Correlation Between Female Genital Tract Levels of Interleukin-6 ...

Egyptian Journal of Medical Microbiology, October 2008
Vol. 17, No. 4
Correlation Between Female Genital Tract Levels of Interleukin-6 and
Interleukin-8 in Recurrent Miscarriage Associated with Bacterial
Vaginosis
Sahar Taher 1 and Mohamed El-Negeri 2
(1) Department of Medical Microbiology and Immunology, Faculty of Medicine, Mansoura
University. (2) Department of Obstetric and Gynecology, Faculty of Medicine, Mansoura
University.
ABSTRACT:
Bacterial vaginosis (BV) is the most common cause of abnormal vaginal discharge among women in childbearing period. It
develops when vaginal lactobacilli are replaced by an overgrowth of Gardnerella vaginalis, anaerobes, and mycoplasma. BV
is associated with subclinical endometritis and may be correlated with recurrent miscarriages. Miscarriage refers to the loss a
pregnancy before 24 weeks. It was reported that there is a relation between IL-6 and IL-8 in cervical and amniotic fluid and
microbial invasion of chorioamniotic membranes. Our aim is to determine the value of genital tract levels of IL-6 and IL-8 in
women with recurrent miscarriages in the presence or absence of BV. The present study included 94 women, 54 of them had
history of recurrent miscarriage (patients group) and 40 females with normal pregnancy (control group). Samples were
collected by vaginal swabs and cervicovaginal lavages(CVL) and examined physically, microscopically and were graded by
Gram stain for BV, cytokine levels measured by ELISA kits.
There was an increase in frequency of miscarriage with age. BV was detected in 37% of patients group and in 20% of
control group. Gardnerella vaginalis was found in 33% and 15% of patients and control group respectively. When
comparing patients group versus control group both cytokines levels were elevated, the mean (± SD) of IL-6 was
66.88±60.00 vs 11.14 ±13.69 pg/ml and that of IL-8 was 2265.47 ±3328.92 vs 195.12 ±203.59 pg/ml, P-value was highly
significant 4.5) and can experience a vaginal
homogenous discharge which oven has a
distinctive fishy odor. The addition of potassium
hydroxide to a drop of vaginal secretion on a
slide may produce or accentuate the odor and
these three signs in addition to the presence of
"Clue cells" on a wet mount smear comprise the
criteria. (3)
The prevalence varies widely from 5 to 51%
in different populations and about 24.6% of
women having in-vitro fertilization (IVF)
treatment. (4) Bacterial vaginosis is associated
with complications of pregnancy ,including
miscarriage, preterm birth post-partum
endometeritis. It is though that infection ascends
from the lower genital tract through the cervix
leading to deciduitis, chorioamnionitis and
amniotic fluid and fetal infection. (5)
Miscarriage refers to the loss of a pregnancy
before 24 weeks. It is a common complication of
pregnancy, occurring in 12-30% of all clinical
pregnancies. (6)
Miscarriage is a common event that causes
considerable morbidity for the mother. The
second trimester loss is less common, occurring
in 1-2% of pregnancies. (1)
Bacterial vaginosis is associated with
subclinical endometritis, (7) which may create an
adverse endometrial milieu for developing
embryo or fetus .This link might partially
account for the reported association between
bacterial vaginosis or intermediate flora and 1 st
trimester miscarriage. (2)
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Egyptian Journal of Medical Microbiology, October 2008
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Many women are asymptomatic, but some
experience frequent symptomatic episodes of the
condition. Current antibiotic treatments restore
the normal lactobacillus dominated flora in the
short term, but approximately 30% relapse
within one month. (8)
Bacterial vaginosis is so named because it is
not a vaginitis as some studies reported that there
is no polymorph nuclear leucocytes (PMNL). A
proteinacious toxin was isolated from
Gardnerella vaginalis with activity against
neutrophils, human erythrocytes and endothelial
cells, preventing the purulent response in BV
infection. (9)
Gardnerella vaginalis an important organism
of BV, was found to interfere with adherence of
lactobacilli to vaginal epithelial cells in vitro.
