Redirection of CD4+ and CD8+ T lymphocytes via an anti-CD3 × anti-CD19 bi-specific antibody combined with cytosine arabinoside and the efficient lysis of patient-derived B-ALL cells.

Abstract

BACKGROUND:

B-acute lymphoblastic leukemia (B-ALL) is derived from B cell progenitors. Recently, the development of appropriate combinations of chemotherapy and immunotherapy represents a promising approach for eliminating cancer. We previously constructed an anti-CD3 × anti-CD19 bi-specific antibody in a diabody configuration and its disulfide-stabilized format (ds-diabody). The combination of the diabody or ds-diabody and Ara-C was highly effective in enhancing the cytotoxicity of T cells against the CD19+ human leukemia cell-line, Nalm-6, both in vitro and in vivo. This study verified whether B-ALL patient-derived cells were sensitive to the diabody or ds-diabody and low-dosage Ara-C combination.

METHODS:

This study aimed to detect the B7 family members B7.1 (CD80) and B7.2 (CD86) that were expressed in B-ALL patient-derived cells pre-treated by Ara-C (0.25 μM) and to determine the targeted killing ability of T cell subtypes induced by the diabody or ds-diabody combination with Ara-C both in vitro and in vivo. We also determined the levels of the cytokines that were released by activated CD4+ or CD8+ T cells during therapy.

RESULT:

Low-dose Ara-C enhanced CD80 and CD86 expression in nearly 50 % of specimens of B-ALL patient-derived cells. A combination of diabody or ds-diabody and Ara-C enhanced T cell against B-ALL cells in vitro and in vivo. Both CD8+ and CD4+ T cells were potently activated. Expression of CD25 and CD69 was augmented equally by CD4+ or CD8+ T cells. However, CD8+ T cells made the major contribution by redirecting target cell lysis in a granzyme B and perforin-dependent mechanism. CD4+ T cells played an important immunomodulatory role by secreting IL2. Consequently, IL3, IL6, TNFα, and IFNγ were also released by CD4+ or CD8+ T cells following diabody-mediated T cell activation.

CONCLUSION:

T cell therapy induced by diabody or ds-diabody combined with low dose of Ara-C was effective against cancer cell-lines and in clinical trials. In vivo, the ds-diabody was more efficient than its parent diabody due to its enhanced stability.

• Authors: Dongmei Fan, Wei Li, Yuqi Yang, Xiaolong Zhang, Qing Zhang, Yan Yan, Ming Yang, Jianxiang Wang, Dongsheng Xiong
• Journal: Journal of hematology & oncology (J Hematol Oncol) Vol. 8 Pg. 108 (Oct 06 2015) ISSN: 1756-8722 [Electronic] England
• PMID: 26444983 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Bispecific
• Antigens, CD19
• CD3 Complex
• Cytokines
• Cytarabine
• Perforin
• Granzymes

Topics

• Animals
• Antibodies, Bispecific (administration & dosage, immunology, pharmacology)
• Antigens, CD19 (immunology)
• Antineoplastic Combined Chemotherapy Protocols (immunology, pharmacology)
• CD3 Complex (immunology)
• CD4-Positive T-Lymphocytes (immunology, metabolism)
• CD8-Positive T-Lymphocytes (immunology, metabolism)
• Cell Line, Tumor
• Cells, Cultured
• Cytarabine (administration & dosage, immunology, pharmacology)
• Cytokines (genetics, immunology, metabolism)
• Cytotoxicity, Immunologic (drug effects, immunology)
• Flow Cytometry
• Gene Expression Regulation, Leukemic (drug effects, immunology)
• Granzymes (immunology, metabolism)
• Humans
• Lymphocyte Activation (drug effects, genetics, immunology)
• Mice, Inbred NOD
• Mice, SCID
• Perforin (immunology, metabolism)
• Precursor B-Cell Lymphoblastic Leukemia-Lymphoma (drug therapy, genetics, immunology)
• Reverse Transcriptase Polymerase Chain Reaction
• Tumor Burden (drug effects, immunology)
• Tumor Cells, Cultured
• Xenograft Model Antitumor Assays