Aging and innate immune cells
Timothy P. Plackett,*,†,‡ Eric D. Boehmer,* Douglas E. Faunce,†,§,¶
and Elizabeth J. Kovacs*,†,§,¶,1
Departments of *Cell Biology, Neurobiology, and Anatomy and ¶Surgery, §Immunology and Aging Program, †Burn
and Shock Trauma Institute, Loyola University Chicago, Maywood, Illinois; and ‡Chicago College of Osteopathic
Medicine, Midwestern University, Downers Grove, Illinois
Abstract: The innate immune system serves an
important role in preventing microbial invasion.
However, it experiences significant changes with
advancing age. Among the age-associated changes
are: Aged macrophages and neutrophils have im-
paired respiratory burst and reactive nitrogen in-
termediates as a result of altered intracellular sig-
naling, rendering them less able to destroy bacte-
ria. Aged neutrophils are also less able to respond
to rescue from apoptosis. Aged dendritic cells (DC)
are less able to stimulate T and B cells. The altered
T cell stimulation is a result of changes in human
leukocyte antigen expression and cytokine produc-
tion, and lower B cell stimulation is a result of
changes in DC immune complex binding. Natural
killer (NK) cells from the elderly are less capable of
destroying tumor cells. NK T cells increase in num-
ber and have greater interleukin-4 production with
age. Levels of various complement components are
also altered with advancing age. J. Leukoc. Biol.
76: 291–299; 2004.
Key Words: immunosenescence 䡠 macrophage 䡠 dendritic cell
䡠 polymorphonuclear neutrophil
INTRODUCTION
Clinical data have demonstrated that the ability of the elderly
to respond to infection is diminished in comparison with young
adults. As a result, aged individuals make up a disproportion-
ate number of infectious disease patients and are at greater risk
of contracting more severe and longer lasting infections. In
particular, they have higher rates of respiratory tract infections,
which are more likely to be caused by atypical organisms [1, 2].
Moreover, elderly individuals are more prone to developing
invasive Staphyloccus aureus infections and tetanus [3, 4] and
have poor responses to influenza vaccination [5, 6]. The aged
population also has a higher incidence of infections and sepsis
after a traumatic injury than young adults [7]. An age-associ-
ated decline in immunity is implicated as the primary cause of
these problems. This review will examine the effects of aging
on the innate immune system, in particular, examining neutro-
phils, dendritic cells (DC), macrophages, natural killer (NK)
cells, NK T (NKT) cells, and complement.
For the purposes of this review, any study using a human
population 60 years of age or greater is considered elderly, and
groups between 18 and 59 years of age are considered young.
The SENIEUR protocol, which designates young as 25–34
years old and aged as 65 and above, is not exclusively used as
a result of its limitations [8]. The SENIEUR protocol has
extensive exclusion criteria, limiting studies to only the most
healthy individuals. As a consequence, as much as 85% of the
aged population is not included by these studies [9, 10]. These
subjects are excluded based on the concern that any altered
immune responses may be a result of underlying infection or
pathology; however, it is equally possible that this group has an
aging-related, altered immune system that predisposed them to
developing infection or pathology. Given our inability to re-
solve these two possibilities, we have noted when information
is based on studies using the strict SENIEUR protocol guide-
lines.
Concerning animal studies, no concise definition of aged has
been developed. However, Miller and Nadon [11] have outlined
multiple considerations relevant to such a definition; in light of
these factors, rodents, aged 16 months and older, are consid-
ered aged for the purposes of this review. Aging studies that
specifically used animals with cancer, autoimmune disorders,
or other conditions are excluded from this discussion.
POLYMORPHONUCLEAR NEUTROPHILIC
LEUKOCYTES (PMN)
PMN quickly respond to the site of infection and begin to
phagocytose opsonized particles. Consequently, they are
among the first cells to produce bacteriostatic and bacteriocidal
products, as well as influence adaptive immunity. As the body’s
first response to microbial invasion and injury, the chemotactic
migration, adhesion, and phagocytic capabilities of PMN are
critical to their proper functioning. In vivo, examination of skin
abrasion exudate demonstrated equal migration of PMN into
the site of injury in young and old, although there was a greater
variation in migration amongst the elderly subjects meeting the
SENIEUR protocol guidelines compared with the young [12].
When young and old volunteers were compared in an experi-
1
Correspondence: Department of Cell Biology, Neurobiology, and Anatomy,
Department of Surgery, Loyola University Chicago, Stritch School of Medicine,
Building 110, Room 4237, 2160 South First Avenue, Maywood, IL 60513.
E-mail: ekovacs@lumc.edu
Received November 25, 2003; accepted February 16, 2004; doi: 10.1189/
jlb.1103592.
Journal of Leukocyte Biology Volume 76, August 2004 291
mental gingivitis model, an equal number of PMN were de-
tected in the junctional epithelium 7 days after injury [13]. In
vitro research has yielded more conflicting results. Corberand
et al. [14] reported chemotaxis to be significantly decreased in
people over the age of 80 years, and there was no significant
difference in 60- to 70-year-olds compared with young volun-
teers. The very opposite was true in a study by Niwa et al. [15],
which found a correlation between 60- to 70-year-old volun-
teers with diminished PMN chemotaxis and respiratory burst
and their mortality 7 years after the initial study. However, the
study demonstrated no difference between people older than 80
years old and the young. They suggested that the lack of
significant difference between the over-80-year-old population
and the youth was a result of a selective pressure that left only
those individuals with the “healthiest” PMN surviving into the
oldest age group.
Once PMN have migrated to the site of injury or bacterial
entry, they must adhere to the endothelium to diapedis out of
the vasculature. Adhesion, however, is not impaired in the
elderly. When stimulated by f-Met-Leu-Phe (fMLP), opsonized
zymosan, phorbol myristate acetate (PMA), or calcium iono-
phore A23187, human PMN obtained from young and aged
subjects adhere equally to plastic, gelatin, and bovine aortic
endothelium [12, 16].
Phagocytosis is also unimpaired in the elderly. Studies show
that the amount of opsonized paraffin oil or yeast cells engulfed
is equal in PMN from young and aged volunteers [15, 17]. It is
interesting that another study with yeast cells showed an in-
crease in phagocytosis by the elderly in comparison to 20- to
29-year-olds; however, no difference between the elderly and
30- to 60-year-olds [14] was noted. These combined results
indicated that there is no impairment in the ability of PMN
from aged subjects to traffic to tissues and engulf microbes.
In contrast to phagocytosis, the microbiocidal capacity of
PMN is significantly decreased with advancing age. The ability
of stimulated and unstimulated PMN to kill Candida albicans
is attenuated by 10 –50% in the elderly [14, 18], and Esche-
ricihia coli killing is 44% lower than that of young subjects
[19]. Contributing to this depressed killing ability is a signif-
icant decrease in the production of reactive oxygen species
(ROS) by PMN. Following stimulation with fMLP, interferon-␥
(IFN-␥), granulocyte/monocyte-colony stimulating factor (GM-
CSF), or lipopolysaccharide (LPS) [18 –22], the production of
superoxide was greatest by PMN from young rather than aged
humans and rodents. Likewise, nitroblue tetrazolium dye-re-
duction tests have yielded higher rates of oxidative dye reduc-
tion by a mixture of young, mature, human neutrophilic gran-
ulocytes and macrophages [14].
Impaired intracellular signaling has been implicated as a
leading cause for the altered oxygen-free radial production in
the aged. Intracellular Ca2⫹ is decreased in stimulated PMN
from elderly volunteers, suggesting that there is an impairment
in Ca2⫹ flux during cell signaling [18, 23]. Also, actin, which
may play a role in cell-surface receptor movement and expres-
sion, has been suggested to contribute to the altered free-
radical production. Using actin-stabilizing and -disrupting
agents, Piazzolla et al. [20] demonstrated that the release of
O2– by PMN from elderly volunteers is more sensitive to these
agents, yielding significantly lower superoxide release. Addi-
292 Journal of Leukocyte Biology Volume 76, August 2004
tionally, actin polymerization is significantly diminished after
stimulation of PMN from aged subjects with fMLP or PMA,
relative to young [24, 25]. These age-associated differences in
actin correlate with altered cell-surface markers. In particular,
there is lower surface expression of the activation-inducer
molecule CD69 and chemotactic peptide receptor for N-formyl-
Nle-Leu-Phe,Tyr-Lys hexapeptide on aged PMN after stimula-
tion, and no differences are noted in CD11b or complement
receptor b [25, 26]. These results, taken in their totality,
suggest that the diminished production of ROS may be caused
partially by impaired intracellular signaling, resulting from an
inability of actin to adequately transport the appropriate cell-
surface receptors to the cell membranes of PMN from the aged.
