Immunopharmacology and Immunotoxicology, 2012; 34(3): 477–483

© 2012 Informa Healthcare USA, Inc.

ISSN 0892-3973 print/ISSN 1532-2513 online
DOI: 10.3109/08923973.2011.630008

Research Article

Immunomodulatory activity of diethylcarbamazine on

humoral, cellular cytokine response and respiratory
burst in BALB/c mice
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Carlos E. Medina-De la Garza1,2, Graciela Guerrero-Ramírez1, Marisela García-Hernández1,2,

M. Angeles Castro-Corona1,2, Ernesto Torres-López1,2, Norbert W. Brattig3, and
Mario C. Salinas-Carmona1
Immunology Department, School of Medicine, 2Center of Research and Development in Health Science,
1

Universidad Autónoma de Nuevo León (UANL), Monterrey, México, and 3Bernhard Nocht Institute for Tropical
Medicine, Hamburg, Germany

Abstract
Diethylcarbamazine (DEC) is an anthelmintic piperazine derivative drug with putative immunomodulating properties,
including increased platelet and granulocyte adhesion to parasites and enhanced production of cytokines. To further
For personal use only.

analyse these properties in a well-established animal model, we evaluated the effect of DEC on antibody, cellular
cytokine response and respiratory burst in BALB/c mice. Animals were challenged with a thymus-dependent (tetanus
toxoid, (TT)) and with a thymus-independent (lipopolysaccharide, (LPS)) antigen and treated with DEC for seven
days with two different doses (50 mg/day and 500 mg/day). Serum was assessed for antibody production at 0, 4, 7,
14, 21 and 28 days after stimulation and at 0, 24 and 48 h for IL-2, IFN-γ, IL-10 and IL-12 release. Respiratory burst of
neutrophils and monocytes from peripheral blood was measured by flow cytometry. We found low-dose treatment
with DEC enhanced cytokine production vs. TT and antibody production vs. LPS, whereas a higher dose enhanced
significantly the respiratory burst of both polymorphonuclear leukocytes and monocytes, with a significant higher
effect on the former. Our results suggest a stimulating, dose-dependent immunomodulatory effect of DEC with a
higher effect on the phagocytic cells.
Keywords:  Diethylcarbamazine, immunomodulation, cytokines, antibody production, respiratory burst

Introduction and deficient on nitric oxide synthase showed a lesser

The piperazine derivative diethylcarbamazine citrate
(DEC, N,N-dyethyl-4-methylpiperazine-1-carboxamide)
is a synthetic organic antifilarial drug which has been
extensively used since 1947 for the treatment for
onchocerciasis and is currently used against Wuchereria,
Brugia malayi and B. timori. The drug is effective in vivo
against microfilaria (mf ) of the parasites. Nevertheless,
its mechanisms of antiparasitic action remain poorly
understood.(1) Interference with the metabolism of
arachidonic acid and breakdown of mf membranes,
organelle damage and apoptosis among others, have
been proposed.(2–4) DEC inhibits leukotriene production
via 5-lipoxigenase and mice infected with W. bancrofti
effect of the drug on the parasite as is also observed
with the inhibition of cyclooxygenase-1.(5,6) The involve-
ment of phagocytes and immunologic effectors in the
increased in vitro adherence and killing of Onchocerca
mf by eosinophils, increased activation of neutrophils
and enhanced adherence of platelets to parasites, sug-
gested DEC would be effective through the immune
system of infected host.(7–9) Based on the evidence of
DEC interaction with the humoral and cellular effec-
tors, putative immunomodulatory properties have been
investigated: an increased antibody response and 10-fold
increased peripheral monocyte counts in cats after a
vaccination with feline leukemia virus vaccine has been

Address for Correspondence:  Dr. Carlos E. Medina-De la Garza, Immunology Department, School of Medicine, UANL. Monterrey, N.L.
64460, México. Tel: +52(81) 83294211. Fax: +52(81) 83331058. E-mail: carlos.medina@uanl.mx
(Received 31 May 2011; revised 08 September 2011; accepted 03 October 2011)

