193

Journal of Oral Science, Vol. 53, No. 2, 193-202, 2011

Original

Association of interleukin-1 polymorphisms

with periodontitis in Down syndrome
Ahmed Khocht1), Kevin Heaney2), Malvin Janal3) and Bobby Turner4)
1)Department

of Periodontology, School of Dentistry, Temple University, Philadelphia, PA, USA

2)Private Practice, Hackensak, NJ, USA
3)Department of Epidemiology and Health Promotion, New York University, New York, NY, USA
4)Department of Dentistry, East Central Regional Hospital, Gracewood, GA, USA
(Received 11 November 2010 and accepted 28 March 2011)

Abstract: This study examined the association of

IL1 genetic polymorphisms (IL-1A +4845, IL-1B +3954
& IL-1RN +2018) with periodontal disease status of
Down syndrome (DS) individuals. Fifty-four DS patients
(18-56 yr, 48.15% male, 77.78% Caucasians) were
recruited from the Georgia Regional Hospital (GRH)
health care system. Two comparable groups (71
mentally retarded patients and 87 control subjects)
were also recruited. All subjects were nonsmokers.
Periodontal evaluations (plaque index, gingival index,
bleeding-on probing and clinical attachment levels
(AL)), personal and professional dental care habits
were recorded. Blood was collected by a venipuncture.
The IL-1A +4845, IL-1B +3954 & IL-1RN +2018 loci
were genotyped by the TaqMan assay. No statistically
significant differences were noted in the distribution
of IL-1 gene polymorphisms between the three groups.
The IL-1 variant genotypes varied by race; for both IL1A and IL-1RN, the variant gene was significantly
more prevalent among whites than non-whites (ps > 0.1).
ANCOVA, which also adjusted for age, showed a 3-way
interaction among dental visits, gene variation and
Down status [(F(1, 179) = 3.96, P = 0.048 in White
subjects and F(1, 241) = 2.96, P = 0.087 in all subjects).
Post-hoc t-tests confirmed lower levels of AL in IL-1RNvariant Down subjects receiving more frequent dental
visits (P < 0.05). ANCOVA, which also adjusted for age,
Correspondence to Dr. Ahmed Khocht, Department of
Periodontology, School of Dentistry, Temple University, 3223
North Broad Street, Philadelphia, PA 19140, USA
Tel: +1-215-707-1114
Fax: +1-215-707-2439
E-mail: [email protected]

showed an interaction between IL-1A/B gene variation

and Down status (F(1, 174) = 3.04, P = 0.083 in White
subjects and F(1, 235) = 3.72, P = 0.055 in all subjects).
Post-hoc t-tests confirmed lower levels of AL in IL-1A/Bvariant Down subjects (P < 0.05). The distribution of
variant IL-1 genes in DS subjects was not different from
the general population. However the association
between the carriage of the IL-1 rare alleles and
periodontitis differed between the Down and nonDown subjects. The carriage of the IL-1 rare alleles in
the Down subjects tended to confer a protective effect
against loss of periodontal attachment. (J Oral Sci 53,
193-202, 2011)

Keywords: Down syndrome; genetic markers;

interleukin-1; periodontitis; genetic
polymorphism.

Introduction
Down syndrome (DS) is a genetic disease known as
trisomy 21. The condition is associated with an extra
chromosome #21 in affected individuals, giving them a total
chromosome count of 47. It is generally produced by an
abnormal segregation of chromosomes during cell division
(maternal meiotic nondisjunction). It is the most common
chromosomal disorder, with an estimated prevalence of 9.2
cases per 10,000 live births in the United States (1,2).
Different racial groups are equally affected (3). The
condition is associated with characteristic dysmorphic
features, mental retardation, congenital heart defects, and
altered immune responses. In developed nations, life
expectancy of individuals with DS has improved with

194

average life expectancy reaching 56-60 years (4,5).