Anaerobes play a synergistic role in flourishing
of Gardnerella vaginalis by raising vaginal pH
through amines production. (10)
Established causes of recurrent miscarriage
are abnormal chromosomes in either partner,
particularly translocations, anti-phospholipid
antibodies, however half of all cases are
unexplained. (11) Women experiencing recurrent
miscarriage show evidence of excessive type 1
immune parameters and reduced type 2 and T
regulatory immune parameters, both
systematically and in endometrial tissues. (12)
Immunohistochemical and flow cytometric
analysis suggest that, in non- pregnant state,
predisposition to miscarriage is characterized by
changes in the balance between endometrial
lymphocyte populations. (13)
Infection of intrauterine tissue, the decidua
and/or amnion, results in macrophage activation
and secretory production of inflammatory
cytokines, including interleukin-1 (IL-1), tumour
necrosis factor (TNF), and IL-6. (14) Elevated
amniotic fluid IL-6 has been found to be
associated with acute inflammatory lesions,
histologic chorioamnionitis, pre-term labor. (15) In
addition the cervical IL-8 concentrations have
been reported to be associated with vaginal
bacteria in pregnancy and intrauterine infections
in patients with pre-term labor, providing an
indication for treatment to prevent pre-term
birth. (16)
Recently, a relationship between IL-6 and
IL-8 in cervical and amniotic fluid and microbial
invasion of chorioamniotic membranes was
reported. (17)
It is well known that the frequency of preterm
birth is high in successfully treated patients
suffering recurrent spontaneous miscarriage
.This means that these might be a group of
heterogeneous unexplained recurrent
miscarriages whose miscarriages are caused by
inflammation. (18) Bacterial vaginosis or disrupted
vaginal flora may alter the local mucosal
immune environment and the concentration of
cytokines present in genital secretions. BV was
found to be associated with increased levels of
various cytokines. (19)
In humans, the immune system is deviated
towards the Th2 immune response during normal
pregnancy, while women with recurrent
spontaneous abortions(RSA) have a bias toward
Th1 immune response. (20) Failure to unable the
shifting from a pro-inflammatory(Th1) to an
anti-inflammatory(Th2) pattern, as evidenced by
a persisting or dominant Th1 immune response,
is associated with complication such as multiple
implantation failure, RSA(21),pre-eclampsia,
pre-term delivery, and intra-uterine growth
restriction. (22)
The aim of the present work was to
determine the genital tract levels of antiinflammatory
cytokines IL-6 and IL-8 in women
with recurrent miscarriage in the presence or
absence of BV.
PATIENTS, MATERIALS, AND
METHODS:
Patients with two or more consecutive first
or second trimester miscarriages were enrolled to
the present study. All were managed at
Mansoura University Hospital Obstetrics
Clinics. Hesterosalpingoraphy, chromosome
analysis for both parents, immunologic tests
parameters such as β2glycoprotein I-dependent
anticardiolipin antibodies and lupus
anticoagulant, blood tests for hyperthyroidism,
diabetes mellitus, hyperprolactinemia and luteal
phase defects were performed before the next
pregnancy. All the above investigations were
done as a routine to exclude all these factors
from the study group.
After getting approval consent from ethical
research committee of Ob/Gyn Department,
Mansoura University Hospital. A total of 94
women joined the present study. Fifty four
women with history of recurrent abortion served
as patients group and 40 women with no history
of abortion attending the clinic for follow up
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Egyptian Journal of Medical Microbiology, October 2008
Vol. 17, No. 4
were served as control group, from the period of
January 2007 to January 2008. All of them were
pregnant at the time of the study. All women
gave informed written consent to undergo
speculum examination.
Samples collection
1.Vaginal swab specimens were collected by
using Dacron swabs and immediately processed
for routine laboratory testing.
2.Cervicovaginal lavage(CVL) specimens: were
obtained by instilling 10 ml of sterile saline into
the vaginal vault directing the steam of fluid at
the cervical os. The fluid was left for about 30
seconds and then aspirated. The fluid was then
transferred to a sterile 15 ml conical test tube
and stored at -70C◦.
Laboratory methods
A macroscopic examination of the vaginal
swab was performed to record the aspect, smell,
abundance, pH of secretions. Yeast, Tricomonas
vaginalis, and clue cells were identified, and
epithelial cells, red blood cells, and leucocytes
were counted by direct microscopic examination
(×400 amplification) of vaginal secretion wet
mount smears. A Gram’s stain of the vaginal
smear then was done to identify G. vaginalis,
Mobiluncus species, and lactobacilli and for
scoring of the vaginal flora.
