Table 1.

CpG ODN as adjuvants in infectious disease trials.

Trial no.	Infection	Treatment	Route	Phase	Date	Results	Reference
NCT00344539	Malaria	AMA1-C1/Alhydrogel® (20, 80 μg) ± CpG 7909 (564 μg) n = 75 in 3 groups days: 0, 28, 56	IM	I	04/05–03/07	AMA-1 + CpG vaccination supported an 11–14 fold increase in anti-AMA1 Ab titers (p < 02), PF GIA was 3–4 fold higher (p < 0.0001) than AMA-1 alone, but similar growth inhibitory activity against PF in standardized AMA1-IgG titer.	[76,135]
NCT00414336	Malaria	AMA1-C1/Alhydrogel® (80 μg) ± CpG 7909 (564 μg) n = 24 in 2 groups days: 0, 28	IM	I	10/07–11/07	AMA-1 + CpG vaccination of malaria exposed adults increased AMA1-specific Abs by 2–3 fold vs. AMA-1 alone; no significant differences in Ag specific memory B cells or GIA between groups.	[74,136]
NCT00320658	Malaria	AMA1-C1/Alhydrogel® (20, 80 μg) + MSP142 -C1/Alhydrogel® (80 μg) ± CpG 7909 (564 μg) n = 24 in 2 groups days: 0, 28, 56	IM	I	07/06–07/07	CpG 7909 enhanced significantly the appearance or AMA1-C1-specific memory B cells (2–10 fold at days 35, 56, 59; p < 0.01) and MSP42 -specific memory B cells (7–10 fold at day 3 and 84 after 3rd vaccination) in PBMC.	[75,126]
NCT00984763	Malaria	AMA1-C1/Alhydrogel® (80 μg) ± CpG 7909 (564 μg) n = 10 in 2 groups days: 0, 28, 56	IM	I/IIa	07/09–09/10	Inclusion of CpG significantly increased GIA titers (63% versus 13% p < 0.01) and correlated with lower PMR.	[78]
NCT00889616	Malaria	BSAM2/Alhydrogel® (40, 160 μg) + CPG 7909 (564 μg) n = 30 in 2 groups days: 0, 56, 180	IM	I	06/09–03/10	Both doses of BSAM2/Alhydrogel® + CpG 7909 were immunogenic: increase of anti-AMA1 (86 and 113 μg/mL) and anti-MSP42 titers (57 and 76 μg/mL) Ab at day 194 with boosting effect after the second vaccination; GIA increased 7–41%.	[137]
NA	Influenza	Fluarix® (0.5, 0.05 mL) ± CpG 7909 (1 mg) n = 60 in 4 groups Single dose	IM	Ib	08/99–10/99	CpG 7909 did not increase anti-HI Ab titers vs. vaccine alone (p > 0.05). Upon Ag restimulation, PBMC at 4 week showed significantly higher IFN-γ production for A/Beijing/262/95 (p = 0.048) and B/Harbin/7/94 (p = 0.0057) in 1/10th dose Fluarix® + CpG 7909 compared to 1/10th dose Fluarix® alone.	[82]
NCT00562939	Pneumococcus in HIV-infected adults	PCV7 (Prevnar®), PPV-23 (PneumoNovum®) (1 mL) ± CPG 7909 (1 mg) n = 97 in 2 groups 0, 3, 9 months	IM	Ib/IIa	01/08–01/09	CpG group showed a mean reduction in proviral DNA of 12.6% vs. a 6.7% increase in the non-GpG group (p = 0.02). No significant changes in HIV specific Ab or T cell immunity. PBMC from the CpG group produced significantly more IL-1β (p = 0.0046), IL-6 (p = 0.0051), IFN-γ (p = 0.0047), MIP-β (p = 0.0086), IL-2R (p = 0.0062) levels. PBMC of CpG vaccinated patients stimulated with LPS showed elevated production of L-1β (p = 0.038) and IFN-γ (p = 0.019) vs. vaccine alone.	[93–95]
NCT00722839	Cytomegalovirus (CMV)	PADRE-CMV (1.5, 2.5, 10 mg), Tetanus-CMV fusion peptide, ±CpG 7909 (1 mg) n = 63 in 4 groups days: 0, 21, 42, 63	SQ	Ib	05/10–10/10	CMV alone induced no response. CD8 T cell responses induced in 30% of CpG plus PADRE-CMV and 70% of Tetanus-CMV plus CpG groups (p = 0.002). pp65 specific CD8 T cells increased 5–10 fold in tetramere binding assay compared to vaccine alone (p = 0.004) and 4–5 fold vs. prevaccination levels (p = 0.008).	[138]