Periodontal disease is a serious and morbid oral condition
among individuals with DS (6). Gingivitis and periodontitis
start early and their severity increases with age (6). Fiftyeight percent of DS individuals younger than 35 years old
have periodontitis (7). In addition, periodontitis is an
important cause of tooth loss among individuals with DS
(8).
A multitude of factors may be involved in the increased
susceptibility to periodontitis among individuals with DS
(9). Factors previously investigated included mental
retardation (10), subgingival plaque composition,
immune/inflammatory responses and others (9).
Microbiological studies showed that individuals with DS
have significantly higher levels of periodontopathic bacteria
including Porphyromonas gingivalis and Tannerella
forsythensis (11). Neutrophil chemotaxis in individuals with
DS was significantly impaired and alveolar bone loss was
inversely proportional to the chemotactic index (12). Other
immune defects associated with periodontitis in DS
included lymphocyte dysfunction (13) and altered antibody
production (14). Inflammatory mediators (prostaglandin
E2 and leukotriene B4) and degrading enzymes (matrix
metalloproteinase-9) were also increased in gingival
crevicular fluid from patients with DS (15).
The role of the IL-1 family of proinflammatory cytokines
in the pathogenesis of periodontitis is well documented (16).
A cluster of three genes located on the long arm of
chromosome 2q13 code for and regulates IL-1 production.
The IL-1 gene cluster is comprised of IL-1A, IL-1B and
IL-1RN genes that code for IL-1a, IL-1b and IL-1 receptor
antagonist (IL-1ra) respectively. IL-1a and IL-1b are
involved in initiating and propagating immune and
inflammatory reactions. IL-1ra is involved in terminating
the action of both IL-1a and IL-1b by blocking the IL-1
receptors. Polymorphic variations in the IL-1 genes are not
uncommon and are associated with an altered/exaggerated
inflammatory response (17). Single nucleotide
polymorphisms at the following IL-1 gene loci IL-1A
+4845, IL-1B +3954 and IL-1RN +2018 have been
implicated in increased severity and susceptibility to many
inflammatory diseases including periodontitis (17,18).
The simultaneous occurrence of both polymorphic alleles
at the IL-1A +4845 and IL-1B +3954 loci (referred to as:
composite IL-1 genotype positive) has been specifically
associated with increased severity of periodontitis (18).
The prevalence of IL-1 polymorphic alleles and their
association with periodontitis in DS individuals has not
been previously investigated. We hypothesized that DS
subjects would be more likely than non-DS subjects to carry
IL-1 allelic variants associated with increased susceptibility

to periodontitis. The aim of the present study was to

investigate the distribution of IL-1 genotypes in a Down
syndrome subject population and examine the association
of IL1 genetic polymorphisms at the following loci IL-1A
+4845, IL-1B +3954 and IL-1RN +2018 with their
periodontal disease status.

Materials and Methods

Subject recruitment and clinical evaluations were
previously described (10).

Study sites
This study was done in cooperation with the Georgia
Department of Human Resources/Georgia Regional
Hospitals (GRH) in Atlanta, Savannah and Augusta. The
study protocol and consent forms were approved by the
Georgia Regional Hospital Institutional Review Board. The
study included three subject groups, Down syndrome
group (DS), mental retardation non-Down group (MR) and
a mentally normal control group (C). Both the DS and MR
subjects were recruited from the GRH healthcare systems
in Atlanta, Savannah and Augusta, Georgia. All DS and
MR subjects were patients of record at the three hospital
locations; some were institutionalized while others were
outpatients living in group-homes or with their families.
All DS and MR subjects were receiving periodic dental
care at one of the three GRH locations.

Inclusion criteria
The study inclusion criteria implemented for DS subjects
were: confirmed diagnosis of Trisomy 21, receiving
periodic dental care, age 18 years or older, a minimum of
10 teeth present, no other medical conditions known to
affect periodontal status (e.g. diabetes mellitus), no
antibiotic treatment in the past 3 months prior to entry in
the study, no history of cigarette smoking and being able
to cooperate with the study examiners. Study inclusion
criteria for the MR subjects were similar to the DS subjects
except for a confirmed diagnosis of mental retardation
without Trisomy 21. Study inclusion criteria for the C
subjects were also similar to the DS subjects except for
absence of mental retardation.

Subject recruitment
The attending dentist in charge of the dental clinic at
each of the three GRH sites reviewed the available records
and identified dentulous DS patients who would meet the
study criteria and would be able to participate in a dental
examination. Then MR patients matched to the previously
identified DS patients on age, race and gender were
identified from the same hospital records. The matched MR

195

patients were selected based on their ability to cooperate

and sit for the dental examination without need for sedation.
Their mental retardation was secondary to head trauma at
birth. The C subjects were recruited from the general
population living in the vicinity of the GRH locations
used. All C subjects were under care of private dentists
and they also were matched on gender, race and age to the
DS subjects.

Subject screening and enrollment

A total of 289 subjects were screened for the study, 26
were disqualified for medical reasons, 46 completed
portions of the study evaluations and were not able to
return to complete the remaining portions, and 217
completed most of the study evaluations. This report will
focus on a subgroup of 212 subjects with IL-1 genotype
data. Fifty-four of those were DS, 71 were MR and 87 were
C.

Subject characteristics
All subjects in the three groups were adults 18 years or
older, mostly Caucasian and the distribution of males to
females was equal. While groups in the initial cohort were
matched on age, this was no longer true after attrition. None
of the subjects in all groups smoked cigarettes, had diabetes,
or was on a medication known to influence periodontal
status.

