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INTRODUCTION
Autoimmune diseases including rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE)
are relatively common disorders.1 Although the underlying etiologies of these illnesses are still
elusive, they arise in the context of a break in the immune tolerance to self.2,3 The mechanisms for
abrogation of immune self-tolerance appear to be multifactorial, including genetic and environmental,
that work in concert to initiate the eventual hallmarks of disease: unregulated immune activation
against self-antigens and subsequent tissue destruction. Immune activation against self-antigens is
clinically manifest by the presence of autoantibodies and autoreactive T cells.1,2 Based upon their
autoantigenic targets, autoimmune diseases can be classified into organ-specific and systemic
autoimmune processes. For example, Grave’s disease, with its autoantibodies against the thyroid-
stimulating hormone (TSH) receptor, is a typical example of organ-specific autoimmune disease, as
is type I diabetes mellitus (DM) with its autoantibodies and autoreactive T cells directed against
components of pancreatic β cells, whereas SLE with its characteristic autoantibodies against
ubiquitous nuclear antigens is a good example of a systemic autoimmune disease. Although adaptive
immune cells such as B cells and T cells recognize self-antigens and hence dominate the phenotype of
the patient with autoimmunity, other immune components including antigen-presenting cells (APC)
and complement are involved in various steps from initiation of the autoimmune response to tissue
destruction.4–6 In this chapter, the contributions of these components to the development of
autoimmune diseases will be discussed, focusing on the latest discoveries.
GENETICS
AUTOIMMUNE DISEASES ARE POLYGENIC
Autoimmune diseases are polygenic, involving both major histocompatibility complex (MHC) and
non-MHC genes.7,8 Yet, the concordance rate of autoimmune diseases in monozygotic twins is not
100%, ranging from 10% to 50%, including in RA and SLE,7,9 indicating a significant role for
nongenetic factors in the development of these disorders. Nevertheless, genetic influences are strong.
For instance, the contribution of alleles of MHC to the development of autoimmunity has been known
for more than 30 years. An increased frequency of HLA (human leukocyte antigen)-DR4 has been
observed in some patients with RA.8 Using more sophisticated molecular techniques, the sequence of
genetic alleles encoding HLA loci have been typed, with the resultant demonstration that the HLA-
DRB1 gene is highly polymorphic and such polymorphisms can affect the binding of peptides to HLA
molecules and the contacts between the T-cell receptor (TCR) and the HLA molecule.8 Thus, the
associations between autoimmune diseases and particular HLA molecules can be explained by a
model where disease susceptibility is determined by differences in the ability of different HLA
alleles to present autoantigenic peptides to autoreactive T cells.2,8 However, the genetic effect of
MHC on disease propensity is broader. As an example, a recent study genotyped a panel of 1,472
single nucleotide polymorphisms (SNPs) across the 3.44 megabase (Mb) classic MHC region in
10,576 DNA samples from patients with autoimmune diseases including SLE, RA, and multiple
sclerosis, and from appropriate controls.10 The results of this study showed multiple risk alleles
across MHC class I, II, and III in these autoimmune diseases, indicating complex, multilocus effects
that span the entire region.
AUTOIMMUNE REGULATOR
Recently, a gene called autoimmune regulator (AIRE) gene has been identified as a candidate gene
responsible for the development of autoimmune diseases.12,13 AIRE is a transcription factor that
regulates the ectopic expression of proteins, normally expressed in peripheral tissues, in the thymus,
allowing for thymic expression of the latter and subsequent negative selection of self-reactive
thymocytes before they migrate as mature T cells to the secondary lymphoid organs such as the spleen
and lymph nodes. An alteration in thymic expression of AIRE can lead to increased generation of
autoreactive T cells due to their impaired negative selection.14 Indeed, patients with mutations of the
AIRE gene develop a syndrome called autoimmune polyendocrinopathy candidiasis ectodermal
dystrophy (APECED) or autoimmune polyglandular syndrome 1 (APS-1) that is characterized by
chronic mucocutaneous candidiasis, hypoparathyroidism, and Addison’s disease.15 In addition,
patients with this condition often have other organ-specific autoimmune diseases including type 1
DM, autoimmune thyroid diseases, gonadal failure, vitiligo, alopecia, dystrophy of dental enamel and
nails, and pernicious anemia.15
CYTOKINE RECEPTORS
Polymorphisms in several cytokine receptors are associated with autoimmunity.11 The IL-23 receptor
complex consists of IL-23R and IL-12Rβ1 subunits, with the latter shared with the IL-12 receptor
complex that is composed of IL-12Rβ1 and IL-12Rβ2 subunits. The IL23R receptor gene is
associated with inflammatory bowel disease and psoriasis.31,32 This is an intriguing point since IL-23
is involved in regulating the development of T helper 17 (Th17) cells that produce the potent
proinflammatory cytokine IL-17. In fact, IL-17 is found in psoriatic skin lesions.33 Likewise, the
IL12B gene that encodes IL-12β (the p40 subunit of IL-12) has been found to associated with
psoriasis.34
Figure 154-1 Mechanisms for autoimmune diseases. The figure shows potential mechanisms for
initiation of autoimmune diseases including genetic and environmental factors that work in concert to
initiate breakage of immune tolerance to self-antigens. Subsequently, unregulated immune activation
and tissue damage arise, leading to the development of autoimmune disease with activation of
autoreactive T cells and B cells with autoantibody production. Abbreviations: AIRE (autoimmune
regulator), CTLA-4 (cytotoxic T-lymphocyte antigen-4), PD-1 (programmed cell death 1), PTPN22
(protein tyrosine phosphatase 22), and FcgR (receptor for immunoglobulin G (IgG) Fc portion). See
details in text.
