Journal: Current Research in Microbial Sciences

Article Title: A multi-glycoprotein synergized recombinant virus system resolved the research dilemma of Epstein-Barr virus entry and facilitated the development of entry-specific drugs or vaccines

doi: 10.1016/j.crmicr.2026.100572

Figure Lengend Snippet: Biological and physical features of rEBV-VSV. (A) ELISA detecting the expression of glycoproteins (gp350, gp42, gH/gL, gB) on EBV Epi , EBV B , and the vector control. Two-way ANOVA was performed (p-values labelled in titles) followed by Tukey test among groups. * p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001. (B) TEM photographs of EBV Epi , EBV B , and the vector control. Scale bars: 500 nm. (C) Particle size distribution of EBV Epi , EBV B , and the vector control with Z-average Size (Z-Ave) and Polydispersity Index (PDI).

Article Snippet: Then, BSA was washed, and serially diluted antibodies (anti-gp350 pAbs (Sino Biological, 40373-RP01), anti-gp42 mAb 3E8, anti-gH/gL mAb AMMO1, and anti-gB mAb 3A5) were added as the primary antibodies and incubated at 37 °C for 1 h. After washing, HRP-conjugated anti-IgG secondary antibodies of corresponding species were added and incubated at 37 °C for 1 h. Then, the secondary antibodies were washed, and HRP was developed by TMB Chromogen Solution (Sangon Biotech, E661007) and terminated by ELISA Stopping Solution (Sangon Biotech, E661006).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Plasmid Preparation, Control

Journal: Current Research in Microbial Sciences

Article Title: A multi-glycoprotein synergized recombinant virus system resolved the research dilemma of Epstein-Barr virus entry and facilitated the development of entry-specific drugs or vaccines

doi: 10.1016/j.crmicr.2026.100572

Figure Lengend Snippet: mAbs’ entry-inhibiting assay based on rEBV-VSV. (A) The flowchart describing the process of this entry-inhibiting assay. (B) Purified anti-EBV mAbs used in this assay with the mAbs order same as the order in (C). (C) The table summarizing mAbs’ targets and their effects on epithelia and B cells. (D-G) Entry-inhibiting activities of anti-gp350 (D), anti-gp42 (E), anti-gH/gL (F), and anti-gB (G) mAbs that listed above with IC 50 +95 % CI.

Article Snippet: Then, BSA was washed, and serially diluted antibodies (anti-gp350 pAbs (Sino Biological, 40373-RP01), anti-gp42 mAb 3E8, anti-gH/gL mAb AMMO1, and anti-gB mAb 3A5) were added as the primary antibodies and incubated at 37 °C for 1 h. After washing, HRP-conjugated anti-IgG secondary antibodies of corresponding species were added and incubated at 37 °C for 1 h. Then, the secondary antibodies were washed, and HRP was developed by TMB Chromogen Solution (Sangon Biotech, E661007) and terminated by ELISA Stopping Solution (Sangon Biotech, E661006).

Techniques: Purification

Journal: Current Research in Microbial Sciences

Article Title: A multi-glycoprotein synergized recombinant virus system resolved the research dilemma of Epstein-Barr virus entry and facilitated the development of entry-specific drugs or vaccines

doi: 10.1016/j.crmicr.2026.100572

Figure Lengend Snippet: Mutated rEBV-VSV for the studying of EBV entry mechanisms. (A-B) Infectivity of mutated EBV B and EBV Epi on Raji (A) and C666-1 (B) cells. The infectivity was quantified by the infection fold ( %) to wild type EBV B and EBV Epi . One-way ANOVA was performed followed by Tukey test. * p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001. (C) The infectivity of wild type EBV Epi and EBV Epi with gH L65A+L69A mutation in different combinations of gp350 and gp42. Unpaired t-test was performed, and the increasing folds with p-values were listed above columns. (D) Entry-inhibiting activities of E1D1 against wild type EBV Epi or mutated EBV Epi (gH L65A+L69A) in different combinations of gp350 and gp42.

Article Snippet: Then, BSA was washed, and serially diluted antibodies (anti-gp350 pAbs (Sino Biological, 40373-RP01), anti-gp42 mAb 3E8, anti-gH/gL mAb AMMO1, and anti-gB mAb 3A5) were added as the primary antibodies and incubated at 37 °C for 1 h. After washing, HRP-conjugated anti-IgG secondary antibodies of corresponding species were added and incubated at 37 °C for 1 h. Then, the secondary antibodies were washed, and HRP was developed by TMB Chromogen Solution (Sangon Biotech, E661007) and terminated by ELISA Stopping Solution (Sangon Biotech, E661006).

Techniques: Infection, Mutagenesis

Journal: Oncology Letters

Article Title: EBV-positive large-cell and small-cell neuroendocrine carcinoma of the nasopharynx: A case report and review of the literature

doi: 10.3892/ol.2022.13584

Figure Lengend Snippet: Immunohistochemical features of the tumor. The tumor was immunopositive for (A) CD56, (B) synaptophysin and (C) CK and negative for (D) NSE and (E) TTF-1. (F) The tumor was positive for EBER according to the result of in situ hybridization (magnification, ×400; scale bars, 25 µm). CK, cytokeratin; NSE, neuron-specific enolase; TTF-1, thyroid transcription factor-1; EBV, Epstein-Barr virus; EBER, Epstein-Barr virus-encoded RNA.

Article Snippet: In situ hybridization for Epstein-Barr virus (EBV)-encoded RNA (EBER) , which was performed with an EBER detection kit (ISH-7001; OriGene Technologies, Inc.) according to the manufacturer's instructions, indicated that the sample was positive for EBER ( ).

Techniques: Immunohistochemical staining, In Situ Hybridization, Virus

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Role of IL-6-IL-27 Complex in Host Antiviral Immune Response.

doi: 10.4049/jimmunol.2100179

Figure Lengend Snippet: FIGURE 1. Interactions between the subunits of IL-6 and IL-27 in vivo and in vitro. (AC) CoIP and immunoblotting of serum samples were performed with the indicated Abs. IL-6 interacted with IL-27A (A), IL-6 interacted with EBi3 (B), and IL-27A interacted with EBi3 (C). (DG) HEK293T cells were transfected with the indicated plasmids for 24 h. CoIP and immunoblotting were performed with the indicated Abs. IL-6-3HA interacted with IL-27A-V5 (D), IL-6-3HA interacted with EBi3-Myc (E), IL-27A-V5 interacted with EBi3-Myc (F), and IL-6-3HA interacted with IL-27-V5 (G). (H) Colocalization of IL-6 and IL-27 was detected by confocal microscopy. A549 cells were transfected with IL-6-eGFP-KDEL and IL-27-DsRed-KDEL for 24 h, and the nuclei were stained with DAPI before observation using confocal microscopy. All experiments were repeated at least three times with consistent results.

Article Snippet: Abs against V5 tag (66007-1-Ig), EBi3 (12371-1-AP), myxovirus resistance A (MxA; 13750-1-AP), MAVS (14341-1-AP), TGFb-activated kinase 1 (12330-2-AP), IRF3 (11312-1-AP), NF-kB p65 (10745- 1-AP), NF-kB p50 (14220-1-AP), Lamin A/C (10298-1-AP), GAPDH (60004-1-Ig), and b-actin (60008-1-Ig) were purchased from ProteinTech Group (Wuhan, China).

Techniques: In Vivo, In Vitro, Western Blot, Transfection, Confocal Microscopy, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Role of IL-6-IL-27 Complex in Host Antiviral Immune Response.

doi: 10.4049/jimmunol.2100179

Figure Lengend Snippet: FIGURE 2. Construction of the IL-6IL-27 complex and its antiviral activity. (A) A schematic of the IL-6IL-27 construction via flexible self-cleaved linker is shown. Immunoblotting analysis of IL-6, EBi3, IL-27A, and b-actin of HEK293T cells transfected with empty vector pcDNA3.1(1) or IL-6IL-27 complex expression plasmids for 24 h. (B) A549 cells were transfected with indicated plasmids for 24 h followed by H1N1 infection (MOI = 1) for 12 h. The NP-specific mRNA, cRNA, and vRNA levels were measured using QRT-PCR. (C) RD cells were transfected with the indicated plasmids for 24 h fol- lowed by EV71 infection (MOI = 1) for 24 h. The VP1 mRNA levels were measured using QRT-PCR. (D) Vero cells were (Figure legend continues)