Ethical issues
All subjects were able to communicate and understand
spoken English. Prior to commencing the study, consent
was obtained and documented for all subjects. The GRH
dentist in charge personally contacted the family or
caretaker of each potential DS or MR subject, explained
the study protocol and obtained their consent to enroll the
subject in the study. In addition to obtaining the family or
caretaker consent, prior to commencing the study
examination, the study protocol was explained to the DS
or MR subject and their personal consent was also obtained
and witnessed. The C subjects consented on their own
behalf.

Oral/periodontal assessments
All subjects received a comprehensive oral/periodontal
evaluation including probing measurements. Two
experienced dental hygienists blinded to the objectives of
the study performed all dental exams under supervision
of investigator AK. The examiners were calibrated and
standardized in the use of the clinical evaluation measures
employed in the study. Standardization sessions were
performed periodically to recalibrate examiners throughout

the study period. The examiners recorded the Loe and

Silness gingival index (GI) (19) around all teeth present.
Each tooth was scored at six sites, mesiobuccal, buccal,
distobuccal, mesiolingual, lingual and distolingual. The
teeth were then disclosed with D and C Red No. 28 dye
(Sunstar Americas, Inc. Chicago, IL, USA) and the Quigley
Hein plaque index (20) on the same six surfaces was
determined. Surfaces with large restorations and teeth
with crowns were not scored.
A conventional periodontal probe with Williams
markings (PQ-OW, Hu-Friedy, Chicago, IL, USA) was used
for all probing measurements. Probing depth (PD) was
taken on six sites per tooth, mesiobuccal, buccal,
distobuccal, mesiolingual, lingual and distolingual. The
probe was inserted parallel to the long axis of the tooth
on the buccal and lingual surfaces. Interproximally, the
probe was placed with slight angulation, as close to the
contact area as possible. At the gingival margin the reading
was taken to the nearest millimeter. The position of the
gingival margin (GM) to the cementoenamel junction was
recorded at the same six sites per tooth to the nearest
millimeter. A (-) sign was given when the gingival margin
was coronal to the cementoenamel junction and a (+) sign
when it was apical. Attachment levels were calculated
according to the formula AL = PD + GM. Periodontitis
was defines as 5% or higher of teeth scored exhibiting
attachment loss 5 mm. For all the aforementioned
examinations, only fully erupted teeth were used, except
third molars were not included. Caries and missing teeth
were also recorded. The demographic and clinical data of
the entire subject populations were previously presented
(10).
Blood sample collection: Blood was collected by a
venipuncture for IL-1 genotyping and other analysis. For
IL-1 genotyping, a few drops of the collected blood sample
were immediately placed on an AmpliCard (Chemicon
International Inc., Temecula, CA, USA). AmpliCard was
allowed to dry at room temperature then placed in an
envelope and stored at -70C for later analysis.

Analysis of genetic polymorphism

Within a three-month time-period from date of sample
collection, AmpliCards were sent to Interleukin Genetics,
Inc. (Boston, MA, USA) for human DNA extraction and
determination of IL-1 polymorphism. Genotyping was
performed without knowledge of clinical status. All samples
were genotyped for the biallelic markers IL-1A(+4845),
IL-1B(+3954), and IL-1RN(+2018). The wild (common)
allele was designated as (1) and the variant (rare) allele
was designated as (2). DNA was isolated from AmpliCards
using the Whatman FTA Purification reagent (Whatman

196

Inc., Clifton, NJ, USA).

Genotyping was performed using the TaqMan assay
(Perkin-Elmer, Foster City, CA, USA). The TaqMan assay
utilizes the 5-3 nuclease activity of Taq DNA polymerase
to cleave a fluorogenic probe specific for one of two alleles
at the target polymorphism site. Quantitation of
fluorescence was made by comparing each samples
fluorescent activity with that of samples of known genotype,
blank standards containing no DNA, and a background dye
present in the reaction buffer.
Sequences and conditions used in TaqMan genotyping:
IL-1A (+4845)
Probe 1: 5-C(FAM)AA GCC TAG GTC ATC ACC TTT
TAG CTT CTT T(TAMRA)-3
Probe 2: 5-C(TET)AA GCC TAG GTC AGC ACC TTT
TAG CTT CTT T(TAMRA)-3
Forward: 5-ACC CCC TCC AGA ACT ATT TTC CCT-3
Reverse: 5-TGT AAT GCA GCA GCC GTG AGG TAC-3
Cycling: [95C for 2 min] 1; [94C for 1 min, 65C for
1 min, 72C for 1 min] 40; [94C for 12 min, 65C for
2 min, 72C for 5 min] 1
IL-1B (+3954)
Probe 1: 5-A(FAM)CC TAT CTT CTT TGA CAC ATG
GGA TAA CGA T(TAMRA)-3
Probe 2: 5-A(TET)CC TAT CTT CTT CGA CAC ATG

