Journal: Nature Communications

Article Title: Asthma reduces glioma formation by T cell decorin-mediated inhibition of microglia

doi: 10.1038/s41467-021-27455-6

Figure Lengend Snippet: a Schematic representation of the journey of T cells from the periphery (blood) into the optic nerve and draining through the cervical lymph node. b Increased Ccl4 mRNA expression was observed in optic nerves ( n = 4) of Nf1 OPG mice relative to blood ( n = 5) and draining cervical lymph nodes ( n = 5). One-way ANOVA with Bonferroni post hoc correction. c Schematic representation of the interactions between T cells and microglia in the optic nerve and draining cervical lymph node. d No change in Ccl4 mRNA expression was found in the optic nerves of Nf1 OPG mice treated with OVA ( n = 4) or HDM ( n = 3) relative to vehicle-treated ( n = 7) mice. One-way ANOVA with Bonferroni post hoc correction. e Decreased optic nerve Ccl5 mRNA expression was observed in both OVA- ( n = 4) and HDM- ( n = 3) treated mice relative to PBS-treated controls ( n = 7). One-way ANOVA with Bonferroni post hoc correction. f While Ccl4 gene expression was increased in the cervical lymph nodes of Nf1 OPG mice relative to controls (WT, n = 3; Nf1 OPG , n = 3), Two-tailed Student’s t test. ( g ) No change in cervical lymph node Ccl4 mRNA expression was observed between OVA- or HDM-treated mice (OVA, n = 4; HDM, n = 3) relative to controls ( n = 7). One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. Exact P values are indicated within each panel (ns, not significant). From left to right in each panel: b P < 0.0001, P = 0.0460; d ns, ns; e P = 0.0069, P = 0.0359; f P = 0.0058; g ns.

Article Snippet: Recombinant mouse midkine (R&D Systems, 9760-MD-050) was added, and Ccl4 levels were quantitated by ELISA (R&D Systems, MMB00).

Techniques: Expressing, Gene Expression, Two Tailed Test

Journal: Nature Communications

Article Title: Asthma reduces glioma formation by T cell decorin-mediated inhibition of microglia

doi: 10.1038/s41467-021-27455-6

Figure Lengend Snippet: a Schematic representation of the mouse Nf1 OPG neuron-immune-cancer cell axis in the setting of asthma. b No difference in optic nerve Mdk RNA expression was observed between OVA- ( n = 3) or HDM- ( n = 3) treated mice and controls ( n = 6). Bar graphs represent the means ± SEM of independent biological samples. One-way ANOVA with Bonferroni post hoc correction. c Ccl4 levels are similarly induced with increasing midkine concentrations (0–100 ng/ml) in CD3 + T cells from OVA- and PBS-treated mice. Data are presented as the means ± SEM of n = 3 independent biological samples. One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. d , e Similarly increased CD8 + , relative to CD4 + , T cell content was detected in the optic nerves of Nf1 OPG mice treated with PBS ( n = 8), OVA ( n = 8) or HDM ( n = 8). One-way ANOVA with Bonferroni post hoc correction. Activated T cell medium from ( f ) OVA- ( n = 4) and g HDM- ( n = 4) treated mice induced lower levels of microglial Ccl5 relative to PBS-treated Nf1 OPG mice. Two-tailed Student’s t test. Similar reductions of Ccl5 expression were observed in CD4 + and CD8 + T cells from ( h ) OVA- ( n = 3) and ( i ) HDM- ( n = 3) treated mice relative to PBS-treated controls ( n = 3). One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. d Scale bars, 40 µm. From left to right in each panel: b ns; c ns; ns; ns; ns; e P < 0.0001, P = 0.0022, P = 0.0011; f P = 0.0008; g P = 0.0015; h P = 0.0186, P = 0.0054; i P = 0.0224, P = 0.0007.

Article Snippet: Recombinant mouse midkine (R&D Systems, 9760-MD-050) was added, and Ccl4 levels were quantitated by ELISA (R&D Systems, MMB00).

Techniques: RNA Expression, Two Tailed Test, Expressing

Journal: Nature Communications

Article Title: Asthma reduces glioma formation by T cell decorin-mediated inhibition of microglia

doi: 10.1038/s41467-021-27455-6

Figure Lengend Snippet: a Decorin (800 pg/ml) attenuated activated TCM (act-Tm)-mediated microglia Ccl5 production ( n = 3). Two-tailed Student’s t test. Exact P values are indicated within each panel (ns, not significant). b Decorin reduces microglia Ccl5 production in response to Ccl4 exposure (6 ng/ml) over a physiologic dose range in vitro ( n = 4). One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. c TCM (activated T cell-conditioned media) induced microglial Ccl5 production was significantly attenuated by decorin. The addition of Dcn with CCR5 has the greatest inhibition similar to the combination of CCR5 and CCR8 inhibitors ( n = 6). One-way ANOVA with Bonferroni post hoc correction. Data are presented as the means ± SEM. d Schematic representation of decorin binding to the CCR8 receptor on microglia, culminating is reduced microglial Ccl5 production by inhibiting NFκB activation. e Schematic representation of decorin treatment of Nf1 OPG mice. Nf1 OPG mice were treated between 4 and 6 weeks of age with decorin, while control Nf1 OPG mice received PBS only. Isolated optic nerves were analyzed at 12 weeks of age. f Decorin treatment has no change on optic nerve volume, but ( g ) decreased proliferation (%Ki67 + cells) of Nf1 OPG mice relative to the vehicle-treated Nf1 OPG controls (PBS, n = 8, Decorin, n = 8). No difference in microglia (%Iba1 + cells) or T cell (CD3 + cells) content was observed between decorin-treated mice and their respective controls. Two-tailed Student’s t test (ns, not significant). g Scale bars, 40 µm. From left to right in each panel: a P = 0.0134, b P < 0.0001, P = 0.0035; c P < 0.0001, ns, P = 0.0086, ns; f ns; g P = 0.0010, ns, ns.

Article Snippet: Recombinant mouse midkine (R&D Systems, 9760-MD-050) was added, and Ccl4 levels were quantitated by ELISA (R&D Systems, MMB00).

Techniques: Two Tailed Test, In Vitro, Inhibition, Binding Assay, Activation Assay, Control, Isolation

Journal: Nature Communications

Article Title: Asthma reduces glioma formation by T cell decorin-mediated inhibition of microglia

doi: 10.1038/s41467-021-27455-6

Figure Lengend Snippet: a Nf1 OPG mouse optic nerves ( n = 3) have increased Iκbα phosphorylation relative to WT mice ( n = 3). b The increased Iκbα phosphorylation in PBS-treated Nf1 OPG mouse optic nerves ( n = 3) was reduced by OVA treatment ( n = 3). c Activated TCM (act-Tm) increased Iκbα phosphorylation in microglia, which was reduced following the addition of decorin (800 pg/ml). d Decorin (800 pg/ml) blocks the increased p65-NFκB Ser 536 phosphorylation induced by activated TCM (act-Tm) treatment ( n = 3 ). e Decorin (800 pg/ml) blocks the increased total p65-NFκB expression, ( f ) as well as the nuclear localization of p65-NFκB, induced by activated TCM (act-Tm) treatment ( n = 3). β-actin and HDAC are used as controls for total protein expression and the nuclear fractions, respectively. g Schematic representation of the NFκB inhibitor treatment used. Nf1 OPG mice were treated between 4 and 6 weeks of age with the CAPE NFκB inhibitor ( n = 8), whereas control Nf1 OPG mice received PBS only ( n = 8). Isolated optic nerves were analyzed at 12 weeks of age. h NFκB inhibitor treatment reduced optic glioma volume and ( i ) proliferation (%Ki67 + cells), as well as microglia (%Iba1 + cells) and T-cell (CD3 + ) content within the optic nerves of Nf1 OPG mice relative to vehicle-treated controls. Two-tailed Student’s t test. j Reduced Ccl5 RNA expression was observed in the optic nerves from Nf1 OPG mice treated with the CAPE NFκB inhibitor (NFκB-IN) ( n = 5). Two-tailed Student’s t test. k Proposed model of asthma-induced decorin suppression of the Nf1 OPG neuron-immune-cancer cell axis. Asthma induces T cell production of decorin, which reduces T cell Ccl4-mediated microglia Ccl5 expression through inhibition of NFκB signaling. Data are presented as the means ± SEM. Exact P values are indicated within each panel. i Scale bars 40 µm. From left to right in each panel: h P = 0.0288; i P = 0.0003, P = 0.0196, P = 0.0025; j P = 0.0018.

Article Snippet: Recombinant mouse midkine (R&D Systems, 9760-MD-050) was added, and Ccl4 levels were quantitated by ELISA (R&D Systems, MMB00).

Techniques: Phospho-proteomics, Expressing, Control, Isolation, Two Tailed Test, RNA Expression, Inhibition

Journal: Frontiers in Immunology

Article Title: Low perforin expression in CD8+ T lymphocytes during the acute phase of severe SARS-CoV-2 infection predicts long COVID

doi: 10.3389/fimmu.2022.1029006

Figure Lengend Snippet: Absence of correlation between the frequency of perforin-positive T4 cells and T8 cells and the plasma level of 15 biomarkers of COVID-19 severity.

Article Snippet: Biomarker levels (IL-1-beta, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-17A, GM-CSF, IFN-alpha, IFN-gamma, MIP-1alpha, MIP-1beta, TNF-alpha, IP-10) were measured by Luminex/xMAP immunoassays (ProcartaPlex, ThermoFisher Scientific) in 1:2 diluted plasma according to the manufacturer’s instructions.

Techniques: Clinical Proteomics

Journal: Cancers

Article Title: Ex Vivo Mitochondrial Respiration Parallels Biochemical Response to Ibrutinib in CLL Cells

doi: 10.3390/cancers13020354

Figure Lengend Snippet: Ibrutinib has similar effects on mitochondrial respiration profiles, CCL3 and 4, β-2 M, lactate dehydrogenase (LDH), Bruton tyrosine kinase (BTK) signaling and caspase-3 cleavage in chronic lymphocytic leukemia (CLL) patients independent of dose. The effect of ibrutinib treatment on basal respiration ( A ), maximal respiration ( B ), spare respiratory capacity ( C ), and respiratory control ratio ( D ) in primary CLL cells from patients pre-treatment and on ibrutinib treatment with low (blue) or standard (grey) dose. N = 14 for low dose and N = 5 for standard dose of ibrutinib. Values are mean ± S.D. **** p < 0.0001 (paired two-tailed Student’s t -test). The reduction of mitochondrial respiration parameters with low and standard dose of ibrutinib treatment is summarized for basal respiration ( E ) and maximal respiration ( F ) in freshly isolated CLL cells. N = 14 for low dose and N = 5 for standard dose of ibrutinib. Values are mean ± S.D. *** p < 0.0005, NS: non-significant (unpaired two-tailed Student’s t -test). In parallel, there is a reduction of chemokine ligands CCL3 ( G ) and CCL4 ( H ) in ibrutinib treated CLL patient plasma samples by dose. Decreased levels of β-2 M and LDH in ibrutinib treated patients compared to pre-treatment ( I , K ) and the decrease of β-2 M or LDH was similar to low or standard dose ibrutinib CLL patients ( J , L ). Demonstration of effects on pBTK, tBTK ( M ), and caspase-3 cleavage ( N ) are independent of dose. OCR: Oxygen consumption rate; resp: Respiratory; IB: Ibrutinib; TX: Treatment; Uncl. caspase-3: Uncleaved caspase-3; Cl. caspase-3: Cleaved caspase-3.

Article Snippet: Plasma CCL3 and CCL4 levels were measured pre and post treatment where matched plasma was available (Meso Scale Diagnostics, Rockville, MD [ , ].

Techniques: Control, Two Tailed Test, Isolation, Clinical Proteomics