Vaginal flora typing and bacterial vaginosis
diagnosis
Vaginal flora was scored by a Gramed
stained smear according to the following
classification scheme; exclusively lactobacilli
(type I), predominantly lactobacilli (type II),
presence of lactobacilli but predominance of
other gram- positive and gram-negative bacteria
(type III), and absence of lactobacilli and
presence of a mono-or polybacterial flora (type
IV). Both type III and IV classifications
represent flora in which non lactobacilli
morphotypes exceed lactobacilli. (23) Therefore,
BV was determined by (1) the presence of clue
cells in a vaginal smear confirmed by the
identification of G.vaginalis in a Gram stained
smear,(2) an increased vaginal pH (pH >4.5) and
(3) a lacto deficient vaginal flora (types III- IV).
Cytokine assays
Frozen CVL samples were thawed and
centrifuged at 500X g to remove debris; they
were then centrifuged at 2000Xg and
supernatants were harvested. The resulting
supernatant was divided in two aliquots, and
cytokine concentrations were measured using
commercial quantitative sandwich enzyme
immunoassay (Quantikine, R&D systems,
Minneapolis, MN). All samples run in duplicate
and results were reported in pg/ ml. Samples
were diluted 1:4 for IL-6 and 1:10 for IL-8
according to manufacturer’s directions. Briefly,
immunoplates coated with monoclonal
antibodies to the studied cytokines were
incubated with samples and standards. After
washing, a biotin-cojugated polycolonal
antiserum to each cytokine was added, the plates
were then incubated and washed. Next, avidinperoxidase
was added. Following washing, the
color reaction was developed with tetramethyle
bencidine. SO4H2 was added a stopper and
absorbencies were measured at 450 nm by
means of an ELISA plate reader. Values were
calculated from standard curves based on
prepared dilutions of recombinant cytokines.
Statistical analysis
It was performed using SPSS version 11
using windows XP. Qualitative data were
presented as frequency and percentage.
Quantitative data were presented as mean and
standard deviation. Kolmogorov-Smirnov test
was used to determine the distribution of data.
All data were non-normally distributed. X-
square and Fisher exact test were used to
determine significance among qualitative data
while Mann-Whitney test determine significance
among quantitative data. Odds ratio determines
the relation between genital tract levels of IL-6
and IL-8 and development of further
miscarriage. P-value is significant when < 0.05.
RESULTS
Of the total 94 women undergoing the
present study, 54 patients experienced 2 or more
consecutive miscarriages and 40 women with
normal pregnancy attending Obstetric Clinic for
follow up. The median age of the participants
was 26 years (range, from 19-42 years), all
women were pregnant at the time of the study.
Table (1) describes age distribution and
miscarriage frequency in patients group, the
frequency of miscarriage increases with age
while table (2) demonstrates characteristics of
BV among studied groups, BV was detected in
37.03% and 20% of patients and control group
respectively. Gardnerella vaginalis was found in
33.33% of patients and 15% of control group.
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Egyptian Journal of Medical Microbiology, October 2008
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Table (3) illustrates the correlation between BV
status and genital tract levels of IL-6 and IL-8 in
the studied groups. When comparing the
cytokine levels between the patients group and
control group, p- value was highly significant
less than 0.0001. On the other hand, BV positive
patients versus BV negative patients, for IL-6 p-
value was non significant =0.069 and for IL-8
was moderately significant =0.004. On
comparison of patients group BV positive versus
control group negative, p-value was moderately
significant for IL-6, p=0.004 and significant for
IL-8 p=0.037. Table (4) shows the diagnostic
efficiency of the combination of IL-6 and IL-8
levels in the genital tract for the risk of further
miscarriages. For IL-6 Odds ratio (OR) was 5.19
and 95% confidence interval (CI) ranged from
2.45-10.99 and that of IL-8 OR was 3.08 and CI
ranged from 1.74-5.30. The risk estimate of IL-6
for further miscarriage is 5 and that of IL-8 is 3
times.
Table (1): Age distribution and miscarriage frequency in patients group
Miscarriage frequency (n.) Patients (n.) % Age ( Mean ± SD)
2 18 33.33% 26 ± 5.1
3 26 48.15% 28 ± 3.8
4 2 3.70% 30 ± 6.3
More than 4 8 14.81% 37 ± 5.0
Table (2): Characteristic of BV in participating women.
