IL-7 During Antigenic Stimulation Using Allogeneic Dendritic Cells Promotes Expansion of CD45RA-CD62L+CD4+ Invariant NKT Cells With Th-2 Biased Cytokine Production Profile

Abstract

Invariant natural killer T (iNKT) cells are innate-like T lymphocytes cells that recognize glycolipid antigens associated with CD1d, non-classical antigen presenting proteins. They can drive either pro-inflammatory (Th-1) or anti-inflammatory (Th-2) immune microenvironment through the production of both Th-1 and Th-2 type cytokines upon activation, thus play a vital role in cancer, infection, and autoimmune diseases. Adoptive cell therapy using ex vivo expanded iNKT cells is a promising approach to enhance anti-tumor immunity or immunosuppression. However, overcoming phenotypic and functional heterogeneity and promoting in vivo persistency of iNKT cells remains to be a challenge. Here, we compared various methods for ex vivo expansion of human iNKT cells and assessed the quality of expansion, phenotype, and cytokine production profile of expanded iNKT cells. While a direct stimulation of iNKT cells in peripheral blood mononuclear cells with agonist glycolipid led to the expansion of iNKT cells in varying degrees, stimulation of enriched iNKT cells by irradiated autologous peripheral blood mononuclear cells or allogeneic dendritic cells resulted in consistent expansion of highly pure iNKT cells. Interestingly, the mode of antigenic stimulation influenced the dominant subtype of expanded iNKT cells. Further, we evaluated whether additional IL-7 or IL-15 during antigenic stimulation with allogeneic dendritic cells can improve the phenotypic heterogeneity and modify cytokine production profile of iNKT cells expanded from 18 consecutive donors. The presence of IL-7 or IL-15 during antigenic stimulation did not affect the fold of expansion or purity of expanded iNKT cells. However, IL-7, but not IL-15, led to a better expansion of CD4+ iNKT cells, enhanced Th-2 type cytokine production of CD4+ iNKT cells, and maintained the expansion of central memory (CD45RA-CD62L+) CD4+ iNKT cells. Our results suggest the addition of IL-7 during antigenic stimulation with allogeneic dendritic cells can promote the expansion of CD62L+Th-2+CD4+ human iNKT cells that can be used as novel immunotherapeutic to control excessive inflammation to treat various autoimmune diseases.

Methylation changes induced by a glycodendropeptide immunotherapy and associated to tolerance in mice

“…Comparing the Desensitized and Anaphylactic groups, 5,596 unique DMPRs were found, from which several upmethylated DMPRs could influence genes linked to DC activation. Among them, we found toll-like receptors ( Tlr4 , Tlr3 , Tlr8 , Tlr5 ), and other genes linked to its function including Cd180 and Tirap ( 54 ); genes involved in IL-2 production from DC like Cd247 , Irf4 , and Pde4b ( 55 ); and other cytokines such as Th1 promoting Il27 ( 56 ), Th2 enhancing Il7 ( 57 ), and pro-cytotoxic Il15 ( 58 , 59 ). Assuming a canonical relation between CpG methylation and expression, these changes would result in a reduction of DC proinflammatory activity.…”

Harnessing invariant natural killer T cells to control pathological inflammation

“…Alternatively, if human iNKT cells generally retain functional plasticity, it may be important to identify methods to specifically promote their regulatory functions. To this end, a recent study found that the presence of IL-7 during in vitro expansion of human iNKT cells resulted in a CD4 + population with enhanced T H2 cytokine production ( 106 ), while exposure to short chain fatty acids, palmitate, or the mTOR inhibitor rapamycin may induce a regulatory phenotype ( 27 , 37 , 107 ). Another important consideration is that iNKT immunotherapy that engages T H2 pathways would likely be contraindicated in certain inflammatory diseases, including asthma, chronic obstructive pulmonary disease, and ulcerative colitis, where T H2 cytokine production by iNKT cells has been associated with disease-exacerbating effects (reviewed in ( 108 – 110 )).…”

Towards a better understanding of human iNKT cell subpopulations for improved clinical outcomes

“…treatment with a combination of IL-2 and IL-21 (but not IL-2 alone) downregulated the proapoptotic protein BIM in CD62L + iNKT cells, thus protecting them from activation-induced cell death (111,112). While these studies have been conducted in mice, evidence suggests that human iNKT cells can also express CD62L (113).…”