Journal: The Journal of experimental medicine

Article Title: Regulation of pulmonary plasma cell responses during secondary infection with influenza virus.

doi: 10.1084/jem.20232014

Figure Lengend Snippet: Figure 2. IFNγ-producing NK cells and CD4 T cells support secondary IgG+ PC responses. (A) Top, experimental layout. Bottom, representative flow cytometry plots showing frequencies of NK cells (DX5+ CD3−), CD4+, and CD8+ T cells within the pool of IFNγ+ cells. (B) IFNγ+ fraction of NK cells, CD4+, and CD8+ T cells in resting memory phase mice, and 1 day after rechallenge in mice treated with control or CLL liposomes, as indicated. (C) Left, experimental layout. Right, quantification of intracellular IFNγ expression in DX5+ CD3−NK cells, DX5−CD3+ CD8+ T cells, or DX5−CD3+ CD4+ T cells analyzed by flow cytometry. (D–F) Analysis of PC response at day 4 after rechallenge in the absence of CD4, CD8 T, or NK cells. (D) Experimental layout. Lungs were analyzed by flow cytometry. The figure shows counts of (E) IgG+ PCs and (F) IgA+ PCs. Data in A, B, and C are each pooled from two independent experiments with n = 3–4 per experiment. Each circle represents one mouse. Data in E and F are pooled from two to three independent experiments with n = 3–4 per experiment. Each circle represents one mouse. Statistical comparisons throughout the figure were made using unpaired Student’s t test (ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: For depletion of CD4 and CD8 T cells during rechallenge, cells were depleted using anti-CD4 antibodies (Clone GK1.5, BE0003-1; BioXCell), anti-CD8β antibodies (Clone 53-5.8, BE0223; BioXCell), or control rat IgGb (Clone RTK4530, cat. 400667; Biolegend).

Techniques: Flow Cytometry, Control, Liposomes, Expressing

Journal: The Journal of experimental medicine

Article Title: Regulation of pulmonary plasma cell responses during secondary infection with influenza virus.

doi: 10.1084/jem.20232014

Figure Lengend Snippet: Figure 3. Accumulation of BRM cells near infected sites during rechallenge is regulated by NK cells. (A–D) Effect of NK and CD4 T cell depletion on AlvPC positioning in rechallenged lungs. (A) Experimental layout. (B) Representative images from thick sections using static TPLSM. The white line highlights an approximate border between areas that are highly versus poorly infected. Boxes focus on uninfected (box 1) and infected (box 2) areas and are displayed as magnified images in the lower panel. Dotted white circles mark AlvPCs. Each image represents one tiled of three independent experiments with n = 3 per group per time point. (C) Quantification of AlvPC density, calculated using high-volume static TPLSM imaging from images as above. Each point represents cells quantified from a large tiled image analyzed from three independent experiments. n = 3 per group per time point. (D) PC density at day 4 rechallenge, in infected and uninfected areas of control Ig-treated, anti-CD4–, or anti-NK1.1–treated mice. On the right, data displayed as the ratio of PC density in infected

Article Snippet: For depletion of CD4 and CD8 T cells during rechallenge, cells were depleted using anti-CD4 antibodies (Clone GK1.5, BE0003-1; BioXCell), anti-CD8β antibodies (Clone 53-5.8, BE0223; BioXCell), or control rat IgGb (Clone RTK4530, cat. 400667; Biolegend).