477
 478  C. E. Medina-De la Garza et al.
Table 1.  Experimental groups of BALB/c mice.
Group Treatment
1 Control (untreated)
2 TT 0.03 IU/mouse
3 LPS 10 µg/mouse
4 Control 25 µg DEC
5 TT 0.03 IU/mouse + 25 µg DEC
6 LPS 10 µg/mouse + 25 µg DEC
7 TT 0.03 IU/mouse + 250 µg DEC
8 LPS 10 µg/mouse + 250 µg DEC
DEC, diethylcarbamazine; LPS, lipopolysaccharides; TT, tetanus
toxoid.
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reported.(10) Similarly, kittens infected with the virus and
treated with DEC were prevented or delayed to develop
lymphopenia.(11) Some other studies with variant meth-
odology have reported diverse degree of putative immu-
nomodulatory effects, including the increased clearance
of bacteria and fungi in mice under treatment with the
drug and lack of growth of Mycobacterium tuberculosis-
infected human blood supplemented with the drug.(12) So
far, different animal models and notably variant experi-
mental approaches have been used for the assessment of
DEC as a potential immunomodulator.
Therefore, a definite evaluation of DEC as immuno-
modulator deserves a methodological continuity that
allows a comprehensive assessment of the drug’s poten-
For personal use only.
tial. In this study we investigated the effect of DEC in an
immunologically well-defined model of healthy BALB/c,
analysing the antibody production, cytokine response
and oxidative metabolism of phagocytes.
Materials and methods
Agents
DEC citrate was purchased from Sigma (Sigma, St Louis,
Mo., USA). Antigens used to inoculate mice were: as thy-
mus-dependent antigen, tetanus toxoid (TT) in solution
(Behring-Aventis, Mexico) and as thymus-independent
antigen, lipopolysaccharides (LPS) from Escherichia coli
(serotype 0111:B4) (Sigma L2630). For the standardization
of an ELISA-test to determine antibodies against these
antigens, we used anti-mouse peroxidase conjugated anti-
bodies of isotypes IgG (Sigma A2554) and anti-mouse poly-
valent immunoglobulins (G,A, and M) (Sigma A0412).
Mice, immunization and treatments
This study was performed in accordance with the Studies
Division Posgrade and Ethical Committee of School
of Medicine, University Autonomous of Nuevo León
(U.A.N.L.). All animal protocols were approved by the
rules and regulation of our Institution for animal care.
Experiments were performed on BALB/c female and
male mice, between 6 and 12 weeks old. We used eight
groups of seven BALB/c mice each, for the different
experimental conditions: Group 1, non-immunized and
untreated mice as control; Group 2, mice immunized
with TT 0.03 IU/mouse and Group 3: mice immunized
 Immunopharmacology and Immunotoxicology
with LPS 10 µg/mouse, both subcutaneously (s.c.) on the
leg as appropriate as antigen control. Group 4, pharma-
cological control, mice treated with DEC in a daily dose
of 25 µg/mouse for 7 days. Group 5, mice immunized
with 0.03 IU of TT/mouse and treated with a daily dose
of DEC 25 µg/mouse (equivalent of a 50 mg/day human
dose) for seven days; Group 6, mice immunized with 10
µg of LPS/mouse and treated with a daily dose of DEC
25 µg/mouse. Group 7: mice immunized with 0.03 IU of
TT/mouse and treated with a daily dose of DEC 250 µg/
mouse (equivalent to the highest human dose, 500 mg/
day) for 7 days. Group 8: mice immunized with 10 µg/
mouse of LPS, and treated with a daily dose of DEC 250
µg/mouse for 7 days (Table 1).
For stimulation, all antigens were injected subcutane-
ously without additional adjuvant. However, mice of TT
group were also immunized 7 days prior to start of trial
(treatment) with a dose of 0.03 IU. For oral administra-
tion of DEC, we used 1 mL syringes and a gastrointesti-
nal tube. DEC solutions were prepared of 0.25 mg/mL,
0.85 mg/mL, 2.5 mg/mL and 1.7 mg/mL in sterile saline
solution 0.9% PiSA) for the administration to mice. DEC
was administered on a dose of 25 µg/mouse/day (Groups
3 and 4), and with 250 µg/mouse a day (Group 5). This
treatment was given for 7 days.
Sera for antibody and cytokine determination
All groups of mice were bled with Pasteur pipettes through
puncture of retro-orbital plexus on days 0, 7, 10, 14, 21 and
28 for serum to determine antibodies, and on days 0, 24
and 48 h for serum to determine concentration of cytok-
ines. An amount of approximately 500 µL of whole blood
from each mouse was retrieved; blood samples were
immediately placed in a centrifuge (Beckman, Allegra
64R) with refrigeration at 4°C and 12,000 rpm/10 min.
Those for determination of antibodies were stored at
−20°C and for determination of cytokines were aliquoted
and stored at −70°C.
Control sera
For negative and positive control sera vs. LPS, five mice
were immunized subcutaneously with 10 µg/mouse LPS
with Freund’s complete adjuvant (Sigma F5881) on day
0, and with incomplete Freund’s adjuvant (Sigma F5506)
on days 7, 14, 21 and 28. The mice were bled on days
0 and 28. The pool of serum from day 0 was labeled as
negative, and serum pools from day 28 were labeled as
positive controls. For control sera to TT, five mice were
immunized by subcutaneous injection of 0.3 IU/mL of TT
with Freund’s complete adjuvant (Sigma F5881) on day 0,
and with incomplete Freund’s adjuvant (Sigma F5506) on
days 7, 14, 21 and 28. The mice were bled on days 0, and
28. The pool of serum from day 0 was labeled as negative
control and sera from day 28 as positive control.
Determination of antibodies to TT by ELISA
ELISA plates and assays for IgG and total antibodies vs.
TT by ELISA were prepared as described elsewhere.(13)