Characteristic Patient (No.) % Control (No.) %
(1) Bacterial flora :
-Type I
9 16.67 18 45
-Type II
11 20.37 9 22.5
-Type III
20 37.04 7 17.5
-Type IV
14 25.92 6 15
(2)Vaginal pH:
4.5
21 38.89 8 20
(3) Bacterial vaginosis:
Positive
20 37.03 8 20
Negative
34 62.97 32 80
(4) Gardnerella Vaginalis:
Positive
18 33.33 6 15
Negative
36 66.67 34 85
Type I, exclusively lactobacilli; type II, predominantly lactobacilli; type III, presence of lactobacilli
but predominance of other gram positive and gram negative bacteria; type VI, absence of lactobacilli
and presence of a mono- or polymicrobial flora.
Table (3): Correlation between BV status and genital tract levels of IL-6 and IL-8 in
studied groups
Bacterial vaginosis status
Cytokine levels in cervico-vaginal lavage (mean ± SD )
IL-6 (pg / ml)
IL-8 (pg / ml)
(1) Patients (54):
BV positive (20)
BV negative (34)
(2) Control (40):
BV positive (8)
66.88 (±60.000)
86.81 (± 68.75)
55.75 (± 52.15)
11.14 (±13.69)
21.97 (±26.15)
BV negative (32) 8.49(±6.64)
• Patients group vs. control P

Egyptian Journal of Medical Microbiology, October 2008
Vol. 17, No. 4
-IL-6 P = 0.069 non significant
-IL-8 P = 0.004**
• Patients group BV positive vs. control group negative :
-IL-6 P = 0.004**
-IL-8 P = 0.037*
Table (4):Diagnostic efficiency of the combination of IL-6 and IL-8 levels in genital for the
risk of further miscarriages.
Cytokine
level (pg/ml)
IL-6:
>18
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Egyptian Journal of Medical Microbiology, October 2008
Vol. 17, No. 4
(OR) was 5.19 and 95%confidence interval (CI)
ranged from 2.45-10.99 for IL-6 while OR was
3.08 and 95% CI ranged from 1.74-5.30 for IL-
8. The risk estimates of IL-6 for further
miscarriage is 5 times and that of IL-8 is 3 times.
Similarly, Jun et al., (26) found that pre-term
labor may be due to intrauterine infection and
IL-6 in cervical fluid was earlier found to have
diagnostic and prognostic value for both preterm
rupture of membranes and preterm
delivery. Furthermore, bacterial vaginosis is
reported to cause late miscarriage through
inflammation and production of IL-1α
prostaglandin. (27) Complement 3 and 4, which are
elevated with non-specific inflammatory
phase. (28)
It is possible that any inflammation might be
localized to the uterus and not reflected by blood
parameters. (29) Cervical inflammatory cytokines
may thus be needed to be assessed and we
therefore paid attention to IL-6 and IL-8
concentrations cervical secretion as possible
predictors of further miscarriage in recurrent
cases.
Cytokines which are secreted in response to
pathogens mainly by macrophages, are a
structurally diverse group of molecules and
include IL-1, IL-6 and TNF-α, and IL-8 are
involved in local inflammatory responses,
helping to attract neutrophiles to sites of
infection. Thus, chemokines direct the migration
of the leukocytes along a gradient that increases
in concentration toward site of infection. (30)
It was shown that production of Th2
cytokines including IL-6 by peripheral blood
mononuclear (PBMCs) are significantly elevated
with miscarriage. (31) However, it has also been
reported that Th1 cytokines might not be
predominant in recurrent miscarriage patients (32)
and it should be borne in mind that cytokines
also act locally, with T helper cells constituting
only a minor population within the secretory
endometrium and first trimester-decidua tissue.
It is known that cytokines can be produced by
the epithelial and stromal cells of the
endometrium and decidual and cytotrophoblast
cells of placenta. Epithelial cells are major
source of cytokines in non-pregnant
endometrium, but after decidualization, there is
increased production by various populations of
stromal cells. (33) The degree of interaction
between the two systems is yet to be determined
and cytokine shift theory is still controversial. (34)
All bacteria produce proteins with an aminoterminal
N-formylated methionine, and the f-
Met-Leu Phe (fMLP) peptide is a potent
chemotactic factor for inflammatory cells,
especially neutrophils. The fMLP receptors for
chemokines and for the complement fragments
C5a, C3a, and C4a. Thus, there is a common
mechanism for attracting neutrophils, whether by
complement, chemokines, or bacterial
products. (30)
Hattori et al., (34) found that endometrial
expression of Il-6 and IL-1α are reduced in a
group of women experiencing unexplained
recurrent miscarriage (RM). On the other hand,
the finding of reduced IL-6 synthesis in
endometrial tissue from women experiencing
(35, 36)
RM is consistent with previous reports.
