Journal: Frontiers in Immunology

Article Title: PD-1 blockade does not enhance alloimmunization after allogeneic dendritic cell vaccination in cancer patients

doi: 10.3389/fimmu.2026.1763434

Figure Lengend Snippet: Expression of HLA Class I and Class II molecules by PDC*line cells. HLA Class I (HLA-ABC, HLA-A2, HLA-B7) and Class II molecules (HLA-DR/DP/DQ, HLA-DRB1, HLA-DP, HLA-DQ) were quantified on PDC*line cells. (A, B) Representative histograms showing PDC*line cell labelling with FITC-conjugated anti-HLA Class I (A) and -HLA Class II (B) antibodies. Antibodies specific to cognate isotypes were used as control. (C) The means +/- SD of 3 to 5 experiments measuring the HLA antigen density at the PDC*line cell surface are shown.

Article Snippet: To assess Complement-Dependent Cytotoxicity (CDC), 25,000 to 100,000 PDC*line or control B cells were incubated with patient serum or control anti-HLA antibodies (anti-pan Class I, ref 824101 from Bio-Rad; anti-pan Class II, ref hla-c2 from Invivogen) in the presence of functional complement (from healthy donors serum pool or patient serum) for 1 h at 37 °C.

Techniques: Expressing, Control

Journal: Frontiers in Immunology

Article Title: PD-1 blockade does not enhance alloimmunization after allogeneic dendritic cell vaccination in cancer patients

doi: 10.3389/fimmu.2026.1763434

Figure Lengend Snippet: Kinetics of anti-HLA humoral response in patients from cohorts A2 and B2. The detections of anti-HLA Class I (A, B) and -HLA Class II (C, D) antibodies were performed over time in the sera of patients of cohort A2 (n=12; red) and B2 (n=48; blue). The most represented HLA molecules are shown HLA-B*07:02 (A) , HLA-B*44:02 (B) , HLA-DRB1*01:03 (C) , HLA-DRB1*08:01 (D) . V, visit; FU, Follow-Up.

Article Snippet: To assess Complement-Dependent Cytotoxicity (CDC), 25,000 to 100,000 PDC*line or control B cells were incubated with patient serum or control anti-HLA antibodies (anti-pan Class I, ref 824101 from Bio-Rad; anti-pan Class II, ref hla-c2 from Invivogen) in the presence of functional complement (from healthy donors serum pool or patient serum) for 1 h at 37 °C.

Techniques:

Journal: Frontiers in Immunology

Article Title: PD-1 blockade does not enhance alloimmunization after allogeneic dendritic cell vaccination in cancer patients

doi: 10.3389/fimmu.2026.1763434

Figure Lengend Snippet: Complement-dependent cytotoxicity (CDC) mediated by control anti-HLA antibodies against control B cells and PDC*line cells. (A, B) . Binding of anti-HLA class I and class II control antibodies (red) on primary B cells (A) and PDC*line cells (B) . Secondary antibody alone (blue) and unstained cells (grey) were used as negative control. (C, D) Percentage of B cell and PDC*line cell death after CDC experiments with anti-HLA class I or II control antibodies in the presence of human serum pool as source of functional complement. Human serum pools without anti-HLA control antibodies were used as negative controls (Control sera). In C, the results gathered experiments done with three healthy donors (HD#01-03) of B cells. Decomplemented human sera were used as negative control in each experiment (not shown). The horizontal dotted line indicates the 20% positivity threshold. (E) Fold change of the cell death percentage in complete versus decomplemented human donor serum conditions. The horizontal dotted line indicates the twofold positivity threshold. In (C) (n=20), (D) (n=25), and (E) , the bars show the median values. One-way ANOVA with multiple comparisons was used to statistically compare the conditions (control sera, versus +anti-HLA I and +anti-HLA II); ****(p<0.0001), ns (non-significant).

Article Snippet: To assess Complement-Dependent Cytotoxicity (CDC), 25,000 to 100,000 PDC*line or control B cells were incubated with patient serum or control anti-HLA antibodies (anti-pan Class I, ref 824101 from Bio-Rad; anti-pan Class II, ref hla-c2 from Invivogen) in the presence of functional complement (from healthy donors serum pool or patient serum) for 1 h at 37 °C.

Techniques: Control, Binding Assay, Negative Control, Functional Assay

Journal: Frontiers in Immunology

Article Title: PD-1 blockade does not enhance alloimmunization after allogeneic dendritic cell vaccination in cancer patients

doi: 10.3389/fimmu.2026.1763434

Figure Lengend Snippet: Kinetics of complement-dependent cytotoxicity (CDC) activity according to anti-HLA levels. The sera of two patients (P) from Cohort A2 (A) and B2 (B) were collected at different timepoints during the treatments (see Figure 1B ). The line graphs present the mean fluorescence intensity (MFI) of anti-HLA antibodies. The levels of anti-HLA Class I (B7 and B44) and Class II (DR103, DR8, DQB04, DQB05, and DPB02) matching HLA typing of the indicated cell type, are shown. CDC activity was evaluated against primary B cells from two allogeneic healthy donors (HD#01 and HD#02) and PDC*line cells. The bar plots show the cell death percentage in untreated (S) and decomplemented serum (dS) conditions (means +SD; n=1-3). The horizontal dotted line indicates the 20% positivity threshold. The “xn” sign above the bar plots indicates the fold change in dead cells between the untreated serum and decomplemented serum conditions for the corresponding patient.

Article Snippet: To assess Complement-Dependent Cytotoxicity (CDC), 25,000 to 100,000 PDC*line or control B cells were incubated with patient serum or control anti-HLA antibodies (anti-pan Class I, ref 824101 from Bio-Rad; anti-pan Class II, ref hla-c2 from Invivogen) in the presence of functional complement (from healthy donors serum pool or patient serum) for 1 h at 37 °C.

Techniques: Activity Assay, Fluorescence, Immunopeptidomics