Microplates were coated with antigen adding 50 µL of a
solution of 15 IU of TT diluted in PBS buffer pH 7.4 and
to a final concentration of 0.03 IU per well and incu-
bated overnight at 2–8°C. The content was discarded and
washed thrice with PBS–Tween Buffer 1:1000 (PBS–T).
After washing, 50 µL/well blocking solution of PBS–T 5%
skim milk was added, and incubated 1 h. After washing
thrice, the microplate was stored at 2–8°C until later use.
For determining total antibodies, 50 µL of the antibody
conjugate were added (Anti-mouse polyvalent immuno-
globulins (G,A, and M) antibody conjugated with peroxi-
dase, Sigma A0412) diluted 1:10 000 in blocking solution
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PBS−T and 1% skim milk. For IgG determination, 50
µL of conjugate were added (anti-mouse IgG-antibody
conjugated with peroxidase, Sigma A2554) diluted 1:70
000 in blocking solution and incubated for 1 h/37°C and
washed. 50 µL substrate o-phenyldiamine dihydrochlo-
ride (OPD, Sigma P9187) was added and incubated in
darkness at room temperature for 30 min. Reaction was
stopped with 0.1 M H2SO4 and proceeded immediately to
determine the absorbance at 495 nm wavelength.
Determination of antibodies to LPS by ELISA
Briefly, for IgG and total antibodies vs. LPS, antigen was
fixed as 200 µL of a solution of 50 mg of LPS in 1 mL of
carbonate buffer pH 9, with a final concentration of 0.3
µg/well. Subsequently the microplate was incubated at
For personal use only.
2–8°C overnight. The content was discarded and the plate
washed thrice with wash solution (PBS−T). Once washed,
it was added 200 µL/well of blocking solution (PBS–T and
5% skim milk), and incubated 1 h. It was washed again
as described and stored at −20°C. For total antibodies
determination, 200 µL of antibody conjugate were added
(anti-mouse polyvalent immunoglobulins (G,A,M) anti-
body conjugated with peroxidase, Sigma A0412) 1:10 000
diluted in blocking solution (PBS–T and 1% skim milk).
For IgG antibodies determination, 200 µL of anti-mouse
IgG-antibody conjugated with peroxidase (Sigma A2554)
diluted 1:70 000 in blocking solution was added and
incubated for 1 h at 37°C. After incubation and wash-
ing, 100 µL substrate (OPD) was added and incubated in
darkness at room temperature for 30 min. Reaction was
stopped with 0.1 M H2SO4 and proceeded immediately to
determine the absorbance at 495 nm wavelength propor-
tional to antibodies amount.
Determination of cytokines
Measurement of cytokines was made after the inocula-
tion of TT or LPS and concomitant treatment with DEC
at 25 µg/mouse/day (low-dose) and 250 µg/mouse/
day (high-dose) throughout the 48 h. Sera from mice so
treated were obtained at 0, 24, and 48 h after the inocula-
tion and start of treatment, and stored at 70°C. These sera
were thawed for determination of following cytokines
IL-2, IL-10, IL-12 and IFN-γ by a commercial ELISA kit
accordingly to the manufacturer’s specifications for each
cytokine. The kits used were: IL-2 (mouse Interleukin-2,
EMIL22, Pierce, USA), IL-10 (mouse interleukin-10
© 2012 Informa Healthcare USA, Inc.
Immunomodulatory activity of diethylcarbamazine  479
ELISA, EM2IL10, Pierce), IL-12 (Mouse Interleukin12 P70
ELISA, EMIL122, Pierce), IFN-γ (Mouse IFN-γ gamma
ELISA, EM10012, Pierce).
Determination of respiratory burst in neutrophils and
monocytes
To determine respiratory burst, two control groups
and four test groups of 5 mice each were formed. The
first group was untreated mice and second group mice
were treated with 0.1 mL of saline. To conform the test
groups, two variables were considered: (1) doses of DEC
and (2) time of testing after DEC intake. Therefore, two
groups were set using a treatment dose of 85 µg/mouse
(200 mg/day) of DEC in 0.1 mL of saline solution, where
one was measured at 1 h and the other at 2.5 h. Other two
groups were set using a treatment dose of 170 µg/mouse
(400 mg/day) of DEC in 0.1 mL of saline solution, where
one was measured at 1 h and the other at 2.5 h. Doses
used for respiratory burst experiments were based on
pharmacologic studies using the range corresponding to
a one-dose scheme(14) and are different from those used
as stimulant for humoral and cellular response, where
DEC is administered upon several days. The timing of
measurement was based upon reported serum high-
est peak on rodents (1 h)(15,16) and human (2.5 h).(14,17)
Mice leukocytes were obtained using 700 µL of blood
with EDTA. The blood was placed into tubes and mixed
with 7 mL of lysis solution, previously incubated at
room temperature (FACS, Becton Dickinson). The tubes
mixed with the lysis solution were incubated for 10 min
at 37°C and centrifuged at 1300 rpm at 4°C by 5 min.
(centrifuge, Eppendorf 5403). The supernatant was
discarded, and the pellet was washed and centrifuged
twice with PBS pH 7.4 at the same conditions. Finally,
the cells were resuspended in 7 mL of PBS pH 7.4 for
each tube (approximately 2.5 × 106 cells/mL), and ready
to use in the assay. The cells were analysed under dif-
ferent conditions with the dyhydrorhodamine 123 tech-
nique (123-DHR, Sigma D1054) using zymosan (ZYM;
Zymosan A Saccharomyces cerevisiae, Sigma Z4250)
and PMA (Phorbol-myristate acetate, Sigma P8139)
as stimulus.(18) Measurement of mean fluorescence
intensity (MFI) of neutrophils was conducted by flow
cytometry (Becton Dickinson FACsort) using Cellquest
software. We analysed the region for granulocytes and
for monocytes. The results were obtained by printing
histograms containing values and statistical analysis of
the MFI and percentage of cells that were detected for
both cell populations.
Statistical analysis
For the comparison of the results obtained and deter-
mine significant differences among studied groups
tested for antibodies, cytokines and respiratory burst, a
nonparametric Wilcoxon test for multiple comparisons
was applied, using the statistical packages SPSS 10.0, R
software for statistical computing and graphs version
2.10.0 and Excel.
 480  C. E. Medina-De la Garza et al.

