Immunoglobulin G antibodies against Porphyromonas gingivalis or Aggregatibacter actinomycetemcomitans in cardiovascular disease and periodontitis

Figures & data

Figure 1. Analysis of antigen preparations from A. actinomycetemcomitans (A.a.) and P. gingivalis (P.g.) as used for coating of ELISA plates.

Lanes of sodium dodecyl sulfate polyacrylamide (7%) gels were loaded with amounts of antigens equal to those used for coating of one 96-well plate. Staining with Coomassie Blue (polysaccharides and LPS) are not visible. M_r markers in kilo Daltons.

Figure 2. Validation of assay sensitivity and specificity to antigen preparations from A. actinomycetemcomitans.

A) Homologous titration curves for serum from a patient with aggressive periodontitis colonized with the JP2 clone (serotype b) of A. actinomycetemcomitans in wells saturated with disintegrated serotype b bacterial antigen (●) and whole, formalin-treated serotype b bacteria (■), respectively. B) Comparative titration of sera from four microbiologically and clinically characterized individuals in an ELISA plate coated with an equivalent mixture of antigens from three strains of A. actinomycetemcomitans representing serotypes a, b, and c. The four sera were from a patient with aggressive periodontitis colonized with the JP2 clone of A. actinomycetemcomitans (●), a healthy individual colonized by a non-JP2 clone of A. a. (∎), two healthy individuals without detectable A.a. (▲,▼). C) Comparative titrations of a serum from a patient with aggressive periodontitis colonized with the JP2 clone (serotype b) of A. actinomycetemcomitans in ELISA wells coated with antigens from either A. actinomycetemcomitans serotype b (●), serotype a (∎), or serotype c (▲).

Table 1. Distribution of study population exposure- and covariables by cardiovascular disease (CVD) and periodontitis.

	All	CVD	Periodontitis
All	576 (100%)	200 (35%)	208 (36%)
Age (mean [SD]), years	54 (14)	62 (10)	61 (9)
Sex
Male	311 (54%)	95 (31%)	104 (33%)
Female	265 (46%)	105 (40%)	104 (39%)
Smoking status
Never	252 (44%)	68 (27%)	60 (24%)
Previous	221 (38%)	106 (48%)	96 (43%)
Current	103 (18%)	26 (25%)	52 (50)
Alcohol consumption, units/week
None	84 (15%)	30 (36%)	27 (32%)
1–7 (women)/1–14 (men)	334 (58%)	110 (33%)	109 (33%)
8–14 (women)/15–21 (men)	101 (18%)	36 (36%)	39 (39%)
>14 (women)/>21 (men)	57 (10%)	24 (42%)	33 (58%)
Diabetes*
HbA1c <6.5%	555 (96%)	186 (34%)	199 (36%)
HbA1c ≥6.5%	21 (4%)	14 (67%)	9 (43%)
ISCED
≤10 years	91 (16%)	37 (41%)	44 (48%)
11–12 years	117 (20%)	41 (35%)	52 (44%)
13–14 years	126 (22%)	33 (26%)	36 (29%)
≥15 years	242 (42%)	89 (37%)	76 (31%)
BMI (mean [SD])	25 (4)	26 (4)	26 (4)
CAL mm (mean [SD])	2.52 (1.02)	2.72 (1.06)	3.48 (1.09)
Antibody titers (median [range])
A. actinomycetemcomitans	5 (0–1011)	8 (0–1,011)	7 (0–1,011)
P. gingivalis	82 (2–25,600)	104 (5–25,600)	158 (7–25,600)

BMI, body mass index; CAL, clinical attachment level; CVD, cardiovascular disease; HbA_1c, glycosylated hemoglobin; ISCED, International Standard Classification of Education; SD, standard deviation.

*Defined as HbA_1c ≥6.5 in peripheral blood.

Table 2. Periodontitis: univariable logistic regression analyses.

	p-value	OR (95% CI)
Age (decades)	5.6 × 10^−19	2.25 (1.89–2.70)
Sex (male)	0.150	1.29 (0.91–1.81)
Smoking status
Previous smoker	7.5 × 10^−6	2.46 (1.66–3.66)
Current smoker	1.6 × 10^−6	3.26 (2.02–5.31)
Alcohol consumption, units/week
0	0.930	0.98 (0.58–1.62)
8–14 (women)/15–21 (men)	0.270	1.30 (0.81–2.05)
>14 (women)/>21 (men)	3.6 × 10^−4	2.84 (1.61–5.08)
Diabetes*	0.510	1.34 (0.54–3.23)
ISCED group
1 (≤10 years)	0.004	2.04 (1.25–3.35)
2 (11–12 years)	0.016	1.75 (1.11–2.75)
3 (13–14 years)	0.580	0.87 (0.54–1.39)
BMI	0.057	1.04 (1.00–1.09)
Log A. actinomycetemcomitans Abs	4.3 × 10^−4	1.31 (1.13–1.52)
Log P. gingivalis Abs	2.0 × 10^−10	1.48 (1.31–1.67)
CVD	0.001	1.79 (1.26–2.55)

Bold values indicate statistical significance (p < 0.05).