Given the short life-span of a PMN, alterations in apoptotic
cell death may also predispose the elderly to an increased risk
of infection. In the absence of stimulation, the amount of
apoptotic cell death of human PMN is unaltered by aging
[27–29], and there is no difference in the expression of CD95,
APO1/Fas, an apoptotic ligand [30]. In contrast, there are
significant variations in apoptosis following cell stimulation.
Under normal conditions, in the young, inflammatory mediators
are able to prevent apoptosis [31–34]. This response to inflam-
mation helps guarantee the continued involvement of PMN in
preventing the spread of microbes, while other cell populations
traffic to the site of injury or insult. However, unlike with the
young, interleukin (IL)-2, LPS, GM-CSF, or G-CSF do not
rescue PMN from the elderly [28, 29]. This is paralleled by
alterations in the Janus tyrosine kinase (Jak)2-signal trans-
ducer and activator of transcription (STAT)5 signaling pathway
in PMN from the elderly. Ultimately, this results in aged PMN
producing increased Bax and decreased Mcl-1, creating a
proapoptotic environment [35].
The failure of inflammatory mediators to prevent apoptosis in
the elderly may also reflect an inability of the cells to prevent
reactive oxygen-induced damage. Tortorella et al. [27] demon-
strated that high levels of superoxide dismutase were better
able to inhibit apoptosis in PMN from young volunteers than
from aged; however, the addition of catalase raised the apo-
ptosis inhibition of the aged subjects to a similar level as the
young. As inflammatory mediators not only prevent apoptosis
but also stimulate the production of ROS in the young [36, 37],
these results suggest that in an inflammatory environment, the
PMN from the aged are less able to neutralize free radicals that
escape from phagolysosomes and will be substantially dam-
aged. As a result of this damage, these cells are shifted toward
a proapoptotic environment and instead undergo programmed
cell death.
DC
Between a few hours to several days after PMN migration,
antigen-presenting cells (APCs) enter the site of infection and
inflammation. DC are the most potent, professional APC of the
immune system [38]. With their ability to interact with B and
T cells and their widespread localization, they are a vital
component of the innate immune system.
The follicular DC (FDC) is critical to the formation of plasma
cells in the germinal centers of secondary lymphoid organs and
http://www.jleukbio.org
the subsequent generation of antibodies [39]. Szakal and co-
workers [40, 41] reported that decreases in the cell-surface
receptors for complement, CD21, and Fc␥ in aged mice im-
paired the initial trapping of immune complexes by FDC. This
is further complicated by a decrease in the number of icco-
somes, antigen:antibody complexes that bud off FDC and are
taken up by B cells [40]. The combined effect is less-stable
binding and decreased stimulation of B cells. Deficient anti-
body production can be partially restored in vitro by using FDC
that have a higher percentage of immune complexes and com-
plement bound [40, 41]. In addition, FDC of aged mice often
remained trapped in the subcapsular sinus of lymph nodes and
are unable to reach the follicle and its B cells, resulting in
decreased germinal center formation [40, 42]. These changes
in FDC are partially responsible for the significant modification
of humoral immunity in the elderly [43].
Non-FDC, hereafter referred to simply as DC, are capable of
activating CD4⫹ T cells via their major histocompatibility
complex class II (MHC-II) [44]. DC from young and aged
humans induce similar amounts of T cell proliferation and have
similar levels of human leukocyte antigen (HLA) expression
[45, 46]. However, lower HLA-DR expression has also been
reported [47]. In the former studies, DC were obtained by
stimulating monocytes with GM-CSF, and the latter study
analyzed peripheral blood DC. Differences in these protocols
may account for the opposing results and suggest that there is
a need for careful interpretation of ex vivo experiments. These
different results also suggest that although circulating DC from
the elderly are less prepared to bind antigens, the deficiency
can be corrected in the appropriate environment. Animal mod-
els have also encountered conflicting information. T cell pro-
liferation was attenuated when cultured with DC from aged
DBA or A mice, and there was no difference with BALB/c,
CBA, or C3H/He mouse DC [48]. Reasons for these differences
remain to be explored. Aside from T cell proliferation, aged,
murine DC are also less able to stimulate peripheral blood
mononuclear cell (PBMC) production of IFN-␥, further con-
tributing to the changes in T cell functionality with advancing
age [49].
Within the peripheral blood of the elderly, there is nearly
50% less CD11c⫹B220⫹ plasmacytoid DC [50]. However, it is
unknown if there are differences in the CD11c⫹B220–CD8⫹
lymphoid or CD11c⫹B220–CD8– myeloid DC subsets with
advancing age. This decrease in DC exists despite an increase
in the proliferation of peripheral DC from the elderly after in
vivo GM-CSF incubation when compared with the young [45].
The proliferation studies, however, use the SENIEUR protocol
and did not account for age-related differences in circulating
GM-CSF, whose levels are lower in aged animals; therefore,
stimulating PBMC from young and aged subjects with equiv-
alent concentrations of GM-CSF does not accurately reflect the
natural microenviroment [51, 52]. The lower circulating GM-
CSF of the elderly may result in less DC proliferation. Addi-
tionally, it has been noted that in vitro and in vivo models of
DC function have resulted in divergent results [53]. Conse-
quently, although there is a decrease in absolute number of
DC, further research is needed to determine if DC retained
their proliferative capacity with advancing age and what effects
lower circulating GM-CSF levels may have on DC function.
MACROPHAGES
Macrophages play important roles in the immune response,
capable of not only phagocytosis and destruction of foreign
cells and debris but also acting as APCs. Like DC, they stand
at the crossroads between adaptive and innate immunity.
Macrophages are able to directly destroy microbes through
the products of the respiratory burst and reactive nitrogen-
intermediate pathways. Both of these processes are induced by
IFN-␥ from T cells and NK cells or by components of bacterial
cell walls [54, 55]. The binding of IFN-␥ or bacterial products
to their appropriate receptors results in phosphorylation of
mitogen-activated protein kinase (MAPK) and the subsequent
release of free radicals. Studies using rats have demonstrated a
75% decrease in the ability of macrophages from aged animals
to produce superoxide anion following incubation with IFN-␥
or opsonized zymosan [56]. The impaired superoxide produc-
tion by macrophages from the aged was further investigated by
Ding et al. [57], who demonstrated that caseinate-elicited
peritoneal macrophages from aged mice had undetectable
phosphorylation of MAPK following incubation of the cells with
IFN-␥. Functionally, this abrogation in MAPK activation was
correlated with 50% less hydrogen peroxide production follow-
ing stimulation of macrophages from aged mice with PMA or
opsonized zymosan. Others [58 – 60] have also reported a sim-
ilar decrease in oxygen free-radial production by macrophages
from aged mice. As a consequence of reduced respiratory burst
in the elderly, the intercellular killing of bacteria is hindered
and thus may cause the elderly to have infections of longer
duration [61– 63].
The production of reactive nitrogen intermediates is the
other key mechanism by which macrophages exert their mi-
crobicidal properties; however, studies into the production of
nitrogen compounds in the aged offer conflicting results. Sev-
eral reports demonstrated that inducible nitric oxide synthase
(iNOS) mRNA is decreased in aged mice [64, 65]. Addition-
ally, in the absence of iNOS, aged mice [64 – 66] significantly
decreased the production of NO2– and other intermediates.
Conversely, enhanced iNOS mRNA and nitrite production by
macrophages of the aged have also been reported. Chen et al.
[67] demonstrated an increase in nitrite production in resident
and thioglycollate-elicited peritoneal macrophages of aged
mice after LPS stimulation, as well as increased iNOS mRNA
in thioglycollate-elicited peritoneal macrophages. Others [68]
have also reported similar increases. The conflicting reports
are likely a result of differences in experimental protocols.
Recently, it was demonstrated that thioglycollate-elicited mac-
rophages stimulated with LPS and IFN-␥ display different
age-specific nitrite production patterns based on the dose of
IFN-␥ with which the cells were cultured [69]. At low doses of
IFN-␥, the production of nitrite was higher in young mice than
in old mice. However, at a higher dose, the macrophage from
older mice had a greater production of nitrite than the young
mice. Therefore, interpretation and application of the results
require a consideration of the microenvironment in which the
cells function.
