mhc ii binding peptides  (Sartorius AG)

Journal: Cancer Immunology Research

Article Title: Antitumor CD4 + T Helper 1 Cells Target and Control the Outgrowth of Disseminated Cancer Cells

doi: 10.1158/2326-6066.CIR-24-0630

Figure Lengend Snippet: Intratumoral activation of antitumor CD4Th1 cells inhibits spontaneous metastasis. A, MRI images of BALB-neuT mice treated with or without intramammary gland HER2-DC1 ( n = 6/group). B, Total tumor burden was recorded by MRI for experimental BALB-neuT mice ( n = 6/group). C–E, IHC HER2 and CYT 8/18 double staining for the detection of micrometastases in lung ( C ), liver ( D ), and brain ( E ) sections from BALB-neuT mice treated with or without HER2-DC1 ( n = 3/group). F, Quantification of spontaneous micrometastases in the lung and liver of experimental BALB-neuT mice ( n = 3/group). G, Quantification of metastatic cancer cells seeding in brain of experimental BALB-neuT mice ( n = 3/group). H, Infiltration of CD4T and CD8T cells in mammary gland of HER2-DC1–treated BALB-neuT mice and control analyzed by flow cytometry ( n = 3–4/group). I, Frequency of CD4 T cells infiltration in the mammary gland of HER2-DC1–treated BALB-neuT mice and control determined by IHC ( n = 3/group). J, IFN-γ secretion after coculturing TDLNs (from experimental BALB-neuT mice) with or without HER2-DC1 individually pulsed with p5, p435, and p1209 for 72 hours by automated ELISA ( n = 2/group). K, IFN-γ secretion after restimulating splenocytes of experimental mice with rat HER2/neu peptides p5, p435, and p1209 individually and analyzed by automated ELISA ( n = 4–6/group). L, Tumor growth curves of control and intratumoral HER3-DC1–treated BALB/c mice bearing 4T1 TNBC tumors ( n = 9–10/group). M and N, IHC HER3 and Pan-cytokeratin (Pan-CYT) double staining for the detection and quantification of metastases in lung ( M ) and liver ( N ) sections from 4T1 TNBC spontaneous metastasis model ( n = 3/group). O, Tumor growth curves of control and intratumoral HER3-DC1–treated C57BL/6 mice bearing B16F10 melanoma with spontaneous metastasis ( n = 10/group). P, Tumor growth curves of B16-F10 high tumor burden spontaneous metastasis–bearing mice treated with or without intratumoral HER3-DC1 ( n = 7–10/group). Q, Hematoxylin and eosin staining for the detection and quantification of spontaneous metastatic nodules in lungs from experimental B16F10 melanoma spontaneous metastasis model ( O and P ; n = 5–8/group). All data are presented as mean ± SEM. P values were calculated with one- or two-tailed Student t test. i.t., intratumoral delivery; ROI, region of interest.

Article Snippet: All these MHC class II–binding HER3 multiepitope peptides were purchased from GenScript.

Techniques: Activation Assay, Double Staining, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Two Tailed Test

Journal: Cancer Immunology Research

Article Title: Antitumor CD4 + T Helper 1 Cells Target and Control the Outgrowth of Disseminated Cancer Cells

doi: 10.1158/2326-6066.CIR-24-0630

Figure Lengend Snippet: Antitumor CD4 Th1 cells are primarily responsible for metastatic prevention. A, Total tumor burden was calculated for control and HER2-DC–treated BALB-neuT mice with or without CD4 or CD8 T-cell depletion by MRI ( n = 4–6/group). B, IHC HER2 and cytokeratin 8/18 double staining for micrometastasis detection and quantification in lung and liver sections for experimental BALB-neuT mice ( n = 3/group). C, Quantification of metastatic cancer cells seeding in brain of experimental BALB-neuT mice by IHC HER2 and cytokeratin 8/18 double staining ( n = 3/group). D, Tumor growth curves of control and intratumoral HER3-DC1–treated mice bearing 4T1 tumors depleted with or without CD4 or CD8 T cells ( n = 6–10/group). E and F, Hematoxylin and eosin staining for the detection and quantification of metastatic nodules in lung ( E ) and liver ( F ) from experimental 4T1 TNBC mice model ( n = 3–8/group). G, Tumor growth curves of CD4 knockout mice bearing B16-F10 spontaneous metastasis treated with or without intratumoral HER3-DC1 ( n = 6/group). H, Quantification of spontaneous lung metastatic nodules from CD4 knockout mice model ( G ; n = 5–6/group). I, 4T1 TNBC tumor–bearing mice treated with HER3-DC1 s.c. and intratumoral HER3-DC1, immature HER3-iDC, unpulsed DC1, or HER2-DC1 ( n = 10/group). J, Quantification of spontaneous metastatic nodules in lungs from 4T1 TNBC mice model by hematoxylin and eosin staining ( n = 5–10/group). K and L, Detection ( K ) and frequency ( L ) of CD4 + IFN-γ + T cells in control and intratumoral HER3-DC1–treated 4T1 TNBC tumors analyzed by flow cytometry ( n = 2–3/group). M, Tumor growth curves of 4T1 TNBC tumor–bearing mice treated with intratumoral HER3-DC1 with or without IFN-γ neutralizing antibody ( n = 9–10/group). N, Quantification of metastatic nodules in lungs from 4T1 TNBC spontaneous metastasis mice model determined by hematoxylin and eosin staining ( n = 7–10/group). O, Tumor growth curves of IFN-γ knockout mice bearing B16F10 melanoma spontaneous metastasis treated with or without intratumoral HER3-DC1 ( n = 10/group). P, Frequency of spontaneous metastatic nodules in lungs from IFN-γ knockout mice bearing B16F10 melanoma spontaneous metastasis treated with or without intratumoral HER3-DC1 ( n = 4–5/group). Mean ± SEM represented. P values were determined by one- or two-tailed Student t test. ns, not significant; ROI, region of interest; SSC-A, side scatter area.