Endometrial IL-6 produced by APCs is
implicated as a key immune-deviating cytokine,
acting to promote Th2 differentiation through
induction of IL-4 synthesis to selectively
polarize Th2 cells, and concurrently inhibits Th1
development through upregulating suppressor of
cytokine signaling (SOCS)-1. (37) During the
process of embryo implantation, IL-6 facilitates
tissue remodeling to allow trophoplast invasion
by induction of decidual cell metalloproteinases
(MMP)-14 and MMP-11 (38) and potentially by
regulating decidual angiogenesis. (39) Importantly,
Il-6 a key regulator of early placental
development acting to regulate trophoblast cell
proliferation , differentiation and synthesis of
hCG, IL-1 and Il-6, and expression of integrins
and other markers of trophoblast invasion. (40 )
BV diagnosed in early pregnancy is
undoubtedly associated with preterm labor or
late miscarriage (41, 42) and with delivery of low
(43)
birth – weight infants. There are several
theories to explain this relationship. The first
maintains that protease produced by various
bacteria found in bacterial vaginosis weaken and
destroy the connective tissue framework of the
fetal membranes and thus lead to their rupture.
(44) It has also been suggested that mucinases and
sialidases produced by the bacteria hydrolyse the
protective cervical mucine so allowing the entry
of micro-organisms into the uterus where they
cause infection and inflammation. (45) In support
of this theory is frequent histological evidence of
endometritis in non-pregnant women with BV
and recovery of bacteria associated with BV
from fetal membranes and amniotic fluid
subsequent to premature rupture of membranes
and preterm labor. (46) Furthermore,
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Egyptian Journal of Medical Microbiology, October 2008
Vol. 17, No. 4
prostaglandins as well as endotoxins and
interleukin-1a have been found in high
concentrations in the cervical mucous and
vaginal secretions of pregnant women with BV.
(27)
Ugwumadu et al., (24) showed that treatment
of asymptomatic intermediate abnormal vaginal
flora and bacterial vaginosis in a general
obstetric population reduces the occurrence of
late miscarriage and spontaneous preterm
delivery. Therefore, initiation of systemic
antibiotic treatment early might offer the best
chance of modifying the risks associated with
abnormal vaginal flora bacterial vaginosis.
Finally, the results obtained in the present
work suggest that IL-6 and IL-8 could be
considered potentially eligible for use as
infection predictive markers in recurrent
miscarriages and in evaluation of therapeutic
success in genitourinary infections. Furthermore,
the biological material (vaginal swab and
cervicovaginal -lavage) used in this work is easy
to obtain and non-invasive for the mother or the
fetus, which could be an advantage over the use
of amniotic fluid.
REFERENCES:
1- Hay,P.E.(2004): Bacterial vaginosis and
miscarriage. Curr. Opin. Infect. Dis. ;17:41-44.
2- Ralph, S.G., Rutherford, A.J. and Wilson
J.D.(1999): Influence of bacterial vaginosis on
conception and miscarriage in the first trimester:
cohort study. BMJ ; 319:220-223.
3- Eschenbach, D.A.,Bekassy, S. and Blackwell,
A.(1984): The diagnosis of bacterial vaginosis. In
Mardh,P.A. and Tylor-Robenson, D. (eds), Bacterial
vaginosis. Almqvist& Wiksell International,
Stockholm, pp.260-261.
4- Blackwell, A.L.Thomas, P.D. Wareham , K. and
Emery, S.J.(1993): Health gains from screening for
infection of the lower genital tract in women
attending for termination of pregnancy. Lancet;
342:206-210.
5- Goldenberg,R.L., Hauth, J.C. and Andrews,
W.W.(2000): Intrauterine infection and preterm
delivery. N. Engl. J. 342:1500-1507.
6- Everett, C. (1997): Incidence and outcome of
bleeding before the 20 th week of pregnancy:
prospective study from general practice. BMJ;315:32-
34.
7- Korn,A.P., Bolan, G. and Padian, N. (1995):
Plasma cell endometritis in women with symptomatic
bacterial vaginosis.Obstet.Gynecol;85:387-390.
8- Hay, P.E. (1998): Therapy of bacterial vaginosis. J
Antimicrob Chemother ;41:6-9.
9- Rottini, G. D., Forgiarini, O. and Nordon, E.
(1990): Identification and partial characterization of
cytolytic toxin produced by Gardnerella vaginalis.
