Development of an artificial antibody specific for HLA/peptide complex derived from cancer stem-like cell/cancer-initiating cell antigen DNAJB8.

Abstract

BACKGROUND:

Peptide-vaccination therapy targeting tumour-associated antigens can elicit immune responses, but cannot be used to eliminate large tumour burden. In this study, we developed a therapeutic single-chain variable-fragment (scFv) antibody that recognises the cancer stem-like cell/cancer-initiating cell (CSC/CIC) antigen, DNAJB8.

METHODS:

We screened scFv clones reacting with HLA-A24:20/DNAJB8-derived peptide (DNAJB8_143) complex using naive scFv phage-display libraries. Reactivity and affinity of scFv clones against the cognate antigen were quantified using FACS and surface plasmon resonance. Candidate scFv clones were engineered to human IgG1 (hIgG1) and T-cell-engaging bispecific antibody (CD3xJB8). Complement-dependent cytotoxicity (CDC) and bispecific antibody-dependent cellular cytotoxicity (BADCC) were assessed.

RESULTS:

scFv clones A10 and B10 were isolated after bio-panning. Both A10-hIgG1 and B10-hIgG1 reacted with DNAJB8-143 peptide-pulsed antigen-presenting cells and HLA-A24(+)/DNAJB8(+) renal cell carcinoma and osteosarcoma cell lines. A10-hIgG1 and B10-hIgG1 showed strong affinity with the cognate HLA/peptide complex (KD = 2.96 × 10-9 M and 5.04 × 10-9 M, respectively). A10-hIgG1 and B10-hIgG1 showed CDC against HLA-A24(+)/DNAJB8(+) cell lines. B10-(CD3xJB8) showed superior BADCC to A10-(CD3xJB8).

CONCLUSION:

We isolated artificial scFv antibodies reactive to CSC/CIC antigen DNAJB8-derived peptide naturally present on renal cell carcinoma and sarcoma. Immunotherapy using these engineered antibodies could be promising.

• Authors: Hiroki Tadano, Tomohide Tsukahara, Emi Mizushima, Asuka Akamatsu, Kazue Watanabe, Iyori Nojima, Terufumi Kubo, Takayuki Kanaseki, Yoshihiko Hirohashi, Noriyuki Sato, Toshihiko Torigoe
• Journal: British journal of cancer (Br J Cancer) Vol. 123 Issue 9 Pg. 1387-1394 (10 2020) ISSN: 1532-1827 [Electronic] England
• PMID: 32753678 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Neoplasm
• Cancer Vaccines
• DNAJB8 protein, human
• HLA-A24 Antigen
• HSP40 Heat-Shock Proteins
• Molecular Chaperones
• Nerve Tissue Proteins
• Peptide Fragments
• Peptide Library
• Single-Chain Antibodies

Topics

• Antibody Specificity
• Antibody-Dependent Cell Cytotoxicity
• Antigens, Neoplasm (genetics, metabolism)
• Bone Neoplasms (immunology, pathology, therapy)
• Cancer Vaccines (biosynthesis, therapeutic use)
• Carcinoma, Renal Cell (immunology, pathology, therapy)
• Cell Line, Tumor
• Cell Transformation, Neoplastic (genetics, metabolism)
• HEK293 Cells
• HLA-A24 Antigen (genetics, immunology, metabolism)
• HSP40 Heat-Shock Proteins (genetics, immunology, metabolism)
• HT29 Cells
• Humans
• Immunotherapy (methods)
• Kidney Neoplasms (immunology, pathology, therapy)
• Molecular Chaperones (genetics, immunology, metabolism)
• Neoplastic Stem Cells (immunology, metabolism, pathology)
• Nerve Tissue Proteins (genetics, immunology, metabolism)
• Osteosarcoma (immunology, pathology, therapy)
• Peptide Fragments (immunology, metabolism)
• Peptide Library
• Protein Engineering (methods)
• Single-Chain Antibodies (biosynthesis, therapeutic use)