The effect of age and gender on cytokine production by human peripheral
blood mononuclear cells and markers of bone metabolism
Peter Pietschmanna,b,*, Eva Golloba, Susanne Broschb, Philipp Hahnb, Stephan Kudlacekc,
Martin Willheima, Wolfgang Woloszczukd, Meinrad Peterlika, Karl Heinz Traglb
aDepartment of Pathophysiology, University of Vienna, Waehringer Guertel 18-20, Vienna A-1090, Austria
bLudwig Boltzmann-Institute of Aging Research, Vienna, Austria
cDepartment of Medicine, Hospital Barmherzige Bru ¨der, Vienna, Austria
dLudwig Boltzmann-Institute of Experimental Endocrinology, Vienna, Austria
Received 15 April 2003; received in revised form 16 July 2003; accepted 17 July 2003
Background. Aging has been associated with various alterations of immune functions, the musculoskeletal system and a decline of sex
hormone levels. Estradiol has a central role in the regulation of bone turnover and also modulates the production of cytokines such as
interleukin-1 and -6 and tumor necrosis factor-a. We therefore studied the effect of age and gender on cytokine production by mononuclear
cells and markers of bone metabolism.
Methods. Peripheral blood mononuclear cells were isolated from young and elderly subjects; intracellular detection of cytokine production
after stimulation with ionomycine and PMA (T cells) or LPS (monocytes) was performed by four color flow cytometry. Sex hormone levels
and markers of bone metabolism were measured by RIA or ELISA.
Results. When we compared elderly to young women we found an increased proportion of T cells that were positive for interferon-g,
interleukin-2, -4, -10 and -13. Also the percentage of cells producing interleukin-4 or interferon-g within the CD8þpopulation was higher in
the group of elderly women. In contrast, proportionally fewer monocytes of elderly women were positive for tumor necrosis factor-a or
interleukin-6 than those of young women. In elderly men a higher percentage of T cells produced interleukin-2, -4 and -13. In the group of
aged men we found a higher frequency of cells that produced interleukin-4 within the CD4þor CD8þpopulation. Moreover, within
monocytes ofelderly men we found an increased percentage of cells positive forbothinterleukin-1b andtumor necrosisfactor-a.The data on
markers of bone metabolism indicated an increase of bone turnover in old age.
Conclusion. Our data demonstrate that aging is associated with significant alterations of bone metabolism and cytokine production by T
cells and monocytes. For particular cytokines (interferon-g and interleukin-10 in T cells, interleukin-6 and tumor necrosis factor-a in
monocytes) these changes are gender specific.
q 2003 Elsevier Inc. All rights reserved.
Keywords: Aging; T lymphocytes; Monocytes; Cytokines; Sex hormones
There is a general consensus that various functions of
the immune system decline with age. Clinically, the
consequences of impaired immune functions in the elderly
include an increased susceptibility to infections, malig-
nancies and autoimmunity (Wick and Grubeck-Loeben-
subpopulations with aging have been described in
the literature (Xu et al., 1993; Sansoni et al., 1993;
Hulstaert et al., 1994; Stulnig et al., 1995). We found an
increased percentage of memory CD4þand CD8þ/CD57þ
cells in elderly subjects (Kudlacek et al., 1995); these data
are in line with an expansion of CD282T cells in healthy
aging people (Fagnoni et al., 1996). In addition to
phenotypic data, there is a vast amount of literature on
the decline of T cell functions with aging (Wick and
Grubeck-Loebenstein, 1997; Globerson and Effros, 2000);
these include an impaired response to mitogens (Song et al.,
1993), a decreased production of interleukin-2 (Nagel et al.,
1988) and a relative deficit of the capacity of transendothe-
lial migration (Stohlawetz et al., 1996). Nevertheless, there
0531-5565/$ - see front matter q 2003 Elsevier Inc. All rights reserved.
