Antibody-dependent
Cellular Cytotoxicity
(ADCC)
Jean-Luc Teillaud, Cordeliers Research Center, INSERM U.872, Université Paris-Descartes et
Université Pierre et Marie Curie, Paris, France
Based in part on the previous version of this eLS article ‘Antibody-
dependent Cellular Cytotoxicity’ (2006) by Robert F Graziano and Paul
M Guyre.
Antibody-dependent cell-mediated cytotoxicity (ADCC) is
the killing of an antibody-coated target cell by a cytotoxic
effector cell through a nonphagocytic process, charac-
terised by the release of the content of cytotoxic granules
or by the expression of cell death-inducing molecules.
ADCC is triggered through interaction of target-bound
antibodies (belonging to IgG or IgA or IgE classes) with
certain Fc receptors (FcRs), glycoproteins present on the
effector cell surface that bind the Fc region of immuno-
globulins (Ig). Effector cells that mediate ADCC include
natural killer (NK) cells, monocytes, macrophages, neu-
trophils, eosinophils and dendritic cells. ADCC is a rapid
effector mechanism whose efficacy is dependent on a
number of parameters (density and stability of the anti-
gen on the surface of the target cell; antibody affinity and
FcR-binding affinity). ADCC involving human IgG1, the
most used IgG subclass for therapeutic antibodies, is
highly dependent on the glycosylation profile of its Fc
portion and on the polymorphism of Fcc receptors.
Introduction
Antibody-dependent cell-mediated cytotoxicity (ADCC)
was initially described by Moller in 1967 as the ability of
natural killer (NK) cells – at the time identified only as
lymphocytes – to destroy antibody-coated target cells. A
nonphagocytic mechanism, whereby most leucocytes can
kill target cells in the absence of complement and without
major histocompatibility complex (MHC) restriction (a
mechanism by which specific cytotoxic T cells (CTLs) kill
target cells expressing peptides loaded onto surface MHC
Class I molecules) thus became apparent. The specificity of
eLS subject area: Immunology
How to cite:
Teillaud, Jean-Luc (July 2012) Antibody-dependent Cellular
Cytotoxicity (ADCC). In: eLS. John Wiley & Sons, Ltd: Chichester.
DOI: 10.1002/9780470015902.a0000498.pub2
eLS & 2012, John Wiley & Sons, Ltd. www.els.net
Advanced article
. Introduction
Article Contents
. Fc Receptors are Trigger Molecules for ADCC
. Cellular Effectors of ADCC
. Mechanisms of Cytotoxicity
. Inhibition of Cytotoxicity
. Regulation of ADCC
. Role of ADCC in Host Immunity
. Summary
Online posting date: 16th July 2012
ADCC is conferred by target-bound antibody and can be
induced in vitro by antibody concentrations well below
those needed to activate complement-mediated lysis.
Receptors for the Fc region of Ig (FcR) on the plasma
membrane of the effector cell are necessary but not suf-
ficient for ADCC activity. Once antibody has bound to the
target cell, its interaction with either activating or inhibi-
tory FcR on the cytotoxic effector cell appears to dictate the
precise signalling pathways that are stimulated. When
target-bound antibodies crosslink activating FcR (char-
acterised by the presence of an immunoreceptor tyrosine-
based activating motif, ITAM, in their intracellular (IC)
domain or in the intracellular domain of the associated
transducing unit termed g chain), a cytotoxic pathway is
initiated that results in the release of mediators such as
perforin, granzymes, tumour necrosis factor (TNFa) and
reactive oxygen intermediates (ROI) – which induce target
cell death. Conversely, ligation of inhibitory FcR (char-
acterised by the presence of an immunoreceptor tyrosine-
based inhibitory motif (ITIM) in their IC domain) prevents
effector cell activation, resulting in survival of the target
cell.
