Journal: Translational Cancer Research

Article Title: OX40L and IL-2 combination strategy for gastric cancer immunotherapy

doi: 10.21037/tcr-2025-707

Figure Lengend Snippet: Immunohistochemical and flow cytometric analysis of gastric cancer tissue. (A) H&E histochemical staining of tumor-free area (left), tumor margin (middle), and gastric tumor site (right). (B) Representative IHC images for CD3 + , CD4 + , CD8 + , FOXP3, OX40, and OX40L in tumor-free area (left column), tumor margin (middle column), and gastric tumor site (right column) sections. (C) Immunofluorescence staining for the co-expression of CD3 + and OX40 in gastric cancer with anti-CD3 and anti-OX40 antibodies and DAPI for nuclear staining. (D) Flow cytometry analysis of OX40 expression in CD3 cells obtained from PBMC of gastric cancer patients (upper panel) and OX40 expression on CD3 on TILs in gastric cancer patients (low panel). Statistical analysis of all data were performed using one-way ANOVA. Data are shown as mean ± standard deviation. ns, nonsignificant; *, P<0.05; ***, P<0.001. ANOVA, analysis of variance; DAPI, 4',6-diamidino-2-phenylindole; H&E, hematoxylin & eosin; IHC, immunohistochemistry; OX40L, OX40 ligand; PBMC, peripheral blood mononuclear cells; TILs, tumor-infiltrating lymphocytes.

Article Snippet: Immunohistochemistry (IHC) analyses were performed using specific primary rabbit anti-human CD3 monoclonal antibody (Proteintech Group, Wuhan, China), mouse anti-human OX40 and rabbit anti-human OX40L monoclonal antibodies (CST, Danvers, MA, USA), and rabbit anti-human CD4 (Proteintech Group, Wuhan, China), mouse anti-human CD8 (Proteintech Group, Wuhan, China) antibody, rabbit anti-human FOXP3 monoclonal antibodies (CST, Danvers, MA, USA).

Techniques: Immunohistochemical staining, Staining, Immunofluorescence, Expressing, Flow Cytometry, Standard Deviation, Immunohistochemistry

Journal: Pediatric Rheumatology

Article Title: Fasciitis-panniculitis syndrome with autoantibodies reacting to adipocyte pericellular fibers: a case report

doi: 10.1186/s12969-025-01071-w

Figure Lengend Snippet: Biopsy findings. a : A representative hematoxylin and eosin (HE)-stained section of the patient’s fat tissue showing thickened collagen bundles (×40). b : A HE-stained section of the patient’s en bloc biopsy showing infiltration of inflammatory cells into the adipose tissue and necrotizing vasculitis (×100). c : A HE-stained section showing edema and diffuse infiltration of inflammatory cells into the perifascia with fibrinoid necrosis of the vascular wall (×100). d : A phosphotungstic acid hematoxylin (PTAH)-stained section showing fibrin deposition in the vessel wall (×200). e : Immunohistochemical staining showing infiltration of CD3 + T lymphocytes into the perivascular area of the adipose septa (×100). Anti-human CD3 rabbit antibody was obtained from Cell Signaling Technology (Danvers, MA, USA). HRP-conjugated anti-rabbit IgG Novolink polymer (Leica Biosystems, Nussloch, Germany), was used as a secondary antibody. f : Immunohistochemical staining showing less dominant infiltration of CD19 + B lymphocytes into the perivascular area of the adipose septa (×100). Anti-CD19 rabbit antibody was obtained from Cell Signaling Technology and used along with the above Novolink polymer. g : Immunohistochemical staining showing infiltration of CD68 + macrophages into the perivascular area of the adipose septa (×100). Anti-CD68 antibody [PG-M1] was obtained from abcam. Alkaline phosphatase-conjugated anti-mouse IgG Histofine Simple Stain (Nichirei) was used as a secondary antibody. h : A HE-stained section showing unaffected muscle tissue in the same biopsy (×100)

Article Snippet: Anti-human CD3 rabbit antibody was obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Staining, Immunohistochemical staining, Polymer

Journal: Cell Reports Medicine

Article Title: Miltefosine reinvigorates exhausted T cells by targeting their bioenergetic state

doi: 10.1016/j.xcrm.2024.101869

Figure Lengend Snippet: Miltefosine enhances CAR-T cell efficacy against solid tumors in vivo (A) Experimental timeline of CDX model and the fold change of tumor volume in CAR-T cell-treated CDX mouse model in the presence or absence of miltefosine administration (PBS group, n = 3; other groups, n = 5). (B) Tumor weight of each group at the end of experiment (PBS group, n = 3; other groups, n = 5). (C) The percentages of mesothelin- and hCD3-positive area in tumor tissues (PBS group, n = 2; other groups, n = 5). Scale bar: 200 μm. (D) The fold change of tumor volume in CDX mouse model treated with CAR-T cells derived from another donor, in the presence or absence of miltefosine or anti-PD-1 antibody treatment ( n = 4). (E) The proportion of hCD3 + T cells in the peripheral blood of CDX mouse model ( n = 4). (F) The fold change of tumor volume in CAR-T cell-treated PDX mouse model, in the presence or absence of miltefosine or anti-PD-1 antibody treatment ( n = 4). (G) The proportion of hCD3 + T cells in the peripheral blood of PDX mouse model ( n = 4). Unpaired t test was used in statistical analysis. (H) Experimental timeline of the OT-1-treated melanoma isograft tumor model. (I) The fold change in tumor volume in the OT-1-treated melanoma isograft tumor model in the presence or absence of miltefosine administration ( n = 7). Unpaired t test was used in statistical analysis. NS, not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. All error bars denote SEM. See also Figure S7 .