Experimental Gerontology 38 (2003) 1119–1127
* Corresponding author. Tel.: þ43-1-40400-5122; fax: þ43-1-40400-
E-mail address: email@example.com (P. Pietschmann).
is a lack of influence of donor age on human T cell clone
longevity (Pawelec et al., 2002).
Beside lymphocytes, monocytes as well as dendritic cells
from aged subjects have been studied and their function has
been found to be normal (Schwab et al., 1985; Fagiolo et al.,
1993; Steger et al., 1996; Pietschmann et al., 2000). Other
authors, however, suggested a decreased functional activity
of monocytes from aging individuals (Beckman et al., 1990;
Rich et al., 1993). In a study from our laboratory, we
demonstrated an increased expression of several surface
markers, including ICAM-1, on monocytes from elderly
subjects (Stohlawetz et al., 1998). Interleukin-1 production
of monocytes from elderly subjects has been reported to be
increased (Pacifici et al., 1987; Fagiolo et al., 1993;
O’Mahony et al., 1998), as well as decreased (McLachlan
et al., 1995; Gon et al., 1996).
Aging is associated not only with a deterioration of
functions of the immune system but also with a decline in
the secretion of numerous hormones; these include sex
hormones such as estradiol and testosterone. Estradiol is
involved in the regulation of cytokine production; for
instance, peripheral blood monocytes from premenopausal
or postmenopausal women under hormone replacement
therapy secrete less interleukin-1 than untreated postmeno-
pausal women (Pacifici et al., 1989). Peripheral blood
mononuclear cells from ovarectomized premenopausal
women constitutively secrete more tumor necrosis factor-
a as well as interleukin-1 than age-matched controls
(Pacifici et al., 1991). Beside estrogens, also androgens
modulate the production of bone resorptive cytokines.
Androgens inhibit the production of interleukin-1 by human
synovial macrophages (Cutolo and Masi, 1998). Bellido
et al. (1995) demonstrated in a murine model that male sex
steroids, acting through the androgen-specific receptor,
inhibit the expression of the interleukin-6 gene. These data
are in line with those of Kanda et al. (1996) who found that
testosterone treatment reduced interleukin-6 production by
human monocytes. Thus, the age-associated decrease of sex
steroid levels could alter the production of cytokines.
Therefore, the specific aim of our study was to determine the
gender specificity of cytokine production by monocytes or T
cells in elderly compared to younger women and men. Since
cytokines such as interleukin-1, tumor necrosis factor-a and
interleukin-6 are involved in the regulation of bone turnover
(Gowen et al., 1983; Bertolini et al., 1986; Jilka et al., 1992),
we also measured several markers of bone metabolism
(osteocalcin, bone alkaline phosphatase, c-terminal telopep-
tide of type I collagen).
A total number of 129 subjects was studied after
informed consent had been obtained:
(A) 25 young (age: 30 ^ 4 years) premenopausal healthy
(B) 25 young (age: 31 ^ 5 years) healthy males
(C) 41 elderly (age: 85 ^ 7 years) females according to the
SENIEUR protocol (Ligthart et al., 1984)
(D) 38 elderly (age: 82 ^ 12 years) males according to
the SENIEUR protocol (Ligthart et al., 1984).
In the elderly subjects, the presence of primary/idio-
pathic osteoporosis was neither an inclusion nor an
exclusion criterion. In addition to the exclusion criteria
specified by the SENIEUR protocol (Ligthart et al., 1984),
the following conditions were defined as exclusion criteria
for this study:
† The presence of any disease or condition known to affect
bone metabolism (e.g. hepatic or renal insufficiency,
thyroid disorders and alcohol abuse) with the exception
of primary/idiopathic osteoporosis.
† The presence of a non-osteoporotic osseus disease (e.g.
osteomalacia, hyperparathyroidism, Paget’s disease).
† Any medication known to affect bone metabolism (e.g.
hormone replacement therapy, bisphosphonates, gluco-
corticoids) with the exception of calcium and/or vitamin
The study protocol was approved by the Ethical
Committees of the University of Vienna Medical School
and of the Hospital Barmherzige Bru ¨der, Vienna.
