Journal: Nature immunology

Article Title: Antigen-specific B cells direct T follicular- like helper cells into lymphoid follicles to mediate Mycobacterium tuberculosis control

doi: 10.1038/s41590-023-01476-3

Figure Lengend Snippet: (a) Representative image of inflammatory lesion in lung of NHP LTBI (left) vs ATB (right) stained with antibodies specific for CD3 = white; IRF4 = green; PD1 = red. Circles with yellow dashed lines depict ectopic lymphoid structures and asterix = center of granuloma. The numbers of (b) IRF4+ CD3+ (n = 4 NHPs; 7–21 data points from different fields) and (c) only CD3+ (n=3 NHPs/ group) T cells in NHP ATB vs LTBI lung lesions were estimated by blinded morphometric analysis. (d) Correlation between the number of CD3+CD4+PD1+ Tfh-like cells inside B cell areas with the average area of the GrALT in the lungs of Mtb-infected NHPs (n = 5–6 NHPs/group). (e) Representative human ATB lesion, (f) number of T cells in human lung lesion expressing either PD1, IRF4 or both (n = 3 males and 1 female lung biopsies; age: 29–41 years). Data represents the mean ± SD. Statistical significance was determined with two-sided unpaired t-test (b and c), Pearson’s correlation (d) and Kruskal-Wallis ANOVA with Dunn’s post-hoc test (f). *, p≤0.05; **,p≤0.005; ***, p≤0.0005.

Article Snippet: To detect Tfh-like cells in B cell depleted macaques or control macaques, lung sections were stained with antibodies specific for CD3ε, IRF4, PD1 (Cat. No. 10377-MM23, Sino Biological), and BCL6.

Techniques: Staining, Infection, Expressing

Journal: Nature immunology

Article Title: Antigen-specific B cells direct T follicular- like helper cells into lymphoid follicles to mediate Mycobacterium tuberculosis control

doi: 10.1038/s41590-023-01476-3

Figure Lengend Snippet: FFPE lungs sections from Mtb-infected B6 and gene deficient mice were stained for the presence of Mtb 16S rRNA via in situ hybridization (ISH) at (a) 30, 50, and 75 dpi. FFPE lungs sections from Rag1KO Mtb-infected mice that either received WT T cells:WT B cells or Pd1KO T cells:WT B cells, were stained for (b) the presence of Mtb 16S rRNA via in situ hybridization (ISH) at 30 dpi, (c) presence of GrALT via immunofluorescence; CD3 (red); B220 (white); PD-1 (green).

Article Snippet: To detect Tfh-like cells in B cell depleted macaques or control macaques, lung sections were stained with antibodies specific for CD3ε, IRF4, PD1 (Cat. No. 10377-MM23, Sino Biological), and BCL6.

Techniques: Control, Infection, Staining, In Situ Hybridization, Immunofluorescence

Journal: Nature immunology

Article Title: Antigen-specific B cells direct T follicular- like helper cells into lymphoid follicles to mediate Mycobacterium tuberculosis control

doi: 10.1038/s41590-023-01476-3

Figure Lengend Snippet: (a) Average area of GrALT collated from all mouse models (n = 3–10 mice/group) used in studies at 50 dpi and 128 dpi. (b) Representative images from FFPE lung sections stained with CD3 (red), PD-1 (green) and CD45R/B220 (white) in indicated models at 50 or 128 dpi. Data represent mean ± SD and statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparison test (3 groups), and two-tailed unpaired t-test (2 groups). *, p≤0.05; **,p≤0.005.

Article Snippet: To detect Tfh-like cells in B cell depleted macaques or control macaques, lung sections were stained with antibodies specific for CD3ε, IRF4, PD1 (Cat. No. 10377-MM23, Sino Biological), and BCL6.

