Journal: Nature Cardiovascular Research

Article Title: Progenitor exhausted PD-1 + T cells are cellular targets of immune checkpoint inhibition in atherosclerosis

doi: 10.1038/s44161-025-00713-2

Figure Lengend Snippet: ( a ) Splenocytes from Ifng YFP/YFP Apoe − / − mice were stimulated with PMA/ionomycin with Brefeldin A for 4 hr and analyzed for IFN-γ-YFP and IFN-γ-AF700 expression. ( b-d ) Ifng YFP/YFP Apoe − / − mice were fed a high-cholesterol diet for 12 weeks, and flow cytometry was performed to characterize IFN-γ-producing T cells (n = 11). ( b ) Increased frequency of IFN-γ-YFP expression in Tbet + splenic CD4 (**** p = 6.9E-5) and CD8 T cells (*** p = 0.0004) compared to Tbet − T cells. ( c ) Overlay flow cytometry plots of IFN-γ + (orange) and IFN-γ − (grey) aortic CD4 and CD8 T cells. ( d ) Flow cytometry plots of PD1-PE/Dazzle antibody staining and matched controls, PE/Dazzle-rat IgG2a isotype control (PD1 isotype) and PD1-PE/Dazzle fluorescent minus-one (FMO), of splenic CD4 T cells. ( e ) Representative immunohistochemical staining of PDL1 in aortic root cross-section of Ifng YFP/YFP Apoe − / − mice fed a high-cholesterol diet for 16 weeks (n = 4). ( b ) Bars denote median, analyzed with two-sided Mann-Whitney U test.

Article Snippet: Mice were randomly assigned to receive intraperitoneal injections of either murinized and effector-less anti-PD-1 antibody (clone: RMP1-14, anti-mPD-1-mIgG1e3; InvivoGen, mpd1-mab15-50) or isotype control IgG2a (anti-mPD-1-mIgG1e3, InvivoFit; InvivoGen) biweekly for 6 weeks at 10 mg kg −1 ( n = 12 per group).

Techniques: Expressing, Flow Cytometry, Staining, Control, Immunohistochemical staining, MANN-WHITNEY

Journal: Nature Cardiovascular Research

Article Title: Progenitor exhausted PD-1 + T cells are cellular targets of immune checkpoint inhibition in atherosclerosis

doi: 10.1038/s44161-025-00713-2

Figure Lengend Snippet: a – c , TCR signaling reporter mice ( Nur77 wt/GFP Apoe − / − ) were generated and fed an HCD for 12 weeks, and, at euthanization, T cell phenotype was analyzed ( n = 9). a , Flow cytometry plots of Nur77–GFP expression on aortic T cells. b , MFI of Slamf6 (** P = 0.004), PD-1 (** P = 0.0019) and Tim3 (*** P = 0.0002) of Nur77 − and Nur77 + (recent TCR activation) aortic CD4. c , MFI of Slamf6 (**** P = 9.2 × 10 −5 ), PD-1 and Tim3 (* P = 0.01) of Nur77 − and Nur77 + aortic CD8 T cells. d , Flow cytometry plots of CD69 and CD103 expression on aortic CD8 T cells of Ifng YFP/YFP Apoe − / − mice fed an HCD for 14 weeks ( n = 9). e , f , Quantification of percent CD69 + T cells (CD4 **** P = 8.5 × 10 −5 , * P = 0.049; CD8 **** P = 5.6 × 10 −9 , *** P = 0.0004) and CD69 + CD103 + T cells (CD4 * P = 0.012, CD8 * P = 0.011) within PD-1 subsets in the aorta. g , h , Flow cytometry analyzing CD69 and Nur77–GFP expression within PD-1 subsets of aortic CD4 T cells from Ifng YFP/YFP Nur7 wt/GFP Apoe − / − mice fed an HCD for 10 weeks ( n = 6). Quantification of non-activated CD4 T RM -like cells (CD69 + Nur77–GFP − ) and recently activated CD4 T cells (CD69 − Nur77–GFP + and CD69 + Nur77–GFP + ). i – j , Flow cytometry analyzing CD69 and Nur77–GFP expression within PD-1 subsets of aortic CD8 T cells. k – m , In a separate cohort, Ifng YFP/YFP Apoe − / − mice were fed an HCD for 10 weeks before receiving intraperitoneal injections of anti-IL-2 antibodies or control IgG ( n = 9 per group) for 2 weeks. k , l , Flow cytometric analysis of aortic memory T cells to assess composition of PD-1 subsets and subsets of cells expressing Slamf6 and/or Tim3. m , Flow cytometric analysis of IFNγ production by aortic CD44 + CD4 and CD44 + CD8 T cells. b , l , Bars denote median, analyzed with two-sided Mann–Whitney U -test. c , m , Bars denote mean, analyzed with two-sided unpaired t -test. e , f , h , j , Bars denote median, analyzed with two-sided Kruskal–Wallis test. h , j , P values compared to PD-1 − subset (CD4 PD-1 int ** P = 0.0088, CD4 PD-1 high * P = 0.0016, CD8 PD-1 int * P = 0.031, CD8 PD-1 high *** P = 0.0005). act, activated; Ctrl, control.