2.2. Hormones, biochemical parameters of bone metabolism
Serum levels of estradiol, total and free testosterone, sex
hormone binding globulin (SHBG), LH, FSH, TSH, PTH,
25 hydroxy-vitamin D, osteocalcin (a parameter of bone
formation), the bone specific isoenzyme of alkaline
phosphatase (BAP, Ostasew, a parameter of bone for-
mation), and c-terminal telopeptide of type I collagen (CTX,
Cross Lapsw, a serum marker of bone resorption) were
determined by commercial ELISAs or RIAs. Details of
these assays are given in Table 1. The PTH, 25 hydroxy-
vitamin D, estradiol and testosterone data were also part of a
large study on reference values for osteoprotegerin
(Kudlacek et al., 2003).
2.3. Immunophenotyping and intracellular detection
Flow cytometric assessment of cytokine production
was performed essentially as described previously
(Willheim et al., 1997; Pietschmann et al., 2001).
Mononuclear cells were isolated from heparinized
peripheral blood by density gradient centrifugation with
Ficoll-Paque. After washing in PBS, cells were resus-
pended (2 £ 106ml21) in Ultra Culture Medium (Bio-
Whittaker, Walkersville, MD, USA) supplemented with
P. Pietschmann et al. / Experimental Gerontology 38 (2003) 1119–11271120
2 mM L-glutamine (Sigma Chemical Co, St Louis, MO,
USA), 170 mg/l gentamycin sulfate (Sigma Chemical Co,
St Louis, MO, USA) and 5 ml/l mercaptoethanol (Merck,
For the determination of cytokine production of T
cells, peripheral blood mononuclear cells were incubated
for 4 h at 37 8C in culture medium (as described above)
containing in addition phorbol 12-myristate 13-acetate
(10 ng/ml), ionomycin (1.25 mM) and brefeldin A
(10 mg/ml; all from Sigma Chemical Co, St Louis, MO,
USA). Thereafter cells were harvested, washed twice with
phosphate-buffered saline (PBS) and fixed with 2%
formaldehyde in PBS for 20 min at room temperature.
Subsequently cells were washed, resuspended in HBSS
containing 0.3% BSA (Biomol Feinchemikalien, Ham-
burg, Germany) and 0.1% sodium azide (Merck, Darm-
stadt, Germany) and stored at 4 8C.
For the determination of cytokine production of
monocytes, peripheral blood mononuclear cells were
incubated for 4 h at 37 8C in culture medium (as described
(50 ng/ml) and brefeldin A (10 mg/ml; all from Sigma
Chemical Co, St Louis, MO, USA). At the end of the
incubation period cells were harvested and washed twice
with PBS. Since previous experiments from our laboratory
indicated that the antigenicity of CD14 is lost after
fixation with formaldehyde, cells were stained for the
surface expression of CD14 immediately after washing.
Thereafter, cells were fixed and further treated as
For staining of intracellular cytokines, the cells were
washed once with PBS and permeabilized by washing
twice with saponin (0.1%, Sigma Chemical Co.) in PBS.
Subsequently, cells were incubated in the presence of
saponin with saturating concentrations of antibodies for
25 min at room temperature. After washing once with
saponin (0.1%) in PBS, the cells were resuspended and
analyzed by four color flow cytometry. Lymphocytes
and subsequently defined as CD3þ, CD4þor CD8þ
cells. Cells fulfilling both criteria (lymphocyte and
CD4þand/or CD8þ) were further analyzed for their
coexpression of cytokines. Monocytes were gated as
Cytokine positive cells were evaluated in a two
parameter dot plot; quadrants for positive/negative dis-
crimination were set according to a negative control and
adjusted to the visible negative population within the
sample. All data presented reflect percentages of cells
positive for a respective cytokine.