PERIPHERAL TOLERANCE
Despite removing or modifying autoreactive lymphocytes in the thymus and bone marrow by selective
processes, some lymphocytes reactive to self-antigens still mature and enter the peripheral immune
system; i.e., negative selection is not perfect, probably because if it were, potentially beneficial T
and B cells would never make it past central selection in the thymus and bone marrow, respectively.
The autoreactive T and B cells that escape central selection can potentially be activated upon
recognition of self-antigens; thus, peripheral tolerance mechanisms need to be available to avoid the
development of autoimmunity. Such mechanisms are several, and involve minimizing contacts of
autoreactive cells with self-antigens (ignorance of the self-reactive lymphocyte), not providing
appropriate signals for their activation, terminating activated autoreactive cells by regulatory
molecules such as PD-1 or CTLA4, and/or actively suppressing autoimmune responses by other T
cells.2 In critical organs like the brain, eyes, and gonads, immunological ignorance is a principal
mechanism for avoidance of autoreactivity via the provision of anatomical barriers separating the
tissue and lymphocytes. Activation of the latter can be further prevented by the relative paucity of
APCs in these organs and the minimal expression of MHC molecules in these tissues.46,47 These sites
are termed immunologically privileged since even tissue grafts do not elicit immune responses
therein. Immunologically privileged sites have additional mechanisms that prevent immune
activation, including production of TGF-β, which can suppress immune responses such as in the
anterior chamber of the eyes, and constitutive expression of Fas ligand that induces apoptosis of
infiltrating activated lymphocytes.46,47
Nevertheless, since autoantigens are expressed on some tissues and autoreactive T (and B)
lymphocytes are present in the circulation, there is still a reasonable chance for the latter to recognize
autoantigens expressed by MHC molecules on APC. However, such an autoantigenic signal through
potentially self-reactive antigen receptors is not enough to activate lymphocytes, as additional signals
(second signals) provided by APCs, such as CD80 and CD86, to CD28 on T cells are required for
proper activation.17,48 When proper costimulation is not provided by antigen presenting cells,
autoreactive T cells that are only signaled through the TCR undergo apoptosis or become anergized.
As CD80 and CD86 upregulation on APCs requires an inflammatory stimulus provided to the APC
such as that provided by a pathogen during infection, presentation of self-antigens in the absence of
inflammation does not lead to autoreactive T cell activation. By contrast, peptides of pathogens lead
to cellular activation as they are presented to the immune system in the appropriate inflammatory
context. Of course, this regulatory control on autoimmunity may be bypassed if in a similar manner
self-antigens are presented at a site of inflammation. Fortunately, this appears to be a relatively
unusual event, and autoimmunity invoked by this mechanism is typically avoided.
Autoreactive lymphocytes that manage to get activated in the periphery can be also deleted by
apoptosis (programmed cell death) and/or their activation terminated by inhibitory molecules
expressed on autoreactive lymphocytes. Fas ligand, or Fas, expressed on T cells is largely
responsible for the former.49,50 The latter process can be achieved through CTLA-4 and PD-1
expressed on activated T cells that inhibit and down-regulate T cell activation.17,21 The potential
roles for CTLA-4 and PD-1 in controlling autoimmunity have been demonstrated by mouse studies
where these genes have been genetically eliminated, for example, with resultant autoimmunity.51,52 In
addition, genetic polymorphisms in these molecules have been reported to be associated with
autoimmune diseases including SLE and RA (see Section “Genetics”). More importantly, there have
been attempts to treat autoimmune diseases by modulating CTLA-4 and PD-1 inhibitory
pathways.53,54 Indeed, abatacept, a recombinant fusion protein comprising the extracellular domain of
human CTLA4 and a fragment of the Fc domain of human IgG1, is available for the treatment of
RA.53 This drug, like CTLA4, competes with CD28 for CD80 and CD86 binding; however, in
contrast to CTLA4 engagement that leads to downregulation of T cell activation, it blocks T cell
activation and thus selectively modulates T cell activation.