Article Snippet: Mice were randomly assigned to receive intraperitoneal injections of either murinized and effector-less anti-PD-1 antibody (clone: RMP1-14, anti-mPD-1-mIgG1e3; InvivoGen, mpd1-mab15-50) or isotype control IgG2a (anti-mPD-1-mIgG1e3, InvivoFit; InvivoGen) biweekly for 6 weeks at 10 mg kg −1 ( n = 12 per group).

Techniques: Generated, Flow Cytometry, Expressing, Activation Assay, Control, MANN-WHITNEY

Journal: Nature Cardiovascular Research

Article Title: Progenitor exhausted PD-1 + T cells are cellular targets of immune checkpoint inhibition in atherosclerosis

doi: 10.1038/s44161-025-00713-2

Figure Lengend Snippet: ( a - c ) Atherosclerotic dual-reporter mice ( Ifng YFP/YFP Nur77 wt/GFP Apoe − / − ) were generated, fed a high-cholesterol diet for 10 weeks, and flow cytometry was performed to characterize Nur77 expression on IFN-γ-producing T cells in spleen, iliac aortic-draining lymph node (aLN), and aorta (n = 6). ( a ) Flow cytometry plots and ( b-c ) quantification of association between Nur77-GFP expression and PD1 expression within IFN-γ-YFP + CD4 and CD8 T cells in the aorta, iliac aortic-draining lymph nodes (aLN), and spleen. ( d ) Representative flow cytometry gating for identifying follicular T helper cells (Tfh; CXC5 + PD1 high CD4 + ) in Ifng YFP/YFP Apoe − / − mice. ( e ) Experimental design for anti-IL-2 study. Ifng YFP/YFP Apoe − / − mice were fed a high cholesterol diet (HCD) for 10 weeks before receiving i.p . injections (3 doses, 5 days apart) of anti-IL-2 antibodies or control IgG (n = 9/group) for 2 weeks.

Article Snippet: Mice were randomly assigned to receive intraperitoneal injections of either murinized and effector-less anti-PD-1 antibody (clone: RMP1-14, anti-mPD-1-mIgG1e3; InvivoGen, mpd1-mab15-50) or isotype control IgG2a (anti-mPD-1-mIgG1e3, InvivoFit; InvivoGen) biweekly for 6 weeks at 10 mg kg −1 ( n = 12 per group).

Techniques: Generated, Flow Cytometry, Expressing, Control

Journal: Nature Cardiovascular Research

Article Title: Progenitor exhausted PD-1 + T cells are cellular targets of immune checkpoint inhibition in atherosclerosis

doi: 10.1038/s44161-025-00713-2

Figure Lengend Snippet: a – k , o – u , Ifng YFP/YFP Apoe − / − mice were fed an HCD for 10 weeks before administering biweekly intraperitoneal injections of a murine blocking anti-PD-1 antibody or isotype IgG control ( n = 12 per group). Mice received a total of 12 injections over 6 weeks and were euthanized after a total of 16 weeks of diet. b , Quantification of average plaque area of aortic subvalvular cross-sections. c , d , Quantification and representative immunohistochemical staining of T cells (CD3 + ) in aortic root plaques (**** P = 5 × 10 −5 ). e – i , Representative flow cytometry plots and quantification of frequency and numbers of PD-1 subsets of aortic memory CD4 (*** P = 0.0001) and CD8 ( h **** P = 7.4 × 10 −7 , i **** P = 5.2 × 10 −6 ) T cells. j , k , Flow cytometric analysis of frequency of Tim3 + PD-1 + memory CD4 and CD8 aortic T cells (** P = 0.0024) and frequency of IFNγ production of memory CD4 and CD8 aortic T cells (** P = 0.0023, 0.0022). l , m , Representative immunohistochemical images of and quantification of presence of at least one T cell foci in the plaque and/or surrounding adventitia (* P = 0.024). For study details of 3-week anti-PD-1 cohort, see Extended Data Fig. (ctrl IgG n = 12, anti-PD-1 n = 13). n , Impact of presence of tumor (MC38 tumor) during anti-PD-1 therapy ( n = 7 per group) on T cell (CD3 + ) infiltration into subvalvular aortic plaques (** P = 0.0079, * P = 0.026) compared to ctrl IgG ( n = 5 per group), determined by immunohistochemistry. o – q , Quantification and representative immunohistochemical staining of neutrophils (Ly6G + ) in aortic root plaques (** P = 0.0070) and adventitia (**** P = 9.5 × 10 −5 ) in Ifng YFP/YFP Apoe − / − mice treated with PD-1 blockade or isotype IgG ctrl for 6 weeks ( n = 12 per group). r , Quantification of immunohistochemical staining of macrophages (CD68 + ) in aortic root plaques. s , Gene expression analysis of aortic subvalvular cross-sections of ctrl IgG-treated or anti-PD-1-treated mice. Genes are reported as upregulated or downregulated in anti-PD-1-treated mouse compared to isotype IgG control. Adjusted P value is adjusted for FDR. t , u , Concentration of plasma cytokines CXCL9 (** P = 0.0063) and CXCL10 (* P = 0.037). b , c , i , n , p – r , u , Bars denote median, analyzed with two-sided Mann–Whitney U -test. t , Bars denote mean, analyzed with two-sided unpaired t -test. f – h , j , k , Bars denote median, analyzed with two-sided Mann–Whitney U -test or two-sided unpaired t -test. m , Analyzed with one-sided Fisher’s exact test. adj., adjusted; Adv., adventitia; Avg., average; ctrl, control; FC, fold change; i.p., intraperitoneal; w, week.

Article Snippet: Mice were randomly assigned to receive intraperitoneal injections of either murinized and effector-less anti-PD-1 antibody (clone: RMP1-14, anti-mPD-1-mIgG1e3; InvivoGen, mpd1-mab15-50) or isotype control IgG2a (anti-mPD-1-mIgG1e3, InvivoFit; InvivoGen) biweekly for 6 weeks at 10 mg kg −1 ( n = 12 per group).

