 weeks ( n = 5). B) The representative FACS plots show the abundance of T RM cells (CD8 + CD44 + CD69 + CD62L − ) in the mouse pancreas. The right panel is the quantification of the percentage of T RM cells ( n = 5). C) Diabetes incidence of mice ( n = 5). D) Scoring of histological grades of insulitis in mice ( n = 5). E) Representative images of immunofluorescent co‐staining for insulin‐positive β cells (green) and TUNEL‐positive apoptotic cells (red) in mouse pancreas (scale bar 20 µm). The lower panel is the quantification of the number of apoptotic cells within the islet area ( n = 5). F) The mRNA abundance of inflammatory cytokines and chemokines in mouse pancreas ( n = 5). G) The abundance of CXCL10 protein in mouse pancreatic islets ( n = 4). H) Representative immunoblots of CXCL10 and HSP90 in mouse pancreas. The lower panel is the quantification of the band intensity of CXCL10 relative to HSP90 ( n = 4). I) The abundance of total cytotoxic T cells (CD8 + CD44 + ), antigen‐experienced T cells (CD8 + CXCR3 + ), and IFN γ or Granzyme B expressing CD8 + T cells infiltrated in the pancreatic islets of the above mice ( n = 5). Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. ANOVA followed by Sidak's test was used in (B), (E), (F), (G), (H), and (I). The log‐rank test was used in (C). The Pearson's chi‐square test was used in (D).

Article Snippet: To assess the alarming function of FABP4 +/+ T RM and FABP4 −/− T RM cells and the involvement of CXCL10, the conditioned medium of respective cells were added to the lower wells. and supplemented with recombinant murine CXCL10 (2 g mL −1 , PeproTech) or IgG as control.

Techniques: Injection, Staining, TUNEL Assay, Western Blot, Expressing

Journal: Advanced Science

Article Title: Islet‐Resident Memory T Cells Orchestrate the Immunopathogenesis of Type 1 Diabetes through the FABP4‐CXCL10 Axis

doi: 10.1002/advs.202308461

Figure Lengend Snippet: T RM cells orchestrate the immunopathogenesis of type 1 Diabetes through FABP4‐CXCL10 axis.

Article Snippet: To assess the alarming function of FABP4 +/+ T RM and FABP4 −/− T RM cells and the involvement of CXCL10, the conditioned medium of respective cells were added to the lower wells. and supplemented with recombinant murine CXCL10 (2 g mL −1 , PeproTech) or IgG as control.

Techniques: