Journal: Scientific reports

Article Title: A nonadjuvanted HLA-restricted peptide vaccine induced both T and B cell immunity against SARS-CoV-2 spike protein.

doi: 10.1038/s41598-024-71663-1

Figure Lengend Snippet: Fig. 4. In vivo immune responses of injected UC peptides and complement activity of immunized serum. (a) BALB/c mice were intramuscularly administered 3 doses of 50 µg of UC-100 (n = 3) or 400 µg of UC-152 (n = 4) 14 days apart. (b–d) The serum of UC-100 mice was collected pre-(Day-1) and posttreatment (Day41) and tested in duplicate and represented as the mean for the levels of (b) total IgG; (c) specific IgG against UC-100 peptide; (d) specific IgG against SARS-CoV-2 spike trimer protein. (e–g) The serum of UC-152 mice was collected pre-(Day-1) and posttreatment (Day41) and tested in duplicate and represented as the mean for the levels of (e) total IgG; (f) specific IgG against UC-152 peptide; (g) specific IgG against SARS-CoV-2 spike trimer protein. (h,i) Antibody-mediated complement activity was tested in duplicate and is represented as the mean for pre- and postserum of mice vaccinated with UC-100 and UC-152 peptides, respectively. The complement activity was expressed as the IC50 of cytotoxicity of the (h) UC-100-vaccinated group and (i) UC-152-vaccinated group. Significant differences are indicated by asterisks (unpaired one-tailed Student’s t test; *p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: A final concentration of 10% young rabbit complement (CL3441-S50-R, Cedarlane, Ontario, Canada) was added to each well and incubated for an additional 30 min at 37 °C and 5% CO2.

Techniques: In Vivo, Injection, Activity Assay, One-tailed Test

Journal: Scientific reports

Article Title: A nonadjuvanted HLA-restricted peptide vaccine induced both T and B cell immunity against SARS-CoV-2 spike protein.

doi: 10.1038/s41598-024-71663-1

Figure Lengend Snippet: Fig. 5. In vivo immune responses of oral UC peptides and complement activity of immunized serum. (a) BALB/c mice (n = 5) were orally administered 3 doses of 200 µg UC-100 14 days apart. (b–d) The serum of UC-100 mice was collected pre-(Day-1) and posttreatment (Day 41) and tested in duplicate and represented as the mean for the levels of (b) total IgG; (c) specific IgG against UC-100 peptide; (d) specific IgG against SARS-CoV-2 spike trimer protein; (e) The antibody-mediated complement activity was tested in duplicate and represented as the mean for pre- and postserum of mice vaccinated with UC-100. The complement activity was expressed as the IC50 of cytotoxicity. (f,g) The serum of UC-100 mice was collected pre-(Day-1) and posttreatment (Day 41) and tested in duplicate and represented as the mean for the levels of (f) total IgA and (g) specific IgA against UC-100 peptide. Significant differences are indicated by asterisks (unpaired one-tailed Student’s t test; *p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: A final concentration of 10% young rabbit complement (CL3441-S50-R, Cedarlane, Ontario, Canada) was added to each well and incubated for an additional 30 min at 37 °C and 5% CO2.

Techniques: In Vivo, Activity Assay, One-tailed Test

Journal: Scientific reports

Article Title: A nonadjuvanted HLA-restricted peptide vaccine induced both T and B cell immunity against SARS-CoV-2 spike protein.

doi: 10.1038/s41598-024-71663-1

Figure Lengend Snippet: Fig. 6. Summary of the SARS-CoV-2 HLA-restricted peptide T-cell vaccine. The possible mechanistic immune response to HLA-restricted peptides has been elucidated. The 9-mer UC peptides can be primed onto both HLA class I and II molecules, and the immune response is triggered for both humoral and cellular immunity. Through HLA class I molecule binding, CTLs can be activated, and an increase in IFN-γ cytokines and specific lytic activity have been observed. Through HLA class II molecule binding, B-cells are activated, and specific antibodies are produced in accordance with complement cytotoxicity activity.

Article Snippet: A final concentration of 10% young rabbit complement (CL3441-S50-R, Cedarlane, Ontario, Canada) was added to each well and incubated for an additional 30 min at 37 °C and 5% CO2.