Results 24 h (267 ± 170 pg/mL; p ≤ 0.05) when compared to nega-

tive control (Figure 3). The other test groups showed very
Effect of in vivo DEC treatment on the production of low cytokine concentrations at both, 24 and 48 h with
antibodies to TT and LPS in BALB/c mice almost no differences. Production of IFN-gamma was not
The effect of a DEC treatment on the production of
IgG antibodies to TT in mice is shown in Figure 1. A
pronounced antibody response (OD = 0.54 ± 0.07) was
obtained with a high-dose DEC (500 mg/day) treatment
when compared with the untreated control group at day
28. DEC at low dose (50 mg/day) presented values similar
to untreated control at day 28 (OD = 0.34 ± 0.15). The dif-
ferences, however, were not statistically significant. The
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determination of total antibody response to TT (data not

shown) had a similar shape as the IgG production.
Regarding antibody response to LPS, only the group
with low-dose DEC (50 mg/day) showed a higher value
constantly from day 14 on 28 days after immunization.
The antibody response to LPS is significantly higher
in animals treated with low concentration of DEC
(OD = 0.62 ± 0.1) compared with the untreated control
(OD = 0.23 ± 0.05) (p ≤ 0.05) (Figure 2). The OD values for
the total anti-LPS antibodies were similar to those of IgG
anti-LPS antibodies (data not shown).