GGA TAA CGA T(TAMRA)-3

Forward: 5-GCT CAG GTG TCC TCC AAG AAA TC-3
Reverse: 5-GTG ATC GTA CAG GTG CAT CGT-3
Cycling: [95C for 2 min, 62C for 1 min, 72C for 1 min]
2; [95C for 1 min, 62C for 1 min, 72C for 1 min] 27;
[94C for 1 min, 62C for 1 min, 72C for 5 min] 3
IL-1RN (+2018)
Probe 1: 5-C(FAM)AA CCA ACT AGT TGC TGG
ATA CTT GCA AG(TAMRA)-3
Probe 2: 5-C(TET)AA CCA ACT AGT TGC CGG ATA
CTT GCA AG(TAMRA)-3
Forward: 5-AAG TTC TGG GGG ACA CAG GAA G-3
Reverse: 5-ACG GGC AAA GTG ACG TGA TG-3
Cycling: [96C for 1 min] 1; [94C for 1 min, 63C for
1 min, 70C for 1 min] 35; [63C for 5 min, 70C for 5
min] 1.

Statistical analysis
Analysis of variance (parametric data) and chi-square
analysis (non-parametric data) were used to examine the
differences between the groups. Analysis of covariance
(ANCOVA) was used to examine the effect of IL-1 variant
genes on periodontal disease measures after adjusting for
age, gender, plaque levels and dental visits.

Results
Demographic and clinical data of this subset of subjects
with IL-1 genotyping data are summarized in table 1. In

Table 1 Summary of demographic and clinical data of a subset of subjects with IL-1
genotype data presented in this report

197

summary all three groups were matched on race and

gender. The MR group was older than the DS and C group
(P = 0.0001). Number of institutionalized subjects in the
MR group was higher than the DS group (0.001). Clinically,
subjects in both DS and MR groups showed higher levels
of GI (P = 0.0001), PI (P = 0.0002) and missing teeth (P
= 0.0001) than the control group. The DS subjects showed
greater loss of clinical periodontal attachment (P = 0.002)
and higher percentage of teeth with AL 5mm (P = 0.001)
than both the MR and the C groups. Percentage of subjects
with periodontitis was similarly distributed among the
three groups, DS 72%, C 71% and MR 80%.
The IL-1A, IL-1B and IL-1RN genotype and allele
frequencies were essentially similar between the three
groups. Also no statistical differences were found between
the three groups in the distribution of IL-1 composite
genotype (simultaneous carriage of variant alleles in both
the IL-1A and IL-1B genes. Thus the carriage of the IL1 gene polymorphisms in the Down subjects was similar
to the non-Down subjects. Table 2 summarizes the
distribution of the alleles for all subjects combined.
On the other hand significant differences were found in
the distribution of IL-1 genotypes between Whites and nonWhites. Table 3 shows that the IL-1 variant (rare) genotypes
varied by race. For both IL-1A and IL-1RN, the variant
gene was significantly more prevalent among Whites than
non-Whites. For IL-1B and composite genotype IL-1A/B,
the variant gene also trended higher in Whites, but did not

reach statistical significance. Thus, data generally suggest

that variant forms of the IL-1 genes are more likely to be
found among White than Non-white individuals.
We next investigated the effect of the IL-1 gene
polymorphisms on gingival inflammation. ANCOVA,
which also adjusted for age, plaque and group differences,
showed that in individuals positive for IL-1B (+3954)
gene polymorphism had less GI scores (F(1, 242) = 5.70,

Table 2 Distribution of the IL-1 alleles for all subjects

combined

Table 3 IL-1 variant genotype distribution by race

198

P = 0.01). GI for IL-1 +3954 positive individuals averaged

0.75, while IL-1 +3954 negative individuals averaged
0.88. The model showed no differences in effect on GI
between Down and non-Down subjects. This analysis
suggests that the variant form of IL-1B genotype appears
to lessen gingival inflammation scores. No effects were
found for IL-1A or IL-1RN or IL-1 composite positive
genotype on gingival inflammation scores.
We next investigated whether professional dental visits

Fig. 1 Bar chart of adjusted AL mean (SEM) and IL1-RN

polymorphism by group and frequency of dental visits.
The bars represent the AL mean after adjusting for age.
The whiskers indicate the standard error of the mean.
PDV- = professional dental visits once or less per
year. PDV+ = professional dental visits more than
once per year. Significance of differences among
groups was sought using ANCOVA.

