Branched multipeptide immunotherapy for glioblastoma using human leukocyte antigen-A*0201-restricted cytotoxic T-lymphocyte epitopes from ERBB2, BIRC5 and CD99

Abstract

We investigated the use of cytotoxic T-lymphocyte (CTL) epitopes in peptide immunotherapy for glioblastoma. Three peptides (ERBB2, BIRC5 and CD99) were selected based on their peptide-T2 cell binding affinities and combined in a multipeptide cocktail or a branched multipeptide synthesized with mini-polyethylene glycol spacers. Dendritic cells (DCs) pulsed with the multipeptide cocktail or branched multipeptide were compared based on their immunophenotype and cytokine secretion. FACS analysis of alpha-type 1 polarized dendritic cells (αDC1s) revealed that both groups highly expressed CD80, CD83 and CD86, indicating that both treatments efficiently generated mature αDC1s with the expected phenotype. Production of IL-12p70, IL-12p40 and IL-10 also increased upon αDC1 maturation in both groups. CTLs stimulated by either αDC1 group (“DC-CTLs”) included numerous IFN-γ-secreting cells against T2 cells loaded with the corresponding multipeptides. Large numbers of IFN-γ-secreting cells were observed when human glioblastoma cell lines and primary cells were treated with multipeptide-pulsed DC-CTLs. Both multipeptide-pulsed DC-CTL groups exhibited cytotoxic activity of 40-60% against the U251 cell line and 60-80% against primary cells. Branched multipeptide from ERBB2, BIRC5 and CD99 stably bound with T2 cells, and its cytotoxicity toward target cells was similar to that of the multipeptide cocktail. Thus, branched multipeptides could be promising candidates for immunotherapeutic glioblastoma treatment.

CD99: A Cell Surface Protein with an Oncojanus Role in Tumors

“…CD99 ligation by antibodies can increase natural killer cell-mediated tumor lysis by inducing HSP70 expression [ 93 ]. In addition, peptides from CD99 have been described as promising candidates for immunotherapeutic glioblastoma treatment [ 101 ]. Branched multipeptides from ERBB2, BIRC5, and CD99 stably bound with T2 cells, and multipeptide-pulsed denditric cells–cytotoxic T lymphocytes exhibited remarkable cytotoxic activity against primary glioblastoma cells.…”

Identification of Key Genes and Signaling Pathways Associated with the Progression of Glioblastoma multiform

“…In pathway enrichment analysis for up regulated genes was performed. Enriched genes such as SOD2 [68], RPS11 [69], RPL9 [70], MYC (MYC proto-oncogene, bHLH transcription factor) [71], SEC61G [72], BIRC5 [73], NEK2 [74], CDK2 [75], AURKB (aurora kinase B) [76], RPS3 [77], MGP (matrix Gla protein) [78], AEBP1 [79], CTHRC1 [80], COL1A1 [81], COL3A1 [82], TNC (tenascin C) [83], POSTN (periostin) [84], IGFBP2 [85], IGFBP3 [86], IGFBP4 [87], SRPX2 [88], LAMB1 [89], ESM1 [90], TGFBI (transforming growth factor beta induced) [91], ITGA5 [92], RAP1B [93], CAV1 [94], HMOX1 [95] and LOX (lysyl oxidase) [96] were linked with progression of GBM. GPX7 was important for advancement of gastric cancer [97], but this gene was identified first time in GBM and may be liable for progression of GBM.…”

Identification of core differentially methylated genes in glioma

“…The ERBB2 proto-oncogene (also known as HER2 ) is one of the most useful molecules for classification and prognosis in cancer, and its activation and downstream signals are highly dependent on dimerization with EGFR (58). ERBB2 , in combination with survivin and CD99 molecule, has the ability to form branched multipeptides, which stably bind with T2 cells to exhibit 40–60 and 60–80% cytotoxic activity against the U251 GBM and primary cells, respectively; therefore, they are promising candidates for immunotherapeutic GBM treatment (59). Caspases are well-renowned proteases that play a central role in initiating and executing cell apoptosis (60).…”