Macrophages from the aged can also contribute to the dys-
regulation seen in other immune cell populations (Fig. 1). The
production of PGE2 by macrophages is enhanced in aged mice,
Plackett et al. Innate immunity in aging 293

Fig. 1. Changes in macrophage signaling, enzymatic processes, and output
with advancing age. Arrows (1 and 2) indicate increased or decreased levels
in the elderly compared with young. TNF-␣, Tumor necrosis factor ␣; JNK,
c-jun NH2-terminal kinase; TLR4, Toll-like receptor 4; COX, cyclooxygenase;
PGE2, prostaglandin E2.
which correlates with an increase in COX-2 mRNA and en-
zyme levels [70]. PGE2 is able to alter DC function by blocking
the production of IL-12, increasing production of IL-10, and
decreasing MHC-II expression in mice [71–73]. It also exerts
effects on T cells. The arachidonic acid metabolite is capable
of decreasing IL-2 production and subsequently lowering T
cell-proliferative responses [74, 75], two well-documented
changes seen with advancing age [76 –78]. Up-regulation of T
helper cell type 2 (Th2) cytokines, which are known to be
elevated in elderly human patients and in animal models of
aging [76, 79, 80], is also a result of mouse splenocyte stim-
ulation with PGE2 [74].
Macrophage-derived cytokines also affect the immune re-
sponse. Several recent reports have suggested that there is a
decrease in the production of proinflammatory cytokines by
macrophages from aged humans and mice. Renshaw et al. [81]
showed that LPS stimulation of splenic and thioglycollate-
elicited peritoneal macrophages resulted in less IL-6 and
TNF-␣ secretion by macrophages from aged mice compared
with young mice. Furthermore, the study suggested that lower
proinflammatory cytokine differences may be a result of lower
TLR4 mRNA in macrophages of aged mice, rendering the
macrophages less responsive to LPS. Lower IL-6 and TNF-␣
production by LPS-stimulated macrophages is also associated
with decreased, activated p38 and JNK MAPKs [82]. Beharka
et al. [83] demonstrated a decrease in the production of IL-6 by
human PBMC cultured with autologous plasma. Most litera-
ture, however, suggests that circulating levels of proinflamma-
tory cytokines are elevated in the aged [84 – 87]. Macrophages
have been assumed to be the primary producer of these cyto-
kines. Several studies have demonstrated that in fact, under-
lying inflammatory disease and poor nutrition may actually be
responsible for this circulating, proinflammatory state, rather
than the natural aging process [88 –91]. Also, although mac-
rophages have been assumed to be a primary producer of the
proinflammatory cytokines in the elderly, other cell popula-
tions may contribute to cytokine production as well. In light of
the more recent findings, further research is needed to deter-
294 Journal of Leukocyte Biology Volume 76, August 2004
mine which cell population is responsible for potential height-
ened inflammatory states.
Wound healing is dependent on macrophage function, as
macrophages are able to keep the wound bed free from infec-
tion and promote angiogenesis [92]. With its dual roles, mac-
rophage migration to the site of injury can be critical in the
post-injury immune response. After injury, chemokines, in-
cluding monocyte chemoattractant protein-1 (MCP-1), are pro-
duced to promote the migration of monocytes and macrophages
into the wound bed. Following excisional wounding, macro-
phages begin to infiltrate into the wound bed within 24 h [93].
The infiltration is significantly greater in aged mice and cor-
relates with increased MCP-1 in wound homogenates 12 h after
injury, relative to young mice. However, burn injury is associ-
ated with decreased MCP-1 by aged, injured mice, relative to
young, injured mice, 24 h after injury. However there was
equal macrophage accumulation in the region immediately
adjacent to the necrotic, burn-injured tissue in young and aged,
injured mice [94]. The differences between the results of these
two studies may reflect variations in the experiment models.
Burn injury is associated with significant increases in proin-
flammatory cytokines in aged mice [85], which may alter
macrophage migration. Studies of excisional wound healing in
humans demonstrate a delay in monocyte and macrophage
infiltration with age [95]. The slow infiltration is associated
with a decrease in intercellular adhesion molecule-1 and vas-
cular adhesion molecule-1 expression, suggesting an impair-
ment in the ability of the cells to bind to the endothelium to
leave the circulation [95].
Once at the site of injury, macrophages can promote angio-
genesis and wound healing through the production of vascular
endothelial growth factor (VEGF). This angiogenic mediator is
present at lower levels in excisional wounds from aged mice,
which correlates with 37% less VEGF production by stimu-
lated peritoneal macrophages from aged mice [96]. The dimin-
ished VEGF production and angiogenesis may delay wound
closure, allowing more time for bacteria to circumvent the
natural skin barrier and develop into a full infection. With only
isolated studies exploring the role of macrophages in wound
healing of the elderly, further research is needed before defin-
itive conclusions can be reached.
NK CELLS
Although PMN, DC, and macrophages are capable of seeking
out and eliminating extracellular pathogens, NK cells are crit-
ical to removal of intracellular pathogens. In addition to their
importance in viral infections, they also possess vital tumori-
cidal activities.
There is an increase in the relative percentage of NK cells
with advancing age when comparing SENIEUR protocol-qual-
ified elderly and young individuals [97–99]. The rise in per-
centage of NK cells in the elderly is related to a decrease in the
number of T cells and an increase in the absolute number of
NK cells [30, 97, 100 –102]. Aside from an increase in number,
NK cells obtained from the aged are more likely to have a
mature phenotype, having a higher percentage and absolute
number of CD56dim cells and a smaller CD56bright population
http://www.jleukbio.org
[100, 103]. Additionally, NK cells from the aged may be less
prepared to carryout their prescribed function, as there is a
significant increase in the number of agranular cells in humans
[97] and a decrease in the adherence to tumor cells in mice
[104].
Conflicting reports exist as to the cytolytic ability of NK cells
in the elderly. Independent studies demonstrated that NK cells
in the elderly have normal [105] or enhanced activity [106]
compared with young individuals when the elderly population
is selected based on strict adherence to the SENIEUR protocol.
However, when elderly patients were selected for good health
but did not fully meet the SENIEUR protocol, the NK cell
activity against tumor cells was significantly decreased com-
pared with young adults. Other human studies, which more
loosely followed the suggestion of the SENIEUR protocol, also
demonstrated an age-specific decrease in NK cell cytotoxicity
toward tumor cells [98, 107, 108], and similar results in animal
models used LPS or concanavalin A as cell stimulants [109 –
112]. These reports indicate that although fully functional NK
cells may be desirable for optimal health, the majority of the
elderly population will experience some deficit in NK cell
activity. This also highlights the need for careful interpretation
and application of SENIEUR protocol-based information.
Alterations in intracellular signaling are key contributors to
the tumoricidal deficiency [102]. In particular, there is a delay
in phosphatidlyinositol biphosphate hydrolysis, coupled with a
failure of inositol triphosphate to increase above basal levels
[102]. However, these results were specific to spontaneous
cytolytic activity directed toward tumor cells, as antibody-
dependent, cell-mediated cytotoxicity (ADCC), assayed by an-
ti-CD16 antibody stimulation of the Fc receptor, was intact in
the aged subjects. Likewise, the production of insositol phos-
phate activity was comparable with the young during ADCC.
Others have also reported an intact ADCC [113, 114]. The
impaired, spontaneous cytolytic activity allows for continued
growth and expansion of neoplastic cells and is likely a con-
tributor to the increased incidence of cancer among the elderly
[115, 116].
In addition to decreased toxicity, the NK cells from the
elderly are unable to properly proliferate following IL-2 stim-
ulation. In human- and murine-derived NK cells, the prolifer-
ative response to IL-2 is decreased by 40 – 60% among the
elderly [100, 104]. The decreased, proliferative response is
paralleled by an age-specific decrease in Ca2⫹ mobilization
[100]. The diminished Ca2⫹ prevented the up-regulation of
CD69 on elderly human NK cells, an early activator of NK cell
proliferation and cytotoxicity. The combined effect of dimin-
ished proliferation responses and cytotoxic activity of NK cells
is that the elderly are more prone to developing longer lasting
infections and a decreased ability to rid the body of tumor cells
[117, 118].
NKT CELLS
NKT cells are a minor cell population found in most lymphoid
organs. They are characterized as CD1d-restricted cells [119]
and have been implicated as a significant contributor to post-
injury responses [120], microbial clearance [121], autoimmune
disease [122], and other immune responses [123, 124].