Infect. Immunol., 58:3751-3758.
10- Wood, J.R., Sweet, R.L. and Catena, A. (1985):
In vitro adherence of lactobacillus species to vaginal
epithelial cells. Am. J. Obstet. Gynecol., 153: 740-
743.
11- Herz-Picciotto, I. and Samuels, S. J. (1998):
Incidence of early pregnancy and loss of pregnancy.
N. Engl. J. Med.; 319:1483-1484.
12- Saito, S., Saaki, Y. and sakai, M., (2005): CD4
(+)CD25 high regulatory T cells in human pregnancy.
J. Reprod. Immunol. 65:111-120.
13- Laird, S.M., Tuckerman, E.M., Cork, B.A.,
LRinjawi, S.,Blackmore,A.I. and Li,T.C.(2003): A
review of immune cells and molecules in women with
recurrent miscarriage. Hum. Reprod. Update. 9:163-
174.
14- Gravett, M.G., Witkin, S.S., Haloska, G.J.,
Edwards, J.L., Cook, M.J., and Novy,M.J. (1994):
An experimental model for intra-amniotic infection
and preterm labor in rhesus monkeys. Am. J. Obstet.
Gynecol. 171:1660-1667.
15- Lockwood, C.J., Ghidini, A. ,Wein, R.,
Lapinski, R., Casal, D. and Berkowitz,R.L.(1994):
Increased interleukin-6 concentrations in cervical
secretions are associated with preterm
delivery.Am.J.Obstet.Gynecol; 171:1097-1102.
16- Sakai,M.,Sasaki,Y., Yoneda,S., Kasahara,T.,
Arai,T., Okada,M., Hosokawa,H., Kato,K.,
Soeda,Y. and Saito,S., (2004): Elevated interleukin-
8 in cervical mucous as an indicator for treatment to
prevent premature birth and preterm, pre-labor
rupture of membranes: a prospective study.
Am.J.Reprod.Immunol; 51:220-225.
17- Jacobsson,B., Mattsby-Baltzer,I. and
Hagberg,H. (2005): Interleukin-6 and interleukin-8
in cervical and amniotic fluid: relationship to
microbial invasion of chorioamniotic membranes.
BGOG;112:719-724.
18- Jivraj,S., Anstie,B., Cheong Y.C., Fairlie F.M.,
Laird,S.M. and Li,T.C.(2001): Obstetric and
neonatal outcome in women with a history of
recurrent miscarriage: a cohort study.Hum.Reprod;
16:102-106.
19- Wennerholm,U.B., Holm,B. and Mattsby-
Baltzer,I. (1998): Interleukin-1 alpha, interleukin-6
and interleukin-8 in cervical-vaginal secretion for
screening of preterm birth in twin gestation.
Acta.Obstet.Gynecol.Scand; 77:508-514.
643

Egyptian Journal of Medical Microbiology, October 2008
Vol. 17, No. 4
20- Raghupathy,R., Makhseed,M., Azizieh,F.,
Hassan.,N., Al-Azemi,M.and AlShamali,E.(1999):
Maternal Th1- and Th2-type reactivity to placental
antigens in normal human pregnancy and unexplained
recurrent spontaneous abortions. Cell Immunol;
196:122-130.
21- Choudhury,S.R. and Knapp,L.A.(2001):
Human reproductive failure 1: Immunological factors.
Hum. Reprod. Update; 7:113-134.
22- Marzi,M., Vigano,A., Trabattoni, D., Villa,
M.L. and Clerici,M. (1996): Characterization of
type 1 and type 2 cytokine production profile in
physiologic and pathologic pregnancy. Clin.Exp.
Immunol; 106:127-133.
23- Carbonnelle, B., Denis, F., Marmonier, A.,
Pinon, G. and Vargues, R. (1987):Bacteriologie
medicale: techniques usuelles . Paris: SIMEP.
24- Ugwumadu, A., Manyonda, I., Reid, F. and
Hay, P. (2003): Effect of early oral clindamycin on
late miscarriage and preterm delivery in
asymptomatic women with abnormal vaginal flora
and bacterial vaginosis: a randomized controlled trial.
Lancet, 361: 983-988.
25- Riduan, J.M., Hiller, S.L. and Utomo, B.
(1993): Bacterial vaginosis and prematurity in
Indonesia: association in early and late pregnancy.
Am. J. Obstet. Gynecol., 169: 175-178.