Article Snippet: Abs against V5 tag (66007-1-Ig), EBi3 (12371-1-AP), myxovirus resistance A (MxA; 13750-1-AP), MAVS (14341-1-AP), TGFb-activated kinase 1 (12330-2-AP), IRF3 (11312-1-AP), NF-kB p65 (10745- 1-AP), NF-kB p50 (14220-1-AP), Lamin A/C (10298-1-AP), GAPDH (60004-1-Ig), and b-actin (60008-1-Ig) were purchased from ProteinTech Group (Wuhan, China).

Techniques: Activity Assay, Western Blot, Transfection, Plasmid Preparation, Expressing, Infection, Quantitative RT-PCR

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Role of IL-6-IL-27 Complex in Host Antiviral Immune Response.

doi: 10.4049/jimmunol.2100179

Figure Lengend Snippet: FIGURE 5. The IL-6IL-27 complex interacts with MAVS. (AC) A549 cells were transfected with increasing amounts of the indicated expression plas- mids (wedge; 100, 200, and 300 ng) of the IL-6IL-27 complex expression plasmid and luciferase reporter plasmids containing the IFN-b (A), NF-kB (B), and ISRE (C) promoter for 24 h followed by VSV infection (MOI = 1) for 24 h. Reporter assays were performed. pRL-TK was used as an internal control. (DI) HEK293T cells were transfected with the indicated plasmids for 24 h. CoIP and immunoblotting were performed with the indicated Abs. IL-6-3HA interacted with Flag-MAVS (D), IL-27A-V5 interacted with Flag-MAVS (E), EBi3-Myc interacted with Flag-MAVS (F), IL-6-HA interacted with Flag- TBK1 (G), IL-27A-V5 interacted with Flag-TBK1 (H), and EBi3-Myc interacted with Flag-TBK1 (I). All experiments were repeated at least three times with consistent results.

Article Snippet: Abs against V5 tag (66007-1-Ig), EBi3 (12371-1-AP), myxovirus resistance A (MxA; 13750-1-AP), MAVS (14341-1-AP), TGFb-activated kinase 1 (12330-2-AP), IRF3 (11312-1-AP), NF-kB p65 (10745- 1-AP), NF-kB p50 (14220-1-AP), Lamin A/C (10298-1-AP), GAPDH (60004-1-Ig), and b-actin (60008-1-Ig) were purchased from ProteinTech Group (Wuhan, China).

Techniques: Transfection, Expressing, Plasmid Preparation, Luciferase, Infection, Control, Western Blot

Journal: Virology Journal

Article Title: A study of Epstein-Barr virus infection in the Chinese tree shrew( Tupaia belangeri chinensis )

doi: 10.1186/s12985-017-0859-5

Figure Lengend Snippet: Expression of EBV genes in PBMCs of Ts1–4. In Ts1 and Ts2, only EBNA1 and LMP1 were detected at week 4, respectively. In Ts3, BZLF1 was expressed from weeks 6 to 10, and LMP1 was observed only at week 6.In Ts4, BZLF1 was expressed from weeks 2 to 4, and the expression of LMP1 persisted throughout the observation period with the exception of week 4

Article Snippet: EBV antibodies in tree shrew serum were measured using an enzyme-linked immunosorbent assay (ELISA).The anti-EBV capsid antigen(VCA) IgG,EBV nuclear antigen (EBNA) IgG, and early antigen (EA) IgG were measured with the Epstein-Barr virus VCA IgG ELISA Kit(Abnova,Taiwan, China),a human Epstein-Barr virus nuclear antigen (EBNA1) antibody (IgG) ELISA kit (CUSABIO, Wuhan, China), and a human Epstein-Barr virus early antigen (EBEA) antibody (IgG) ELISA kit (CUSABIO).Tree shrew serum diluted 1:10 with dilution buffer was placed into each plate, and bound antibodies were subsequently detected using rabbit anti-tree shrew IgGsecondary antibody synthesized by Sangon Biotech Co.,Ltd. (Shanghai, China).

Techniques: Expressing

Journal: Virology Journal

Article Title: A study of Epstein-Barr virus infection in the Chinese tree shrew( Tupaia belangeri chinensis )

doi: 10.1186/s12985-017-0859-5

Figure Lengend Snippet: Expression of EBV genes in PBMCs of Ts7–10. In Ts7, the expression of LMP1 persisted throughout the observation period, excluding week 2, BZLF1 appeared at weeks 2 to 4,EA was detected intermittently from weeks 4 to 18, and EBNA1 was observed only at week 18. In Ts8, EBNA1, EBNA2 and LMP1were expressed at week 2. In Ts9, LMP1was detected at weeks 2, 6 and 21, and EA and EBNA1were detected only at weeks 2 and 16, respectively. In Ts10, the expression of BZLF1 persisted throughout the observation period,and LMP1 was detected at week 1

Article Snippet: EBV antibodies in tree shrew serum were measured using an enzyme-linked immunosorbent assay (ELISA).The anti-EBV capsid antigen(VCA) IgG,EBV nuclear antigen (EBNA) IgG, and early antigen (EA) IgG were measured with the Epstein-Barr virus VCA IgG ELISA Kit(Abnova,Taiwan, China),a human Epstein-Barr virus nuclear antigen (EBNA1) antibody (IgG) ELISA kit (CUSABIO, Wuhan, China), and a human Epstein-Barr virus early antigen (EBEA) antibody (IgG) ELISA kit (CUSABIO).Tree shrew serum diluted 1:10 with dilution buffer was placed into each plate, and bound antibodies were subsequently detected using rabbit anti-tree shrew IgGsecondary antibody synthesized by Sangon Biotech Co.,Ltd. (Shanghai, China).

Techniques: Expressing

Journal: Virology Journal

Article Title: A study of Epstein-Barr virus infection in the Chinese tree shrew( Tupaia belangeri chinensis )

doi: 10.1186/s12985-017-0859-5

Figure Lengend Snippet: Levels of EBV antibody in the serum of tree shrews post-infection. The level of VCA IgG increased to varying degrees in all tree shrews in which the EBV copy number increased; EBNA1 IgG increased in only1 tree shrew (Ts7),and EA IgG was not elevated ( a ). The levels of these antibodies were not increased in the EBV-infection-negative tree shrews ( b ) or in any of the 3 tree shrews that served as negative controls (data not shown)

Article Snippet: EBV antibodies in tree shrew serum were measured using an enzyme-linked immunosorbent assay (ELISA).The anti-EBV capsid antigen(VCA) IgG,EBV nuclear antigen (EBNA) IgG, and early antigen (EA) IgG were measured with the Epstein-Barr virus VCA IgG ELISA Kit(Abnova,Taiwan, China),a human Epstein-Barr virus nuclear antigen (EBNA1) antibody (IgG) ELISA kit (CUSABIO, Wuhan, China), and a human Epstein-Barr virus early antigen (EBEA) antibody (IgG) ELISA kit (CUSABIO).Tree shrew serum diluted 1:10 with dilution buffer was placed into each plate, and bound antibodies were subsequently detected using rabbit anti-tree shrew IgGsecondary antibody synthesized by Sangon Biotech Co.,Ltd. (Shanghai, China).

Techniques: Infection

Journal: Virology Journal

Article Title: A study of Epstein-Barr virus infection in the Chinese tree shrew( Tupaia belangeri chinensis )

doi: 10.1186/s12985-017-0859-5

Figure Lengend Snippet: Western blotting for the detection of EBV gene expression in the livers and spleens. EBNA1 was detected in the spleens of Ts2, Ts3, Ts9 and Ts10. No EBNA1 expression was detected in the livers of any of the tree shrews. B95–8 cells and Ts11–13 were used as positive and negative controls, respectively. Ts5 and Ts6 were EBV-infection-negative. No EBNA1expression was detected inTs5, Ts6 or Ts11–13

Article Snippet: EBV antibodies in tree shrew serum were measured using an enzyme-linked immunosorbent assay (ELISA).The anti-EBV capsid antigen(VCA) IgG,EBV nuclear antigen (EBNA) IgG, and early antigen (EA) IgG were measured with the Epstein-Barr virus VCA IgG ELISA Kit(Abnova,Taiwan, China),a human Epstein-Barr virus nuclear antigen (EBNA1) antibody (IgG) ELISA kit (CUSABIO, Wuhan, China), and a human Epstein-Barr virus early antigen (EBEA) antibody (IgG) ELISA kit (CUSABIO).Tree shrew serum diluted 1:10 with dilution buffer was placed into each plate, and bound antibodies were subsequently detected using rabbit anti-tree shrew IgGsecondary antibody synthesized by Sangon Biotech Co.,Ltd. (Shanghai, China).