Absolute numbers and relative percentages of NKT cells
increase in number with advancing age. The increase in NKT
cells appears to be a global rise, as increases of 150 – 400%
have been reported in circulating, splenic, hepatic, mesenteric
lymph node, and peripheral lymph node counts in humans and
rodents [125–128]. Cytokine production by NKT cells has also
varied with aging. Dubey et al. [129] showed an increase in
production of IL-4 mRNA and protein by CD4⫹ NKT cells
from aged B.10 mice and a concurrent decrease in IFN-␥
mRNA and protein after stimulation of the cells with ␣CD3 and
IL-2; however, the aged group included mice as young as 12
months old [129]. In contrast, Poynter et al. [128] found that
␣CD3 stimulation of NKT cells leads to lower IL-4 and IL-2
mRNA in aged C57BL/6 mice. Although both studies con-
cluded that there is a decrease in Th1 cytokine mRNA, the
discrepancy in IL-4 remains to be resolved.
Expression of Fas ligand following cell stimulation displays
an age-related effect as well. Younger mice express lower
levels of Fas ligand after injection of ␣-GalCer into the mice
[130]. Studies with young Fas-deficient and wild-type mice
have demonstrated that injection of ␣-GalCer results in an
increase in NKT cell apoptosis and that the cell death is
Fas-dependent [131]. However, whether the increased Fas
ligand in aged mice exerts an effect on NKT apoptosis is
unknown.
COMPLEMENT
Most studies examining the relationship of age and comple-
ment focused on the role of C1q in Alzheimer’s disease [132–
134]; however, a limited number of studies compare the levels
of complement proteins and their functionality in the young
and aged. When considering the classical complement path-
way, serum levels of C4 were increased in aged volunteers
[135–137] or unchanged between the young and old [138].
These differences in C4 protein may reflect the use of the
SENIEUR protocol by Bellavia et al. [138]. In an isolated
study, there was an increase in the plasma levels of C4-binding
protein (C4BP) with age [139]. However, interpretation of this
datum is limited, as the study does not state which age groups
were compared. The role that the age-associated increase in
C4BP has on immunity is also uncertain, as the increase is
attributed to a rise in C4BP␤. The C4BP␤ subunit interacts
with the coagulation cascade, whereas C4BP␣ specifically
binds to C4b and inhibits complement activation [140]. Using
a rat model, Lavery and Goyns [141] described a decrease in
the expression of the C4BP␣ in the livers of aged animals;
however, it is unknown whether this observation is organ- or
species-specific.
Description of age-related changes in the alternative com-
plement pathway has also yielded conflicting results. Serum C3
levels were reported to be increased [135, 137] in the elderly
or equivalent in young and aged [136, 138]. Additionally,
Factor B levels in elderly humans are either decreased [135] or
not significantly different from the young [136].
Plackett et al. Innate immunity in aging 295
 TABLE 1. Significant Changes with Advancing Age
Cell population Changes with aging References
Neutrophils 2 Reactive oxygen species [19,
1 Apoptosis [33,
Dendritic cells 2 B cell stimulation [40,
2 Number of plasmacytoid DC [50]
Macrophages 2 Reactive oxygen species [56,
2 Reactive nitrogen intermediates [65]
1 Prostaglandin E2 [70]
2 Interleukin-6 [81,
Natural killer
cells 1 Number of NK cells [30]
2 Tumor-killing activity [108, 109]
Natural killer T
cells 1 Number of NKT cells [125, 126]
As with the components unique to the classic and alternative
complement pathways, research into the common pathway
components is also extremely limited and fraught with contra-
dictions. Cannon et al. [142] found equivalent rises of C3a
anaphylatoxin in plasma obtained from young and old humans
after exercise-induced tissue damage, whereas Kyrkanides et
al. [143] showed an increase in C3a1 mRNA in aged mice after
a cortical stab injury. The ultimate significance of either of
these results remains to be demonstrated.
Functional studies of complement activity indicate normal
functioning or a decrease in activity with advancing age. Haz-
lett et al. [144] reported a 50% decrease in alternative path-
way-induced hemolysis by aged Swiss ICR mice. The impaired,
alternative pathway was also associated with a decrease in the
number of PMN undergoing phagocytosis of Pseudomonas
aeruginosa. However, there was no difference in the total
amount of bacteria remaining after the incubation. Using the
SENIEUR protocol, Bellavia et al. [138] found no difference in
the ability of the alternative or classical pathway to induce
hemolysis in aged versus young individuals. Given the limited
number of studies examining complement functionality, further
research is needed before definitive conclusions can be drawn.
CONCLUSION
This review has provided an in-depth examination of the cur-
rent knowledge concerning aging and its effects on innate
immunity. Contrary to prior beliefs, the innate immune system
is not free from the effects of aging. Although the age-associ-
ated decrease in T and B cell populations allows for an in-
crease in the percentage of innate cell, like their adaptive cell
counterparts, the innate cells do not function normally (Table
1). The production of ROS and nitrogen species is significantly
impaired in neutrophils and macrophages from the aged, pos-
sibly affecting bacterial destruction, and NK cells are less able
to destroy tumor cells. There is a significant increase in the
number of NKT cells in the aged, but further research is
needed to determine the functional significance. Likewise,
complement research remains a vastly understudied area of
aging research. Finally, the innate immune system also con-
tributes to altered acquired immunity with aging, via interac-
296 Journal of Leukocyte Biology Volume 76, August 2004
tions between DC and macrophages with T and B cells. Thus,
the effect of aging on innate immunity is not merely an increase
in relative percentage of innate immune cells, but it is a
20, 22] dynamic system reflecting vast changes in all of its many
35] components.
43]
60]
ACKNOWLEDGMENTS
82]
We thank Dr. Pamela Witte, Director of the Immunology and
Aging Program (Loyola University, Maywood, IL), for her crit-
ical review of the manuscript and thoughtful discussion. NIH
R01 AG18859 supported this work.
REFERENCES
1. Gotfried, M. H. (2001) Epidemiology of clinically diagnosed community-
acquired pneumonia in the primary care setting: results from the 1999 –
2000 respiratory surveillance program. Am. J. Med. 111, 25S–29S.
2. Ruiz, M., Santiago, E., Marcos, M. A., Martinez, J. A., Arancibia, F.,
Mensa, J., Torres, A. (1999) Etiology of community-acquired pneumonia:
impact of age, comorbidity, and severity. Am. J. Respir. Crit. Care Med.
160, 397– 405.
3. Laupland, K. B., Church, D. L., Mucenski, M., Sutherland, L. R., Davies,
H. D. (2003) Population-based study on the epidemiology of and the risk
factors for invasive Staphylococcus aureus infections. J. Infect. Dis. 187,
1452–1459.
4. Pascual, F. B., McFinley, E. L., Zanardi, L. R., Cortese, M. M., Murphy,
T. V. (2003) Tetanus surveillance–United States, 1998 –2000. MMWR
Surveill. Summ. 52, 1– 8.
5. Bridges, C. B., Harper, S. A., Fukuda, K., Uyeki, T. M., Cox, N. J.,
Singleton, J. A., Advisory Committee on Immunization Practices. (2003)
Prevention and control of influenza. Recommendations of the Advisory
Committee on Immunization Practices (ACIP). MMWR Recomm. Rep.
52, 1–34.
6. Potter, J. M., O’Donnell, B., Carman, W. F., Roberts, M. A., Stott, D. J.
(1999) Serological response to influenza vaccination and nutritional and
functional status of patients in geriatric medical long-term care. Age
Ageing 28, 141–145.
7. Linn, B. S. (1980) Age differences in the severity and outcomes of burns.
J. Am. Geriatr. Soc. 28, 118 –123.
8. Ligthart, G. J., Corberand, J. X., Fournier, C., Galanaud, P., Hijmans,
W., Kennes, B., Muller-Hermelink, H. K., Steinmann, G. G. (1984)
Admission criteria for immunogerontological studies in man: the
SENIEUR protocol. Mech. Ageing Dev. 28, 47–55.
9. Ligthart, G. J., Corberand, J. X., Geertzen, H. G., Meinders, A. E.,
Knook, D. L., Hijmans, W. (1990) Necessity of the assessment of health
status in human immunogerontological studies: evaluation of the
SENIEUR protocol. Mech. Ageing Dev. 55, 89 –105.
10. Traill, K. N., Schonitzer, D., Jurgens, G., Bock, G., Pfeilschifter, R.,
Hilchenbach, M., Holasek, A., Forster, O., Wick, G. (1985) Age-related
changes in lymphocyte subset proportions, surface differentiation antigen
density and plasma membrane fluidity: application of the Eurage
SENIEUR protocol admission criteria. Mech. Ageing Dev. 33, 39 – 66.
11. Miller, R. A., Nadon, N. L. (2000) Principles of animal use for geronto-
logical research. J. Gerontol. A Biol. Sci. Med. Sci. 55, B117–B123.