Techniques: Infection, Imaging, Control

Journal: The Journal of experimental medicine

Article Title: Regulation of pulmonary plasma cell responses during secondary infection with influenza virus.

doi: 10.1084/jem.20232014

Figure Lengend Snippet: Figure 4. CD4 T cells are critical for PC development in iBALTs. (A) Experiment layout. (B–D) (B) Counts of GC B cells, (C) counts of IgG and IgA PCs, and (D) counts of IgG and IgA BRM cells analyzed by flow cytometry 1 week after CD4 T cell depletion in memory phase mice. (E) Representative images from lungs of memory phase BAT reporter mice 1 week after treatment with control or anti-CD4 antibodies, highlighting BRM distribution through the alveoli. Analysis was performed on thick sections using static TPLSM. BRM cells (white) and PCs (orange) are highlighted using spots created by Imaris. (F) Quantification of BRM density in lungs of mice treated as in A. (G) Distribution of BRM cells in control and anti-CD4 treated mice. Left, plots display clustering (Ripley’s L) values of observed data (black) versus complete spatial random simulation (simulation n = 1,000; red). Right, a summary of L values of observed BRM distribution at r = 200 µm. Data in B–D are representative of three independent experiments with n = 3–5 per experiment. Each point in B–D represents one mouse. Data in E–G are pooled from two independent experiments, total n = 3. Each point in F and G represents values measured from one analyzed tiled image. Statistical comparisons were made using Mann–Whitney U tests (B–D) or unpaired Student’s t test (F and G) (ns, not significant, *P < 0.05, **P < 0.01).

Article Snippet: For depletion of CD4 and CD8 T cells during rechallenge, cells were depleted using anti-CD4 antibodies (Clone GK1.5, BE0003-1; BioXCell), anti-CD8β antibodies (Clone 53-5.8, BE0223; BioXCell), or control rat IgGb (Clone RTK4530, cat. 400667; Biolegend).

Techniques: Flow Cytometry, Control, MANN-WHITNEY

Journal: Pathogens

Article Title: IFN-Gamma-Dependent and Independent Mechanisms of CD4 + Memory T Cell-Mediated Protection from Listeria Infection

doi: 10.3390/pathogens7010022

Figure Lengend Snippet: CD4 + memory T cells induce rapid clearance following Lm-gp61 rechallenge. Naïve or lymphocytic choriomeningitis virus (LCMV)-immune (>day 30 post-infection) B6 mice were infected with Lm-gp61. Additionally, some LCMV-immune mice were treated with anti-CD4 antibody to deplete CD4 + T cells prior to challenge. Bacterial burden in the ( A ) spleen (CFU/spleen) and ( B ) liver (CFU/g) was measured 3 days post-challenge; ( C ) Bacterial burden in the spleen of naïve mice with and without CD4 depletion, infected with Lm-gp61 was measured 3 days post-challenge; ( D ) Total numbers of CD4 + and CD8 + T cells in the spleen at day 3 post-challenge of naïve and LCMV-immune mice; ( E ) At day 3 post-challenge, splenocytes were stimulated ex vivo with the LCMV peptide GP 61–80 in the presence of Brefeldin A, then fixed and permeabilized and stained with antibodies to detect the presence of intracellular cytokines. Representative flow plots are gated on total CD4 + T cells. n = 4–5 mice/group, data are representative of two separate experiments.

Article Snippet: 200 μg anti-CD4 depleting antibodies (BioXCell, Clone GK1.5) were given i.p. on day-2 and -1 prior to infection with Lm-gp61.

Techniques: Virus, Infection, Ex Vivo, Staining

Journal: Pathogens

Article Title: IFN-Gamma-Dependent and Independent Mechanisms of CD4 + Memory T Cell-Mediated Protection from Listeria Infection

doi: 10.3390/pathogens7010022

Figure Lengend Snippet: Protection mediated by CD4 + memory T cells is heavily dependent on TNF but only partly dependent on IFNγ. Naïve and LCMV-immune mice were challenged with Lm-gp61 and some groups of mice were additionally treated with neutralizing antibodies to IFNγ or TNF. Bar graphs indicate bacterial load in the ( A ) spleen and ( B ) liver day 3 following primary or secondary challenge ( n = 4–7 mice/group); ( C , D ) Bar graphs indicate bacterial load in the spleen and liver, as indicated, on day 3 following primary Lm-gp61 infection with or without TNF neutralization ( n = 4 mice/group); ( E ) Bar graph shows bacterial burden in the spleen of LCMV-immune LysM Cre /Ifngr1 fl/fl mice day 3 after rechallenge with Lm-gp61 ( n = 4 mice/group). Data are representative of at least two separate experiments.