Techniques: Western Blot, Gene Expression, Expressing, Infection

Journal: Frontiers in Immunology

Article Title: Characteristics of tertiary lymphoid structures in prostate cancer and the impact of neoadjuvant therapy on their formation and maturation

doi: 10.3389/fimmu.2025.1663396

Figure Lengend Snippet: Clinicopathological features and prognostic significance of tertiary lymphoid structures (TLS) (A) The distribution of clinical features and immature density for the 27 patients in cohort 1. (B) Kaplan-Meier survival curves for progression-free survival (PFS) of 27 patients with no treatment before surgery stratified based on the median density of TLSs (B) , density of intra-/peri- TLSs (C) , maturation of TLSs (D) and the existence of SFL-TLSs (In SFL-TLS, CD21+ follicular dendritic cells form dense clusters within the core germinal center) (E) . B–E: Differentiate Kaplan-Meier curves with distinct colors for better visual clarity. 3D & 3E: Clarify the difference between “mature TLS” and “SFL-TLS.”.

Article Snippet: Then, slides were treated by microwave to induce antigen retrieval using citric acid solution for 15 min. 6 primary antibodies were used, including CD4 (1:500, #Ab67480, Abcam), CD8 (1:300, #66868-1-Ig, proteintech), CD20 (1:1000, #60271-1-Ig, proteintech), CD21 (1:200, #24374-1-AP, proteintech), Ki67(1:1000, #HA721115, HuaBio), pan-cytokeratin (1:2000, # HA601094 , HuaBio), DAPI (1:500, #C1002, Beyotime).

Techniques:

Journal: Frontiers in Immunology

Article Title: Characteristics of tertiary lymphoid structures in prostate cancer and the impact of neoadjuvant therapy on their formation and maturation

doi: 10.3389/fimmu.2025.1663396

Figure Lengend Snippet: Characteristics of tertiary lymphoid structures (TLS) in treatment-naive group and neoadjuvant hormone therapy (NHT) group in cohort 2. (A) mIHC of immature and mature TLS in NHT group and treatment-naive group in cohort 2; (B) Comparison of the ratio of CD4 + cells, CD8+ cells, CD20+cells and CD21+cells in tumor microenvironment (TME) in the NHT group and treatment-naive group. (C) Proportion of different TLSs maturation stage in the NHT group and treatment-naive group. (D) Proportion of different density of TLS in the NHT group and treatment-naive group. (E) Proportion of different location of TLS in the NHT group and treatment-naive group. P values are denoted as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ***p < 0.0001.

Article Snippet: Then, slides were treated by microwave to induce antigen retrieval using citric acid solution for 15 min. 6 primary antibodies were used, including CD4 (1:500, #Ab67480, Abcam), CD8 (1:300, #66868-1-Ig, proteintech), CD20 (1:1000, #60271-1-Ig, proteintech), CD21 (1:200, #24374-1-AP, proteintech), Ki67(1:1000, #HA721115, HuaBio), pan-cytokeratin (1:2000, # HA601094 , HuaBio), DAPI (1:500, #C1002, Beyotime).

Techniques: Comparison

Journal: Frontiers in Immunology

Article Title: Characteristics of tertiary lymphoid structures in prostate cancer and the impact of neoadjuvant therapy on their formation and maturation

doi: 10.3389/fimmu.2025.1663396

Figure Lengend Snippet: Characteristics and changes of immune cells in pre- and post- neoadjuvant hormone therapy (NHT) groups in GSE111177 . (A) Supervised clustering of PCa specimens by NHT (n = 24), displaying ssGSEA scores ( GSE111177 ). (B) The biological process of DEGs identified in pre-/post- NHT groups ( GSE111177 ). (C) Comparison of the expression of CD4, CD8A and CD8B, CD20, FOXP3, CD21, GZMB, PRF1 in PCa patients before and after NHT using GSE111177 cohort (n = 24). (D) Comparison of GSVA scores utilizing four TLS signatures in 24 paired pre-NHT and post-NHT samples from GSE111177 .