12. Biasi, D., Carletto, A., Dell’Agnola, C., Caramaschi, P., Montesanti, F.,
Zavateri, G., Zeminian, S., Bellavite, P., Bambara, L. M. (1996) Neutro-
phil migration, oxidative metabolism, and adhesion in elderly and young
subjects. Inflammation 20, 673– 681.
13. Fransson, C., Mooney, J., Kinane, D. F., Berglundh, T. (1999) Differ-
ences in the inflammatory response in young and old human subjects
during the course of experimental gingivitis. J. Clin. Periodontol. 26,
453– 460.
14. Corberand, J., Ngyen, F., Laharrague, P., Fontanilles, A. M., Gleyzes, B.,
Gyrard, E., Senegas, C. (1981) Polymorphonuclear functions and aging in
humans. J. Am. Geriatr. Soc. 29, 391–397.
15. Niwa, Y., Kasama, T., Miyachi, Y., Kanoh, T. (1989) Neutrophil chemo-
taxis, phagocytosis and parameters of reactive oxygen species in human
aging: cross-sectional and longitudinal studies. Life Sci. 44, 1655–1664.
http://www.jleukbio.org
16. Damtew, B., Spagnuolo, P. J., Goldsmith, G. G., Marino, J. A. (1990)
Neutrophil adhesion in the elderly: inhibitory effects of plasma from
elderly patients. Clin. Immunol. Immunopathol. 54, 247–255.
17. Fulop, T., Foris, G., Leovey, A. (1984) Age-related changes in cAMP and
cGMP levels during phagocytosis in human polymorphonuclear leuko-
cytes. Mech. Ageing Dev. 27, 233–237.
18. Seres, I., Csongor, J., Mohacsi, A., Leovey, A., Fulop, T. (1993) Age-
dependent alterations of human recombinant GM-CSF effects on human
granulocytes. Mech. Ageing Dev. 71, 143–154.
19. Fu, Y-K., Arkins, S., Li, Y. M., Dantzer, R., Kelley, K. W. (1994)
Reduction in superoxide anion secretion and bactericidal activity of
neutrophils from aged rats: reversal by combination of ␥ interferon and
growth hormone. Infect. Immun. 62, 1– 8.
20. Piazzolla, G., Tortorella, C., Serrone, M., Jirillo, E., Antonaci, S. (1998)
Modulation of cytoskeleton assembly capacity and oxidative response in
aged neutrophils. Immunopharmacol. Immunotoxicol. 20, 251–266.
21. Tortorella, C., Ottolenghi, A., Pugliese, P., Jirillo, E., Antonaci, S. (1993)
Relationship between respiratory burst and adhesiveness capacity in
elderly polymorphonuclear cells. Mech. Ageing Dev. 69, 53– 63.
22. Tortorella, C., Polignano, A., Piazzolla, G., Serrone, M., Jirillo, E.,
Antonaci, S. (1996) Lipopolysaccharide-, granulocyte-monocyte colony
stimulating factor- and pentoxifylline-mediated effects on formyl-methio-
nyl-leucine-phenylalanine-stimuated neutrophil respiratory burst in the
elderly. Microbios 85, 189 –198.
23. Fulop, T., Varga, Z., Jacob, M. P., Robert, L. (1997) Effect of lithium on
superoxide production and intracellular free calcium mobilization in
elastin peptide (␬-elastin) and FMLP stimulated human PMNs. Effect of
age. Life Sci. 60, 325–332.
24. Rao, K. M. K. (1986) Age-related decline in ligand-induced actin poly-
merization in human leukocytes and platelets. J. Gerontol. 41, 561–566.
25. Rao, K. M. K., Currie, M. S., Padmanabhan, J., Cohen, H. J. (1992)
Age-related alteration in actin cytoskeleton and receptor expression in
human leukocytes. J. Gerontol. 47, B37–B44.
26. Noble, J. M., Ford, G. A., Thomas, T. H. (1999) Effect of aging on CD11b
and CD69 surface expression by vesicular insertion in human polymor-
phonuclear leukocytes. Clin. Sci. 97, 323–329.
27. Tortorella, C., Piazzolla, G., Spaccavento, F., Jirillo, E., Antonaci, S.
(1999) Age-related effects of oxidative metabolism and cyclic AMP
signaling on neutrophil apoptosis. Mech. Ageing Dev. 110, 195–205.
28. Tortorella, C., Piazzolla, G., Spaccavento, F., Pece, S., Jirillo, E., An-
tonaci, S. (1998) Spontaneous and Fas-induced apoptotic cell death in
aged neutrophils. J. Clin. Immunol. 18, 321–329.
29. Fulop, T., Fouquet, C., Allaire, P., Perrin, N., Lacombe, G., Stankova, J.,
Rola-Pleszczynski, M., Gagne, D., Wagner, J. R., Khalil, A., Dupuis, G.
(1997) Changes in apoptosis of human polymorphonuclear granulocytes
with aging. Mech. Ageing Dev. 96, 15–34.
30. Di Lorenzo, G., Balistreri, C. R., Candore, G., Cigna, D., Colombo, A.,
Romano, G. C., Colucci, A. T., Gervasi, F., Listi, F., Potestio, M., Caruso,
C. (1999) Granulocyte and natural killer activity in the elderly. Mech.
Ageing Dev. 108, 25–38.
31. Colotta, F., Re, F., Polentarutti, N., Sozzani, S., Mantovani, A. (1992)
Modulation of granulocyte survival and programmed cell death by cyto-
kines and bacterial products. Blood 80, 2012–2020.
32. Brach, M. A., de Vos, S., Gruss, H. J., Herrmann, F. (1992) Prolongation
of survival of human neutrophils by granulocyte-macrophage colony-
stimulating factor is caused by inhibition of programmed cell death.
Blood 80, 2920 –2924.
33. Pericle, F., Liu, J. H., Diaz, J. I., Blanchard, D. K., Wei, S., Forni, G.,
Djeu, J. Y. (1994) Interleukin-2 prevention of apoptosis in human
neutrophils. Eur. J. Immunol. 24, 440 – 444.
34. Lee, A., Whyte, M. K. B., Haslett, C. (1993) Inhibition of apoptosis and
prolongation of neutrophil functional longevity by inflammatory media-
tors. J. Leukoc. Biol 54, 283–288.
35. Fulop, T., Larbi, A., Linteau, A., Desgeorges, S., Douziech, N. (2002)
The role of Mcl-1 and Bax expression alteration in the decreased rescue
of human neutrophils from apoptosis by GM-CSF with aging. Ann. N. Y.
Acad. Sci. 973, 305–308.
36. Power, C., Wang, J. H., Sookhai, S., Wu, Q. D., Redmond, H. P. (2001)
Proinflammatory effects of bacterial lipoprotein on human neutrophil
activation status, function, and cytotoxic potential in vitro. Shock 15,
461– 466.
37. Mansfield, P. J., Minkovska-Galcheva, V., Carey, S. S., Shayman, J. A.,
Boxer, L. A. (2002) Regulation of polymorphonuclear leukocyte degran-
ulation and oxide production by ceramide through inhibition of phospho-
lipase D. Blood 99, 1434 –1441.
38. Li, J., Schuler-Thurner, B., Schuler, G., Huber, C., Seliger, B. (2001)
Bipartite regulation of different components of the MHC class I antigen-
processing machinery during dendritic cell maturation. Int. Immunol.
13, 1515–1523.
39. Zhang, X., Li, L., Jung, J., Xiang, S., Hollmann, C., Choi, Y. S. (2001)
The distinct roles of T cell-derived cytokines and a novel follicular
dendritic cell-signaling molecule 8D6 in germinal center-B cell differ-
entiation. J. Immunol. 167, 49 –56.
40. Szakal, A. K., Taylor, J. K., Smith, J. P., Kosco, M. H., Burton, G. F.,
Tew, J. J. (1990) Kinetics of germinal center development in lymph nodes
of young and aging immune mice. Anat. Rec. 227, 475– 485.
41. Aydar, Y., Balogh, P., Tew, J. G., Szakal, A. K. (2002) Age-related
depression of FDC accessory functions and CD21 ligand-mediated repair
of co-stimulation. Eur. J. Immunol. 32, 2817–2826.
42. Holmes, K. L., Schnizlein, C. T., Perkins, E. H., Tew, J. G. (1984) The
effect of age on antigen retention in lymphoid follicles and in collagenous
tissue of mice. Mech. Ageing Dev. 25, 243–255.
43. Szakal, A. K., Aydar, Y., Balogh, P., Tew, J. G. (2002) Molecular
interactions of FDCs with B cells in aging. Semin. Immunol 14, 267–
274.