Effect of DEC on in vivo cytokine response to TT and LPS

To evaluate the effect of DEC on a Th1 cytokine response,
we analysed the release of IL-2, IFN-γ, and IL-12 at day
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0, 1 (24 h) and 2 (48 h) after stimulus. Concerning the
release of IL-2 in response to TT, the mice treated with the
low-dose of DEC showed a higher cytokine production at
Figure 2.  Production of IgG antibodies induced by lipopolysac­
charides (LPS). Five groups of BALB/c mice (n = 7) were used in this
study. Blood was collected at days 0, 7, 10, 14, 21 up to 28. Serum
was separated and used to the estimation of antibody production
using LPS as antigen. OD values represent mean ± S.D. There was a
significant difference at day 28 (*p ≤ 0.05) comparing the low dose of
diethylcarbamazine (DEC) to antigen alone and to high-dose DEC.

Figure 1.  Production of IgG antibodies induced by tetanus toxoid

(TT). Five groups of BALB/c mice (n = 7) were used in this study.
Blood was collected at days 0, 7, 10, 14, 21 up to 28. Serum was
separated and used to the estimation of antibody production
using TT as antigen. Values represent mean ODs ± S.D. There was
no significant difference.
Figure 3.  Effect on IL-2 production induced by TT. Cytokine
expression was followed up to 48 h (n = 5). Values represent mean ±
S.D. There was a significant difference at 24 h (*p ≤ 0.05) comparing
the low dose of DEC to antigen alone and to high-dose DEC.

 Immunopharmacology and Immunotoxicology

 Immunomodulatory activity of diethylcarbamazine  481
stimulated in any group and any time (data not shown). but significantly higher than the DEC-low-dose at both
IL-10 was slightly stimulated, but again there were no time conditions, using PMA and PMA/ZYM (p ≤ 0.05). In
significant differences between any group at any time or all cases, both, neutrophils and monocytes showed dose-
doses. In difference, the production of IL-12 was signifi- dependent reactions with a highest effect recorded at 1 h
cantly enhanced at 24 h by low-dose DEC (379 ± 13 pg/mL) after intake of DEC.
compared to the untreated control (281 ± 63 pg/mL) and
the high-dose of DEC (235 ± 62; p ≤ 0.05) (Figure 4).
Equally to the response to TT, after stimulation with
LPS, all groups showed similar IL-2 and IFN-gamma lev-
els. Interleukin-10, however, showed at 24 h significantly
higher concentrations after treatment with both DEC
doses than the control (p ≤ 0.05); at 48 h, the production
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of the cytokine was decreased (Figure 5). The levels of

IL-12 were significantly higher in the group treated with
low-dose DEC (p ≤ 0.05) than the antigen control, DEC
alone and high-dose DEC at 48 h (Figure 6).

In vivo effect of DEC on the respiratory burst of

neutrophilic granulocytes and monocytes
One hour after oral intake of DEC at high-dose (170 µg/
mouse), peripheral neutrophilic granulocytes (neu-
trophils) reacted with a more pronounced respiratory
burst stimulated with PMA, ZYM and both PMA/ZYM
and expressed by mean fluorescence intensity (MFI)
(Figure 7A). A comparatively much lower response was
observed when measured 2.5 h after intake of the drug.
The MFI of peripheral monocytes was much lower
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(Figure  7B). The high-dose DEC effect measured at 1 h

Figure 5.  Effect on IL-10 production induced by LPS with DEC
showed a significant difference to low-dose DEC and treatment. The cytokine expression was followed up to 48 h (n = 5).
unstimulated controls (p ≤ 0.05). MFI measurements Values represent mean ± S.D. There was a significant difference at
with high-dose DEC at 2.5 h were lower than that of 1 h, 24 h (*p ≤ 0.05) comparing both doses of DEC to antigen alone.

Figure 4.  Effect on IL-12 production induced by TT. Cytokine
expression was followed up to 48 h (n = 5). Values represent
mean ± S.D. There was a significant difference at 24 h (*p ≤ 0.05)
comparing the low dose of DEC to antigen alone and to high-dose
DEC.
Figure 6.  Effect on IL-12 production induced by LPS with DEC
treatment. The cytokine expression was followed up to 48 h (n = 5).
Values represent mean ± S.D. There was a significant difference at
48 h (*p ≤ 0.05) only vs. antigen alone, DEC alone and high-dose
DEC.

© 2012 Informa Healthcare USA, Inc.