Techniques: Blocking Assay, Control, Immunohistochemical staining, Staining, Flow Cytometry, Immunohistochemistry, Gene Expression, Concentration Assay, Clinical Proteomics, MANN-WHITNEY

Journal: Nature Cardiovascular Research

Article Title: Progenitor exhausted PD-1 + T cells are cellular targets of immune checkpoint inhibition in atherosclerosis

doi: 10.1038/s44161-025-00713-2

Figure Lengend Snippet: Ifng YFP/YFP Apoe − / − mice were fed a high-cholesterol diet for 10 weeks before administration of bi-weekly i.p . injections of a murine blocking anti-PD1 antibody or isotype IgG control (n = 12/group) for 6 weeks. ( a ) Quantification of CD3 + cells in adventitia of aortic-root sections. Adventitia designated as 100 μm from smooth muscle layer of aortic valve. ( b ) Isotype IgG staining for CD3 immunohistochemical analysis of aortic root cross-sections. ( c ) Quantification of T cells (TCRβ + CD45 + ) per whole-aorta digest analyzed by flow cytometry (* p = 0.010). ( d-h ) In the short-term anti-PD1 cohort, Ifng YFP/YFP Apoe − / − mice were fed a high-cholesterol diet for 21 weeks before administration of bi-weekly i.p . injections of anti-PD1 antibody or isotype IgG control (n = 11-12/group). Mice received a total of six injections and were terminated after a total of 24 weeks of diet. ( e ) Average plaque area determined by Oil Red O staining of aortic subvalvular cross-sections. ( f-g ) Quantification and representative image of immunohistochemical analysis of CD3 + cells in aortic root plaques (*** p = 0.0002). ( h ) Representative immunohistochemical staining images and confocal images of CD3 and CD19 staining of advanced adventitial foci. ( i ) Measurements of MC38 tumor diameter in HCD-fed Ifng YFP/YFP Apoe − / − mice with (n = 5) and without (n = 7) PD1 blockade. Data presented as mean values +/- SD. ( j-k ) Representative images of immunohistochemical staining of neutrophils (Ly6G) and macrophages (CD68) in Ifng YFP/YFP Apoe − / − mice treated with PD1 blockade or isotype IgG control for 6 weeks (n = 12/group). ( l-p ) Concentration of plasma cytokines CCL4 (* p = 0.023), IL-6, TNF-α, IL-4 (*** p = 0.0009), and CCL2 (** p = 0.0013) in Ifng YFP/YFP Apoe − / − mice treated with anti-PD1 or isotype IgG control for 6 weeks. ( a , c , f , m - p ). Bars denote median, analyzed with two-sided Mann-Whitney U test. ( e, l ) Bars denote mean, analyzed with two-sided unpaired t test.

Article Snippet: Mice were randomly assigned to receive intraperitoneal injections of either murinized and effector-less anti-PD-1 antibody (clone: RMP1-14, anti-mPD-1-mIgG1e3; InvivoGen, mpd1-mab15-50) or isotype control IgG2a (anti-mPD-1-mIgG1e3, InvivoFit; InvivoGen) biweekly for 6 weeks at 10 mg kg −1 ( n = 12 per group).

Techniques: Blocking Assay, Control, Staining, Immunohistochemical staining, Flow Cytometry, Concentration Assay, Clinical Proteomics, MANN-WHITNEY

Journal: Nature Cardiovascular Research

Article Title: Progenitor exhausted PD-1 + T cells are cellular targets of immune checkpoint inhibition in atherosclerosis