Antibodies used for the determination of surface
phenotype and cytokine production are listed in Table 2.
gatedby theirlight scattercharacteristics
Data are given as means ^ SD. Kruskal–Wallis test and
Kendall-tau correlation coefficient were used for statistical
RIAs and ELISAs used for the determination of hormones and markers of bone metabolism
Company Country Detection limit Intra-assay CV (%)Inter-assay CV (%)
CIS bio international
CIS bio international
CIS bio international
Malmo ¨, Sweden
BAP: bone specific isoenzyme of alkaline phosphatase; CTX: c-terminal telopeptide of type I collagen.
Monoclonal antibodies used for the determination of the cell surface
phenotype and cytokine production, respectively
Antibody Conjugation CloneManufacturer
P. Pietschmann et al. / Experimental Gerontology 38 (2003) 1119–11271121
3.1. Hormones and markers of bone metabolism
Serum levels of hormones, markers of bone metab-
olism and its regulators in all groups are summarized in
Table 3. As expected, serum estradiol levels were
significantly lower in the elderly than in the young
women; nevertheless, estradiol levels were similar in the
elderly and young men. Total and free testosterone levels
were significantly decreased in the elderly men and
Hormones and markers of bone metabolism in young and elderly women and men. Data are means ^ SD
Young women ðn ¼ 22–25Þ
Elderly women ðn ¼ 20–28Þ
Young men ðn ¼ 25Þ
Elderly men ðn ¼ 34–35Þ
Total testosterone (ng/ml)
Free testosterone (pg/ml)
25-Hydroxy-vitamin D (ng/ml)
106.5 ^ 57.2
0.44 ^ 0.34
4.5 ^ 3.3
4698 ^ 2046
12.2 ^ 12.3
7.9 ^ 2.9
1.9 ^ 1.1
23.3 ^ 9.6
43.5 ^ 14.6
20.2 ^ 6.5
5.01 ^ 1.83
2850 ^ 1715
19.3 ^ 9.1**
0.27 ^ 0.44**
2.49 ^ 2.8**
4894 ^ 1875
21.1 ^ 14.9*
46.0 ^ 20.3**
1.9 ^ 2.0
49.2 ^ 30.1**
20.2 ^ 15.1**
22.2 ^ 8.6
11.7 ^ 6.43**
5519 ^ 2422**
35.3 ^ 12.4
4.2 ^ 1.73
14.6 ^ 9.2
2067 ^ 705
5.2 ^ 2.5
5.0 ^ 1.5
1.4 ^ 0.6
21.6 ^ 11.0
35.3 ^ 9.8
22.6 ^ 4.8
7.7 ^ 3.16
3611 ^ 1961
32.2 ^ 13.6
1.80 ^ 0.98**
6.6 ^ 4.8**
4441 ^ 2085**
13.9 ^ 11.1**
18.9 ^ 19.2**
1.3 ^ 0.9
49.6 ^ 29.3**
19.9 ^ 12.2**
19.5 ^ 8.7*
11.2 ^ 5.6**
5341 ^ 3244*
BAP: bone specific isoenzyme of alkaline phosphatase; CTX: c-terminal telopeptide of type I collagen. *p , 0:05 vs. young population; **p , 0:001 vs.
Fig. 1. Cytokine production by T cells in young women ðn ¼ 12–20Þ;
elderly women ðn ¼ 30–32Þ; young men ðn ¼ 11–23Þ and elderly men
ðn ¼ 16–27Þ: Data are means ^ SD. *p , 0:05.
Fig. 2. Cytokine production by T cells in young women ðn ¼ 12–20Þ;
elderlywomenðn ¼ 27–33Þ youngmenðn ¼ 11–23Þ andelderlymenðn ¼
16–27Þ: Data are means ^ SD. *p , 0:05, **p , 0:01, ***p , 0:001.
