Transient blocking of NK cell function with small molecule inhibitors for helper dependant adenoviral vector-mediated gene delivery

Abstract

One major challenge in gene therapy is the host immune responses against viral vectors. Previous studies indicate the involvement of NK cells in stunted gene expression in viral vector mediated gene therapy. To understand the problem of the immune responses, we have developed an in-vitro co-culture system with human NK cell line, macrophages and airway epithelial cells. We showed that small molecule blockers, CAPE and ruxolitinib, for NF-κB and JAK-STAT pathways, respectively, significantly inhibited cytokine secretion by macrophages. When NK cells are co-cultured with helper-dependent adenoviral (HD-Ad) vector activated macrophages, IFN-γ cytokine expression by NK cells increased significantly, which was inhibited effectively by ruxolitinib and CAPE, and there was an additive effect when both inhibitors were used. We demonstrated that NK cells activated by cytokines produced by HD-Ad-activated macrophages kill HD-Ad vector transduced bronchial epithelial cells. This cell killing activity was significantly reduced by CAPE and ruxolitinib. Combination of these two inhibitors had an additive effect on inhibiting NK cell mediate killing of gene transduced cells. Transient inhibition of NK cell response at its peak may enhance sustained gene expression. Our data suggest that combination of CAPE and ruxolitinib may help in protecting gene transduced airway epithelial cells to prolong transgene expression.Electronic supplementary materialThe online version of this article (doi:10.1186/s13578-015-0023-0) contains supplementary material, which is available to authorized users.

Mechanisms Underlying the Anti-inflammatory and Immunosuppressive Activity of Ruxolitinib

“…Lastly, Ankathatti and Hu (58) showed, in a study with human cell lines, that small molecule blockers, including ruxolitinib, for JAK-STAT pathway, significantly inhibited cytokine secretion by macrophages activated by viral infectious trigger. Usually, macrophages respond to viral infection by releasing soluble mediators, helping in the recruitment of innate, and adaptative effector cells.…”

NK Cells in Myeloproliferative Neoplasms (MPN)

“…Later studies confirmed that IL-12 was completely blocked by ruxolitinib, as well as IL-15 and the phosphorylation of STAT5, resulting in a functional impairment of IFN-γ production by NK cells (Figure 3, ruxolitinib) [131]. An inhibition of cytokine secretion by macrophages was also reported, possibly effecting NK cell recruitment, maturation, and killing activity [132]. The reported impairment of DC function also resulted in the impaired induction of antigen specific T cell responses [130].…”

COVID-19: High-JAKing of the Inflammatory “Flight” by Ruxolitinib to Avoid the Cytokine Storm

“…However, some specific concerns should be taken into account: as other JAK inhibitors, ruxolitinib impairs the capability of antigen presenting cells (such as macrophages and dendritic cells) to produce, among others, type I interferons, Il-12, IL-15, and IL-23. This event negatively influences NK activation, antigen presentation and Th1/Th17 polarization, with consequent alterations in antigen-specific T cells response, including viral clearance, providing a possible mechanistic explanation for the increase of infection rates in patients with MPN undergoing longterm ruxolitinib treatment (36,(79)(80)(81)(82)(83)(84)(85)(86)(87).…”