NA	HBV	HBsAg (20 μg) + 1018 ISS (300, 650, 1000, 3000 μg) n = 48 in 6 groups 0, 8 wk	IM	I	NA	Seroprotective titers (>10 mIU/L) were reached by 0, 25, 75 and 87% of patients after 1st vaccination vs. 62, 100, 100, 100% after 2nd vaccination with increasing CpG ODN doses. GMA anti-HBsAg were 1.22, 5.78, 24.75, 206.5 after 1st and 65.37, 877.6, 1545, 3045 mIU/mL after 2nd vaccination. Injection site reactions were more frequent in CpG group (p ≤ 0.05) but all doses were well tolerate.	[62]
NA	HBV	rHBsAg (20 μg) + 1018 ISS (3 mg), Engerix-B® n = 99 in 2 groups 0, 8, 24 weeks	IM	II	NA	CpG increased the percentage of participants achieving seroprotection after 1st, 2nd and 3rd vaccination compared to Engerix-B® alone (79% vs. 12%; 100% vs. 18%; 100% vs. 64%; p < 0.001), GMC of anti-HBsAg Ab were higher in CpG group at week 1, 4, 8 and 16 after the 1st and 2nd dose (p < 0.001).	[63]
NA	HBV	rHBsAg (20 μg) + 1018 ISS (3 mg), Engerix-B® (1 mL) n = 2415 in 2 groups 0, 4 or 0, 4, 24 weeks	IM	III	12/06–07/07	Higher anti-HBV titers and improved seroprotection was achieved in the CpG group at 4, 8, 24 and 28 weeks post vaccination (23% vs. 3%; 88% vs. 26%; 98% vs. 32%; 97% vs. 81%). GMC were >20-fold higher after the 2nd vaccination and at similar levels as Engerix-B® group after the 3rd vaccination.	[64]
NA	HBV	rHBsAg (20 μg) + 1018 ISS (3 mg), n = 41 in 2 groups 0, 4 or 0, 8 week	IM	III	NA	Seroprotective anti-HBsAg Ab titers (≥ 10 mIU/mL) were higher in the CpG adjuvanted group (94% vs. 69%) at week 4. GMC levels were 15-fold higher at 4 week (243 vs. 16 mIU/mL; p < 0.001). Difference between groups diminished with boosting but persisted through week 32 (863 vs. 439 mIU/mL; p = 0.038).	[139]
NA	HBV in adults 40–70 years	rHBsAg (20 μg) + 1018 ISS (3 mg), Engerix-B® (1 mL) n = 409 in 2 groups 0, 4, 8, 24 weeks	IM	III	NA	Seroprotection achieved in more CpG recipients at 4 and 28 week (96% vs. 24%; 100% vs. 73%; p < 0.0001). Titers of anti-HBsAg >100 mIU/mL were induced in 97% vs. 58% at week 28 and in 90% vs. 43% at week 50 compared to Engerix-B® (p < 0.0001) as well as significantly higher GMC were seen at these time points (p < 0.0001).	[65]
NA	HBV	Engerix-B® (20 μg HBsAg) ± CpG 7909 (0.125, 0.5, 1 mg) n = 56 in 4 groups 0, 4, 24 weeks	IM	I/II	04/99–06/01	Inclusion of CpG accelerated the development of protective anti-HBsAg Ab titers (p < 0.001) vs. vaccine alone. The 0.5 mg dose of CpG 7909 was most effective (2-fold higher than 0.125 or 1.0 mg, p < 0.05).	[61]
NA	HBV in HIV-infected adults	Engerix-B® (40 μg HBsAg) + CpG 7909 (0.5, 1 mg) n = 58 in 4 groups (n = 38 in 2 groups) 0, 4, 8 week	IM	Ib/IIa	01/01–08/02	Inclusion of CpG significantly increased anti-HBsAg Ab titers from 6 to 48 weeks. T helper cells showed higher proliferative response to HBsAg ex vivo at week 8 (p = 0.042) and 48 (p = 0.024). No changes in CD4 or CD8 T cell subsets through 5 years of follow up. The CpG group retained seroprotective Anti-ABsAg titers at 60 months (80% vs. 40%, p = 0.004).	[90–92]
NA	Anthrax	Bio-Thrax® (0.5 mL), ±CpG 7909 (1 mg) n = 69 in 3 groups days: 0, 14, 28	IM	I	NA	BioThrax® + CpG 7909 induced 6 fold higher IgG anti-PA Ab titers (1465 μg/mL vs. 232 μg/mL, p < 0.001) that peaked earlier (day 21 vs. 40, p < 0.001) vs. vaccine alone. Toxin neutralizing capacity was 8.8 fold higher (p < 0.001) and also developed earlier (day 22 vs. 46, p < 0.001).	[53]
NCT01263691	Anthrax	Bio-Thrax® (0.5 mL), AV7909: AVA (0.25–0.5 mL) + CpG 7909 (0.25–0.5 mg) n = 105 in 4 groups days: 0, 14	IM	I	NA	Inclusion of any dose of AV 7909 yielded 2-fold higher GMT in TNA NF50 (50% neutralization factor). CpG accelerated reaching peak titer (day 28 vs. 35).	[54]