Fig. 2 Bar chart of adjusted AL mean (SEM) and IL1 A/B

composite genotype by group. The bars represent the
AL mean after adjusting for age. The whiskers indicate
the standard error of the mean. Significance of
differences among groups was sought using ANCOVA.

and variation in the IL-1 genotypes might mediate the

association between AL and Down status. Figure 1 shows,
for IL-RN, AL of just over 2 mm for non-Down subjects,
regardless of visits or gene variation. On the other hand,
Down subjects with lower levels of dental care or higher
levels of dental care and the wild genotype averaged near
3 mm of AL. Interestingly, however, Down subjects with
higher levels of dental care and the variant IL-RN genotype,
averaged near 2 mm of AL, similar to non-Down subjects.
ANCOVA, which also adjusted for age, showed a 3-way
interaction among dental visits, gene variation and Down
status [(F(1, 179) = 3.96, P = 0.048 in White subjects and
F(1, 241) = 2.96, P = 0.087 in all subjects). While this
interaction was stronger among White subjects, race did
not produce a statistically significant difference in this
interaction]. Post-hoc t-tests confirmed lower levels of
AL in IL-1RN-variant Down subjects getting higher than
lower levels of dental care (P < 0.05). Thus, AL appears
preventable in one subgroup of Down subjects, those that
receive more professional care and also evidence the
variant form of the IL-1RN genotype. Stated differently,
this analysis suggests that the wild form of this gene
appears to put Down individuals at increased risk for
periodontal disease.
We next investigated whether professional dental visits
also interacted with the IL-1A and IL-1B genotypes to
mediate the association between AL and Down status.
However, while no effects were found for either IL-1A or
IL-1B, a simpler effect was noted for the IL-1A/B
composite measure. Figure 2 shows, for IL-1 composite,
AL of just over 2 mm for non-Down subjects, regardless
of visits or gene variation. On the other hand, Down
subjects showing the wild genotype averaged near 3 mm
of AL., while Down subjects with the variant IL-RN
genotype, averaged near 2.5 mm of AL. ANCOVA, which
also adjusted for age, showed an interaction between IL1A/B gene variation and Down status (F(1, 174) = 3.04,
P = 0.083 in White subjects and F(1, 235) = 3.72, P = 0.055
in all subjects). Post-hoc t-tests confirmed lower levels of
AL in IL-1A/B-variant than IL-1A/B-wild Down subjects
(P < 0.05). Thus, AL appears less severe in the subgroup
of Down subjects that evidence the variant form of the IL1A/B genotype. As above, this analysis suggests that the
wild form of the IL1-A/B genotype appears to put Down
individuals at increased risk for periodontal disease.

Discussion
The objectives of this study were to compare the
distribution of IL-1 genotypes between Down and nonDown subjects and examine the association between
presence of IL-1 variant (rare) alleles with AL in Down

199

and non-Down subjects. Our data showed no difference

in the distribution of IL-1 genotypes between Down and
non-Down subjects, however the association between IL1 polymorphic genotypes and periodontitis differed between
the Down and non-Down subjects. Presence of the variant
alleles of the IL-1 gene family in the Down subjects tended
to be protective against loss of periodontal attachment.
The distribution and frequency of IL-1 alleles and
positive IL-1 composite genotype is known to vary by race
and ethnic groups. In European populations the frequency
of a positive IL-1 composite genotype ranges from 29%
to 46% (21-23). In the United States the frequency of a
positive IL-1 composite genotype in Whites ranges from
29% to 38% (18,24-26) and in African Americans 14%
(27). Frequency of the positive IL-1 composite genotype
in Whites in our study (34.95%) falls within the range
previously reported in White populations. However the
positive IL-1 composite genotype frequency (23.08%) in
our non-White population (predominantly African
Americans) was higher than previously reported by Walker
et al. in African Americans (27). The higher frequency of
the IL-1 variant alleles in the non-Whites in our study may
be due to the diverse ethnic backgrounds of non-White
subjects.
There is mounting evidence linking the presence of IL1 gene polymorphisms with various inflammatory diseases
including periodontitis (17,18). The premise behind this
association may be attributed to increased IL-1 cytokines
production and delayed termination of their action thus
resulting in an exaggerated and uncontrolled inflammatory
response. Research (28) showed that monocytes from
polymorphic IL-1B positive individuals produce higher
amounts of IL-1b than monocytes from individuals with
the wild IL-1B genotype. Engebertson et al. (29) reported
increased IL-1b cytokine levels in gingival fluid in IL-1
composite genotype positive subjects. Shirodaria et al.
(30) reported that polymorphisms in the IL-1A gene are
associated with increased levels of IL-1a protein in gingival
fluid. On the other hand, IL-1RA genotype 2 is associated
with reduced mucosal concentrations of IL1ra cytokine
proteins (31); thus in affected individuals the action of both
IL-1a and IL-1b may be prolonged.
The finding that IL-1B (+3954) positive individuals
had less GI scores than IL-1B (+3954) negative individuals
contradicts with the presumed exaggerated inflammatory
response associated with presence of IL-1 polymorphic
genes. Muller et al. (32) reported similar findings in young
adults. The type of gingivitis being investigated may
explain this contradiction. Steady-state gingivitis (a
longstanding gingival inflammation, as investigate in this
study) is characterized by a dampened inflammatory