Techniques: Control, Staining, Comparison, Two Tailed Test

Journal: Nature immunology

Article Title: Antigen-specific B cells direct T follicular- like helper cells into lymphoid follicles to mediate Mycobacterium tuberculosis control

doi: 10.1038/s41590-023-01476-3

Figure Lengend Snippet: Irf4fl/fl, CD4creIrf4fl/fl, CD19creIrf4fl/fl, Bcl6fl/fl, CD4creBcl6fl/fl, CD19creBcl6fl/fl, iABfl/fl, CD19creiABfl/fl, Blimp1fl/fl, CD19creBlimp1fl/fl, Il10fl/fl, CD19creIl10fl/fl, C57BL/6 and IghelMD4 mice were infected with Mtb HN878 (n = 3–10 mice/group). Lungs were collected and processed at 50 dpi. Mtb-infected B6, IghelMD4 and IghelMD4 (recipient) mice were euthanized, and lungs were collected and processed at 128 dpi. Formalin-fixed lung lobes were sectioned and stained with immunofluorescent antibodies to determine the (a) percentage of PD1+CD3+ Tfh-like cells within B cell areas. Uninfected or Mtb-infected (30 dpi) C57BL/6 mice (n = 5 mice/group) were euthanized, and lung cells were subjected to flow cytometric analysis for PD-L1 (CD247) expression on (b) total B cells and (c) FO B cells. B cells isolated from the spleens and lymph nodes of C57BL/6 mice and T cells isolated from the spleens and lymph nodes of either C57BL/6 or Pd1KO mice were co-transferred to Rag1KO mice, 1–2 days prior to infection with Mtb HN878 (n = 4–5 mice/group). Mice were euthanized 30 dpi. (d) Experimental scheme. (e) activated CD4+ T cells, (f) CD4+ Tfh-like cells (g) IFNγ+CD4+ Tfh-like cells and (h) IFNγ mean fluorescent intensity in activated CD4+ T cells in the lungs were determined by flow cytometry. Uninfected or Mtb-infected C57Bl/6 mice were euthanized (30 dpi) to collect lung (n = 5 mice/group) and mediastinal lymph node (MLN) (n = 4 mice/group). (i) Activated CD4+ T cells, (j) CD4+IFNγ+ T cells in both MLN and lungs; (k) Tfh like cells; MFIs for transcription factors (l) Tbet and (m) Bcl6 in both MLN and lungs activated CD4+ T cells were determined by flow cytometry. Data represents mean ± SD. Statistical significance was calculated with one-way ANOVA with Tukey’s multiple comparison test for group of three and two-sided unpaired t-test for group of two (a-c and e-h), and two-sided Mann-Whitney U-test (i to m). *, p≤0.05; **,p≤0.005; ***, p≤0.0005.

Article Snippet: To detect Tfh-like cells in B cell depleted macaques or control macaques, lung sections were stained with antibodies specific for CD3ε, IRF4, PD1 (Cat. No. 10377-MM23, Sino Biological), and BCL6.

Techniques: Infection, Staining, Expressing, Isolation, Flow Cytometry, Comparison, MANN-WHITNEY

Journal: Nature immunology

Article Title: Antigen-specific B cells direct T follicular- like helper cells into lymphoid follicles to mediate Mycobacterium tuberculosis control

doi: 10.1038/s41590-023-01476-3

Figure Lengend Snippet: Macaques were aerosol exposed to MtbΔsigH prior to challenge with virulent Mtb CDC1551 and received either CD20 depleting (n = 4 NHP) or IgG isotype control antibodies (n = 2 NHP). Clinical samples were collected, and macaques were monitored for clinical signs of disease throughout the study. (a) Experimental scheme. Bacterial burden was determined in (b) bulk lung tissue and (c) individual granulomas at necropsy. (d) CXCR5+CD4+ T cells, and (e) PD1+CD4+ T cells were detected by flow cytometry at the time of necropsy. (f) Representative images of a B cell follicle in lungs of a macaque which received IgG control isotype antibodies (top panels) and images of lung tissue from a macaque which received CD20 depleting antibodies (bottom panels); CD3-red, PD1-green, BCL6-white; dotted yellow line indicate germinal center, white arrows indicate CD3+BCL6+PD1+ Tfh-like cells, orange asterix indicates the bronchus. (g) The number of BCL6+ Tfh-like cells with the GrALT. Data represents mean ± SD. Statistical significance was calculated with tow-sided Mann-Whitney U-test (b to e); no statistics calculated as anti-CD20 treated NHP had all ‘zero’ values (g). *, p≤0.05, ***, p≤0.0005.

Article Snippet: To detect Tfh-like cells in B cell depleted macaques or control macaques, lung sections were stained with antibodies specific for CD3ε, IRF4, PD1 (Cat. No. 10377-MM23, Sino Biological), and BCL6.