Article Snippet: Then, slides were treated by microwave to induce antigen retrieval using citric acid solution for 15 min. 6 primary antibodies were used, including CD4 (1:500, #Ab67480, Abcam), CD8 (1:300, #66868-1-Ig, proteintech), CD20 (1:1000, #60271-1-Ig, proteintech), CD21 (1:200, #24374-1-AP, proteintech), Ki67(1:1000, #HA721115, HuaBio), pan-cytokeratin (1:2000, # HA601094 , HuaBio), DAPI (1:500, #C1002, Beyotime).

Techniques: Comparison, Expressing

Journal: Frontiers in Immunology

Article Title: Characteristics of tertiary lymphoid structures in prostate cancer and the impact of neoadjuvant therapy on their formation and maturation

doi: 10.3389/fimmu.2025.1663396

Figure Lengend Snippet: Characterization of tumor immune microenvironment (TME) and tertiary lymphoid structures (TLS) before and after neoadjuvant hormone therapy (NHT). (A) Comparison of the ratio of CD4 + cells, CD8+ cells, CD20+cells and CD21+cells in TME before and after NHT. (B) Comparison of the TLS presence in TME before and after NHT. (C) Comparison of the intra-tumor TLS presence in TME before and after NHT. (D) Comparison of the TLS maturation stage in TME before and after NHT. (E) Evaluation of TLS maturation stage using mIHC staining of CD4, CD8 CD20, CD21 before and after NHT. P values are denoted as follows: **p < 0.01, ***p < 0.001, NS p>0.05.

Article Snippet: Then, slides were treated by microwave to induce antigen retrieval using citric acid solution for 15 min. 6 primary antibodies were used, including CD4 (1:500, #Ab67480, Abcam), CD8 (1:300, #66868-1-Ig, proteintech), CD20 (1:1000, #60271-1-Ig, proteintech), CD21 (1:200, #24374-1-AP, proteintech), Ki67(1:1000, #HA721115, HuaBio), pan-cytokeratin (1:2000, # HA601094 , HuaBio), DAPI (1:500, #C1002, Beyotime).

Techniques: Comparison, Staining

Journal: International Immunopharmacology

Article Title: IL-35 interferes with splenic T cells in a clinical and experimental model of acute respiratory distress syndrome

doi: 10.1016/j.intimp.2018.12.024

Figure Lengend Snippet: (A, E) Pretreatment with IL-35 neutralizing antibodies (anti-P35 or anti-Ebi3) aggravated LPS- and CLP-induced ARDS. Lungs from each experimental group were processed for histological examination after H&E staining. Compared with the LPS + IgG group, thickened alveolar wall, alveolar hemorrhage and collapse, inflammatory cells in filtration were more severe and pretreated with IL-35 neutralizing antibodies prior to LPS challenge. (B, D) Lung injury scores were estimated by the method of Mikawa, which is from the following four indicators of lung injury score: alveolar congestion; bleeding; gap or vascular wall neutrophil infiltration or aggregation; alveolar septal thickening or transparent membrane formation. 0 marks: no or very slight damage, 1 marks: mild injury, 2 marks: moderate injury, 3 marks: severe injury, 4 marks: very severe damage, the cumulative increase in the number of lesions of the total score is the pathological score of the ARDS. Pretreatment with IL-35 neutralizing antibodies lungs injury were more severe than LPS + IgG group. **p < 0.01, ***p < 0.001, by the two-way ANOVA followed by LSD multiple comparisons test, compared with the LPS + IgG group.

Article Snippet: Neutralization assays were performed by giving 50 μg anti-mouse IL-35 EBI3 (Rockland Immunochemicals) or anti-mouse IL-12A p35 (Abcam Systems) 30 min after CLP or LPS.