Article Snippet: 200 μg anti-CD4 depleting antibodies (BioXCell, Clone GK1.5) were given i.p. on day-2 and -1 prior to infection with Lm-gp61.

Techniques: Infection, Neutralization

Journal: Scientific Reports

Article Title: CD8 + T-cells restrict the development of peritoneal metastasis and support the efficacy of hyperthermic intraperitoneal chemotherapy (HIPEC)

doi: 10.1038/s41598-024-72826-w

Figure Lengend Snippet: Comparison of patient characteristics between CD8 + T-cell high and CD8 + T-cell low . The data is.

Article Snippet: CD-4 + and CD8+-T-cell depletion in mice was achieved by the intraperitoneal injection of 100ug CD4 or CD8a depletion antibody (BioXCell, USA; clone GK 1.5 for CD4 + T-cells and clone YTS 169.4 was used for CD8 + T-cells) 1 day prior to tumor cells injection and 1 day prior to the treatment.

Techniques: Comparison, Cell Differentiation, Mutagenesis, Adjuvant

Journal: Scientific Reports

Article Title: CD8 + T-cells restrict the development of peritoneal metastasis and support the efficacy of hyperthermic intraperitoneal chemotherapy (HIPEC)

doi: 10.1038/s41598-024-72826-w

Figure Lengend Snippet: Assessment of CD8 + T cells in patient samples. ( a and b ) analysis of CD8 + T cells in stroma and epithelium of the primary tumor of 19 patients ( a ) and the PM lesions of 37 patients. The graphs illustrate the number of CD8 + T-cells normalized to the corresponding area of stroma or epithelium. ( c ).The bar graph shows the distribution of intraepithelial CD8 + T-cells normalized by area in PM-lesions among the 37 patients. The dotted line indicates the median and divide the cohort into CD8 + T-cell high and low. ( d ) The scanned histological slide of PM lesions. The upper left picture presents an example with high CD8 + T-cell infiltration and the corresponding HALO classified picture below. The left upper example is CD8 + T-cell low. The classified pictures show the different areas of the tumor (green: stroma, violet: necrosis, red: intraepithelial, yellow: white space). DFS ( e ) and OS ( f ) based on intraepithelial CD8 + T-cell counts of PM lesions. 18 patients belong to the CD8 + T-cell high group and 19 patients to the CD8 + T-cell low group. DFS ( g ) and OS ( h ) based on the stroma content and CD8 + T-cell distribution. 15 patients with a CD8 + T-cell high PM lesion were associated with low stromal content (continuous line), 9 patients with a CD8 + T-cell low PM-lesion had a rich stroma (fine dotted line),10 patients with a CD8 + T-cell low PMlesion had a poor stroma (bold dotted line) and 3 patients with a CD8 + T-cell high PM-lesion were stroma rich (dotted line). Error bars represent the median and the lines the interquartile range. Each dot represents a patient. **** = p ≤ 0.0001, *** = p ≤ 0.001, ** = = p ≤ 0.01, * = p ≤ 0.05, ns = p > 0.05.

Article Snippet: CD-4 + and CD8+-T-cell depletion in mice was achieved by the intraperitoneal injection of 100ug CD4 or CD8a depletion antibody (BioXCell, USA; clone GK 1.5 for CD4 + T-cells and clone YTS 169.4 was used for CD8 + T-cells) 1 day prior to tumor cells injection and 1 day prior to the treatment.