response associated with low IL-1b levels in gingival

crevicular fluid (33). Thus other late-stage intervening
mechanisms involved in controlling the gingival
inflammatory responses may have masked the hyperinflammatory effect of the IL-1 gene polymorphisms.
The initial study (18) that investigated the relation
between presence of IL-1 polymorphic genes and
periodontitis reported that in White non-smokers, the
simultaneous presence of variant IL-1 alleles at the IL-1A889 (currently +4845) and IL-1B+3953 (currently +3954)
loci are associated with increased severity of periodontitis.
When both IL-1 polymorphic alleles were present, the
affected individual was referred to as composite genotypepositive. Presence of IL-1 gene polymorphisms at a single
locus did not associate with periodontal disease. Other
studies in White populations confirmed similar associations
between the IL-1 composite genotype and severity of
chronic periodontitis (26,34) and aggressive periodontitis
(22). However other studies reported contradictory findings
disputing the association between the presence of the IL1 composite genotype and periodontitis (23,27,35). In our
study, in the non-Down subjects there was no association
between the presence of the variant alleles of IL-1A, IL1B, IL-1RN alone or in-combination (when simultaneously
present) and loss of periodontal attachment. However in
the Down subjects, the presence of the IL-1 composite
genotype or the presence of IL-1RN variant allele inversely
associated with loss of periodontal attachment. It is of
interest to note that presence of IL-1RN variant allele
needed the added benefit of more frequent preventive
dental visits to exert its beneficial effect on the periodontal
status of Down subjects.
The lack of association between the IL-1variant alleles
and periodontitis in the non-Down subjects in our study
may be explained by the fact that all subjects were receiving
periodic preventive dental care. Perhaps in the general
population periodic preventive dental care can circumvent
and negate the adverse genetic influence of the IL-1 variant
alleles on the periodontium. These findings agree with other
studies that reported successful outcome of both nonsurgical and surgical periodontal therapy in IL-1 composite
genotype positive individuals (36-39).
The inverse association noted in this study between the
presence of the IL-1 variant alleles and loss of periodontal
attachment in Down subjects is contradictory to previous
reports in the general population. It suggests that the
presence of the IL-1 variant alleles in Down individuals
bestows protection against periodontitis. This was a
surprising and unexpected finding that refuted our
hypothesis for increased susceptibility to periodontitis in
Down subjects. Perhaps the altered/exaggerated inflam-

200

matory reaction associated with the presence of the variant

IL-1 alleles compensates for other immune deficiencies
in the Down individuals and helps them cope better with
their periodontal problems.
Even though the initial report associating IL-1 genotyping
and periodontal disease excluded cigarette smokers from
the analysis (18), other studies showed that both IL-1
genotyping and smoking are relevant independent risk
factors for periodontal disease and they also represent a
gene-environmental interaction in periodontitis (24,25,40).
In the present study the interaction of smoking with IL-1
genotyping was not investigated because none of our
Down subjects smoked. Both our controls and MR subjects
were selected to match the Down group and thus none of
the subjects smoked.
In conclusion, it is well documented that Down syndrome
individuals are more susceptible to periodontal loss of
clinical attachment than non-Down individuals. Our study
showed that even though the IL-1 genotype distribution
in the Down subjects is similar to the non-Down subjects,
the association between the carriage of the IL-1 rare alleles
and periodontitis differed between the Down and nonDown subjects. The carriage of the IL-1 rare alleles in the
Down subjects tended to bestow a protective effect against
loss of periodontal attachment. This suggests that the
pathogenisis of periodontitis in Down subjects is different
from the general population.

Acknowledgements
This study was supported by the National Institute of
Dental and Craniofacial Research, Bethesda, Maryland
(NIDCR: DE15012-02). We are very grateful to Dr.
Kenneth Kornman (Interleukin Genetics, Inc.) for his
guidance, support and help with genotyping of the samples.