44. Granucci, F., Feau, S., Zanoni, I., Pavelka, N., Vizzardelli, C., Raimondi,
G., Ricciardi-Castagnoli, P. (2003) The immune response is initiated by
dendritic cells via interaction with microorganisms and interleukin-2
production. J. Infect. Dis. 187, S346 –S350.
45. Steger, M. M., Maczek, C., Grubeck-Loebenstein, B. (1996) Morpholog-
ically and functionally intact dendritic cells can be derived from the
peripheral blood of aged individuals. Clin. Exp. Immunol. 105, 544 –
550.
46. Lung, T. L., Saurwein-Teissl, M., Parson, W., Schonitzer, D., Grubeck-
Loebenstein, B. (2000) Unimpaired dendritic cells can be derived from
monocytes in old age and can mobilize residual function in senescent T
cells. Vaccine 18, 1606 –1612.
47. Pietschmann, P., Hahn, P., Kudlacek, S., Thomas, R., Peterlik, M. (2000)
Surface markers and transendothelial migration of dendritic cells from
elderly subjects. Exp. Gerontol. 35, 213–214.
48. Komatsubara, S., Cinader, B., Muramatsu, S. (1986) Polymorphism of
age-related changes in stimulatory capacity of murine dendritic cells.
Mech. Ageing Dev. 37, 163–173.
49. Looney, R. J., Falsey, A. R., Walsh, E., Campbell, D. (2002) Effects of
aging on cytokine production in response to respiratory syncytial virus
infection. J. Infect. Dis. 185, 682– 685.
50. Shodell, M., Siegal, F. P. (2002) Circulating, interferon-producting plas-
macytoid dentritic cells decline during human ageing. Scand. J. Immu-
nol. 56, 518 –521.
51. Gon, Y., Hashimoto, S., Hayashi, S., Koura, T., Matsumoto, K., Horie, T.
(1996) Lower serum concentrations of cytokines in elderly patients with
pneumonia and the impaired production of cytokines by peripheral blood
monocytes in the elderly. Clin. Exp. Immunol. 106, 120 –126.
52. Cai, N. S., Li, D. D., Cheung, H. T., Richardson, A. (1990) The expres-
sion of granulocyte/macrophage colony-stimulating factor in activated
mouse lymphocytes declines with age. Cell. Immunol. 130, 311–319.
53. Grewe, M. (2001) Chronological ageing and photoageing of dendritic
cells. Clin. Exp. Dermatol. 26, 608 – 612.
54. Bogdan, C., Nathan, C. (1993) Modulation of macrophage function by
transforming growth factor ␤, interleukin-4, and interleukin-10. Ann.
N. Y. Acad. Sci. 685, 713–739.
55. Crawford, R. M., Leiby, D. A., Green, S. J., Nacy, C. A., Fortier, A. H.,
Meltzer, M. S. (1994) Macrophage activation: a riddle of immunological
resistance. Immunol. Ser. 60, 29 – 46.
56. Davila, D. R., Edwards III, C. K., Arkins, S., Simon, J., Kelley, K. W.
(1990) Interferon-␥-induced priming for secretion of superoxide anion
and tumor-necrosis factor-␣ declines in macrophages from aged rats.
FASEB J. 4, 2906 –2911.
57. Ding, A., Hwang, S., Schwab, R. (1994) Effect of aging on murine
macrophages. Dimished response to IFN-␥ for enhanced oxidative me-
tabolism. J. Immunol. 153, 2146 –2152.
58. Hayakawa, H., Sato, A., Yagi, T., Uchiyama, H., Ide, K., Nakano, M.
(1995) Superoxide generation by alveolar macrophages from aged rats:
improvement by in vitro treatment with IFN-␥. Mech. Ageing Dev. 80,
199 –211.
59. Lavie, L., Weinreb, O., Gershon, D. (1992) Age-related alterations in
superoxide anion generation in mouse peritoneal macrophages studied by
repeated stimulation and heat shock treatment. J. Cell. Physiol. 152,
382–388.
60. Alvarez, E., Machado, A., Sobrino, F., Santa Maria, C. (1996) Nitric
oxide and superoxide anion production decreases with age in resident
and activated rat peritoneal macrophages. Cell. Immunol. 169, 152–
155.
61. Shepherd, V. L. (1986) The role of the respiratory burst of phagocytes in
host defense. Semin. Respir. Infect. 1, 99 –106.
Plackett et al. Innate immunity in aging 297
62. Johnston Jr., R. B., Kitagawa, S., Edwards III, C. K., Channon, J. Y.,
Suzuki, H., Pabst, M. J. (1988) The respiratory burst in activated mac-
rophages: studies of its molecular basis and evidence for downregulation
in chronic infection. Adv. Exp. Med. Biol. 239, 63–72.
63. Kumar, P., Pai, K., Pandey, H., Sundar, S. (2002) NADH-oxidase,
NADPH-oxidase and myeloperoxidase activity of visceral leishmaniasis
patients. J. Med. Microbiol. 51, 832– 836.
64. Khare, V., Sodhi, A., Singh, S. M. (1996 –1997) Effects of aging on the
tumoricidal functions of muring peritoneal macrophages. Nat. Immun.
15, 285–296.
65. Lu, Q., Ceddia, M. A., Price, E. A., Ye, S-M., Woods, J. A. (1999)
Chronic exercise increases macrophage-mediated tumor cytolysis in
young and aged mice. Am. J. Physiol. 276, R482–R489.
66. Kissen, E., Tomasi, M., McCartney-Francis, N., Gibbs, C. L., Smith,
P. D. (1997) Age-related decline in murine macrophage production of
nitric oxide. J. Infect. Dis. 175, 1004 –1007.
67. Chen, L-I., Pace, J. L., Russell, S. W., Morrison, D. C. (1996) Altered
regulation of inducible nitric oxide synthase expression in macrophages
from senescent mice. Infect. Immun. 64, 4288 – 4298.
68. Beharka, A. A., Wu, D., Serafini, M., Meydani, S. N. (2002) Mechanism
of vitamin E inhibition of cyclooxygenase activity in macrophages from
old mice: role of peroxynitrite. Free Radic. Biol. Med. 32, 503–511.
69. Rollo, E. E., Denhardt, D. T. (1996) Differential effects of osteopontin on
the cytotoxic activity of macrophages from young and old mice. Immu-
nology 88, 642– 647.
70. Wu, D., Hayek, M. G., Meydani, S. (2001) Vitamin E and macrophage
cyclooxygenase regulation in the aged. J. Nutr. 131, 382S–388S.
71. Harizi, H., Juzan, M., Pitard, V., Moreau, J-F., Gualde, N. (2002)
Cycloxygenase-2-issued prostaglandin E2 enhances the production of
endogenous IL-10, which down-regulates dendritic cell function. J. Im-
munol. 168, 2255–2263.
72. Harizi, H., Grosset, C., Gualde, N. (2003) Prostaglandin E2 modulates
dendritic cell function via EP2 and EP4 receptor subtypes. J. Leukoc.
Biol 73, 756 –763.
73. Rieser, C., Papesh, C., Herold, M., Bock, G., Ramoner, R., Klocker, H.,
Bartsch, G., Thurnher, M. (1998) Differential deactivation of human
dendritic cells by endotoxin desensitization: role of tumor necrosis fac-
tor-␣ and prostaglandin E2. Blood 91, 3112–3117.
74. Hilkens, C. M. U., Snijders, A., Snijdewint, F. G. M., Wierenga, E. A.,
Kapsenberg, M. L. (1996) Modulation of T-cell cytokine secretion by
accessory cell-derived products. Eur. Respir. J. Suppl. 22, 90s–94s.
75. Woods, J. J., Grbic, J. T., Rodrick, M. L., Jordan, A., Mannick, J. A.
(1987) Suppression of interleukin-2 production in an animal model of
thermal injury is related to prostaglandin synthesis. Arch. Surg. 122,
179 –184.
76. Plackett, T. P., Schilling, E. M., Faunce, D. E., Choudhry, M. A., Witte,
P. L., Kovacs, E. J. (2003) Aging enhances lymphocyte cytokine defects
after injury. FASEB J. 17, 688 – 689.
77. Adolfsson, O., Huber, B. T., Meydani, S. N. (2001) Vitamin E-enhanced
IL-2 production in old mice: naı̈ve but not memory T cells show in-
creased cell division cycling and IL-2 producing capacity. J. Immunol.
167, 3809 –3817.
78. Haynes, L., Linton, P. J., Eaton, S. M., Tonkonogy, S. L., Swain, S. L.
(1999) Interleukin 2, but not other common ␥ chain-binding cytokines,
can reverse the defect in generation of CD4 effector T cells from naı̈ve T
cells of aged mice. J. Exp. Med. 190, 1013–1024.