The activation of autoreactive lymphocytes is also actively suppressed through other mechanisms.
The most appealing such mechanism is immune suppression through subsets of T cells called
regulatory T cells (Tregs), discussed below.
REGULATORY T CELLS
The concept that certain T cells can regulate immune responses dates back to early 1970s. These
cells, that Gershon initially called “suppressor cells,”55 remained largely undefined and their
mechanism of suppression not identified. After their original discovery, they remained out of vogue
until over a decade later when others including Sakaguchi reported modulation of immune responses
by CD4+ T cells expressing CD25 (IL-2 receptor alpha chain)56 as well as by T cells producing
TGF-β and IL-10.57–59 Cells with immune regulatory functions are now called regulatory T cells
(Tregs). Several subsets of Tregs have been identified thus far, including: (1) naturally occurring
CD4+ CD25+ Tregs expressing the transcription factor protein forkhead box P3 (Foxp3); (2) CD4+ T
cells producing IL-10 called type 1 regulatory T cells (Tr1); (3) CD4+ T cells producing TGF-β
named T helper 3 cells (Th3); and (4) CD8+ T cells producing IL-10 or TGF-β.60,61
CD4+ CD25+ REGULATORY T CELLS
Among the best characterized Tregs are naturally occurring CD4+ CD25+ Foxp3+ cells (FOXP3+
Treg) that are produced in the thymus. The potential role for these Tregs in controlling the
development of autoimmunity was initially demonstrated by studies where mouse T cell suspensions
were transferred into congenic athymic nude mice. If the latter animals received such suspensions
depleted of CD4+ CD25+ T cells, they developed an autoimmune syndrome including thyroiditis,
gastritis, insulitis, sialoadenitis, adrenalitis, oophoritis, glomerulonephritis, and polyarthritis.56 By
contrast, athymic nude mice that received whole T-cell suspensions did not develop autoimmunity,
indicating a role for CD4+ CD25+ T cells in preventing autoimmune diseases.56 Subsequently, the
inhibitory effects of CD4+ CD25+ Tregs in different types of animal models of autoimmune diseases
including collagen-induced arthritis (CIA) and autoimmune diabetes were documented.62,63 Yet, not
all CD4+ CD25+ T cells have immunoregulatory functions as CD25 is upregulated upon T-cell
activation. Thus, the search for other markers that specifically identify these cells has been extensive,
and has led to the identification of CD39, CD103, CTLA-4, glucocorticoid-induced TNF receptor
(GITR), lymphocyte activation gene 3 (LAG-3), interleukin 7 receptor, and Foxp364,65 as potential
markers. The most promising of this group is Foxp3. Disruption of its gene in mice results in
autoimmune diseases,66,67 whereas overexpression of Foxp3 in T cells induces expansion of CD4+ T
cells with immunosuppressive function.66–68 Furthermore, CD4+ CD25- T cells transfected with
FoxP3 acquire immunosuppressive function.66,67 Although it was originally thought that FoxP+ Tregs
develop only in the thymus, studies indicate that CD25- CD4+ T cells in the periphery can become
FoxP3+ Treg-like cells with immune regulatory function in the presence of T-cell receptor triggering,
IL-2, and TGF-β.69 The latter cells are now called adaptive or inducible FoxP3+ Treg (FOXP3+
iTreg) whereas the former cells are called naturally occurring FoxP3 Treg (nTreg).69
Although the exact mechanism(s) by which naturally occurring FoxP3+ Tregs suppress
autoreactive T cells is not fully understood, cell-to-cell contact is necessary and CTLA-4 appears to
be required.70,71 CTLA-4 expressed on Tregs interacts with CD80 and CD86 expressed on APC as
well as on the target T cells72,73; while CD80 and CD86, ligands for CTLA-4, are conventionally
expressed on APC as noted above, they have also been found on activated CD4+ T cells.72,73 These
molecules likely interact with CTLA-4 expressed on FoxP3+ Treg, providing “outside-in” signaling
leading to immune suppression. IL-2 appears to be involved in inducing and maintaining these
cells.71 IL-2 can upregulate FoxP3 expression through activating STAT5.74,75 Of interest, CD25, the
IL-2 receptor α chain, could be involved as a suppressive mechanism of FoxP3+ Tregs by absorbing
IL-2 and subsequently reducing the availability of this cytokine to other T cells.76 These Tregs also
express CD39 (ectonucleoside triphosphate diphosphohydrolase 1) and CD73 (ecto-5-nucleotidase).
