3168002b  (fluidigm)

Journal: Nature Immunology

Article Title: Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function

doi: 10.1038/s41590-025-02381-7

Figure Lengend Snippet: a , sjTREC quantification in HDs (black dots), heterozygous donors (gray dots) and individuals with CBL -LOH (colored dots), as determined by qPCR of whole-blood DNA; WBCs, white blood cells. b , Recent thymic emigrant CD4 + and CD8 + T cells quantified in peripheral fresh blood by mass cytometry and gating of CD31 + cells among naive T cells; data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. c , Quantification of the indicated T cell subsets in the peripheral blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. In b and c , controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2) and heterozygous individuals ( n = 3). d , Percentage of dead cells in cultures of activated fresh PBMCs from HDs, the heterozygous father and participants (P1–P3) after 5 days of TCR stimulation, as determined by dead cell marker staining and flow cytometry; n = 3 HDs and n = 3 patient; data are shown as mean ± s.d. e , f , Cell division index of CD4 + ( e ) and CD8 + ( f ) T cells of HDs, the heterozygous father and participants (P1–P3) after 5 days of the indicated TCR stimulation, as determined by dilution of CFSE; n = 3 HDs and n = 3 patients; data are shown as mean ± s.d. g , Cytokine response by STAT5 phosphorylation (left) and cytokine production (right) by T cell blasts that are homozygous (red), heterozygous (gray) and homozygous WT for CBL Ub LOF variants following the indicated stimuli. The bars show the mean of the displayed data points (one for each T blast line); NS, not significant; TEMRA, terminally differentiated effector memory T cells; MAIT, mucosal-associated invariant T cells; MFI, median fluorescence intensity; PHA, phytohemagglutinin; PMA, phorbol 12-myristate 13-acetate.

Article Snippet: 168Er , CD8 , SK1 , 3168002B , Fluidigm.

Techniques: Mass Cytometry, MANN-WHITNEY, Marker, Staining, Flow Cytometry, Phospho-proteomics, Fluorescence

Journal: Nature Immunology

Article Title: Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function

doi: 10.1038/s41590-025-02381-7

Figure Lengend Snippet: ( a ) CytoF immunophenotyping of patients total CD3 + T cells, NK cells and indicated NK cell subsets. Controls 0–3 y.o. (n = 2), Controls 4–15 y.o. (n = 9), Controls 16–100 y.o. (n = 28), pediatric patients (LOH) (n = 5), adult patients (LOH) (n = 2), heterozygous individuals (n = 3), mean ± s.d. Statistical significance was assessed with multiple Mann Whitney tests adjusted for multiple testing. *p<0.05. ( b ) Assessment of T cell proliferation of patients P1, P2 and P3. (top) gating strategy, (bottom) CFSE dilution plots. ( c ) Intracellular TNF production of CD4 + and CD8 + T cell blasts in homozygous (n = 4), heterozygous (n = 2) and wildtype (n = 5) state of CBL Ub LOF variants. Line indicates the mean of the displayed datapoints. ( d,e ) Intracellular and extracellular cytokine production by patients T fh cells stimulated with CD2/CD3/CD28 under T h 0 conditions for 5 days. ( d ) Intracellular staining of the indicated cytokines. Healthy donors (n = 17), CBL-LOH patients (n = 4). Mann-Whitney testing with correction for multiple testing did not reveal significant (p< 0.05) differences between healthy donors and CBL-LOH patients. Line indicates the mean of the displayed datapoints. ( e ) Extracellular detection of cytokines by ELISA. Healthy donors (n = 6), CBL-LOH patients (n = 3). Mann-Whitney testing with correction for multiple testing did not reveal significant (p< 0.05) differences between healthy donors and CBL-LOH patients. Line indicates the mean of the displayed datapoints.

Article Snippet: 168Er , CD8 , SK1 , 3168002B , Fluidigm.

Techniques: MANN-WHITNEY, Staining, Enzyme-linked Immunosorbent Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Preclinical and early clinical studies of a novel compound SYHA1813 that efficiently crosses the blood-brain barrier and exhibits potent activity against glioblastoma.

doi: 10.1016/j.apsb.2023.09.009

Figure Lengend Snippet: Figure 5 Combination of SYHA1813 and anti-PD-1 antibody produces synergistic effects against glioma in immunocompetent mouse allograft model. (A) SYHA1813 suppressed the in vivo DBT tumor growth. BALB/c mice bearing with established DBT tumors were orally administered with or without SYHA1813 (10 mg/kg, twice daily). The RTVs are expressed as the mean SEM. (B) The expression of CD163 and CD8 in the tumor tissue was examined using IHC. (C) Diagram of combination study of SYHA1813 with PD-1 antibody in DBT mouse model. (D) SYHA1813 enhanced the anti-tumor activity of the anti-PD-1 antibody. The RTVs are expressed as the mean SEM, n Z 6. (E) The CD206þF4/ 80þ macrophage in tumor tissue in control or administrated groups was detected by CyTOF. Data were analyzed using t-test, *P < 0.05, **P < 0.01, ***P < 0.001. (F, G) The expression of Granzyme B (F) in CD8þ T cell, Treg (CD4þCD25þFOXP3þ) (G) in the tumor tissue were also detected by CyTOF. Data were analyzed using a t-test, **P < 0.01, ***P < 0.001, n Z 6.

Article Snippet: Two million cells were taken for surface staining, CD45-89Y, CD11b110Cd, CD3-152Sm, F4/80-146Nd, iNOS-161Dy, CD206169 Tm, CD4-145Nd, CD25-151Eu, CD8-168Er, PDL1-153Eu, and PD1-159 Tb antibodies were added, respectively, which were purchased from Fluidigm (San Francisco, CA, USA), and incubated on ice for 30 min.

Techniques: In Vivo, Expressing, Activity Assay, Control