P. Pietschmann et al. / Experimental Gerontology 38 (2003) 1119–11271122
women as well. Significantly elevated LH and FSH levels
were seen both in the elderly women and men. SHBG
levels were similar in young and elderly women, but were
significantly elevated in the elderly men. Regarding the
serum levels of TSH no significant differences were
observed. In both sexes, serum 25-hydroxy-vitamin D
levels were decreased and serum PTH levels were
increased in the elderly when compared to the young
subjects. The levels of the bone specific isoenzyme of
alkaline phosphatase (BAP) were elevated in the elderly
men and women, whereas concentrations of osteocalcin,
another parameter of bone formation, were decreased in
the elderly men and unchanged in the elderly women. In
both sexes, a significant increase of bone resorption as
reflected by serum CTX levels was seen in the elderly
when compared to young subjects.
3.2. Cytokine production by T cells
We next examined the production of several cytokines
produced by T cells in young and elderly women (Figs. 1
and 2). In elderly women, a significantly higher proportion
of CD3þcells were positive for interferon-g. Among the
CD8þsubset of T cells the frequency of interferon-g
producing cells was more than twice as high in the elderly
than in the young women. For interferon-g in CD4þcells
a trend towards an increased production in the elderly was
observed. Moreover, in T cells of elderly women an
increased frequency of interleukin-2 positive cells was
observed whereas no significant alterations with age were
seen for tumor necrosis factor-a (Fig. 1). The production
of interleukin-4 was significantly increased both in CD3þ
and CD8þcells from elderly women. In CD4þcells
interleukin-4 production tended to be higher in old age
(Fig. 2). Comparing elderly to young women we also
found an increase of T cells that were positive for
interleukin-10 and -13.
When compared to young men, a significantly increased
proportion of T cells that produced interleukin-2 was
observed in elderly men (Fig. 1). In contrast to women, no
significant alteration of interferon-g production was
observed in men. Interleukin-4 production by CD3þ,
CD4þand CD8þcells was significantly higher in elderly
vs. young men (Fig. 2). Regarding the frequency of CD3þ
cells positive for tumor necrosis factor-a and interleukin-10
no differences between young and elderly men were seen.
Nevertheless, in the elderly men interleukin-13 was
Fig. 3. Cytokine production by monocytes in young women ðn ¼ 20Þ; elderly women ðn ¼ 38–39Þ; young men ðn ¼ 25Þ and elderly men ðn ¼ 29–30Þ: Data
are means ^ SD. *p , 0:05, **p , 0:01.
P. Pietschmann et al. / Experimental Gerontology 38 (2003) 1119–11271123
3.3. Cytokine production by monocytes
In contrast to T cells, the production of several cytokines
was lower in elderly compared to young women (Fig. 3).
Within monocytes of elderly women, the percentage of cells
positive for tumor-necrosis-factor-a or interleukin-6 was
lower than within those of young women. Moreover, in the
elderly women a decreased proportion of CD14þcells
coexpressed both interleukin-1b and tumor necrosis factor-
a. In contrast, in monocytes of elderly men (vs. young men)
an increased frequency of cells positive for both interleukin-
1b and tumor necrosis factor-a was seen (Fig. 3).
3.4. Correlation analyses
Among elderly women a significant correlation of the
percentage of monocytes positive for interleukin-10 both
with the levels of osteocalcin ðt ¼ 0:364; p ¼ 0:025Þ and
those of CTX ðt ¼ 0:354; p ¼ 0:03Þ was found. In these
subjects, CTX levels correlated also with interleukin-4
production by CD8þcells ðt ¼ 0:404; p ¼ 0:018Þ whereas a
negative correlation of 25 hydroxy-vitamin D with the
percentage of CD8þcells that produced interferon-g ðt ¼
20:411; p ¼ 0:014Þ was seen. Within the group of elderly
men a negative correlation of BAP with interleukin-4 in
CD8 positive cells ðt ¼ 20:529; p ¼ 0:007Þ could be
established. In the elderly men, PTH levels correlated
with the production of tumor necrosis factor-a by mono-
cytes ðt ¼ 0:357; p ¼ 0:012Þ as well as that of interferon-g
by CD4þcells ðt ¼ 0:328; p ¼ 0:05Þ: Among the elderly
women or men no further significant correlations between
bone markers and cytokine production were found.