26- Jun, J.K., Yoon, B.H., Romero, R., Kim, M.,
Moon, G.B., Ki, S.H. and Park, J.S.(2000):
premature rupture of membranes. Am. J. Obstet.
Gynecol.; 183:868-873.
27- Platz-Christensen, J.J., Mattsby-Baltzer, I.,
Thomsen, P. and Wiqvist, N. (1993): Endotoxin and
interleukin-1 alpha in the cervical mucous vaginal
fluid of pregnant women with bacterial vaginosis.
Am. J. Obstet. Gynecol.; 169:1161-1166.
28- Suguira-Ogasawara, M., Nozawa, K.,
Nakanishi, T., Hattori, Y. and Ozaki, Y. (2006):
Complement as a predictor of further miscarriage in
couples with recurrent miscarriages. Hum. Reprod;
21:2711-2714.
29- Makhseed, M., Raghupathy, R., Azizieh, F.,
Farhat, R., Hassan, N. and Bander, A. (2000):
Circulating cytokines and CD30 in normal human
pregnancy and recurrent spontaneous abortions.
Hum.Reprod.,15:2011-2017.
30- Janeway, C.A., Travers, P., Walport, M. and
Shlomchik, M. (2001): Immunology 5 th edn. New
York and London, Garland Science.
31- Makhseed, M., Raghupathy, R., Azizeh, F.,
Omu, A., Al-Shamali, E. and Ashkanani,
L.(2001):Th1 and Th2 cytokine profiles in recurrent
aborters with successful pregnancy and with
subsequent abortions. Hum. Reprod.16:2219-2226.
32- Zenclussen, A.C., Fest, S., Busse, P., Joachim,
R., Klapp, B.F.and Arck, P.C. (2002): Questioning
the Th1/Th2 paradigm in reproduction: peripheral
levels of Il-12 are down- regulated miscarriage
patients. Am. J. Reprod. Immunol., 48:245-251.
33- Laird, S.M., Tuckermann,E.M. and Li,
T.C.(2006): Cytokine expression in the endometrium
of women with implantation failure and recurrent
miscarriage. Reprod. Biomed. Online. 13:13-23.
34- Hattori, Y., Nakanishi, T., Ozaki, Y., Nozawa,
K., Sato, T. and Sugiura-Ogasawarw, M. (2007):
Uterine cervical inflammatory cytokines, interleukin-
6 and -8, as predictors of miscarriage in recurrent
cases. Am. J. Reprod. Immunol.,58:350-357.
35- Lim, K. J., Odukoya, O.A., Ajjan, R.A., Li,
T.C., Weetman, A.P. and Cooke, I.D.(2000): The
role of T-helper cytokines in human reproduction.
Fertil. Steril. 73:136-142.
36- Von Wolff, M., Thaler, C.G., Strowitzki, T.,
Broome, J., Stolz, W. and Tabibzadeh, S. (2000):
Regulated expression of cytokines in human
endometrium throughout the menstrual cycle:
dysregulation in habitual abortion. Mol. Hum.
Reprod., 6:627-634.
37- Diehl, S. and Rincon, M. (2002): The two faces
of Il-6 on Th1/Th2 differentiation. Mol. Immunol.
53:531-536.
38- Hirata, M., Sato, T., Tsumagari, M., Shimada,
A., Nakano, H., Hashizume, K. and Ito, A. (2003):
differential regulation of the expression of matrix
metalloprotienases and tissue inhibitors of
metalloprteinases by cytokines and growth factors in
bovine endometrial stromal cells and trophoblast cell
line BT-1 in vitro. Biol. Reprod., 68:1276-1281.
39- Sunderkotter, C., Steinbrink, K., Goebeler, M.,
Bhardwaj, R. and Sorg, C. (1994): Macorphage and
angiogenesis. J. Leukoc. Biol., 55:410-422.
40- Bowen, J.M., Chamley, L., Mitchell, M.D. and
Keelan, J.A. (2002): Cytokines of the placenta and
extra-placetal membranes: biosynthesis, secretion and
roles in establishment of pregnancy in women.
Placenta, 23:239-256.
41- McGregor, G.A., French, J.I. and Richter, R.
(1990): Antenatal microbiologic and maternal risk
factors associated with prematurity. Am. J. Obstet.
Gynecol., 163:1465-1473.
42- Hay, P. E.,Lamont, R. F. and Taylor-
Robinson, D. (1994): Abnormal bacterial
colonization of the genital tract and subsequent
preterm delivery and late miscarriage. Br. Med. J.,
308:295-299.