Article Snippet: All these MHC class II–binding HER3 multiepitope peptides were purchased from GenScript.

Techniques: Control, Double Staining, Staining, Knock-Out, Flow Cytometry, Two Tailed Test

Journal: Cancer Immunology Research

Article Title: Antitumor CD4 + T Helper 1 Cells Target and Control the Outgrowth of Disseminated Cancer Cells

doi: 10.1158/2326-6066.CIR-24-0630

Figure Lengend Snippet: Antitumor CD4 Th1 cells target DCCs. A, Infiltration of CD4 T cells in the BM of experimental BALB-neuT mice analyzed by flow cytometry ( n = 3–5/group). B and C, HER2 + CYT8/18 + Ki-67 + DCCs in BM ( B ) and lung ( C ) of experimental BALB-neuT mice were determined by flow cytometry ( n = 2–6/group). D and E, SA-β-gal staining for the detection ( D ) and percentage of senescent DCCs ( E ) in the BM of experimental BALB-neuT mice ( n = 5–8). Scale bars, 1 mm. F, Immunoblots for HER2, NR2F1, Wnt4, and Twist proteins in BM DCCs of control and HER2-DC1–treated BALB-neuT mice. G, Immunofluorescence staining of HER2 + CYT8/18 + Ki-67 + DCCs in BM of experimental BALB-neuT mice. Scale bars, 100 μm. H, Percentage of senescent DCCs induced by tumor antigen–specific CD4 Th1 cells secreting IFN-γ on DCCs from the BM of control BALB-neuT mice ( n = 5–10). I, Frequency of CD45 − EpCAM + DCCs (among the 100% of CD45-negative cell population) in the BM of mice bearing 4T1 TNBC spontaneous metastasis receiving various treatment analyzed by flow cytometry ( n = 3/group). J and K, Detection ( J ) and frequency ( K ) of CD4 + IFN-γ + T cells in the BM of control and intratumoral HER3-DC1–treated mice bearing 4T1 TNBC spontaneous metastasis determined by flow cytometry ( n = 2–3/group). L, Tumor growth curves of NSG mice ( n = 4/group). M and N, IHC HER2 and CYT8/18 double staining for the detection of micrometastasis in lung ( M ) and liver ( N ) sections from L ( n = 4). All data are presented as mean ± SEM represented. P values were calculated by one- or two-tailed Student t test and one-way ANOVA with Tukey multiple comparisons test.

Article Snippet: All these MHC class II–binding HER3 multiepitope peptides were purchased from GenScript.

Techniques: Flow Cytometry, Staining, Western Blot, Control, Immunofluorescence, Double Staining, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Amino acid polymorphisms in human histocompatibility leukocyte antigen class II and proinsulin epitope have impacts on type 1 diabetes mellitus induced by immune-checkpoint inhibitors

doi: 10.3389/fimmu.2023.1165004

Figure Lengend Snippet: Binding affinities of the peptides derived from human proinsulin to HLA-DP5 (HLA- DPA1*02:02 , HLA- DPB1*05:01 ) or HLA-DP15 (HLA -DPA1*02:02 , HLA- DPB1*15:01 ) molecules are shown as %RANK in silico (A) , and shown as REVEAL score in vitro ( B ) . Horizontal numbers indicate peptide numbers used in the study (shown in Table 4 ). Note that lower %RANK means predicted strong binder in silico , and higher REVEAL score indicates peptide with high affinity in vitro . Cluster 1 (peptides 6-10) and Cluster 2 (peptide 31-34) were shown with blue and red line, respectively.

Article Snippet: The synthesized peptides were subjected to an in vitro peptide binding assay (ProImmune REVEAL ® MHC class II-peptide binding assay: (ProImmune, Oxford, UK) using recombinant HLA-DP5 and HLA-DP15 proteins.Detection of binding peptides is based on the presence or absence of the native conformation of the MHC-peptide complex in an immunoassay.

Techniques: Binding Assay, Derivative Assay, In Silico, In Vitro