References
1. Forrester MB, Merz RD (2002) Epidemiology of
Down syndrome (Trisomy 21), Hawaii, 1986-97.
Teratology 65, 207-212.
2. Khoshnood B, Wall S, Pryde P, Lee KS (2004)
Maternal education modifies the age-related increase
in the birth prevalence of Down syndrome. Prenat
Diagn 24, 79-82.
3. Siffel C, Correa A, Cragan J, Alverson CJ (2004)
Prenatal diagnosis, pregnancy terminations and
prevalence of Down syndrome in Atlanta. Birth
Defects Res A Clin Mol Teratol 70, 565-571.
4. Glasson EJ, Sullivan SG, Hussain R, Petterson BA,
Montgomery PD, Bittles AH (2002) The changing
survival profile of people with Downs syndrome:
implications for genetic counselling. Clin Genet

62, 390-393.
5. Yang Q, Rasmussen SA, Friedman JM (2002)
Mortality associated with Downs syndrome in the
USA from 1983 to 1997: a population-based study.
Lancet 359, 1019-1025.
6. Reuland-Bosma W, van Dijk J (1986) Periodontal
disease in Downs syndrome: a review. J Clin
Periodontol 13, 64-73.
7. Amano A, Kishima T, Akiyama S, Nakagawa I,
Hamada S, Morisaki I (2001) Relationship of
periodontopathic bacteria with early-onset
periodontitis in Downs syndrome. J Periodontol 72,
368-373.
8. Lpez-Prez R, Borges-Yez SA, Jimnez-Garca
G, Maupom G (2002) Oral hygiene, gingivitis,
and periodontitis in persons with Down syndrome.
Spec Care Dentist 22, 214-220.
9. Morgan J (2007) Why is periodontal disease more
prevalent and more severe in people with Down
syndrome? Spec Care Dentist 27, 196-201.
10. Khocht A, Janal M, Turner B (2010) Periodontal
health in Down syndrome: contributions of mental
disability, personal, and professional dental care.
Spec Care Dentist 30, 118-123.
11. Sakellari D, Arapostathis KN, Konstantinidis A
(2005) Periodontal conditions and subgingival
microflora in Down syndrome patients. A casecontrol study. J Clin Periodontol 32, 684-690.
12. Izumi Y, Sugiyama S, Shinozuka O, Yamazaki T,
Ohyama T, Ishikawa I (1989) Defective neutrophil
chemotaxis in Downs syndrome patients and its
relationship to periodontal destruction. J Periodontol
60, 238-242.
13. Shoel DC, Jonsson R, Johannessen AC, Nilsen R
(1995) Gamma/delta T lymphocytes in marginal
periodontitis in patients with Downs syndrome.
Adv Exp Med Biol 371B, 1135-1136.
14. Chaushu S, Chaushu G, Zigmond M, Yefenof E,
Stabholz A, Shapira J, Merrick J, Bachrach G (2007)
Age-dependent deficiency in saliva and salivary
antibodies secretion in Downs syndrome. Arch
Oral Biol 52, 1088-1096.
15. Tsilingaridis G, Yucel-Lindberg T, Moder T (2003)
Enhanced levels of prostaglandin E2, leukotriene B4,
and matrix metalloproteinase-9 in gingival crevicular
fluid from patients with Down syndrome. Acta
Odontol Scand 61, 154-158.
16. Barksby HE, Lea SR, Preshaw PM, Taylor JJ (2007)
The expanding family of interleukin-1 cytokines
and their role in destructive inflammatory disorders.
Clin Exp Immunol 149, 217-225.