One of the major findings of our study was that age
influences the production of specific cytokines differently
in men and women. T cells from elderly women–but not
from elderly men–were more frequently positive for
interferon-g and interleukin-10. Concerning cytokine
production by monocytes, aging in women was specifically
associated with decreased production of tumor necrosis
factor-a and interleukin-6. Thus, at least some of the
aforementioned alterations of the immune system in the
elderly could result from sex hormone deficiencies and not
merely be the consequence of aging per se. Although, sex
hormone levels decrease with aging both in men and
women, the magnitude of this decline is very different. As
also shown in our study, the mean estradiol level of the
elderly women was less than one fifth of that of
premenopausal women, whereas the mean free testosterone
level of the elderly men was about half of that seen in the
young men. It should also be noted that the average
estradiol level was clearly higher in the elderly men than in
the elderly women. The selective increase of interferon-g
production capacity of T cells with aging in women thus
could be a consequence of estrogen deficiency. This notion
is supported by data from Matejuk et al. (2001) who
demonstrated an inhibitory effect of estradiol on the
production of interferon-g in murine lymphocyte cultures.
Moreover, since in hypogonadism serum interleukin-4
levels are elevated and decrease following testosterone
replacement (Yesilova et al., 2000; Kocar et al., 2000), it is
tempting to speculated that the age-associated increase in
interleukin-4 production capacity seen in both sexes could
be due to androgen deficiency.
Despite many data on the effect of age on cytokines
very few authors specifically studied intracellular cytokine
production in elderly subjects. O’Mahony et al. (1998)
reported significantly increased intracellular production of
tumor necrosis factor-a and interleukin-6 by T cells from
older subjects. In this study, levels of interleukin-1b,
tumor necrosis factor-a and interleukin-6 in monocytes of
older subjects tended to be elevated; but the observed
differences were not statistically significant. McNerlan
and coworkers (2002) found an increased percentage of
CD3þcells producing tumor necrosis factor-a and
interferon-g in elderly compared to young subjects.
Since we found increased interferon-g production of T
cells in elderly women (but not in elderly men) it should
be noted that in McNerlan’s study the group of elderly
subjects consisted predominantly of women whereas the
young control group was composed of three males and
three females only. Our result of a significantly increased
proportion of T cells coexpressing interleukin-2 in the
elderly is in line with McNerlan and coworkers who
found a trend towards a higher interleukin-2 production
with old age. Also Sakato-Kaneko et al., (2000) observed
an augmented interleukin-2 production by CD4þcells in
subjects aged 55–60 years compared to young subjects.
Taken together, several independent investigations using
flow cytometry for cytokine detection demonstrate that
interleukin-2 production capacity increases with age.
These findings challenge the notion of a decline of
interleukin-2 secretion with age as suggested by earlier
studies using traditional methods for cytokine measure-
ment (Nagel et al., 1988; Gillis et al., 1981; Thompson
et al., 1984). Traditional detection in culture supernatants
is likely to be impaired by paracrine consumption of the
cytokine and thus does not reflect the full capacity of
interleukin-2 production (Baroja and Ceuppens, 1987;
Claret et al., 1992).
In a very recent publication, Schwaiger et al. (2003)
demonstrated that IL-4 producing CD8þcells with a
CD62Lbrightphenotype accumulate in a subgroup of older
adults. In subjects more than 60 years of age, IL-4
producing CD8þcells were more frequent in persons who
were capable of raising a humoral immune response
following immunization than in others who failed to
produce protective antibodies after vaccination. The data
of Schwaiger et al. (2003) are in line with our observation
P. Pietschmann et al. / Experimental Gerontology 38 (2003) 1119–1127 1124
of a significantly increased interleukin-4 production by
CD8þcells in old age. In contrast to these findings, in
the above-mentioned study of Sakato-Kaneko et al. (2000)
interleukin-4 production in the elderly was decreased. A
possible explanation for this discrepancy could be, that
our old subjects were more than 20 years older than the
subjects studied by Sakata-Kaneko.
Using traditional methods, several (Kita et al., 1991;
Cakman et al., 1996; Paganelli et al., 1996) but not all
investigators (Sindermann et al., 1993; Born et al., 1995)
reported a decreased interferon-g production in aged
subjects. Moreover, different groups found an enhanced
monokine production in old age (Fagiolo et al., 1993;
Paganelli et al., 1994; Born et al., 1995; Cakman et al.,
1997; Gabriel et al., 2002). The variation of these to our
findings very likely can be explained by differences in the
methods that were used. For instance, the stimulation period
was clearly shorter in our than in the traditional protocol
(4 h vs. 1–6 days). In many studies, different stimuli to
induce cytokine production were used (e.g. phytohemag-
glutinin or concanavalin A). Since the number of CD3þ
cells is decreased in SENIEUR elderly (Sansoni et al., 1993;
Born et al., 1995), whole blood assays may underestimate
cytokine production capacity. In contrast to whole blood
assays (in which a certain volume of blood is analyzed)
cytokine detection by flow cytometry is not influenced by
variations in absolute cell numbers. Moreover, with regard
to the gender differences detected in the present study, it
should be noted that with only one exception (Kita et al.,
1991) in the aforementioned studies results were not
analyzed for women and men separately.
Beside its role as a major regulator of calcium
homeostasis, 1,25(OH)2 vitamin D has also effects on
cells of the immune system (Holick, 2002). Thus, some
alterations of immune functions seen in elderly subjects
may be related to changes of vitamin D levels. For
instance, since 1,25(OH)2 vitamin D has been demon-
strated to decrease interleukin-2 production (Mu ¨ller et al.,
1993; Willheim et al., 1997, 1999), the augmented
interleukin-2 in the elderly could be a consequence of
vitamin D deficiency.
While bone resorption as reflected by serum CTX
increased with age in both sexes, the results from two
bone formation parameters, osteocalcin and BAP were
divergent. Reflecting an overall increase of bone turnover
with aging, BAP levels rose both in elderly men and
women. In contrast, osteocalcin levels were lower in
elderly men and unaffected by age in women. In the
present study, we did not find an increase of ‘classical
bone resorptive cytokines’ interleukin-1b, tumor necrosis
factor-a or interleukin-6 but an augmented production of
interleukin-4, -10 and interferon-g in elderly women.
Noteworthy, the latter three cytokines have been shown to
inhibit the generation and/or activity of osteoclasts
(Peterlik et al., 1985; Watanabe et al., 1990; Xu et al.,
1995; Hong et al., 2000). Thus, our data appear to contrast
those of Pacifici et al. (1987, 1989, 1991) who suggest
that increased production of interleukin-1b and tumor
necrosis factor-a are pathogenic in postmenopausal
osteoporosis. However, in our study the elderly subjects
(although they had evidence of increased bone resorption)
were not specifically selected for the presence of
osteoporosis or osteoporotic fractures. It should also be
noted that in postmenopausal patients with osteoporotic
fractures interleukin-1 production is not significantly
augmented (Pietschmann et al., 2001). Moreover, an
elevated production of bone resorptive cytokines might
be of importance for bone loss in early rather than in late
postmenopausal women. This notion is also supported by
the lack of a correlation between the marker of bone
resorption (CTX) and classical bone resorptive cytokines.
The enhanced production of ‘antiresorptive cytokines’ in
our elderly, late postmenopausal women could reflect
compensatory mechanisms preventing further bone loss.
In conclusion, we demonstrated that aging is associated
with significant alterations of bone metabolism and
cytokine production by T cells and monocytes. For
particular cytokines, these alterations are gender specific
and could be due, at least in part, to effects of estradiol or
testosterone deficiency on the production of the respective
This study was supported by the Jubila ¨umsfonds der
O¨sterreichischen Nationalbank (Project # 8128).
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