Techniques:

Journal: Scientific Reports

Article Title: CD8 + T-cells restrict the development of peritoneal metastasis and support the efficacy of hyperthermic intraperitoneal chemotherapy (HIPEC)

doi: 10.1038/s41598-024-72826-w

Figure Lengend Snippet: Impact of HIPEC treatment in PM mouse model. ( a ) measurement of peritoneal tumor load as PCI. mice were treated with PBS ( n = 9) or heated PBS ( n = 7) or with M/D (n = 10) or with heated M/D (n = 11). ( b ) Staining of tumor tissues for the presence of CD8 + T cells, Granzyme B + cells and macrophages. ( c and d ) Quantification of CD8 + T cells and GZMB + cells. ( e ) PCI of the treated mice with and without CD8 + T-cells. Each dot represents one mouse. Error bars show the mean +/-SD. **** = p ≤ 0.0001, *** = p ≤ 0.001, ** = = p ≤ 0.01,* = p ≤ 0.05, ns = p > 0.05.

Article Snippet: CD-4 + and CD8+-T-cell depletion in mice was achieved by the intraperitoneal injection of 100ug CD4 or CD8a depletion antibody (BioXCell, USA; clone GK 1.5 for CD4 + T-cells and clone YTS 169.4 was used for CD8 + T-cells) 1 day prior to tumor cells injection and 1 day prior to the treatment.

Techniques: Staining

Journal: Scientific Reports

Article Title: CD8 + T-cells restrict the development of peritoneal metastasis and support the efficacy of hyperthermic intraperitoneal chemotherapy (HIPEC)

doi: 10.1038/s41598-024-72826-w

Figure Lengend Snippet: Induction of antigen-specific CD8 + T cells via heated chemotherapy. ( a )Time-line of the experiment. ( b ) flow cytometry data on the maturation state of Mo-DC`s depending on the cancer-cell treatment. ( c )Time-line of the experiment with additional co-culture of CD8 + T-cells. ( d ) Shows flow cytometry data of CD8 + T-cells and their IFN-γ production depending on the cancer-cell treatment. ( e ) Shows the time-line of a similar experiment using splenocytes from OT-I mice, which have a specific TCR for the ovalbumin. ( f ) Presents the ratio of IFN-γ positive CD8 + T-cells after co-culturing with PBS, or M/D or heated M/D treated MC-38-Ova cancer cells with splenocytes from a OT-I mouse. Cancer cell killing by specific CD8 + T-cells. ( g ) time-line of the co-culture experiment. The % of cancer cell killing by the treatment with or without specific CD8 + T-cells is shown in ( h ).The experiments were performed in triplicates. Error bars show the mean +/-SD. **** = p ≤ 0.0001, *** = p ≤ 0.001, ** == p ≤ 0.01, * = p ≤ 0.05, ns = p > 0.05.

Article Snippet: CD-4 + and CD8+-T-cell depletion in mice was achieved by the intraperitoneal injection of 100ug CD4 or CD8a depletion antibody (BioXCell, USA; clone GK 1.5 for CD4 + T-cells and clone YTS 169.4 was used for CD8 + T-cells) 1 day prior to tumor cells injection and 1 day prior to the treatment.

Techniques: Flow Cytometry, Co-Culture Assay

Journal: Scientific Reports

Article Title: CD8 + T-cells restrict the development of peritoneal metastasis and support the efficacy of hyperthermic intraperitoneal chemotherapy (HIPEC)

doi: 10.1038/s41598-024-72826-w

Figure Lengend Snippet: Intraepithelial CD8 + T-cell number after HIPEC in 2 patients. ( a ) Numbers of intraepithelial CD8 + T-cells before first HIPEC and after the first HIPEC treatment of 2 different patient and the DFS and OS of these two patients ( b ).

Article Snippet: CD-4 + and CD8+-T-cell depletion in mice was achieved by the intraperitoneal injection of 100ug CD4 or CD8a depletion antibody (BioXCell, USA; clone GK 1.5 for CD4 + T-cells and clone YTS 169.4 was used for CD8 + T-cells) 1 day prior to tumor cells injection and 1 day prior to the treatment.

Techniques: