Journal: mBio

Article Title: Recovery from Acute SARS-CoV-2 Infection and Development of Anamnestic Immune Responses in T Cell-Depleted Rhesus Macaques

doi: 10.1128/mBio.01503-21

Figure Lengend Snippet: Experimental design and T cell depletions. (a) T cell subset-depleting antibodies were administered on days −7, 0, and +4, as indicated by blue arrows. Infections were done on days 0 and 42, as indicated by red arrows. Blood withdrawals were performed on the days indicated by the black arrows, and flow cytometry was used to determine the lymphocyte subset numbers over time. The flow cytometry gating strategies are shown in Fig. S1b . Each symbol represents a single animal throughout. All CD4-depleted animals except CD4-5 were still more than 90% depleted of CD4 + T cells at 7 dpi. CD4-5 was 78% depleted. CD4 + Th numbers excluded FoxP3 + cells. At 7 days after reinfection (49 dpi), the animals averaged 81% depletion. All CD8-depleted animals were >99% depleted at 7 dpi and remained 78% depleted at 49 dpi. The differences between subset numbers at 0 dpi and 7 dpi were calculated by a two-way paired t test. ns, not significant. P values are shown. Numbers of B cells (d, g, j, and m) were determined by flow cytometry using CD45 and CD20 as markers. The numbers of B cells in the CD4-depleted group were significantly lower over time than those in the controls, as determined by mixed-effects analysis ( P = 0.0118).

Article Snippet: Specific staining was detected using SARS-CoV/SARS-CoV-2 nucleocapsid antibody (Sino Biological cat#40143-MM05) at a 1:1,000 dilution, CD4 antibody (Abcam catalog no. ab133616) at a 1:100 dilution, CD8 antibody (Sino Biological catalog no. 10980-T24) at a 1:500 dilution, and CD20 (Thermo Scientific catalog no. RB-9013) at a 1:250 dilution.

Techniques: Flow Cytometry

Journal: mBio

Article Title: Recovery from Acute SARS-CoV-2 Infection and Development of Anamnestic Immune Responses in T Cell-Depleted Rhesus Macaques

doi: 10.1128/mBio.01503-21

Figure Lengend Snippet: Virus detection from nasal swabs and Broncho-alveolar lavages. (a to d) Each symbol represents the value of viral RNA copies from an individual animal at each time point. The brackets delineate comparisons of cumulative values from the first 2 weeks after infection with those from the 2 weeks after reinfection, and numbers P values from two-way paired t tests showing significantly reduced virus levels following the second infection (b to d) except in the control group (a), for which the difference was marginally nonsignificant. The cumulative RNA titers from the CD4- and CD8-depleted groups for the first 2 weeks after initial infection were not significantly different from those from the controls, but the CD4/CD8-depleted group had significantly higher titers ( P = 0.0362 by one-way analysis of variance (ANOVA) with a Dunnett’s posttest). (e to h) Mean values comparing the total viral RNA data from panels a to d (black lines) with sgRNA results (blue lines). (i to l) Bronchoalveolar lavage fluids were taken at 1 day after infection and reinfection. sgRNA was measured from each animal and showed significantly reduced virus replication upon second infection ( P values from paired t tests are shown).

Article Snippet: Specific staining was detected using SARS-CoV/SARS-CoV-2 nucleocapsid antibody (Sino Biological cat#40143-MM05) at a 1:1,000 dilution, CD4 antibody (Abcam catalog no. ab133616) at a 1:100 dilution, CD8 antibody (Sino Biological catalog no. 10980-T24) at a 1:500 dilution, and CD20 (Thermo Scientific catalog no. RB-9013) at a 1:250 dilution.

Techniques: Infection

Journal: mBio

Article Title: Recovery from Acute SARS-CoV-2 Infection and Development of Anamnestic Immune Responses in T Cell-Depleted Rhesus Macaques

doi: 10.1128/mBio.01503-21

Figure Lengend Snippet: Immunohistochemical staining of cervical lymph nodes for CD4 + and CD8 + T cells and B cells. Representative animals from each experimental group are shown. (a to d) Cervical lymph nodes stained with anti-CD4 antibodies. (e to h) Cervical lymph nodes stained with anti-CD8 antibodies. (i to l) Cervical lymph nodes stained with anti-CD20 antibodies to detect B cells. None of the tissues stained positive for the presence of SARS-CoV-2 at 56 dpi.

Article Snippet: Specific staining was detected using SARS-CoV/SARS-CoV-2 nucleocapsid antibody (Sino Biological cat#40143-MM05) at a 1:1,000 dilution, CD4 antibody (Abcam catalog no. ab133616) at a 1:100 dilution, CD8 antibody (Sino Biological catalog no. 10980-T24) at a 1:500 dilution, and CD20 (Thermo Scientific catalog no. RB-9013) at a 1:250 dilution.

Techniques: Immunohistochemical staining, Staining

Journal: Oncogene

Article Title: Tissue factor overexpression promotes resistance to KRAS-G12C inhibition in non-small cell lung cancer

doi: 10.1038/s41388-023-02924-y

Figure Lengend Snippet: A TCGA datasets were analyzed for TF and KRAS mRNA expression. B OS and PFS of TF high and TF low groups were calculated by Kaplan-Meier method. C The infiltration levels of immune cells in TF high vs TF low groups in TCGA cohort. D KRASmut lung cancer tissue microarrays were subjected to IHC staining. TF expression was analyzed versus the indicated clinical parameters. E OS and PFS in negative/low ( n = 36) and moderate/high ( n = 42) TF expression groups were calculated by Kaplan-Meier method. F IHC was performed for the indicated biomarkers, quantified as described in methods and plotted as expression level heatmap in KRASmut LUAD patients of stage III and IV. G The levels of P-ERK, P-AKT and immune cell markers in TF high/low KRASmut LUAD patients of stage III and IV ( n = 29). H Comparing the correlation between P-ERK, P-AKT, CD206, CD8A + GZMB + and TF expression in KRASmut LUAD patients of stage III and IV. I IHC profile of representative TF-high/low tumors are shown. Loss of tissue during staining was not included in the analysis. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The primary antibodies are as follows: TF (E9M6T, CST), P-ERK1/2 (4370, CST), P-AKT (Ser473) (ab81283, Abcam), CD68 (ab955, Abcam), (24595 S, CST), CD8A (50389-T26, Sino Biological) and GZMB (255598, Abcam).

Techniques: Expressing, Immunohistochemistry, Staining