Techniques: Staining, Filtration, Membrane

Journal: International Immunopharmacology

Article Title: IL-35 interferes with splenic T cells in a clinical and experimental model of acute respiratory distress syndrome

doi: 10.1016/j.intimp.2018.12.024

Figure Lengend Snippet: Interleukin 35 (IL-35) blockade upregulated the production of proinflammatory cytokines and downregulated anti-inflammatory cytokines during ARDS models. Cytokine and chemokine concentrations in BALF and blood specimens from five mice treated with or without anti–Ebi3 or anti–P35 blocking antibodies were determined by mice cytokine/chemokine magnetic bead panel kit assays 24 h after onset of ARDS. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, by the one-way ANOVA followed by LSD multiple comparisons test, compared with mice treated with isotypical IgG control. ARDS featured increased CXCL-1, TNF-α, IL-6, and IL-17A and reduced IFN-γ, IL-10, IL-2, and IL-13 after neutralizing antibody treatment (anti-IL-35 Ebi3 or anti-IL-35 P35) in BALF and serum.

Article Snippet: Neutralization assays were performed by giving 50 μg anti-mouse IL-35 EBI3 (Rockland Immunochemicals) or anti-mouse IL-12A p35 (Abcam Systems) 30 min after CLP or LPS.

Techniques: Blocking Assay, Control

Journal: Oncology Letters

Article Title: Adjuvant immunotherapy plus chemotherapy and maintenance immunotherapy for pulmonary lymphoepithelioma-like carcinoma with hepatitis B virus infection, KRAS mutation and high expression of programmed death ligand 1: A case report

doi: 10.3892/ol.2026.15612

Figure Lengend Snippet: Pathology and IHC of the pulmonary lymphoepithelioma-like carcinoma. (A and B) Histological analysis. (A) heterologous cells were arranged in clusters (magnification, ×40); (B) the nuclei of heterologous cells were vacuolated and the nucleoli were clear (magnification, ×200; H&E staining); the arrows point at heterologous cells. (C-I) IHC analysis. (C) Cytoplasmic CK5/6 was positive, the arrows point at positive CK5/6 cytoplasm; (D) nuclear P40 was positive; the arrows point at positive P40 nuclear; (E) thyroid transcription factor-1 was negative; (F) Napsin A was negative; (G) nuclear EBER was positive; the arrows point at positive EBER nuclear; (H) proliferation index Ki-67 positive ~50%, the arrows point at positive Ki-67 nuclear; (I) PD-L1 detection microscopic image: Tumor proportion score=55% (magnification, ×100), the arrows point at positive PD-L1 nuclear. CK, cytokeratin; PD-L1, programmed cell death ligand 1; EBER, Epstein-Barr virus-encoded small RNA; IHC, immunohistochemistry.

Article Snippet: Hybridization in situ : EBER monoclonal antibody (cat. no. 25080502; dilution, ready-to-use; Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.).

Techniques: Staining, Virus, Immunohistochemistry

Journal: Cell Reports Medicine

Article Title: CD101 genetic variants modify regulatory and conventional T cell phenotypes and functions