doi: 10.1038/s44161-025-00713-2

Figure Lengend Snippet: Ifng YFP/YFP Apoe − / − mice were fed with an HCD for 4 months and treated for the final 3 weeks with isotype control only ( n = 11), depleting anti-PD-1 (RMP1.30, n = 10) only, ctrl IgG for 1 week prior to injection of blocking anti-PD-1 antibodies (ctrl IgG + anti-PD-1, n = 13) or depleting anti-PD-1 (RMP1.30) for 1 week prior to injection of blocking anti-PD-1 antibodies (RMP1.30 + anti-PD-1, n = 13). a , Experimental overview. b , c , Numbers of circulating CD44 + PD-1 int CD4 and CD8 T cells throughout treatment course (days 0, 7 and 21). d , e , Numbers of circulating CD44 + PD-1 high CD4 (** P = 0.0010) and CD8 (** P = 0.0014, * P = 0.017) T cells throughout the treatment course. f , g , Quantification and representative immunohistochemical staining of T cells (CD3 + ) in aortic root plaques (** P = 0.0077, * P = 0.027). h , i , Flow cytometric analysis of counts of progenitor exhausted (CD44 + Slamf6 + Tim3 − PD-1 int ) aortic CD4 (* P = 0.025, ** P = 0.0072) and CD8 (*** P = 0.0004, * P = 0.014, 0.012) T cells. j , k , Flow cytometric analysis of counts CD44 + PD-1 high aortic CD4 (* P = 0.039, 0.011, 0.014) and CD8 (*** P = 0.009, 0.0021, **** P = 2.4 × 10 −6 ) T cells. l , m , Flow cytometric analysis of frequency of IFNγ production by aortic memory CD4 (*** P = 0.0006, ** P = 0.0013, * P = 0.014, 0.026) and CD8 (*** P = 0.0006, 0.0009, ** P = 0.0061, 0.0097) T cells. b – e , Data presented as mean values ± s.e.m., analyzed with mixed-effects multiple comparisons analysis. g – l , Bars denote median, analyzed with Kruskal–Wallis test. m , Bars denote mean, analyzed with one-way ANOVA. P values are reported top-to-bottom and left-to-right. ctrl, control.

Article Snippet: Mice were randomly assigned to receive intraperitoneal injections of either murinized and effector-less anti-PD-1 antibody (clone: RMP1-14, anti-mPD-1-mIgG1e3; InvivoGen, mpd1-mab15-50) or isotype control IgG2a (anti-mPD-1-mIgG1e3, InvivoFit; InvivoGen) biweekly for 6 weeks at 10 mg kg −1 ( n = 12 per group).

Techniques: Control, Injection, Blocking Assay, Immunohistochemical staining, Staining

Journal: Nature Cardiovascular Research

Article Title: Progenitor exhausted PD-1 + T cells are cellular targets of immune checkpoint inhibition in atherosclerosis

doi: 10.1038/s44161-025-00713-2

Figure Lengend Snippet: Ifng YFP/YFP Apoe − / − mice fed HCD-fed for 4 months were treated in the final 3 weeks with either: isotype control only (n = 11), depleting anti-PD1 (RMP1.30, n = 10) only, ctrl IgG for one week prior to injection of blocking anti-PD1 antibodies (ctrl IgG + anti-PD1, n = 13), depleting anti-PD1 (RMP1.30) for one week prior to injection of blocking anti-PD1 antibodies (RMP1.30 + anti-PD1; n = 13). ( a-b ) Quantification of aortic CD44 + PD1 int CD4 (top-down, * p = 0.031, ** p = 0.0044, * p = 0.035) and CD44 + PD1 int CD8 (*** p = 0.0005, * p = 0.019, ** p = 0.0011, * p = 0.033) T cells. Bars denote median, analyzed with two-sided Kruskal-Wallis test.

Article Snippet: Mice were randomly assigned to receive intraperitoneal injections of either murinized and effector-less anti-PD-1 antibody (clone: RMP1-14, anti-mPD-1-mIgG1e3; InvivoGen, mpd1-mab15-50) or isotype control IgG2a (anti-mPD-1-mIgG1e3, InvivoFit; InvivoGen) biweekly for 6 weeks at 10 mg kg −1 ( n = 12 per group).