79. Mascarucci, P., Taub, D., Saccani, S., Paloma, M. A., Dawson, H., Roth,
G. S., Ingram, D. K., Lane, M. A. (2001) Age-related changes in cytokine
production by leukocytes in rhesus monkeys. Aging (Milano) 13, 85–94.
80. Glaser, R., Maccallum, R. C., Laskowski, B. F., Malarkey, W. B.,
Sheridan, J. F., Kiecott-Glaser, J. K. (2001) Evidence for a shift in the
Th1- and Th2-cytokine response associated with chronic stress and
aging. J. Gerontol. A Biol. Sci. Med. Sci. 56, M477–M482.
81. Renshaw, M., Rockwell, J., Engleman, C., Gewirtz, A., Katz, J., Samb-
hara, S. (2002) Cutting edge: impaired Toll-like receptor expression and
function in aging. J. Immunol. 169, 4697– 4701.
82. Boehmer, E. D., Goral, J., Faunce, D. E., Kovacs, E. J. (2004) Age-
dependent decrease in Toll-like receptor 4-mediated proinflammatory
cytokines by production and mitogen-activated protein kinase expres-
sion. J. Leukoc. Biol. 75, 342–349.
83. Beharka, A. A., Meydani, M., Wu, D., Leka, L. S., Meydani, A., Meydani,
S. N. (2001) Interleukin-6 production does not increase with age. J.
Gerontol. A Biol. Sci. Med. Sci. 56, B81–B88.
84. Bruunsgaard, H., Pedersen, B. K. (2003) Age-related inflammatory cy-
tokines and disease. Immunol. Allergy Clin. North Am. 23, 15–39.
85. Kovacs, E. J., Grabowski, K. A., Duffner, L. A., Plackett, T. P., Gergory,
M. S. (2002) Survival and cell mediated immunity after burn injury in
aged mice. J. Amer. Aging Assoc. 25, 3–10.
298 Journal of Leukocyte Biology Volume 76, August 2004
86. Saurwein-Teissl, M., Blasko, I., Zisterer, K., Neuman, B., Lang, B.,
Grubeck-Loebenstein, B. (2000) An imbalance between pro- and anti-
inflammatory cytokines, a characteristic feature of old age. Cytokine 12,
1160 –1161.
87. Franceschi, C., Bonafe, M., Valensin, S., Olivieri, F., De Luca, M.,
Ottaviani, E., De Benedictis, G. (2000) Inflamm-aging: an evolutionary
perspective on immunosenescence. Ann. N. Y. Acad. Sci. 908, 244 –254.
88. Ahluwalia, N., Mastro, A. M., Ball, R., Miles, M. P., Rajendra, R.,
Handte, G. (2001) Cytokine production by stimulated mononuclear cells
did not change with aging in apparently healthy, well-nourished women.
Mech. Ageing Dev. 122, 1269 –1279.
89. Chandra, R. K. (1997) Nutrition and the immune system: an introduction.
Am. J. Clin. Nutr. 66, 460S– 463S.
90. Lesourd, B. M. (1997) Nutrition and immunity in the elderly: modifica-
tion of immune responses with nutritional treatments. Am. J. Clin. Nutr.
66, 478S– 484S.
91. Mazari, L., Lesourd, B. M. (1998) Nutrition influences on immune
responses in healthy aged persons. Mech. Ageing Dev. 104, 25– 40.
92. Barbul, A., Regan, M. C. (1995) Immune involvement in wound healing.
Otolaryngol. Clin. North Am. 28, 955–968.
93. Swift, M. E., Burns, A. L., Gray, K. L., DiPietro, L. A. (2001) Age-related
alterations in the inflammatory response to dermal injury. J. Invest.
Dermatol. 117, 1027–1035.
94. Shallo, H., Plackett, T. P., Heinrich, S. A., Kovacs, E. J. (2003) Monocyte
chemoattractant protein-1 (MCP-1) and macrophage infiltration into the
skin after burn injury in aged mice. Burns 29, 641– 647.
95. Ashcroft, G. S., Horan, M. A., Ferguson, M. W. J. (1998) Aging alters the
inflammatory and endothelial cell adhesion molecule profiles during
human cutaneous wound healing. Lab. Invest. 78, 47–58.
96. Swift, M. E., Kleinman, H. K., DiPietro, L. A. (1999) Impaired wound
repair and delayed angiogenesis in aged mice. Lab. Invest. 79, 1479 –
1487.
97. Vitale, M., Zamai, L., Neri, L. M., Galanzi, A., Facchini, A., Rana, R.,
Cataldi, A., Papa, S. (1992) The impairment of natural killer function in
the healthy aged is due to postbinding-deficient mechanism. Cell. Im-
munol. 145, 1–10.
98. Ogata, K., Yokose, N., Tamura, H., An, E., Nakamura, K., Dan, K.,
Nomura, T. (1997) Natural killer cells in the late decades of human life.
Clin. Immunol. Immunopathol. 84, 269 –275.
99. Mariani, E., Monaco, M. C. G., Cattini, L., Sinoppi, M., Facchini, A.
(1994) Distribution and lytic activity of NK cell subsets in the elderly.
Mech. Ageing Dev. 76, 177–187.
100. Borrego, F., Alonso, M. C., Galiani, M. D., Carracedo, J., Ramirez, R.,
Ostos, B., Pena, J., Solana, R. (1999) NK phenotypic markers and IL2
response in NK cells from elderly people. Exp. Gerontol. 34, 253–265.
101. McNerlan, S. E., Rea, I. M., Alexander, H. D., Morris, T. C. M. (1998)
Changes in natural killer cells, the CD57CD8 subset, and related cyto-
kines in healthy aging. J. Clin. Immunol. 18, 31–38.
102. Mariani, E., Mariani, A. R., Meneghetti, A., Tarozzi, A., Cocco, L.,
Facchini, A. (1998) Age-dependent decrease of NK cell phosphoinosi-
tide turnover during spontaneous but not Fc-mediated cytolytic activity.
Int. Immunol. 10, 981–989.
103. Krishnaraj, R. (1997) Senescence and cytokines modulate the NK cell
expression. Mech. Ageing Dev. 96, 89 –101.
104. Dussault, I., Miller, S. C. (1994) Decline in natural killer cell-mediated
immunosurveillance in aging mice–a consequence of reduced cell pro-
duction and tumor binding capacity. Mech. Ageing Dev. 75, 115–129.
105. Kmiec, Z., Mysliwska, J., Rachon, D., Kotlarz, G., Sworczak, K., Mysliw-
ski, A. (2001) Natural killer activity and thyroid hormone levels in young
and elderly persons. Gerontology 47, 282–288.
106. Mysliwska, J., Bryl, E., Zorena, K., Balon, J., Foerster, J., Mysliwski, A.
(1997) Overactivity of tumor necrosis factor-␣ but not interleukin 6 is
associated with low natural killer cytotoxic activity in the elderly. Ger-
ontology 43, 158 –167.
107. Mysliwska, J., Mysliski, A., Romanowski, P., Bigda, J., Sosnowska, D.,
Foerster, J. (1992) Monocytes are responsible for depressed natural killer
(NK) activity in both young and elderly low NK responders. Gerontology
38, 41– 49.
108. Ravaglia, G., Forti, P., Maioli, F., Bastagli, L., Facchini, A., Mariani, E.,
Savarino, L., Sassi, S., Cucinotta, D., Lenaz, G. (2000) Effects of micro-
nutrient status on natural killer cell immune function in healthy free-
living subjects aged ⱖ90 y. Am. J. Clin. Nutr. 71, 590 –598.
109. De La Fuente, M., Miquel, J., Catalan, M. P., Victor, V. M., Guayerbas,
N. (2002) The amount of thiolic antioxidant ingestion needed to improve
several immune functions is higher in aged than in adult mice. Free
Radic. Res. 36, 119 –126.
http://www.jleukbio.org
110. Mikael, N., Mirza, N. M., Zaharian, B. I., Deulofeut, H., Salazar, M.,
Yunis, E. J., Dubey, D. P. (1994) Genetic control of the decline of natural
killer cell activity in aging mice. Growth Dev. Aging 58, 3–12.
111. Hsueh, C-M., Chen, S-F., Ghanta, V. K., Hiramoto, R. N. (1996) In-
volvement of cytokine gene expression in the age-dependent decline of
NK cell response. Cell. Immunol. 173, 221–229.
112. Plett, A., Murasko, D. M. (2000) Genetic differences in the age-associ-
ated decrease in inducibility of natural killer cells by interferon-␣/␤.
