Chimeric form of tumor necrosis factor-α has enhanced surface expression and antitumor activity

Abstract

Tumor necrosis factor (TNF)-a is a type-II transmembrane protein that is cleaved by TNF-a-converting enzyme (TACE/ADAM-17) to release soluble TNF, a cytokine with potent antitumor properties whose use in clinical applications is limited by its severe systemic toxicity. We found that human cells transfected with vectors encoding TNF without the TACE cleavage site (DTACE-TNF) still released functional cytokine at substantial levels that varied between transfected cell lines of different tissue types. Vectors encoding membrane-associated domains of CD154, another TNF-family protein, conjoined with the carboxyl-terminal domain of TNF, directed higher-level surface expression of a functional TNF that, in contrast to DTACE-TNF, was resistant to cleavage in all cell types. Furthermore, adenovirus vectors encoding CD154-TNF had significantly greater in vivo biological activity in inducing regression of established, syngeneic tumors in mice than adenovirus vectors encoding TNF, and lacked toxicity associated with soluble TNF. As such, CD154-TNF is a novel TNF that appears superior for treatment of tumors in which high-level local expression of TNF is desired.

Tumor Therapy Applying Membrane-bound Form of Cytokines

“…Recently, it was found that the TNFα mutation on ADAM-17 cleavage site was not enough to prevent cleavage and secretion of TNFα. To avoid the effect of secretory TNFα effect completely, Kipps and colleagues expressed a chimeric protein with extracellular domain of TNFα and transmembrane stalk portion of CD154 using adenovirus vector system and tested its antitumor activity ( 27 ). The systemic toxicity was not seen with the adenovirus vector expressing TNFα-CD154 chimera.…”

Membrane-bound form of monocyte chemoattractant protein-1 enhances antitumor effects of suicide gene therapy in a model of hepatocellular carcinoma

“…On the basis of these assumptions, this type of modified cytokine gene therapy has been reported on interleukin-2, 13,19,20 interleukin-12, 21 fractalkine (CX3CL1) 22 and TNF. 23 Indeed, accumulating evidence revealed that the membrane-bound form of cytokine genes can exhibit more antitumor effects than the corresponding soluble ones. Likewise, the current study confirms that the membrane-bound form of MCP-1 can attract more immune cells, including monocytes/macrophages and T lymphocytes, to the tumor sites and can induce the expression of TNF.…”

Alkaline phosphatase-fused repebody as a new format of immuno-reagent for an immunoassay

“…To check this possibility, we chose Mab1 as a capture antibody. Mab1 is a monoclonal antibody against hTNF-a and is widely used as a capture antibody in a sandwich immunoassay together with biotinylated Mab11 as a detection antibody [33]. We optimized the assay conditions with respect to the concentrations of the capture antibody and rB3-mAP fusion protein, and observed that 2 mg mL À1 of Mab1 and 2 mg mL À1 of rB3-mAP fusion protein led to the best assay performance.…”