Article Snippet: Rabbit anti- human CD3 Antibody (clone EP41) , ZSGB-BIO , Cat#ZA-0503.

Techniques: In Vivo, Derivative Assay

Journal: Cell Reports Medicine

Article Title: Miltefosine reinvigorates exhausted T cells by targeting their bioenergetic state

doi: 10.1016/j.xcrm.2024.101869

Figure Lengend Snippet:

Article Snippet: Rabbit anti- human CD3 Antibody (clone EP41) , ZSGB-BIO , Cat#ZA-0503.

Techniques: Functional Assay, Flow Cytometry, Control, Recombinant, Lysis, Activation Assay, Transfection, Luciferase, Viability Assay, SYBR Green Assay, RNA Sequencing Assay, Construct, Plasmid Preparation, Software

Journal: Frontiers in Immunology

Article Title: CD47;Rag2;IL-2rγ triple knock-out mice pre-conditioning with busulfan could be a novel platform for generating hematopoietic stem cells engrafted humanized mice

doi: 10.3389/fimmu.2024.1365946

Figure Lengend Snippet: Immune monitoring of the humanized mouse. Engraftment of human cells was examined by FACS analysis from eight to 48 weeks after hCD34+ HSC injection. Sera were collected by retro orbital bleeding from mice. (A) Gating strategy for flow cytometry analysis. Levels and significant comparisons of (B, C) hCD45 leukocytes, (D, E) hCD19 B cells, (F, G) hCD3 T cells were examined. When hCD3+ cells were taken as a whole (100%), the percentages of (H) hCD4+ cells and (I) hCD8+ cells were calculated. Dot plots for significant comparisons of hCD45 ( Supplementary Figure S1 ), hCD3 and hCD19 ( Supplementary Figure S2 ), hCD4 and hCD8 ( Supplementary Figure S3 ) were provided in supplementary figures. * p <0.05 and ** p <0.01. h, human; FACS, fluorescence-activated cell sorting; RID, Rag2; IL-2rγ double KO NOD mice; RTKO, CD47; Rag2; IL-2rγ triple KO NOD mice; HSC, hematopoietic stem cell; BSF, busulfan; TBI, total body irradiation; wpt, weeks post transplantation.

Article Snippet: The slides were preincubated with blocking serum (Vectastain ABC kit; Vector Laboratories, Burlingame, CA, USA), incubated with rat anti-human CD45 (MA5-17687, 1:100, Invitrogen), rabbit anti-human CD3 antibodies (PA5-32318; 1:1,000, Invitrogen), and mouse anti-human CD19 (14-0199-82; 1:50, Invitrogen).

Techniques: Injection, Flow Cytometry, Fluorescence, FACS, Irradiation, Transplantation Assay

Journal: Frontiers in Immunology

Article Title: CD47;Rag2;IL-2rγ triple knock-out mice pre-conditioning with busulfan could be a novel platform for generating hematopoietic stem cells engrafted humanized mice

doi: 10.3389/fimmu.2024.1365946

Figure Lengend Snippet: Expression of human leukocyte antigens. (A) Immunohistochemical staining of hCD45, hCD3, hCD19 in the lung, liver, kidney, skin, and spleen tissues of the RTKO groups. hCD45 and hCD3 were abundantly expressed in inflammatory cells in perivascular and subcutaneous areas, and some leukocytes were also stained with hCD19. Bar, 50 µm. (B) Western blot analysis of hCD45 and hCD3 proteins using the spleen tissues. Protein concentrations of the RTKO groups were greater than the RID groups, and the significant differences were only observed when comparing hCD3 results of all of the RTKO groups and the RID groups. * p <0.05. h, human; RID, Rag2; IL-2rγ double KO NOD mice; RTKO, CD47; Rag2; IL-2rγ triple KO NOD mice; HSC, hematopoietic stem cell; BSF, busulfan; TBI, total body irradiation.

Article Snippet: The slides were preincubated with blocking serum (Vectastain ABC kit; Vector Laboratories, Burlingame, CA, USA), incubated with rat anti-human CD45 (MA5-17687, 1:100, Invitrogen), rabbit anti-human CD3 antibodies (PA5-32318; 1:1,000, Invitrogen), and mouse anti-human CD19 (14-0199-82; 1:50, Invitrogen).

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot, Irradiation