Techniques: Control, Injection, Blocking Assay

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: LPS/ATP stimulation activates NLRP3/caspase-1/IL-1β axis and promotes MHC-I expression. A Western blot analysis showed increased expression of NLRP3, pro-caspase-1, cleaved caspase-1, pro-IL-1β and mature IL-1β in the Raw 264.7 macrophages under LPS/ATP stimulation. B , C ELISA analysis showed that the expression of IL-1β and TNF-α in the Raw 264.7 macrophages were up-regulated by LPS/ATP. D – G ELISA analysis showed elevated levels of IFNs and MCP-1 produced by the stimulated macrophages. H The mRNA expression of MHC-I in the co-cultured C2C12 cells was increased under LPS/ATP stimulation and peaked at 48 h. I , J The protein expression of MHC-I in the co-cultured C2C12 cells was increased under LPS/ATP stimulation and peaked at 72 h. * p < 0.01, ** p < 0.001, compared with control group or compared between the groups connecting with the line

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Expressing, Western Blot, Analysis, Enzyme-linked Immunosorbent Assay, Produced, Cell Culture, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: Genetic knockdown of NLRP3 suppresses MHC-I overexpression in C2C12 cells. A Raw 264.7 macrophages were transfected with siRNA targeting NLRP3, and the transfection efficiency was measured by western blot analysis. B , C Raw 264.7 macrophages pretreated with NLRP3-siRNA were stimulated with 200 ng/ml LPS + 500 μM ATP, and then the expression of MHC-I in the co-cultured C2C12 cells was significantly reduced compared with those pretreated with NC-siRNA, as revealed by western blot. D , E The stimulated Raw 264.7 macrophages were pretreated with NLRP3-siRNA or NC-siRNA, and the co-cultured C2C12 cells were added with neutralizing IL-1β monoclonal antibody. No differences in the expression of MHC-I in the co-cultured C2C12 cells were found between those groups stimulated with 200 ng/ml LPS + 500 μM ATP or with PBS, as showed by western blot. ** p < 0.001, compared with control group; ¶¶ p < 0.001, compared with before treatment

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Knockdown, Over Expression, Transfection, Western Blot, Analysis, Expressing, Cell Culture, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: Pharmacological inhibition of NLRP3 inflammasome using MCC950 effectively suppresses MHC-I overexpression in C2C12 cells. A , B Cell viability assay revealed no significant cytotoxicity of MCC950 to the Raw 264.7 cells and C2C12 cells, at the dosage from 0.01 to 10 μM. C LPS/ATP stimulated Raw 264.7 macrophages were treated by MCC950 with dosage from 0.01 to 10 μM, and the IL-1β level in the cell supernatant was significantly decreased at the dosage of 10 μM. D , E LPS/ATP stimulated Raw 264.7 macrophages were pretreated by 10 μM MCC950, and the MHC-I expression in the co-cultured C2C12 cells was significantly suppressed as showed by western blot. * p < 0.01, ** p < 0.001, compared with control group; ¶¶ p < 0.001, compared with before treatment

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Inhibition, Over Expression, Cell Viability Assay, Expressing, Cell Culture, Western Blot, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: IL-1β is crucial for MHC-I up-regulation in C2C12 cells. A , B Recombinant IL-1β was added into the C2C12 cells. The expression of MHC-I in C2C12 cells peaked when treated by 1 ng/ml IL-1β for 48 h (n = 5). C Raw 264.7 macrophages were transfected with siRNA targeting IL-1β, and the transfection efficiency was measured by western blot analysis. D , E LPS/ATP stimulated Raw 264.7 macrophages were pretreated with IL-1β-siRNA or NC-siRNA, and the MHC-I expression in the co-cultured C2C12 cells was significantly lower with the former than with the latter pretreatment, as analyzed by western blot. * p < 0.01, ** p < 0.001, compared with control group; ¶¶ p < 0.001, compared with before treatment

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Recombinant, Expressing, Transfection, Western Blot, Analysis, Cell Culture, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: MCC950 alleviates the intensity of inflammation and MHC-I expression in PM model rats. A , B The muscle section samples of PM modal rats showed a substantial reduction in muscle histological inflammation scores after treated by MCC950. C – E Serum CRP, CK, and LDH levels were significantly reduced after MCC950 treatment in PM modal rats. F – I The expression of NLRP3, MHC-I, and IL-1β were effectively reduced in the muscle tissue of PM model rats by MCC950 treatment. ** p < 0.001, compared with control group; ¶ p < 0.01, ¶¶ p < 0.001, compared with before treatment. Scale bars, 50 μm (original magnification, ×400)