Techniques: Binding Assay, Activity Assay, Produced

Journal: Blood Cancer Journal

Article Title: A superantigen-based MHC class II-targeted cancer immunotherapy for the treatment of acute myeloid leukemia

doi: 10.1038/s41408-025-01391-w

Figure Lengend Snippet: A Total and anti-CD33-specific immunoglobin isotypes present in mouse plasma after treatment with M2T-CD33 ( N = 3). Error bars represent SEM. B Flow cytometry of C1498 parental and C1498-hCD33 cells incubated with Naïve, PBS treated, or M2T-CD33 treated mouse plasma. C Flow cytometry of MV4-11 cells incubated with Naïve, PBS treated, or M2T-CD33 treated mouse plasma. D Complement dependent cytotoxicity (CDC) assay with C1498-hCD33 cells incubated with baby rabbit complement and plasma from Naïve or M2T-CD33 treated mice (left). CDC assay with C1498 parental or C1498-hCD33 cells incubated with baby rabbit complement and plasma from M2T-CD33 treated mice (right).

Article Snippet: C1498-hCD33 and C1498 parental cells were incubated with varying dilutions of M2T-CD33 treated or Naïve C57BL/6 J plasma and baby rabbit complement (MP Biomedicals, Cat #642961).

Techniques: Clinical Proteomics, Flow Cytometry, Incubation, CDC Assay

Journal: Scientific Reports

Article Title: A Versatile Safeguard for Chimeric Antigen Receptor T-Cell Immunotherapies

doi: 10.1038/s41598-018-27264-w

Figure Lengend Snippet: In vitro screening, identification and characterization of the CubiCAR architecture. ( a , left panel) Scheme of the 2 nd generation CAR construct used in this study. This construct includes an anti-BCMA ScFV, a CD8 hinge and transmembrane domain, a 4-1BB costimulatory domain and a CD3 activation domain ( a , right panel) Scheme and names of the different engineered extracellular constructs tested. The location of CD20 mimotopes and CD34 epitope are indicated. ( b ) Flow cytometric detection of CAR constructs transiently expressed at the surface of primary T-cells using either the soluble BCMA protein, RTX or QBEND10 as surface markers. The error bars in represent the standard deviation on experimental values computed out of ≥2 biological replicates performed with ≥2 different donors ( c ) Box plot illustrating the median of efficiency of RTX-dependent depletion of primary T-cells transiently expressing CAR constructs. Viability of primary T-cells incubated for 150 min in the presence of 100 µg/mL RTX and complement was determined by flow cytometry and normalized to untreated control (relative viability, see Methods). Relative viability is indicated for each constructs (left panel) or for construct subgroups including those containing 2 consecutive CD20 mimotopes (2 cm) and 2 to 3 separated CD20 mimotopes (2 sm, 3 sm respectively, right panel). The number of independent biological replicates performed is indicated at the top of each box plot. The significance of the differences between subgroups was assessed using a non parametric Mann-Whitney U test (ns, non significant, *p < 0.05, **p < 0.01, ***p < 0.001). ( d ) Schema of the workflow used to characterize primary T-cells steadily expressing the CubiCAR (C14) construct. ( e ) Flow cytometry analysis of CubiCAR T-cells before and after QBEND10 coated beads purification using BCMA soluble protein as surface marker. ( f ) Specific cell lysis activity of unpurified and purified CubiCAR T-cells toward BCMA + and BCMA- tumor cell lines determined at different E/T ratio. ( g ) Kinetic of CubiCAR T-cells depletion by complement and increasing amounts of RTX (10–100 µg/mL). ( h ) Effect of RTX on the specific cell lysis activities of CAR or purified CubiCAR T-cells. Activities were determined after a 30 min long incubation of cells with complement and increasing amounts of RTX. The Error bars in ( f ), ( g ) and ( h ) represent the standard deviation on experimental values (technical triplicate) computed out of 2 biological replicates performed with 2 different donors. The significance of the differences between subgroups in ( f ) and ( h ) was assessed using a one-way ANOVA test (ns, non significant, *p < 0.05, **p < 0.01, ***p < 0.001). Reproduced with permission from Cellectis group.

Article Snippet: To assess the effect of RTX and complement on CAR T-cells viability, 2 × 10 5 CAR T-cells were incubated alone or in the presence of 10–100 μg/mL RTX and complement (diluted 4 times from a 2 ml stock solution of baby rabbit complement, AbD serotec BIO-RAD, cat#C12CA) in a final volume of 400 μL.

Techniques: In Vitro, Construct, Activation Assay, Standard Deviation, Expressing, Incubation, Flow Cytometry, Control, MANN-WHITNEY, Purification, Marker, Lysis, Activity Assay