201

17. Kinane DF, Shiba H, Hart TC (2005) The genetic

basis of periodontitis. Periodontol 2000 39, 91-117.
18. Kornman KS, Crane A, Wang HY, di Giovine FS,
Newman MG, Pirk FW, Wilson TG Jr, Higginbottom
FL, Duff GW (1997) The interleukin-1 genotype as
a severity factor in adult periodontal disease. J Clin
Periodontol 24, 72-77.
19. Le H (1967) The Gingival Index, the Plaque Index
and the Retention Index Systems. J Periodontol 38,
Suppl, 610-616.
20. Quigley GA, Hein JW (1962) Comparative cleansing
efficiency of manual and power brushing. J Am
Dent Assoc 65, 26-29.
21. Hodge PJ, Riggio MP, Kinane DF (2001) Failure
to detect an association with IL1 genotypes in
European Caucasians with generalised early onset
periodontitis. J Clin Periodontol 28, 430-436.
22. Parkhill JM, Hennig BJ, Chapple IL, Heasman PA,
Taylor JJ (2000) Association of interleukin-1 gene
polymorphisms with early-onset periodontitis. J
Clin Periodontol 27, 682-689.
23. Sakellari D, Koukoudetsos S, Arsenakis M,
Konstantinidis A (2003) Prevalence of IL-1A and
IL-1B polymorphisms in a Greek population. J Clin
Periodontol 30, 35-41.
24. McDevitt MJ, Wang HY, Knobelman C, Newman
MG, di Giovine FS, Timms J, Duff GW, Kornman
KS (2000) Interleukin-1 genetic association with
periodontitis in clinical practice. J Periodontol 71,
156-163.
25. McGuire MK, Nunn ME (1999) Prognosis versus
actual outcome. IV. The effectiveness of clinical
parameters and IL-1 genotype in accurately
predicting prognoses and tooth survival. J
Periodontol 70, 49-56.
26. Papapanou PN, Neiderud AM, Sandros J, Dahln
G (2001) Interleukin-1 gene polymorphism and
periodontal status. A case-control study. J Clin
Periodontol 28, 389-396.
27. Walker SJ, Van Dyke TE, Rich S, Kornman KS, di
Giovine FS, Hart TC (2000) Genetic polymorphisms
of the IL-1 and IL-1 genes in African-American
LJP patients and an African-American control
population. J Periodontol 71, 723-728.
28. Pociot F, Mlvig J, Wogensen L, Worsaae H, Nerup
J (1992) A TaqI polymorphism in the human
interleukin-1 (IL-1 ) gene correlates with IL-1
secretion in vitro. Eur J Clin Invest 22, 396-402.
29. Engebretson SP, Lamster IB, Herrera-Abreu M,
Celenti RS, Timms JM, Chaudhary AG, di Giovine
FS, Kornman KS (1999) The influence of interleukin

gene polymorphism on expression of interleukin1 and tumor necrosis factor- in periodontal tissue
and gingival crevicular fluid. J Periodontol 70, 567573.
30. Shirodaria S, Smith J, McKay IJ, Kennett CN,
Hughes FJ (2000) Polymorphisms in the IL-1A
gene are correlated with levels of interleukin-1
protein in gingival crevicular fluid of teeth with
severe periodontal disease. J Dent Res 79, 18641869.
31. Andus T, Daig R, Vogl D, Aschenbrenner E, Lock
G, Hollerbach S, Kllinger M, Schlmerich J, Gross
V (1997) Imbalance of the interleukin 1 system in
colonic mucosa association with intestinal
inflammation and interleukin 1 receptor antagonist
[corrected] genotype 2. Gut 41, 651-657.
32. Mller HP, Barrieshi-Nusair KM (2007) A
combination of alleles 2 of interleukin (IL)-1A(-889)
and IL-1B(+3954) is associated with lower gingival
bleeding tendency in plaque-induced gingivitis in
young adults of Arabic heritage. Clin Oral Investig
11, 297-302.
33. Deinzer R, Weik U, Kolb-Bachofen V, Herforth A
(2007) Comparison of experimental gingivitis with
persistent gingivitis: differences in clinical
parameters and cytokine concentrations. J
Periodontal Res 42, 318-324.
34. Galbraith GM, Hendley TM, Sanders JJ, Palesch Y,
Pandey JP (1999) Polymorphic cytokine genotypes
as markers of disease severity in adult periodontitis.
J Clin Periodontol 26, 705-709.
35. Sakellari D, Katsares V, Georgiadou M, Kouvatsi
A, Arsenakis M, Konstantinidis A (2006) No
correlation of five gene polymorphisms with
periodontal conditions in a Greek population. J Clin
Periodontol 33, 765-770.
36. Christgau M, Aslanidis C, Felden A, Hiller KA,
Schmitz G, Schmalz G (2003) Influence of
interleukin-1 gene polymorphism on periodontal
regeneration in intrabony defects. J Periodontal Res
38, 20-27.
37. Cortellini P, Tonetti MS (2004) Long-term tooth
survival following regenerative treatment of
intrabony defects. J Periodontol 75, 672-678.
38. Knig J, Rhling A, Plagmann HC, Meisel P, Kocher
T (2005) Influence of interleukin (IL)-1 composite
genotype on clinical variables in non-smoking, wellmaintained compliant patients with chronic
periodontitis. Swed Dent J 29, 11-16.
39. Weiss OI, Caton J, Blieden T, Fisher SG, Trafton
S, Hart TC (2004) Effect of the interleukin-1

202

genotype on outcomes of regenerative periodontal

therapy with bone replacement grafts. J Periodontol
75, 1335-1342.
40. Meisel P, Siegemund A, Dombrowa S, Sawaf H,

Fanghaenel J, Kocher T (2002) Smoking and

polymorphisms of the interleukin-1 gene cluster
(IL-1, IL-1, and IL-1RN) in patients with
periodontal disease. J Periodontol 73, 27-32.