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Expressing, Tissue, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: The level of inflammation is increased in the PM patients and the NLRP3/caspase-1/ IL-1β axis is active in their muscle samples. A Immunohistochemistry analysis showed higher percentage of CD68+ cells (solid black arrows) in the muscle biopsies in the PM patients than in the controls. B Immunofluorescence staining showed positive MHC-I staining (grade+ ~ ++) in the PM patients, and negative in the controls (grade−). C Immunohistochemistry analysis of the muscle biopsies showed higher expression of NLRP3, caspase-1 and IL-1β (solid black arrows) in the PM patients than in the controls. ** p < 0.001; scale bars, 50 μm (original magnification, ×400)

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Immunohistochemistry, Analysis, Immunofluorescence, Staining, Expressing

Journal: bioRxiv

Article Title: Dysplastic Epithelial Repair Propagates Chronic Pathology Through the Paracrine Transformation of Pulmonary Fibroblasts

doi: 10.64898/2026.04.02.716135

Figure Lengend Snippet: ( A ) Quantified levels of IL-1α from naïve fibroblast (Naïve fibro), inflammatory fibroblast (Inflam fibro), and ectopic basal cell (Basal) conditioned media as determined by ELISA. ( B ) Saa3 and Ccl2 gene expression by RT-qPCR for fibroblasts cultured on 0.5kPa plates and treated with a 5-fold titration of IL-1α added to control media, including the level determined in (A). ( C ) Graphic illustrating the strategy used to deplete conditioned media of endogenous IL-1α with antibodies, magnetic beads, and column-based separation. ( D ) Saa3 and Ccl2 gene expression by RT-qPCR for fibroblasts cultured on 0.5kPa plates in control or conditioned media depleted using IgG (CTRL IgG or CM IgG) or IL-1α nAb (CTRL nAB, CM nAb) and supplemented with additional IL-1α nAb after depletion (+Spike). [(B) and (D)] Data shows mean fold change ± SD and are pooled from 3 independent experiments (n=3 mice). P values were calculated using either (B) a ratio paired t -test or (D) a one-way ANOVA with post-hoc Turkey multiple comparison test: * P < 0.05, ** P < 0.01, **** P < 0.0001, ns: not significant.

Article Snippet: The working concentration for the following reagents can be found in the text and figure legends: Human IL-1RA Recombinant Protein (IL1Ra) (PeproTech®, 200-01RA), Mouse IL-1 alpha Recombinant Protein, (PeproTech®, 211-11A), Mouse IL-1α Neutralizing Antibody (Invivogen, Anti-mIL-1α-mIgG1 clone 6H7, mIL-1α-mab9-02), Recombinant Mouse IgG1 Isotype Control Antibody (Invivogen, Anti-β-Gal-mIgG1 clone T9C6, bgal-mab9-02), Human TGF-β 1 Recombinant Protein (PeproTech®, 100-21), Human GDF15 Recombinant Protein (PeproTech®, 120-28C), Human VEGF Recombinant Protein (PeproTech®, 450-32), Human OPN Recombinant Protein (PeproTech®, 120-35), Murine CXCL1 Recombinant Protein (PeproTech®, 250-11), Human IGFBP3 Recombinant Protein (PeproTech®, 100-08), Human TNFRSF11B Recombinant Protein (PeproTech®, 450-14), Murine CXCL5 Recombinant Protein (PeproTech®, 250-17), and Muring CCL20 Recombinant Protein (PeproTech®, 250-27).

Techniques: Enzyme-linked Immunosorbent Assay, Gene Expression, Quantitative RT-PCR, Cell Culture, Titration, Control, Magnetic Beads, Comparison