2. Helios in the Immune System
One element in this delicate immune balance is the transcription factor Helios (
Ikzf2 in mice and
IKZF2 in humans), a member of the Ikaros family of transcription factors that appears to fulfill a critical role in maintaining a regulatory profile in the T-cell lineage. Helios deficiency, either in mice lacking Helios in all cells
[34][38] or specifically in FoxP3
+ cells
[60][63] (mostly CD4
+ Tregs), leads to an autoimmune phenotype at 5–6 months of age that is associated with distinct symptoms, reproducing some canonical immunological manifestations of lupus: hypergammaglobulinemia against nuclear antigens, splenomegaly, glomerulonephritis, an enhanced presence of activated Tconvs, and an expansion of follicular T lymphocytes (see below). Indeed, a germline mutation in
IKZF2 was recently described in an SLE patient that was responsible for severe immune dysregulation
[61][64]. This raises the question as to what the role of Helios is in each immune cell population.
2.1. Helios in CD4+ T Cells
Initially, Helios was identified in mouse models as a specific marker of CD4
+ tTregs through a string of discoveries. (1) Until the first week of life, nearly all Tregs (CD4+ FoxP3+ T lymphocytes) that migrate from the thymus express Helios. (2) FoxP3
+ Helios
− CD4
+ T cells only appear from the second week after weaning. (3) In adult animals, around 70% of the CD4
+ Treg compartment expresses Helios. (4) Helios is not expressed in either iTregs or pTreg lymphocytes in an experimental tolerance model
[62][63][65,66]. However, the validity of this latter assumption has become increasingly controversial due to the more recent detection of Helios in CD4
+ pTregs
[64][67], in activated and exhausted CD4
+ Tconvs
[65][66][67][68,69,70], and in iTregs
[68][71]. Nonetheless, few studies have assessed the relative expression of Helios in specific subpopulations of immune cells beyond simply distinguishing between Helios
+ and Helios
− cells. In this sense, the merits of distinguishing up to three biologically relevant levels of Helios expression is currently being explored (Helios
high, Helios
mid, and Helios
low)
[69][70][72,73].
Beyond its proposed function as a unique molecular marker for tTregs, it is apparent that Helios
+ and Helios
− CD4
+ Treg lymphocytes have notable phenotypic, genetic, and functional differences
[71][72][74,75]. CD4
+ Tregs expressing Helios exhibit a more activated phenotype, with a higher percentage of effector (CD44
+ CD62L
−) cells. Moreover, in vitro studies indicate that these Tregs have a stronger immunosuppressive capacity relative to CD4
+ Tregs lacking Helios
[73][76]. Furthermore, experiments on lymphopenic animals have shown that CD4
+ Tregs expressing Helios in vivo express FoxP3 more stably. Indeed, the combined use of FoxP3 and Helios has been proposed to identify bona fide CD4
+ Tregs in humans
[74][77].
Interestingly, the deletion of Helios in mice, both in all cells
[34][38] and specifically in FoxP3
+ cells
[60][63], provokes an autoimmune phenotype at 6 months of age, demonstrating the role of this transcription factor in stabilizing regulatory activity. However, the absence of Helios in the entire CD4
+ T cell repertoire, including both Tconvs and Tregs, does not apparently evoke autoreactivity
[34][62][38,65]. This curiosity may suggest that Helios is involved in both sides of the immune balance and it leads
reus
earchers to draw two hypotheses. (1) Helios expression in CD4
+ Tconvs plays a significant role in the autoimmune reaction; and (2) other cell types with Helios-dependent regulatory activity impede autoreactive immune responses.
2.2. Helios in KIR+/Ly49+ CD8+
If there is one thing that CD4
+ Tregs and CD8
+ Ly49
+ Tregs have in common, it is the expression of Helios as a maker of their regulatory identity. Nevertheless, research on CD8
+ Ly49
+ Tregs is still in its infancy compared to that on CD4
+ Tregs. Evidence for the physiological relevance of Helios in CD8
+ Ly49
+ Tregs and their immunosuppressive function comes from the inability of CD8
+ Ly49
+ Helios
− T lymphocytes to inhibit the follicular T cell response relative to CD8
+ Ly49
+ Helios
+ cells. Moreover, unlike their Helios
+ counterparts, CD8
+ Ly49
+ Helios
− Treg lymphocytes exert a stronger effector phenotype (CD127
low) and worse survival under inflammatory conditions
[34][38]. Independently, weaker Helios expression has also been found in the CD8
+ Ly49
+ Tregs, correlating with autoimmune progression in a murine model that replicates the typical SLE symptomatology from the age of 5 months (including splenomegaly and germinal center reactivity). From this model, with deficient TGF-β signaling, expression of this cytokine may influence the Helios expressed by CD8
+ T cells
[37][41]. In addition,
rwe
searchers found that both the reduction in the proportion of Ly49
+ CD8
+ T cells and the weaker Helios expression by these cells was correlated to disease progression in two mouse models of lupus (MRL/MPJ and MRL/LPR)
[70][73].
A recent article in Science characterized the function and phenotype of the human equivalent of Ly49
+ CD8
+ Tregs in infectious and autoimmune contexts
[7][9]. Although the murine family of Ly49 receptors lacks a genetic human homologue in terms of sequence, the members of the human Killer cell Immunoglobulin-like Receptor (KIR) family are functionally equivalent to these. Both populations, KIR
+ CD8
+ and their murine Ly49
+ CD8
+ T cell counterparts, can eliminate autoreactive CD4
+ T cells in autoimmune and viral contexts, and they are characterized by their Helios
+ phenotype.
As demonstrated, altered Helios expression in the T lymphocyte pool is linked to an autoimmune phenotype that replicates the typical symptoms observed in SLE, an archetypal condition of systemic autoimmunity. Thus, while the “mystery” surrounding CD8
+ regulatory T cells is slowly lifting
[30][34], many questions remain, such as what the origin of these KIR
+/Ly49
+ CD8
+ T lymphocytes is, and what is their relationship to other cells in the immune system, like DCs.
2.3. Helios in Other Immune Cells: Double Negative (DN) T and NK Cells
2.3.1. Helios in DN T Cells
In addition to CD4
+ and CD8
+ T cells, there is a specific subpopulation of T lymphocytes with a TCRαβ
+ profile that expresses neither CD4 nor the CD8 co-receptor. In terms of their possible origin, the model that currently seems to attract the most interest, according to recent studies, suggests that double negative (DN) T cells are derived from autoreactive CD8
+ T cells that have lost their CD8 co-receptor
[75][76][78,79]. In this context, DN T lymphocytes would be characterized by the expression of PD-1 at their surface and by the transient presence of Helios in their nucleus
[75][78]. The pathological role of these PD-1
+ DN T cells is witnessed by their proinflammatory effector phenotype (CD127
low) and by the production of the IL-17 cytokine, which is tightly correlated with the autoimmune phenomenon
[77][78][80,81]. However, the regulatory properties of the DN T repertoire cannot be ignored, and they have also been reported on the other side of the immunological equilibrium, in non-autoimmune contexts and in organ transplantation
[79][80][81][82,83,84]. Again, this dual behavior of DN T cells would reflect the complexity of the immune system, the existence of specific subpopulations characterized by the expression of markers yet to be defined, and the influence of the molecular context (autoimmune or non-autoimmune).
The transcriptional profile of DN T cells from mice was recently analyzed
[82][85], revealing different DN T subpopulations to be distinguished by the Helios gene (
Ikzf2). Indeed, activated and non-activated DN T lymphocytes could be identified through this biomarker. Helios expression is enhanced in naïve DN T cells, whereas activated DN T cells express this transcription factor weakly.
2.3.2. Helios in NK Cells
In humans, after CD4
+ Tregs and mucosal-associated invariant T (MAIT) lymphocytes, NK cells are the immune cell population with the strongest Helios expression (supplemental Figure S3 from
[61][64]). Nevertheless, there are few data regarding Helios expression in the NK compartment. Helios has been proposed to regulate the activity of hyperreactive NK cells in mice resistant to viral infection
[83][86]. Moreover, Helios downregulation was observed in some subsets of “memory-like” NK cells from cytomegalovirus-infected individuals
[84][87], and finally, the proportion of CD16
+ CD56
dim NK cells was reduced in a lupus patient due to germline mutation of the
IKZF2 gene
[61][64]. Together, Helios appears to be an important transcription factor that controls the activity and homeostasis of NK cells, which are involved in autoimmunity
[85][88] and lupus
[86][89]. Further studies will be needed to examine the expression of Helios in autoimmune contexts and in different immune cells.