Mech. Ageing Dev. 112, 197–215.
113. Fernandes, G., Gupta, S. (1981) Natural killing and antibody-dependent
cytotoxicity by lymphocyte subpopulations in young and aging humans.
J. Clin. Immunol. 1, 141–148.
114. Edwards, D. L., Avis, F. P. (1979) Antibody-dependent cellular cytotox-
icity effector cell capacity among normal individuals. J. Immunol. 123,
1887–1893.
115. Edwards, B. K., Howe, H. L., Ries, L. A., Thun, M. J., Rosenberg, H. M.,
Yancik, R., Wingo, P. A., Jemal, A., Feigal, E. G. (2002) Annual report
to the nation on the status of cancer, 1973–1999, featuring implications
of age and aging on U.S. cancer burden. Cancer 94, 2766 –2792.
116. Jemal, A., Murray, T., Samuels, A., Ghafoor, A., Ward, E., Thun, M. J.
(2003) Cancer statistics, 2003. CA Cancer J. Clin. 53, 5–26.
117. Albright, J. W., Albright, J. F. (1998) Impaired natural killer cell
function as a consequence of aging. Exp. Gerontol. 33, 13–25.
118. Ogata, K., An, E., Shioi, Y., Nakamura, K., Luo, S., Yokose, N., Minami,
S., Dan, K. (2001) Association between natural killer cell activity and
infection in immunologically normal elderly people. Clin. Exp. Immunol.
124, 392–397.
119. Exley, M., Garcia, J., Balk, S. P., Porcelli, S. (1997) Requirements for
CD1d recognition by human invariant V␣24⫹ CD4 –CD8 – T cells. J.
Exp. Med. 186, 109 –120.
120. Faunce, D. E., Gamelli, R. L., Choudhry, M. A., Kovacs, E. J. (2003) A
role for CD1d-restricted NKT cells in injury-associated T cell suppres-
sion. J. Leukoc. Biol. 73, 747–755.
121. Gansert, J. L., Kiessler, V., Engele, M., Wittke, F., Rollinghoff, M.,
Krensky, A. M., Porcelli, S. A., Modlin, R. L., Stenger, S. (2003) Human
NKT cells express granulysin and exhibit antimycobacterial activity.
J. Immunol. 170, 3154 –3161.
122. Shao, H., Van Kaer, L., Sun, S. L., Kaplan, H. J., Sun, D. (2003)
Infiltration of the inflamed eye by NKT cells in a rat model of experi-
mental autoimmune uveitis. J. Autoimmun. 21, 37– 45.
123. Sonoda, K. H., Faunce, D. E., Taniguchi, M., Exley, M., Balk, S.,
Stein-Streilein, J. (2001) NK T cell-derived IL-10 is essential for the
differentiation of antigen-specific T regulatory cells in systemic toler-
ance. J. Immunol. 166, 42–50.
124. Moore, K. W., O’Garra, A., de Waal Malefyt, R., Vieira, P., Mosmann,
T. R. (1993) Interleukin-10. Annu. Rev. Immunol. 11, 165–190.
125. Dawson, H. D., Ross, A. C. (1999) Chronic marginal vitamin A status
affects the distribution and function of T cells and natural T cells in aging
Lewis rats. J. Nutr. 129, 1782–1790.
126. Miyaji, C., Wantanabe, H., Toma, H., Akisaka, M., Tomiyama, K., Sato,
Y., Abo, T. (2000) Functinal alterations of granulocytes, NK cells, and
natural killer T cells in Centenarians. Hum. Immunol. 61, 908 –916.
127. Tsukahara, A., Seki, S., Iiai, T., Moroda, T., Watanabe, H., Suzuki, S.,
Tada, T., Hiraide, H., Hatakeyama, K., Abo, T. (1997) Mouse liver T
cells: their change with aging and in comparison with peripheral T cells.
Hepatology 26, 301–309.
128. Poynter, M. E., Mu, H-H., Chen, X-P., Daynes, R. A. (1997) Activation
of NK1.1⫹ T cells in vitro and their possible role in age-associated
changes in inducible IL-4 production. Cell. Immunol. 179, 22–29.
129. Dubey, D. P., Husain, Z., Levitan, E., Zurakowski, D., Mirza, N., Younes,
S., Coronell, C., Yunis, D., Yunis, E. J. (2000) The MHC influences NK
and NKT cell functions associated with immune abnormalities and
lifespan. Mech. Ageing Dev. 113, 117–134.
130. Inui, T., Nakagawa, R., Ohkura, S., Habu, Y., Koike, Y., Motoki, K.,
Kuranaga, N., Fukasawa, M., Shinomiya, N., Seki, S. (2002) Age-asso-
ciated augmentation of the synthetic ligand-mediated function of mouse
NK1.1 Ag⫹ T cells: their cytokine production and hepatotoxicity in vivo
and in vitro. J. Immunol. 169, 6127– 6132.
131. Leite-de-Moraes, M. C., Herbelin, A., Gouarin, C., Koezuka, Y., Schnei-
der, E., Dy, M. (2000) Fas/Fas ligand interactions promote activation-
induced cell death of NK T lymphocytes. J. Immunol. 164, 4367– 4371.
132. McGeer, P. L., Rogers, J., McGeer, E. G. (1994) Neuroimmune mecha-
nisms in Alzheimer disease pathogenesis. Alzheimer Dis. Assoc. Disord.
8, 149 –158.
133. Rogers, J., Schultz, J., Brachova, L., Lue, L. F., Webster, S., Bradt, B.,
Cooper, N. R., Moss, D. E. (1992) Complement activation and ␤-amyloid-
mediated neurotoxicity in Alzheimer’s disease. Res. Immunol. 143,
624 – 630.
134. Lue, L. F., Rydel, R., Brigham, E. F., Yang, L. B., Hampel, H., Murphy
Jr., G. M., Brachova, L., Yan, S. D., Walker, D. G., Shen, Y., Rogers, J.
(2001) Inflammatory repertoire of Alzheimer’s disease and nondemented
elderly microglia in vitro. Glia 35, 72–79.
135. Nagaki, K., Hiramatsu, S., Inai, S., Sasaki, A. (1980) The effect of aging
on complement activity (CH50) and complement protein levels. J. Clin.
Lab. Immunol. 3, 45–50.
136. Oyeyink, G. O., Salimonu, L. S. (1999) Levels of complement compo-
nents, immunoglobulins and acute phase proteins in plasma during aging
in Nigeria. Afr. J. Med. Med. Sci. 28, 177–180.
137. Menzel, E. J., Zlabinger, G. J., Dunky, A., Steffen, C. (1988) Autoim-
munity and T-cell subpopulations in old age. Arch. Gerontol. Geriatr. 7,
249 –260.
138. Bellavia, D., Frada, G., Di Franco, P., Feo, S., Franceschi, C., Sansoni,
P., Brai, M. (1999) C4, BF, C3 allele distribution and complement
activities in healthy aged people and Centenarians. J. Gerontol. A Biol.
Sci. Med. Sci. 54, B150 –B153.
139. Esparza-Gordillo, J., Soria, J. M., Buil, A., Souto, J. C., Almasy, L.,
Blangero, J., Fontcuberta, L., de Cordoba, S. R. (2003) Genetic deter-
minants of variation in the plasma levels of the C4-binding protein
(C4BP) in Spanish families. Immunogenetics 54, 862– 866.
140. Ogata, R. T., Mathias, P., Bradt, B. M., Cooper, N. R. (1993) Murine
C4b-binding protein: mapping of the ligand binding site and the N-
terminus of the pre-protein. J. Immunol. 150, 2273–2280.
141. Lavery, L. W., Goyns, M. H. (2002) Decline in the expression of C4
binding protein ␣-chain gene during ageing of the rat liver. Biogeron-
tology 3, 207–211.
142. Cannon, J. G., Fiatarone, M. A., Fielding, R. A., Evans, W. J. (1994)
Aging and stress-induced changes in complement activation and neutro-
phil mobilization. J. Appl. Physiol. 76, 2616 –2620.
143. Kyrkanides, S., O’Banion, M. K., Whiteley, P. E., Daeschner, J. C.,
Olschowka, J. A. (2001) Enhanced glial activation and expression of
specific CNS inflammation-related molecules in aged versus young rats
following cortical stab injury. J. Neuroimmunol. 119, 269 –277.
144. Hazlett, L. D., Masinick-McClellan, S. A., Barrett, R. P. (1999) Com-
plement defects in aged mice compromise phagocytosis of Pseudomonas
aeruginosa. Curr. Eye Res. 19, 26 –32.
Plackett et al. Innate immunity in aging 299