These molecules generate pericellular adenosine with immunosuppressive activity by catalyzing
extracellular nucleotides.77 FoxP3+ Tregs can suppress T cell function by inducing the production of
the enzyme indoleamine 2,3-dioxygenase (IDO) from DCs. IDO catabolizes conversion of the
essential amino acid tryptophan to kynurenine that is harmful to T cells.71,78 Overall, FoxP3 Tregs
likely employ multiple mechanisms in suppressing other T cell function.
By contrast to animal studies, the data on CD4+ CD25+ Tregs in human autoimmune diseases is
relatively minimal. In patients with type 1 DM, a typical example of an organ-specific autoimmune
disease, the frequency of CD4+ CD25+ Tregs has been shown to be significantly lower than in
healthy controls and patients with type 2 DM, suggesting a role for these cells in disease
development.79 However, in separate studies, no difference in the frequency of CD4+ CD25+ Tregs
and FOXP3 gene allelic variation has been reported in patients with type 1 DM compared to healthy
controls,80,81 although the inhibitory function of such cells was impaired.81 Similarly, there was no
difference in the frequency of CD4+ CD25+ T cells and their functional markers including Foxp3
among patients with APS-II characterized by Addison’s disease, type I diabetes and autoimmune
thyroid disease as well as control patients with single autoimmune endocrinopathies and normal
healthy donors.82 However, CD4+ CD25+ Tregs from APS-II patients were defective in their
suppressive capacity.82 These findings suggest that patients with autoimmune endocrinopathies
including type 1 DM and APS-II may have an alteration in the suppressive function of CD4+ CD25+
Tregs.
A role for CD4+ CD25+ Tregs in the development of SLE and RA, typical systemic autoimmune
diseases, has also been explored. Most studies on CD4+ CD25+ Tregs in human lupus reported a
decreased frequency of this cell subset,83 suggesting their potential role in the development of SLE.
In RA, CD4+ CD25+ Tregs from both peripheral blood and synovial fluid have been studied.84–86
The frequency of CD25+ CD4+ Tregs in the latter fluid was higher than that in peripheral blood,85,86
although there was no significant difference in the numbers of CD4+ CD25+ Tregs in peripheral
blood between patients and controls.85,86 Of interest, compromised function of CD4+ CD25+ Treg
cells in suppressing inflammatory cytokines from CD4+ CD25- T cells was reported in RA.87
Furthermore, such impairment was reversed after treating patients with anti-TNF-α therapy. These
findings suggest that patients with RA have a functional, but not a numerical, defect in CD4+ CD25+
Tregs.
In contrast to CD4+ CD25+ Tregs that naturally occur, other regulatory T cells can be induced in vivo
and in vitro.61 These cells include Tr1, Th3 (CD4+ T cells producing large amounts of TGF-β) and
CD8+ CD28- T cells that develop in the setting of immune stimulation such as that initiated by chronic
infections. For instance, humans chronically infected with hepatitis C or Epstein Barr viruses, or with
M. tuberculosis or the nematode Onchocerca volvulus, may develop pathogen-specific Tr1 cells that
produce large amounts of IL-10 that can suppress antigen-specific or nonspecific T-cell responses.88–
91 These cells can also be generated after ingestion of a foreign antigen via the oral route.57 The
immunosuppressive mechanisms employed by Tr1 and Th3 cells are dependent on IL-10 and TGF-β
production rather than direct cell-to-cell contact as in natural Tregs.61
CD8+ T cells may also have immunosuppressive activity.60 Mice deficient in CD8+ T cells
develop experimental autoimmune encephalomyelitis (EAE), an animal model of multiple
sclerosis.92 However, EAE in the same animals was milder when they received adoptive transfers of
CD8+ CD28- T cells. Moreover, CD8+ CD28-, but not CD8+ CD28+, T cells suppressed in vitro
IFN-γ production by CD4+ T cells specific for myelin oligodendrocyte glycoprotein, an autoantigen
in EAE. Such suppression required cell-to-cell contact and was mediated by inhibition of
upregulation of costimulatory molecules on APCs that led to decreased costimulation of CD4+ T
cells. Of interest, a recent study suggested a potential role for CD8+ T cells with suppressor activity
in improving lupus-like disease in mice.93 Decreased levels of pathogenic anti-dsDNA antibodies
with a resultant increase in survival were found in lupus-prone (NZB ξ NZW)F1 mice when injected
with an artificial peptide called pConsensus (pCons) that was developed based on T cell stimulatory
VH sequences found in (NZB × NZW)F1 anti-DNA antibodies. This protection depended in part on
the generation of peripheral TGFβ- and Foxp3-expressing inhibitory CD8+ T cells, and suggested that
CD8+ T cells with regulatory activity could be a potential way to modulate autoimmune diseases.
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Full reference list available at www.DIGM8.com
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