Fc Receptors are Trigger Molecules
for ADCC
As noted earlier, FcR on the surface of cytotoxic effector
cells are required for ADCC (Fanger et al., 1989). Separate
genes that are members of the immunoglobulin (Ig)
supergene family encode different FcR: FcgR specifically
bind IgG, FcaR bind IgA, FceR bind IgE and Fcm/aR bind
both IgM and IgA (van Egmond et al., 2001a; Dombrowicz
et al., 2000; Kinet and Launay, 2000; Ravetch and Bolland,
2001; van der Poel et al., 2011). In humans, it is clear that
members of the following FcR classes are all capable, when
crosslinked, of activating ADCC: FcgRI (CD64), FcgRIIa
and FcgRIIc (CD32), FcgRIIIa (CD16) (Figure 1), FcaR
(CD89) and FceRI. In mouse, ADCC can be achieved
through the crosslinking of FcgRI, FcgRIII and FcgRIV as
there is no activating FcgRIIa (Nimmerjahn and Ravetch,
2008). In addition, it has also been shown that engagement
1
 Antibody-dependent Cellular Cytotoxicity (ADCC)

FcRI
FcRIIIA FcRIIA/C
(CD64)
(CD16) (CD32)

  ( or )

High affinity Intermediate affinity Low affinity

(PMNC, monocytes/MΦ) (NK, MΦ) (Monocytes, MΦ, NK)

Figure 1 Schematic view of human-activating FcgR. In most cases, human-activating Fcg receptors comprise an IgG binding a chain associated with a
transducing subunit, a g chain (gg) homodimer. In NK cells, the presence of a g/z heterodimer or of a z/z homodimer has also been reported (the z chain is
also part of the T cell receptor (TcR) complex). The associated g or z chain contains an ITAM (immuno-receptor tyrosine-based activating motif) (black
square) that is phosphorylated upon crosslinking of FcgR. Human FcgRIIa is the only activating receptor where the IgG-binding a chain intracellular domain
contains an ITAM, although its association with a transducing g chain homodimer has been debated. Myeloid cells can express both activating and inhibitory
FcgRIIb (not shown in the diagram) and it is considered that cellular activation will result from the fine tuning of the balance between activating and
inhibitory signals. NK cells express activating FcgRIIIa, although the expression of activating FcgRIIc and of inhibitory FcgRIIb by small NK cell subsets has also
been reported. FcgRIIIa is also expressed by some T cells. FcgRIIIb, a GPI-linked surface receptor expressed by neutrophils is not presented. In mouse, there is
no FcgRIIa and inhibitory FcgRIIb is referred only as FcgRII. In contrast, there is another activating FcgR, termed FcgRIV, also associated with a g chain
homodimer.

of mouse inhibitory FcgRII leads to inhibition of ADCC
activation (Clynes et al., 2000). The balance between acti-
vating and inhibitory FcgR thus appears to be a crucial
determinant of the magnitude of ADCC that is induced
when the effector cell engages the antibody-coated target
and expresses both types of Fc receptors (Nimmerjahn and
Ravetch, 2008; Abes et al., 2009). See also: Antibodies;
Antibody Classes; Antibody Function; Fc Receptors;
Hypersensitivity: Antibody-mediated Cytotoxic (Type II)
Cellular Effectors of ADCC
Natural killer cells
NK cells are perhaps the most reliable effectors of ADCC
when tested in cell culture cytotoxicity assays (Perussia,
2000). When freshly isolated from circulating blood, a large
subset of human NK cells (defined as CD56dim CD16+)
express relatively high levels of the type IIIa IgG receptor
(FcgRIIIa or CD16a) and very efficiently mediate ADCC
of tumour cells that are coated with antitumour IgG (Figure
2). NK cells appear not to express other classes of Fc
receptors and, therefore, are not activated for ADCC by
IgA, IgE or IgM. FcgRIIIa is present on both NK cells and
macrophages as a transmembrane receptor that is associ-
ated with the FcR g chain, an activating molecule involved
in signal transduction. This association with the g chain
was found to be essential both for surface expression of
FcgRIIIa and for signal transduction following engage-
ment of this receptor by IgG. In contrast, FcgRIIIb
(CD16b) found on human neutrophils is expressed as a
phosphatidyl inositol glycan-linked receptor that does not
associate with the g chain and does not activate tumour cell
killing. Only FcgRIIIa, and not FcgRIIIb, is found in mice.
The expression of activating FcgRIIc and of inhibitory
FcgRIIb on small subsets of circulating NK cells has been
reported (Metes et al., 1994; Sulica et al., 2001; Dutertre
et al., 2008), but whether these receptors impact NK cell-
mediated ADCC mediated through FcgRIIIa in some
circumstances remains to be elucidated.
Monocytes, macrophages and dendritic cells
Monocytes and macrophages express all three classes of
IgG receptors, FcgRI (CD64), FcgRII (CD32) and
FcgRIII (CD16) (see Figure 1), as well as the receptor for
IgA (CD89) (Ravetch and Bolland, 2001; van Egmond
et al., 2001a; Nimmerjahn and Ravetch, 2008). Each has
been shown to be capable of triggering phagocytosis and
ADCC by monocytes and macrophages. Although only
low levels of FcgRIIIa are expressed on freshly isolated
monocytes, this receptor can be markedly increased when
monocytes mature into macrophages or are cultured with
activating cytokines such as interferon g (IFNg). Cytokine
activation has also been shown to induce expression of
FceRI. It has been noted that activated macrophages have
high FcgRIIIa/IIb ratios, favouring cell activation,
whereas quiescent effectors have low IIIa/IIb ratios, pro-
viding a high threshold for cell activation (Ravetch and
Lanier, 2000; Nimmerjahn and Ravetch, 2008). Human
blood dendritic cells also express IgG FcR and are potent
effectors in antibody-dependent cellular cytotoxicity
(Schmitz et al., 2002). In mouse, a critical role of FcgRIV in
ADCC mediated by monocytes, macrophages and neu-
trophils has been suggested using FcgRIV-deficient mice
(Nimmerjahn et al., 2010).
Neutrophils and eosinophils
Freshly isolated neutrophils do not lyse tumour targets
through FcgR, but can be activated by granulocyte–
macrophage colony-stimulating factor (GM-CSF) to
mediate killing through FcgRIIa, and by IFNg to mediate
killing through FcgRI and FcgRIIa. The type of FcgRIII
expressed by human neutrophils (FcgRIIIb, a GPI-linked
surface receptor) does not mediate cytotoxicity of tumour

2 eLS & 2012, John Wiley & Sons, Ltd. www.els.net


targets, even following cell activation by cytokines. On the
contrary, FcgRIIIa on murine polymorphonuclear neu-
trophil (PMN) does induce ADCC of tumour cells. Cyto-
lysis of red blood cells occurs via FcgRIIa, FcgRIIIa and
FcgRIIIb even when resting human neutrophils are used as
effectors. These observations suggest that erythroid and
tumour targets are killed through different pathways, and
that triggering of FcgRIa or FcgRIIa on neutrophils acti-
vates lytic mechanisms distinct from those initiated via
FcgRIIIb (Fanger et al., 1989). Of particular interest,
neutrophils can efficiently kill human IgA-coated tumour
cells and ADCC is highest when both IgA and IgG recep-
tors are coordinately triggered (van Egmond et al., 2001b).
Furthermore, freshly isolated neutrophils carry out FcaR-
dependent lysis without cytokine preactivation, although
to a low level. Treatment of donors with granulocyte col-
ony-stimulating factor (G-CSF) leads to an upregulation
of FcgRI and enhanced ADCC by neutrophils (Dechant
et al., 2007).
Although freshly isolated eosinophils express FcgRII as
well as low levels of FcgRIII and FceRI, they are not potent
ADCC effectors unless activated with GM-CSF, inter-
leukin 3 (IL-3) or IL-5. After cytokine activation, killing of
tumour or erythrocyte targets is mediated through FcgRII
but not FcgRIII. Thus, like neutrophils, eosinophils
require cytokine activation to perform ADCC through
FcgRII. Also, like neutrophils, freshly isolated eosinophils
are able to mediate ADCC via FcaRI. Eosinophils have
also been shown to mediate ADCC of parasites via FceR
following activation by cytokines (Dombrowicz et al.,
2000) and of tumour cells via FcgRI (Karagiannis et al.,
2007).
Mechanisms of Cytotoxicity
It is now well established that antibodies act as a bridge
between FcR on the effector cell and the target antigen on
the cell that is to be killed. Crosslinking of receptors is
required to activate the cytotoxic event (Figure 2). Even for
high-affinity receptors, binding of monomeric Ig is insuf-
ficient to transmit an activation signal. The nature of the
biological response and the subsequent cytotoxicity initi-
ated by FcR crosslinking depends on the integration of a
variety of signals that are dependent on the class and sub-
class of both the antibody and the FcR involved, on acti-
vation of other signalling molecules, on the effector cell
type, and on the nature of the target cell. Cell surface
molecules other than FcR also may play a role by pro-
moting adhesion between effector and target cells and/or
by providing synergistic signals.
Crosslinking of activating FcR initiates a signalling
cascade that culminates in release of cytoplasmic granules
and the synthesis of surface activation molecules including
CD69 and CD25 (the g chain of the high affinity IL-2
receptor). It often begins with the stimulation of ITAM
sequences (Billadeau and Leibson, 2002; Ravetch and
Lanier, 2000). As discussed earlier, ITAMs have been
eLS & 2012, John Wiley & Sons, Ltd. www.els.net
Antibody-dependent Cellular Cytotoxicity (ADCC)
Tumor antigen
FcR
Tumor cell
FcR+ cell
Anti-tumor mAb
Figure 2 Schematic view of ADCC. The binding of an IgG antitumour
mAb to the target cell (here a tumour cell) allows the recruitment and
crosslinking of activating FcgR expressed by effector cells (NK cells,
neutrophils, etc.). In humans, activating receptors include FcgRI (CD64),
FcgRIIa (CD32) and FcgRIIIa (CD16). In mouse, they include FcgRI, FcgRIII
and FcgRIV. Effector cells are then activated and release molecules that will
lead to the death of tumour cells. The diagram represents a NK cell
containing granules that polarised towards the contact zone with tumour
cell once FcgRIIIa (CD16) has been engaged and crosslinked. The granule
content (perforin and granzyme B) is then released in the close vicinity of
the tumour cell (NK cell ‘synapse’), initiating an apoptotic process that will
lead to cell death.
identified within the cytoplasmic domain of the so-called
alpha or immunoglobulin-binding chains of some FcR
(e.g. FcgRIIa), and also within associated signalling mol-
ecules (e.g. g chain in the case of FcgRI, FcgRIIIa, FcaRI
and FceRI, and mouse FcgRIV). Granule release is
dependent on activation of the mitogen-activated protein
kinase (MAPK) family member ERK (extracellular signal-
related kinase) and includes intermediate interactions
involving the p56lck and many other protein tyrosine
kinases (reviewed by Perussia, 2000).
Many studies have focused on dissecting the mechanisms
of target cell destruction mediated by cytolytic CTL and
NK effector cells. Although the manner in which T cells and
NK cells recognise their targets is different, these cells share
similar cytotoxic mechanisms. Upon recognition of target,
the contents of specialised intracellular granules (also
termed secretory lysosomes) within the effector cell are
released by a calcium-dependent polarised exocytotic
process at the lytic synapse (de Saint Basile et al., 2010). The
NK cell synapse involves the reorganisation of the actin
cytoskeleton and the clustering of some membrane recep-
tors in the NK cell at the contact with the target cell
(Orange, 2007). Perforin, or cytolysin, is one component
that is released from granules. Perforin inserts and poly-
merises within the target cell membrane, in a manner that is
analogous to the membrane insertion of the terminal
components of the complement cascade, resulting in the
formation of a pore in the target cell. Calcium is required
for the insertion and assembly of perforin in the target cell
membrane. A critical role for perforin in host immune
defence has been confirmed in perforin-deficient mice.
Other components of granules released by exocytosis have
also been implicated in the mechanism of CTL and NK
cell-mediated cytotoxicity. One such granule component,
3
 Antibody-dependent Cellular Cytotoxicity (ADCC)
granzyme B, may be involved in deoxyribonucleic acid
(DNA) fragmentation observed in target cells after
engaging a cytotoxic effector cell. However, studies using
granzyme B-deficient mice suggest a less important role for
granzyme B in cytotoxicity. Molecular studies of patients
with inherited defects that impair cytotoxic functions have
also helped to decipher the intimate mechanisms of cyto-
toxicity by NK cells. Notably, it has been shown that the
cytotoxic function requires the cooperation of the lysoso-
mal cytotoxic granule and the endosomal exocytic vesicle.
A rapid polarisation of these two types of granules is
observed, followed by a coalescence near the cell–cell
contact area (Ménager et al., 2007). In vivo imaging
experiments have shown that tumour cells form stable
conjugates with macrophages and neutrophils expressing
FcgRs in tumour-bearing mice treated with a tumour-
specific antibody (Hubert et al., 2011). The contact zone
accumulated actin, FcgR and phosphotyrosines. These in
vivo ADCC synapses appear to be organised in multifocal
supra-molecular activation clusters. Another mechanism
of target cell destruction is the engagement of Fas by Fas
ligand (FasL), molecules expressed, respectively, on the
surface of some target and effector cells. This interaction
initiates a cascade of signals in the target cell resulting in
apoptosis of the target. Crosslinking of FcgRIIIa on NK
cells induces the expression of FasL, enabling them to kill
Fas+ targets. Therefore, FasL/Fas interaction may be an
important mechanism of ADCC mediated by NK cells.
Myeloid cells are able to destroy antibody-opsonised
targets by phagocytosis as well as by extracellular lysis.
Targets ingested phagocytically are destroyed in intracel-
lular phagolysosomes of the effector cell. The mech-
anism(s) by which myeloid cells mediate extracellular lysis
of antibody-opsonised target cells has been controversial.
It is clearly different from ADCC mediated by NK cells. In
one in vitro system, the mechanism of ADCC mediated by
myeloid cells has been shown to be distinct from that
mediated by NK cells based on divalent cation require-
ments (Graziano et al., 1989). ADCC mediated by NK cells
via FcgRIII was Ca2+ dependent and Mg2+ independent.
In contrast, ADCC mediated by monocytes or by IFNg-
activated PMN via FcgRIa or FcgRIIa, or by peritoneal
macrophages via FcgRIa, FcgRIIa or FcgRIIIa was Mg2+
dependent and Ca2+ independent. Thus, even though
macrophages and NK cells both mediate ADCC via
FcgRIIIa, the mechanism by which these two types of
effector cells mediate their killing is distinct. Monocytes,
macrophages and neutrophils release ROI when FcR
are crosslinked, and ROI have been implicated as the
mediators of target cell death. However, other mechanisms
must also exist since neutrophils isolated from patients with
chronic granulomatous disease (CGD), whose cells lack
the ability to form ROI, were able to mediate ADCC as
well as neutrophils isolated from normal donors (Roberts
et al., 1993). Finally, it has been shown that when antibody
binds to an apoptotic trigger molecule on the target
cell, FcR crosslinking of the target-bound immunoglobulin
can induce apoptotic cell death, apparently without a
4 eLS & 2012, John Wiley & Sons, Ltd. www.els.net
requirement for active participation of the effector cell such
as release of toxic mediators by the effector cell. This has
been shown with anti-CD20 antibodies whose binding to
CD20 expressed by lymphoma cells triggers a significant
apoptosis only when a FcgR-mediated crosslinking has
been achieved.
In conclusion, multiple pathways may be involved in
how an effector cell mediates ADCC of an antibody-
opsonised target cell. These include perforin, cytolytic
enzymes, reactive oxygen intermediates and apoptotic
signalling both with dependent and independent of Fas/
FasL interactions. Several mechanisms may combine to
mediate cytotoxicity depending on a variety of factors. The
relative importance of each in vivo and ex vivo will continue
to be dissected using knockout mice and/or effector cells
from individuals deficient in specific effector mechanisms.
Intra-vital imaging and in vitro imaging analyses should
make it possible to better understand the in vivo dynamics
of ADCC and the formation of the ADCC synapse.
Inhibition of Cytotoxicity
Fine tuning between activating and inhibitory pathways is
now also understood to be a critical determinant of ADCC
activity (Billadeau and Leibson, 2002; Ravetch and Lanier,
2000; Nimmerjahn and Ravetch, 2008). FcgRIa (CD64),
FcgRIIa (CD32) and FcgRIIIa (CD16a); FcaR (CD89)
and FceRI are all capable of mediating ADCC when
engaged by their respective ligands and crosslinked. Con-
versely, an inhibitory signal is generated upon crosslinking
of FcgRIIb which contains cytoplasmic ITIM sequences,
aborting cellular activation through ITAM-containing
receptors. The presence or absence of signalling through
other immune inhibitory receptors on ADCC effector cells
also influences cytotoxic activity. These receptors, with
acronyms such as killer inhibitory receptor (KIR) (or
CD158), CD94/NKG2 (NKG2, an activating receptor
for Natural Killer cells is also known as CD159a) and
immunoglobulin-like transcript (ILT) (also called leuco-
cyte inhibitory receptor (LIR or CD85)) associate with
SHP-1 (Src homology region 2 domain-containing phos-
phatase-1) and SHP-2 (Src homology region 2 domain-
containing phosphatase-2) phosphatases to inhibit cell
activation.
The effect of FcgRIIb on ADCC and tumour growth
in vivo was demonstrated in FcgRIIb-deficient mice. Using
models of antibody-mediated tumour treatment, Clynes
et al. (2000) demonstrated that FcgRIIb-deficient mice were
able to inhibit tumour growth and protect from tumour
metastasis as compared with wild-type mice. This finding
confirmed that FcgRIIb acted as an inhibitory receptor.
One interesting development of this observation has been
the generation of antitumour IgG antibodies that weakly
bind the inhibitory FcgRIIb, whereas strongly engaging
activating receptors (Shields et al., 2001; Stavenhagen
et al., 2007).

Regulation of ADCC
The ability of an effector cell to mediate ADCC depends on
a variety of parameters including the type of effector cell,
the class of FcR(s) engaged, the number of FcR per cell and
the state of activation of the effector cell. Treatment of
effector cell populations with cytokines or other physio-
logical mediators may increase or decrease the expression
level of a particular FcR on an effector cell, thereby
enhancing or depressing ADCC through that receptor.
Cytokine treatment may also change the activation state of
an effector cell leading to enhanced or depressed ADCC
without altering the number of FcR on the effector cell. The
expression of other molecules on the effector cell such as
inhibitory receptors or adhesion molecules also impacts the
ability of an effector cell to mediate ADCC. It seems likely
that the specific tissue microenvironment, which dictates
the overall molecular interactions of the FcR-bearing cell,
plays a crucial role in the ultimate level of function that is
triggered by engagement of FcR.
IFNg is a potent modulator of ADCC. Treatment of
monocytes, macrophages or neutrophils with IFNg
increases the expression level of FcgRIa on these cells more
than 10-fold, owing to the presence of interferon-stimu-
lated response elements (ISREs) in the FCGR1 gene pro-
moter. ADCC mediated via FcgRIa is similarly increased,
especially when limited amount of antibody is available on
the target. Interestingly, even though IFNg treatment does
not increase the expression level of FcgRIIa on neutrophils,
it does enable ADCC FcgRIIa, perhaps by altering the
cytotoxic potential of the neutrophil or by changing the
FcgRIIa/FcgRIIb ratio. Furthermore, IFNg induces
higher expression of the Fc receptor g chain that is essential
for ADCC triggered through FcgRIa, and may also par-
ticipate in signal transduction following engagement of
FcgRIIa and FcgRIIIa.
Enhancement of ADCC by cytokines other than IFNg
has also been demonstrated. Both interleukin 2 (IL-2) and
IL-15 that activate NK cells and induce an upregulation of
FcgRIIIa expression stimulate strongly ADCC exerted by
anti-CD20 (rituximab) and anti-epidermal growth factor
(EGFR; cetuximab) antibodies against tumour cells
(Golay et al., 2003; Roberti et al., 2011). GM-CSF and
TNFa enhance ADCC by monocytes cultured in vitro
through FcgRI, FcgRII and FcaRI. ADCC mediated by
PMN via FcgRII is enhanced by in vitro culture in the
presence of G-CSF. Treatment of patients in vivo with G-
CSF or IFNg leads to the upregulation of FcgRI and
FcaRI on neutrophils and enhanced ex vivo ADCC via this
receptor (Valerius et al., 1993; Dechant et al., 2007). This
observation has led to the development of bi-specific anti-
bodies (engineered antibodies directed against a tumour-
associated antigen expressed on tumour cells and against
FcgRI expressed by killer cells) for treating cancer patients
in combination with G-CSF (Michon et al., 1995; Repp
et al., 1995) and, recently, to the development of recom-
binant antibodies directed against EGFR (Dechant et al.,
2007).
eLS & 2012, John Wiley & Sons, Ltd. www.els.net
Antibody-dependent Cellular Cytotoxicity (ADCC)
Role of ADCC in Host Immunity
Early studies showed that tumour-specific antibodies that
were effective in promoting ADCC in vitro were also effi-
cacious in curing tumours in vivo using mouse models
(Herlyn and Koprowski, 1982). This correlative evidence
was confirmed recently in studies using mice deficient in
FcRg. FcRg refers to the g chain-signalling molecule that
was initially described as a subunit of FceRI, but later
shown to be required for optimal expression of – and signal
transduction by – FcRgRIa and FcRgRIIIa (and FcgRIV
in mice). NK cells and macrophages from FcRg–/– mice are
deficient in expression of these activating FcRgR and
ineffective in mediating ADCC in vitro. The physiological
importance of this deficiency has also been demonstrated in
vivo. Functional FcR were required for both passive and
active immunity to tumours. Whereas, human IgG1 and
murine IgG2a antitumour antibodies dramatically
inhibited tumour growth in intact mice, they were without
effect in FcRg –/– mice (Clynes et al., 2000). Similarly, when
the FcR of an antitumour antibody was mutagenised to
eliminate its binding to FcRgRIII, it lost its capacity to
inhibit tumour growth in vivo. Of note, when mice no
longer expressing inhibitory FcgRIIb were used, a strong
improvement in the ability of therapeutic antibodies to
prevent tumour growth was observed, indicating that the
overall balance between activating and inhibitory FcgR is
indeed crucial for the antitumour activity of antibodies
(Clynes et al., 2000).
Evidence implicating the role of ADCC in human host
immunity had been mostly circumstantial until the early
2000s. It was mostly based on several reports correlating
the ability of human effector cells to mediate ADCC, or the
presence of ADCC-mediating antibody titres, with the lack
of disease progression. For example, in one study, ADCC
activity of mononuclear cells isolated from immuno-com-
promised human immunodeficiency virus (HIV) patients
correlated with longer survival in these patients (Forthal
et al., 1999). In a separate study, the levels of ADCC
activity via anti-gp120 antibodies in serum of HIV-1
positive individuals correlated with successful host defence
as defined by rate of progression to acquired immune
deficiency syndrome (AIDS). Furthermore, effector cells
isolated from humans that were capable of mediating
ADCC in conjunction with antibody to herpes simplex
virus (HSV) were able to protect neonatal mice from lethal
infection with HSV. Some bacterial pathogens have
developed mechanisms to disrupt Ig–FcR interactions in
order to circumvent ADCC or phagocytosis. These
mechanisms provide the pathogens an advantage by
interfering with host clearance mechanisms implicating the
importance of ADCC and phagocytosis in host immunity.
The role of ADCC in human host immunity has been
strongly highlighted by a number of reports on the impact
of FcgR polymorphism on the clinical outcome of cancer
patients. FcgRIIIa gene exhibits a polymorphism because
of a single nucleotide change leading to the presence at
position 158 of either a valine or a phenylalanine. The
5
 Antibody-dependent Cellular Cytotoxicity (ADCC)
presence of phenylalanine makes FcgRIIIa a weaker
binder to human IgG1 as compared with FcgRIIIa har-
bouring a valine. It has been demonstrated in vitro that NK
cells derived from Val/Val homozygous donors exhibit a
stronger ADCC against IgG1-coated tumour cells as
compared with Phe/Phe donors. Interestingly, an associ-
ation between the FCGR3A genotype and clinical and
molecular responses of non-Hodgkin lymphomas (NHL)
patients treated with the chimeric human IgG1 anti-CD20
antibody rituximab has been shown (Cartron et al., 2002).
The impact of FcgRIIIa polymorphism, as well as that of
FcgRIIa (where either an histidine or an arginine is present
at position 131, leading to differences in the binding affinity
of both human IgG2 and IgG1; Bruhns et al., 2009) on the
clinical outcome of patients treated with therapeutic anti-
bodies has been further extended, suggesting an important
role of ADCC exerted by these antibodies in vivo (Weng
and Levy, 2003; Zhang et al., 2007; Musolino et al., 2008).
Moreover, a potential role for ADCC in controlling the
initial development of HIV infection has been explored
(Alsmadi and Tilley, 1998; Hessell et al., 2007; Moldt et al.,
2011; Tudor and Bomsel, 2011). The ADCC ability of
antibodies directed against various epitopes of HIV-1
envelope proteins (in particular gp41) through the
recruitment of FcgRIIIa expressed by NK cells (Alsmadi
and Tilley, 1998) or of FcgRI (Tudor and Bomsel, 2011)
and FcgRIIa (Moldt et al., 2011) expressed by monocytes
has been demonstrated. It has been shown that most of the
neutralising high-affinity anti-HIV-1 IgG1 antibodies are
capable of directing ADCC against infected cells (Alsmadi
and Tilley, 1998).
Thus, the critical role of ADCC in vivo has been strongly
supported by the detailed study of the mechanisms of
action of therapeutic antibodies and of antibodies
developed for further therapies in cancer and virus infected
patients. This has led to the emergence of a new generation
of therapeutic antibodies selected and engineered for a
stronger binding to FcgR, in particular FcgRIIIa and with
an enhanced ADCC. These antibodies have been engin-
eered by site-directed mutagenesis (Shields et al., 2001) or
by fine-tuning their glycosylation profile (Shields et al.,
2002; Shinkawa et al., 2003; Sibéril et al., 2006). A low/no
fucose content renders these antibodies high FcgRIIIa
binders and potent ADCC makers (Niwa et al., 2004).
These antibodies are currently tested in clinical trials
(Béliard et al., 2008; Robak, 2009) and may be the best
demonstration that ADCC is indeed critical in the effector
functions of a number of antibodies in patients.
Summary
ADCC occurs when antibodies create a bridge between
antigenic determinants (epitopes) on target cells and FcR
on cytotoxic effector cells. ADCC is activated by the
crosslinking of effector cell FcR, which can trigger the
destruction of erythrocytes, allogeneic cells, virus-infected
cells, tumour cells and parasites. The mechanisms of killing
6 eLS & 2012, John Wiley & Sons, Ltd. www.els.net
include target cell apoptosis as well as effector cell release of
toxic molecules such as perforin, granzyme B, TNFa,
reactive oxygen and reactive nitrogen.
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