Techniques: Aerosol, Control, Flow Cytometry, MANN-WHITNEY

Journal: Anais Brasileiros de Dermatologia

Article Title: METTL3-mediated HPV vaccine enhances the effect of anti PD-1 immunotherapy to alleviate the development of cutaneous squamous cell carcinoma ⋆

doi: 10.1016/j.abd.2023.05.006

Figure Lengend Snippet: Impact of HPV vaccine combined with PD-1 monoclonal antibody for cSCC. (A) Tumor picture; (B) Volume change; (C) Weight change. ***p < 0.001.

Article Snippet: To test this hypothesis, the authors divided 10 mice with cSCC into two groups of 5 mice each and injected PD-1 Abs (10377-M94, SinoBiological) or PD-1Abs+HPV vaccine.

Techniques:

Journal: Anais Brasileiros de Dermatologia

Article Title: METTL3-mediated HPV vaccine enhances the effect of anti PD-1 immunotherapy to alleviate the development of cutaneous squamous cell carcinoma ⋆

doi: 10.1016/j.abd.2023.05.006

Figure Lengend Snippet: Effect of HPV vaccine combined with PD-1 monoclonal antibody on immunofluorescence in mice. (A) Immunofluorescence observation of immune microenvironment differences (CD8, CD4, NK, DC, Macrophage). (B) Image J statistical analysis of the percentage of positive immune cells. ***p < 0.001.

Article Snippet: To test this hypothesis, the authors divided 10 mice with cSCC into two groups of 5 mice each and injected PD-1 Abs (10377-M94, SinoBiological) or PD-1Abs+HPV vaccine.

Techniques: Immunofluorescence

Journal: Anais Brasileiros de Dermatologia

Article Title: METTL3-mediated HPV vaccine enhances the effect of anti PD-1 immunotherapy to alleviate the development of cutaneous squamous cell carcinoma ⋆

doi: 10.1016/j.abd.2023.05.006

Figure Lengend Snippet: Animal-level validation of METTL3 overexpression abolishing HPV vaccine to enhance the sensitivity of cSCC mice treated with PD-1 monoclonal antibody. (A) Tumor picture; (B) volume change; (C) weight change. ***p < 0.001.

Article Snippet: To test this hypothesis, the authors divided 10 mice with cSCC into two groups of 5 mice each and injected PD-1 Abs (10377-M94, SinoBiological) or PD-1Abs+HPV vaccine.

Techniques: Over Expression

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Increased antitumor activities of glypican-3-specific chimeric antigen receptor-modified T cells by coexpression of a soluble PD1–CH3 fusion protein

doi: 10.1007/s00262-018-2221-1

Figure Lengend Snippet: In vitro behavior of CAR-T cells after repetitive stimulation with SK-HEP-1-GPC3 cells. a CAR expression on the T-cell surface after three rounds of antigen-specific stimulation with SK-HEP-1-GPC3 cells. b Each data set is pooled from four independent experiments with individual donors (n = 4, mean ± SEM; ns not significant, P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001); c The ratio of CD8+ versus CD4+ T cells after three rounds of antigen-specific stimulation (n = 3, mean ± SEM; ns not significant, P > 0.05, *P < 0.05, **P < 0.01); d Proliferation of both CAR-T cells after three rounds of antigen-specific stimulation. The two groups of CAR-T cells were prestained with CFSE before the third stimulation, and then the stained CAR-T cells were cocultured with SK-HEP-1-GPC3 at a 1:1 effector-to-target ratio for 96 h. The intensity of CFSE in each group was measured by flow cytometry. e After three rounds of antigen-specific stimulation with SK-HEP-1-GPC3 cells, the expression of exhaustion biomarkers on the T-cell surface, including PD-1, TIM-3 and LAG-3, was determined using flow cytometry with the indicated antibodies; f Each data set is pooled from three independent experiments with individual donors (n = 3, mean ± SEM; ns not significant, P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001); g Cytokine release of the engineered T cells after three rounds of antigen-specific stimulation with SK-HEP-1-GPC3 cells. A total of 1 × 106 engineered T cells were cocultured with 1 × 106 tumor cells for 24 h. The levels of IL-2, IFN-γ and TNF-α in the supernatants were evaluated by ELISA (n = 6, mean ± SEM; ns not significant, P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: After 24 h, 100 µl of supernatant was transferred to a 96-well plate precoated with a PD1/PDCD1/CD279 antibody (Sino Biological, clone: H08H).

Techniques: In Vitro, Expressing, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay