cd30 elisa  (R&D Systems)

Journal: Cancers

Article Title: CD30 Lateral Flow and Enzyme-Linked Immunosorbent Assays for Detection of BIA-ALCL: A Pilot Study.

doi: 10.3390/cancers15215128

Figure Lengend Snippet: Figure 1. Cytospin cytology of fresh seromas from two patients with pathologically diagnosed BIA-ALCL. Giemsa stain of seroma 61 shows monotonous population of anaplastic cells which are CD30+. Apoptotic bodies of dead tumor cells are seen (arrows). Seroma 62 contains a CD30+ large multinucleate tumor cell and a CD30+ large mononuclear tumor cell surrounded by small CD30 negative lymphocytes and many erythrocytes (600×).

Article Snippet: CD30 ELISA was performed using Human CD/TNFRSF8 DuoSet ELISA kit DY612605 and a DuoSet Ancillary Reagent Kit 2 DY008 from R&D Systems, Minneapolis, MN, USA) on samples that were undiluted, and diluted 1:10 and 1:50.

Techniques: Giemsa Stain

Journal: Cancers

Article Title: CD30 Lateral Flow and Enzyme-Linked Immunosorbent Assays for Detection of BIA-ALCL: A Pilot Study.

doi: 10.3390/cancers15215128

Figure Lengend Snippet: Figure 2. LFA of recombinant CD30 protein at different concentrations produces varied intensity test lines corresponding to different intensities on the color scale. CD30 could be detected at concentrations as low as 250 pg/mL.

Article Snippet: CD30 ELISA was performed using Human CD/TNFRSF8 DuoSet ELISA kit DY612605 and a DuoSet Ancillary Reagent Kit 2 DY008 from R&D Systems, Minneapolis, MN, USA) on samples that were undiluted, and diluted 1:10 and 1:50.

Techniques: Recombinant

Journal: Clinical & Translational Immunology

Article Title: Memory stem T cells modified with a redesigned CD30‐chimeric antigen receptor show an enhanced antitumor effect in Hodgkin lymphoma

doi: 10.1002/cti2.1268

Figure Lengend Snippet: T SCM‐like are highly enriched, CD30‐CAR‐transduced and expanded in vitro despite CD30 expression. Naïve T cells from healthy donors ( n = 7) were cultured with CD3/CD28 costimulation in the presence of IL‐7, IL‐15 and IL‐21 during 10 days. (a) Frequencies of CD4 + and CD8 + T‐cell subpopulations at the end of culture. T SCM‐like cells were the most prevalent T‐cell population (mean ± SD). (b) Representative plot of T cells transduced at day 2 with CD30‐CAR‐encoding lentivirus and CD30‐CAR expression and analysed by flow cytometry using an anti‐CD3 and anti‐EGFRt antibodies. (c) Expression of CD30 receptor on CD4 + and CD8 + T‐cell subpopulations in untransduced cells (white bars) and transduced cells (black bars) during the culture (days 4, 7 and 10; mean ± SD). (d) Percentage of T‐cell viability in untransduced cells (grey line) and transduced cells (black line) at days 4, 7 and 10 of culture (mean ± SD). (e) Fold expansion of CD4 + and CD8 + T cells during 10‐day culture in untransduced cells (grey line) and transduced cells (black line; mean ± SD). Data are representative of seven independently repeated experiments.

Article Snippet: Soluble CD30 protein was measured in supernatants of L540 and L428 cell cultures by enzyme‐linked immunoabsorbent assay (Human CD30 PicoKine ELISA Kit; Boster Biological Technology, Pleasanton, California, USA).

Techniques: In Vitro, Expressing, Cell Culture, Flow Cytometry

Journal: Clinical & Translational Immunology

Article Title: Memory stem T cells modified with a redesigned CD30‐chimeric antigen receptor show an enhanced antitumor effect in Hodgkin lymphoma

doi: 10.1002/cti2.1268

Figure Lengend Snippet: CD30‐CAR T SCM‐like enriched culture efficiently eradicates HL in vitro . (a) T SCM‐like enriched cells expressing CD30‐CAR ( n = 7) were exposed to CD30 + target cells (L540 or L428 tumor cells) and control CD30 − cell line (Raji) at different effector:target (E:T) ratios. Specific cytolytic activity was measured at 24 h by bioluminescence assay. Untransduced T SCM‐like cells were used as negative control (mean ± SD). (b) Cytolytic capacity of untransduced and CD30‐CAR T SCM‐like cells ( n = 3) was measured against CD30 + target cells (L540 and L428) at 24 h, in the presence or absence of saturated concentration of soluble CD30 protein (sCD30; 20 μg), at different E:T ratios. Raji cell line (CD30 − ) was used as negative target control (mean ± SD). (c) Cytokine secretion of CD30‐CAR T SCM‐like enriched culture ( n = 3) 24 h after L540, L428 and Raji co‐culture. Untransduced T SCM‐like cells were used as negative control. Cytokine levels of IL‐2, IL‐6, IL‐10, IFN‐γ and TNF‐α were measured by cytometry‐based multiplex analysis (mean ± SD). Data are representative of seven (a) or three (b, c) independently repeated experiments. For each donor, technical duplicate was used (c) .

Article Snippet: Soluble CD30 protein was measured in supernatants of L540 and L428 cell cultures by enzyme‐linked immunoabsorbent assay (Human CD30 PicoKine ELISA Kit; Boster Biological Technology, Pleasanton, California, USA).

Techniques: In Vitro, Expressing, Control, Activity Assay, ATP Bioluminescent Assay, Negative Control, Concentration Assay, Co-Culture Assay, Cytometry, Multiplex Assay

Journal: Clinical & Translational Immunology

Article Title: Memory stem T cells modified with a redesigned CD30‐chimeric antigen receptor show an enhanced antitumor effect in Hodgkin lymphoma

doi: 10.1002/cti2.1268

Figure Lengend Snippet: CD30‐CAR T SCM‐like function persists after CD30 + antigen re‐exposition. (a) Representative graphic of untransduced and CD30‐CAR T SCM‐like enriched cells exposed to CD30 + target cells (L540) at different effector:target (E:T) ratios every 72 h after each antigen re‐exposition (total stimulations: 3). Specific cytolytic activity was measured after 24 h of each antigen stimulation by bioluminescence. (b) Mean fluorescence intensity (MFI) of CD30‐CAR expression and (c) percentage of PD‐1‐TIM‐3 + T cells during antigen re‐exposures (mean ± SD). * P < 0.05; ** P < 0.01. Data are representative of three independently repeated experiments.

Article Snippet: Soluble CD30 protein was measured in supernatants of L540 and L428 cell cultures by enzyme‐linked immunoabsorbent assay (Human CD30 PicoKine ELISA Kit; Boster Biological Technology, Pleasanton, California, USA).

Techniques: Activity Assay, Fluorescence, Expressing

Journal: Clinical & Translational Immunology

Article Title: Memory stem T cells modified with a redesigned CD30‐chimeric antigen receptor show an enhanced antitumor effect in Hodgkin lymphoma

doi: 10.1002/cti2.1268

Figure Lengend Snippet: Therapeutic treatment with CD30‐CAR T SCM‐like enriched cell products induces a potent antitumor response against HL in vivo . NSG mice ( n = 4 for both experimental and control groups) were injected with (a) 2.5 × 10 6 L540 tumor cells (sc) on day 0 and treated 3 days later with 5 or 10 × 10 6 CD30‐CAR T SCM‐like enriched cells (iv), or (b) 2 × 10 6 L428 tumor cells (iv) on day 0 and treated with 5 or 10 × 10 6 CD30‐CAR T SCM‐like enriched cells (iv) 22 days after tumor challenge. In both models, control mice received 10 × 10 6 untransduced (UN) T SCM‐like enriched cells. Mice were monitored every other day for survival, and tumor growth was measured by in vivo bioluminescence. (c) Expression of exhaustion markers (PD1 and TIM3) in CD4 + and CD8 + tumor‐infiltrating T cells found in L540‐bearing mice treated with 5 × 10 6 CD30‐CAR T SCM‐like enriched cells, 48 days after tumor challenge, compared with CD4 + and CD8 + pre‐infused T cells (mean ± SD). * P < 0.05; ** P < 0.01. Data are representative of two independent experiments.

Article Snippet: Soluble CD30 protein was measured in supernatants of L540 and L428 cell cultures by enzyme‐linked immunoabsorbent assay (Human CD30 PicoKine ELISA Kit; Boster Biological Technology, Pleasanton, California, USA).

Techniques: In Vivo, Control, Injection, Expressing

Journal: Clinical & Translational Immunology

Article Title: Memory stem T cells modified with a redesigned CD30‐chimeric antigen receptor show an enhanced antitumor effect in Hodgkin lymphoma

doi: 10.1002/cti2.1268

Figure Lengend Snippet: T SCM‐like enriched cells expressing CD30‐CAR have high persistence in vivo and confer long‐lasting immunity against HL. (a) NSG mice surviving from L428 tumor administration ( n = 4) were rechallenged again with the same tumor dose (2 × 10 6 cells per mouse, iv), 79 days (arrow) after the first tumor challenge (grey discontinued line). An age‐matched mouse group ( n = 4) was injected with 2 × 10 6 L428 tumor cells (iv) as tumor control (pointed line). Black line represents control mice receiving 10 × 10 6 untransduced (UN) T SCM‐like enriched cells. Mice were followed every other day for survival, and tumor growth control was done by in vivo bioluminescence. (b) Bone marrow and lymph nodes from surviving CD30‐CAR‐treated mice were analysed for CAR T cell by flow cytometry (mean ± SD). (c) T‐cell subpopulations were analysed in bone marrow and lymph nodes by flow cytometry (mean ± SD). (d) Expression of exhaustion markers (PD1, TIM3 and LAG‐3) was analysed by flow cytometry in T cells found in lymph nodes and bone marrow (mean ± SD). Data are representative of two independent experiments.

Article Snippet: Soluble CD30 protein was measured in supernatants of L540 and L428 cell cultures by enzyme‐linked immunoabsorbent assay (Human CD30 PicoKine ELISA Kit; Boster Biological Technology, Pleasanton, California, USA).

Techniques: Expressing, In Vivo, Injection, Control, Flow Cytometry

Journal: Clinical & Translational Immunology

Article Title: Memory stem T cells modified with a redesigned CD30‐chimeric antigen receptor show an enhanced antitumor effect in Hodgkin lymphoma

doi: 10.1002/cti2.1268

Figure Lengend Snippet: CD30‐CAR T SCM‐like H products have higher antitumor efficacy and T‐cell persistence than CD30‐CAR T SCM‐like L products. (a) Composition of T SCM‐like H and T SCM‐like L cultures at the day of treatment (mean ± SEM). (b, c) NSG mice ( N = 4/group) were challenged with 2.5 × 10 6 L540 tumor cells (sc) on day 0 and treated when the tumor was well established (day 5) with 5 × 10 6 CD30‐CAR T SCM‐like H (pointed line and ▲) or T SCM‐like L (discontinued line and ■) cells (iv). A group of mice were injected with 2.5 × 10 6 L540 tumor cells (sc) as tumor control (black line and ●). Mice were monitored every other day for (B) tumor growth and (c) survival, measured by in vivo bioluminescence (mean ± SD). (d) Percentage of CD30‐CAR expression in tumor‐infiltrating T cells was analysed in T SCM‐like H and T SCM‐like L treated mice by flow cytometry (e) CD30‐CAR T‐cell detection in blood from mice treated with T SCM‐like H or T SCM‐like L cultures. (mean ± SD). * P < 0.05; ** P < 0.01; *** P < 0.001. Data are representative of two independent experiments.

Article Snippet: Soluble CD30 protein was measured in supernatants of L540 and L428 cell cultures by enzyme‐linked immunoabsorbent assay (Human CD30 PicoKine ELISA Kit; Boster Biological Technology, Pleasanton, California, USA).

Techniques: Injection, Control, In Vivo, Expressing, Flow Cytometry