Abs, serum IgG antibodies; BMI, body mass index; CVD, cardiovascular disease; ISCED, International Standard Classification of Education; log, logarithm transformed.

*Defined as HbA_1c ≥6.5 in peripheral blood.

Table 3. Periodontitis: multivariable logistic regression analyses.

	p-value	OR (95% CI)
Age (decades)	3.96 × 10^−14	2.35 (1.90–2.96)
Sex (male)	0.442	0.84 (0.54–1.30)
Smoking status
Previous smoker	0.001	2.21 (1.39–3.55)
Smoker	3.86 × 10^−7	4.72 (2.61–8.69)
Alcohol consumption, units/week
0	0.936	0.98 (0.53–1.78)
8–14 (women)/15–21 (men)	0.959	0.99 (0.57–1.69)
>14 (women)/>21 (men)	0.137	1.65 (0.86–3.22)
Diabetes*	0.476	0.69 (0.24–1.92)
ISCED group
1 (≤10 years)	0.222	1.42 (0.81–2.52)
2 (11–12 years)	0.351	1.30 (0.75–2.26)
3 (13–14 years)	0.850	1.06 (0.60–1.85)
BMI	0.812	1.01 (0.95–1.06)
CVD/CVD medicine	0.188	0.74 (0.47–1.16)
Log A. actinomycetemcomitans Abs	0.812	1.02 (0.85–1.23)
Log P. gingivalis Abs	1.94 × 10^−5	1.34 (1.17–1.54)

Bold values indicate statistical significance (p < 0.05).

Abs, serum IgG antibodies; BMI, body mass index; CVD, cardiovascular disease; ISCED, International Standard Classification of Education; log, logarithm transformed.

*Defined as HbA1c ≥6.5 in peripheral blood.

Table 4. Cardiovascular disease: univariable logistic regression analyses.

	p-value	OR (95% CI)
Age (decades)	4.4 × 10^−19	2.31 (1.93–2.79)
Sex (male)	0.023	1.49 (1.06–2.11)
Smoking status
Previous smoker	3.0 × 10^−6	2.49 (1.70–3.67)
Current smoker	0.740	0.91 (0.53–1.53)
Alcohol consumption, units/week
0	0.630	1.13 (0.68–1.86)
8–14 (women)/15–21 (men)	0.610	1.13 (0.70–1.79)
>14 (women)/>21 (men)	0.180	1.48 (0.83–2.62)
Diabetes*	0.004	3.97 (1.62–10.62)
ISCED group
1 (≤10 years)	0,520	1.18 (0.72–1.92)
2 (11–12 years)	0,750	0.93 (0.58–1.47)
3 (13–14 years)	0.042	0.61 (0.38–0.97)
BMI	9.6 × 10^−6	1.11 (1.06–1.16)
Log A. actinomycetemcomitans Abs	4.1 × 10^−5	1.37 (1.18–1.60)
Log P. gingivalis Abs	0.006	1.17 (1.05–1.31)
Periodontitis†	0.001	1.79 (1.26–2.55)

Bold values indicate statistical significance (p < 0.05).

Abs, serum IgG antibodies; BMI, body mass index; CVD, cardiovascular disease; ISCED, International Standard Classification of Education; log, logarithm transformed.

*Defined as HbA_1c ≥6.5 in peripheral blood.

†Defined as mean loss of attachment of ≥2.55 mm.

Table 5. Cardiovascular disease: multivariable logistic regression analyses.

	p-value	OR (95% CI)
Age (decades)	5.16×10^−14	2.27 (1.85–2.84)
Gender (men)	0.683	1.09 (0.71–1.67)
Smoking status
Previous smoker	0.002	2.00 (1.28–3.15)
Current smoker	0.941	1.02 (0.55–1.89)
Alcohol consumption, units/week
0	0.278	1.39 (0.76–2.50)
8–14 (women)/15–21 (men)	0.610	0.87 (0.50–1.50)
>14 (women)/>21 (men)	0.809	1.08 (0.56–2.09)
Diabetes*	0.059	2.84 (0.99–8.80)
ISCED group
1 (≤10 years)	0.119	0.63 (0.35–1.12)
2 (11–12 years)	0.038	0.56 (0.32–0.96)
3 (13–14 years)	0.068	0.60 (0.34–1.03)
BMI	0.001	1.09 (1.04–1.15)
Periodontitis†	0.218	0.76 (0.48–1.18)
Log A. actinomycetemcomitans Abs	0.183	1.13 (0.94–1.36)
Log P. gingivalis Abs	0.605	1.04 (0.91–1.18)

Bold values indicate statistical significance (p < 0.05).

Abs, serum IgG antibodies; BMI, body mass index; CVD, cardiovascular disease; ISCED, International Standard Classification of Education; log, logarithm transformed.

*Defined as HbA1c ≥6.5 in peripheral blood.

†Defined as mean loss of attachment of ≥2.55 mm.