 482  C. E. Medina-De la Garza et al.
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For personal use only.
Figure 7.  Effect of DEC on the respiratory burst of phagocytes
from peripheral blood of mice. (A) Neutrophils, the expression
of MFI with high-dose (H) of DEC at 1 h showed a significant
difference compared to control groups and low-dose (L) DEC,
and to DEC (H) at 2.5 h in PMA + DHR and ZYM + DHR (*p ≤ 0.05).
(B) Monocytes, the expression of MFI with high-dose (H) of DEC
at 1 h showed a significant difference compared to control groups
and low dose (L) DEC (*p ≤ 0.05) but not to DEC (H) at 2.5 h. All
values represents mean ± S.D.
Discussion
DEC is a chemically well-characterized drug whose
therapeutic doses and effects have been extensively
studied.(17) DEC functions as antifilarial drug and has
been proposed as candidate for evaluation as immuno-
modulatory drug with therapeutic potential. However,
from our view, some studies to investigate this effect
 Immunopharmacology and Immunotoxicology
have been performed in a non-consistent fashion with
varying drug doses, methodologies and experimental
designs. In the present study, we investigated the in
vivo effect of DEC on humoral and cellular cytokine
responses in BALB/c mice immunized with thymus-
dependent and -independent antigens, and on phago-
cytic effectors. The doses used in antibody and cytokine
production experiments reflected the human thera-
peutic dose (low-dose DEC) and the highest human
dose allowed (high-dose DEC). The respiratory burst
experiments used doses of DEC calculated for a single-
dose scheme. We deliberately avoided experimental
conditions using infra- or supra-therapeutic doses.(10,19)
Positive effects on the antibody production of other
reports was corroborated by our results.(8) The increase
of total antibody classes and IgG anti-TT titers with
high-dose of DEC was demonstrated. The induction of
antibodies against LPS with low-dose DEC produced an
evident effect. These interesting results suggest a poten-
tiating DEC effect on humoral responses that depends
on nature of antigen. A possible adjuvant effect of DEC
must be further investigated. Kitchen et al. observed a
more pronounced effect on the antibody response using
animal model with different vaccines.(19) The increment
of the antibody response against LPS, in particular, sug-
gests a potential use for DEC against LPS-containing
microbes.
Effects on the cytokine production by DEC have
been reported.(20) DEC enhanced the generation of pro-
inflammatory cytokines as IL-6 and IL-2 in mononuclear
cells in vitro.(21) In our study, DEC showed its in vivo
effect on pro- and anti-inflammatory cytokine responses
represented by IL-2, IFN-γ, IL-12 and IL-10. Mice treated
with high-dose DEC and immunized with TT expressed
enhanced IL-2 and IL-12 which relates to the increase
in the antibody response to this antigen. In addition, we
observed a decrease of serum IL-10 level. In contrast, in
sera of the group receiving low-dose DEC and immunized
with LPS, IL-2 was not detected but high levels of IL-10
that correlates with the increasing response of antibodies
to LPS.
We further investigated the phagocytic activity
through burst oxidative. Evident differences of MFI
occurred among all groups. Although intermediary
oxidative species production by phagocytes has been
shown to be enhanced by DEC,(22) the present report
represents the first on the assessment of burst oxida-
tive of phagocytic cells stimulated by DEC treatment
in vivo. Further, these results appear to correlate with
morphological studies, an increase of antifilarial activ-
ity by respiratory burst derivatives from granulocytes in
tissues and ex vivo(2) and also an increase of some other
granulocyte enzymes as myeloperoxidase, elastase,
lactoferrin, and lysosyme.(23) One feature observed was
the time-depending effect observed in the respiratory
burst. Highest DEC concentration has been shown 1 h
after application in cotton rats(16) whereas, for man it
ranges between 2.2 and 2.7 h.(17) The highest effect on

phagocytes at 1 h in BALB/c correlated well with the
pharmacokinetic data from rodents.
Conclusion
The presented results show that DEC could act directly
on phagocytic response and affect the humoral and
cytokine responses. Thus, experimental evidence of DEC
as potential immunomodulator or adjuvant appears jus-
tified for its further assessment as an immunologic modi-
fier for potential use in the treatment of human diseases,
e.g. in chronic pyogenic and insidious infections.
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Acknowledgments
We thank head-librarian Martina-Christine Koschwitz
at Bernhard Nocht Institute, Hamburg, Germany, and
Dr Gloria Molina-Salinas from CIBIN-IMSS, Monterrey,
Mexico, for specific bibliographical material. We thank
the staff of CIMAT, Monterrey-Unit, for support on orga-
nizing data.
Declaration of interest
This work was supported by grant PAICyT SA-611-01
from Universidad Autónoma de Nuevo León, and by the
department of Immunology, School of Medicine, UANL.
For personal use only.
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