43- Hillier, S.L., Nugent, R.P. and Eschenbach,
D.A. for the Vaginal Infections and Prematurity
Study Group (1995): Association between bacterial
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Egyptian Journal of Medical Microbiology, October 2008
Vol. 17, No. 4
vaginosis and preterm delivery of a low birth-weight
infants. N. Engl. J. Med., 333:1737-1742
44- McGregor, J.A., French, J. I. and Lawellin, D.
(1989): In vitro study of bacteria protease-induced
reduction of chorioamionitis membrane strength and
elasticity. Obstet. Gynecol., 69:267-174.
45- Reutter, W.E., Kottgen, E. and Baver, C.
(1982): Biologic significance of sialic acids. In
Shaver, R. (ed.), Sialic acids: Chemistry, metabolism
and function. Springer-Verlag, New York, pp. 263-
305.
46- Paavonen, V., Tersala, K. and Heinone, P.K.
(1987): Microbiologic and histopathologic findings in
acute pelvic inflammatory disease. Br. J. Obstet.
Gynecol., 94:454-460.
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(١)
العلاقة بين مستويات الانترليوآين ٦ و‎٨‎ فى الجهاز التناسلى للسيدات فى حالات الا جهاض المتكرر
المصاحب للبكتيريا المهبلية
٢
١
سحر طاهر و محمد النيجيرى
(٢)
قسم الميكروبيولوجيا والمناعة الطبية آلية الطب جامعة المنصورة قسم النساء و التوليد آلية الطب جامعة
المنصورة
.
تعتبر البكتيريا المهبلية من أشهر أسباب الافرازات المهبلية خاصة خلال فترة الخصوبة ويحدث دلك نتيجة نمو
وتكاثر الجاردنيرللا فاجيناليز بدلا من البكتيريا المهبلية غير الضارة.‏ وقد وجد أنه توجد علاقة بين الاجهاض المتكرر و
تلك الالتهابات التى غالبا ما تكون مصاحبة با رتفاع فى نسب السيتوآينات مثل الانترليوآين تهدف الدراسة الحالية
الى أهمية تحديد نسب الانترليوآين ‎٦‎و‎٨‎ فى حالات الاجهاض المتكرر سواء فى وجود التهاب مهبلى أو عدمه و شملت
٥٤ منهم لديهم تاريخ مرضى للاجهاض المتكرر(مجموعة المرضى)‏ و ٤٠ سيدة لديهم حمل
الدراسة
طبيعى ‏(المجموعة الضابطة)‏ . و قد تم أخذ العينات عن طريق مسحات مهبلية وغسيل مهبلى لعنق الرحم و قد تم فحصها
بواسطة الميكروسكوب و صبغة الجرام ، وتقييم مستويات الانترليوآين ‎٦‎و‎٨‎ بواسطة الاليزا
.
.
‎٦‎و‎٨‎ .
.
.
٩٤ سيدة حامل ،
وقد وجد أن هناك ارتباط بين الاجهاض المتكرر و تقدم سن السيدة آما لوحظ زيادة فرص حدوث الاجهاض المتكرر
في السيدات التى تعانى من الالتهابات المهبلية بنسبة فى مجموعة المرضى و فى المجموعة الضابطة و أن
ميكروب الجاردرينيللا وجد فى نسبة ٣٣% فى مجموعة المرضى و المجموعة الضابطة على التوالى ووجد أن
مستويات الانترليوآين ٦ و‎٨‎ ذات دلالة أحصائية عالية أقل من ٠٫٠٠٠١ بين مجموعة المرضى و المجموعة الضابطة
وأنه باستخدام انترليوآين ٦ التوقع بحدوث الاجهاض المتكرر آان ٥ مرات و فى حالة الانترليوآين
ونستخلص من هده الدراسة أن الالتهابات المهبلية تشكل عاملا من أسباب الاجهاض المتكرر و خاصة فى الحالات الغير
معروف أسبابها وأن هده الالتهابات تكون مصاحبة بارتفاع مستويات السيتوآينات فى الجهاز التناسلى ولذلك ننصح ببدء
علاج هده الالتهابات بواسطة المضادات الحيوية المناسبة أملا فى انخفاض فرص حدوث الاجهاض المتكرر
.
.
٨ آان ٣ مرات .
%٢٠
%٣٧
و ١٥%
646