doi: 10.1016/j.xcrm.2021.100322

Figure Lengend Snippet: T cells from individuals with variants in CD101 have increased potential to express cytokines (A–D) Cytokine-producing CD8 + or CD4 + T cells from EBV-stimulated (A and C) or αCD3/αCD28-stimulated (B and D) whole PBMCs for CD101 Ig-like (N = 42) and cytoplasmic variants (N = 18) compared with no functional CD101 variants (missense, 3′ or 5′ untranslated region, splice site) (N = 40). Live-sorted PBMCs, including lymphocytes and APCs, were recovered and stimulated with EBV lysate or glycine control (A and C) or αCD3/αCD28 or medium control (B and D) for 6 h prior to staining for intracellular cytokine production. The frequencies of CD101 + CD8 + T cells producing IFN-γ after subtracting background values were affected by the presence of Ig-like and cytoplasmic- CD101 variants (A and B). Similarly, production of IFN-γ by CD101 + CD4 + T cells after EBV stimulation was increased significantly for individuals carrying an Ig-like variant, whereas possessing a cytoplasmic variant did not have a significant effect (C). CD8 + T cells co-producing IFN-γ and TNF-α after αCD3/αCD28 stimulation also trended toward an increased frequency in individuals with the cytoplasmic variant (D). Four participants were homozygous for Ig-like variant rs12093834 ; they are denoted by orange symbols. (E–H) Live-sorted PBMCs, including lymphocytes and APCs, were recovered and stimulated with EBV lysate or glycine control (E and G) or αCD3/αCD28 or medium control (F and H) for 6 h prior to staining for intracellular cytokine production. Total CD4 + CD3 + or CD8 + CD3 + T cells were gated by CD101 positivity and then assessed for their ability to co-produce IFN-γ and TNF-α. EBV lysate stimulation of PBMCs isolated from individuals with cytoplasmic variants resulted in an increased frequency of IFN-γ + TNF-α + CD8 + (E) and CD4 + (G) T cells, whereas having an Ig-like variant did not produce a significant effect. Stimulation with αCD3/αCD28 antibodies elicited an increased frequency of IFN-γ + TNF-α + CD8 + (F) and CD4 + (H) T cells in individuals with both Ig-like and cytoplasmic-variants. Matched background control values were subtracted for all participants. Four participants were homozygous for Ig-like variant rs12093834 ; they are denoted by orange symbols. Each data point represents one individual. Adjusted p values were calculated as described in .

Article Snippet: EBV Lysate , East Coast Bio , EV012.

Techniques: Functional Assay, Control, Staining, Variant Assay, Isolation

Journal: Cell Reports Medicine

Article Title: CD101 genetic variants modify regulatory and conventional T cell phenotypes and functions

doi: 10.1016/j.xcrm.2021.100322

Figure Lengend Snippet: Effect of CD101 variation on T cell activation and Treg cell functional characteristics

Article Snippet: EBV Lysate , East Coast Bio , EV012.

Techniques: Activation Assay, Functional Assay, Control, Variant Assay

Journal: Cell Reports Medicine

Article Title: CD101 genetic variants modify regulatory and conventional T cell phenotypes and functions

doi: 10.1016/j.xcrm.2021.100322

Figure Lengend Snippet: CD101 variation diminishes Treg cell-mediated restraint of effector T cells (A) Schematic of the PBMC sorting and stimulation protocol (created with BioRender). PBMCs (N = 100) were sorted into (1) whole live PBMCs and (2) Treg cell-depleted or (3) purified CD3 + T cell fractions. Cells were stimulated with EBV lysate or control or αCD3/αCD28 or control for 6 h. Cells were then analyzed for their expression of cytokines by intracellular cytokine staining (ICS). (B) The frequency of CD4 + and CD8 T cells producing proinflammatory cytokines in response to EBV among Treg-cell-depleted PBMCs was analyzed per case or control. (C and D) Individual trajectory plots (C) and summary results (D) of Δ% IL-2 + CD4 + T cells from Treg cell-depleted and whole PBMCs for CD101 cytoplasmic (N = 18) and Ig-like variants (N = 42) compared with no functional variants (N = 40). Live-sorted “whole” PBMCs, including lymphocytes and APCs, or PBMCs sorted to deplete Treg cells were recovered and stimulated with EBV lysate or glycine control for 6 h prior to staining for intracellular cytokine production. In (C), the frequencies of IL-2-producing CD4 + T cells recovered from the “whole” PBMC fraction and the Treg cell-depleted fraction are plotted as a trajectory plot, and in (D), the difference between the percentage of IL-2 + CD4 + T cells in Treg cell-depleted compared with whole PBMCs is plotted. Results are stratified by the presence of an Ig-like or cytoplasmic variant. Each data point represents one individual. Adjusted p values were calculated as described in .

Article Snippet: EBV Lysate , East Coast Bio , EV012.

Techniques: Purification, Control, Expressing, Staining, Functional Assay, Variant Assay

Journal: Cell Reports Medicine

Article Title: CD101 genetic variants modify regulatory and conventional T cell phenotypes and functions

doi: 10.1016/j.xcrm.2021.100322

Figure Lengend Snippet:

Article Snippet: EBV Lysate , East Coast Bio , EV012.

Techniques: Transmission Assay, Recombinant, DNA Library Preparation, RNA Sequencing, Software, Expressing, Gene Expression