Vaccination with the Chlamydia trachomatis Major Outer Membrane Protein Can Elicit an Immune Response as Protective as That Resulting from Inoculation with Live Bacteria

DOI: 10.1128/IAI.73.12.8153-8160.2005

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FIG. 1.
(A) SDS-PAGE of C. trachomatis MoPn MOMP purified by hydroxyapatite chromatography and stained with silver nitrate. Lane 1, molecular mass standards; lane 2, C. trachomatis MoPn MOMP. (B) Immunoblot of C. trachomatis MoPn EB probed with serum samples collected the day before the genital challenge. Lane 1, molecular mass standards. Probe with serum from mice immunized with the following: MOMP plus CpG plus Montanide (lane 2), ovalbumin plus CpG plus Montanide (lane 3), MOMP plus CpG plus alum (lane 4), ovalbumin plus CpG plus alum (lane 5), C. trachomatis MoPn EB (lane 6), MEM-0 (lane 7), and control monoclonal antibody to the C. trachomatis MoPn MOMP (lane 8).

TABLE 1.

Antibody levels the day before genital challenge

Antigen	Adjuvant	C. trachomatis MoPn-specific ELISA antibody titer									Serum neutralizing titer
		Serum (10³)							Vaginal wash
		IgM	IgG	IgG1	IgG2a	IgG2b	IgG3	IgA	IgA	IgG
MOMP	CpG + Montanide	0.1	409.6	25.6	51.2	204.8	25.6	3.2	160	160	1,250
Ovalbumin	CpG + Montanide	<0.1	<0.1	<0.1	<0.1	<0.1	<0.1	<0.1	<20	<20	<10
MOMP	CpG + alum	0.1	102.4	25.6	6.4	51.2	12.8	0.8	40	20	1,250
Ovalbumin	CpG + alum	<0.1	<0.1	<0.1	<0.1	<0.1	<0.1	<0.1	<20	<20	<10
C. trachomatis		0.1	102.4	1.6	25.6	25.6	3.2	3.2	1,280	320	1,250
MEM-0		<0.1	<0.1	<0.1	<0.1	<0.1	<0.1	<0.1	<20	<20	<10

TABLE 2.

T-cell response the day before genital challenge^a

Antigen	Adjuvant	T-cell proliferation response (10³ cpm) to:					In vitro cytokine production
		T-cell proliferation response (10³ cpm) to:						IFN-γ (pg/ml)			IL-4 (pg/ml)		TNF-α (pg/ml)
				EB^b	ConA^c	Medium	EB^b	ConA^c	EB^b	ConA^c	EB^b	ConA^c
MOMP	CpG + Montanide	10.3 ± 2.3^d	66.5 ± 1.6	0.2 ± 0.05	3,973 ± 1^d	2,510 ± 128	<0.8	58 ± 3	459 ± 41^d	318 ± 0
Ovalbumin	CpG + Montanide	0.9 ± 0.1	81.0 ± 9.1	0.09 ± 0.01	561 ± 14	3,341 ± 34	<0.8	43 ± 2	302 ± 5	288 ± 8
MOMP	CpG + alum	6.1 ± 2.2^d	73.7 ± 9.4	0.3 ± 0.3	2,097 ± 26^d	3,266 ± 0	2 ± 0.7	52 ± 1	372 ± 34^d	310 ± 24
Ovalbumin	CpG + alum	1.7 ± 0.5	40.3 ± 6.2	1.1 ± 0.5	1,361 ± 15	2,575 ± 334	<0.8	66 ± 3	262 ± 9	254 ± 15
C. trachomatis		17.2 ± 2.1^d	57.2 ± 21.3	0.08 ± 0.03	3,081 ± 137^d	2,309 ± 209	5 ± 0.2	75 ± 3	455 ± 14^d	398 ± 10
MEM-0		2.2 ± 0.7	48.4 ± 6.6	0.2 ± 0.08	683 ± 10	2,957 ± 81	<0.8	50 ± 0.1	164 ± 6	236 ± 55

↵ a Results are the means for triplicate cultures (±1 standard deviation). Data correspond to one of the experiments representative of duplicate separate experiments.
↵ b UV-inactivated C. trachomatis MoPn EB were added at a 10:1 ratio to the antigen-presenting cells.
↵ c Concanavalin A (ConA) was added at a concentration of 5 μg/ml.
↵ d P < 0.05 by the Student t test, compared to the ovalbumin- or MEM-0-immunized group.

TABLE 3.

Results of vaginal cultures

Mice immunized with:	Adjuvant	No. of mice that shed^a and no. of C. trachomatis IFU shed^b										Total no. of mice that shed in 6 wk (% positive)
		wk 1		wk 2		wk 3		wk 4		wk 5
		Mice shedding	C. trachomatis IFU shed	Mice shedding	C. trachomatis IFU shed	Mice shedding	C. trachomatis IFU shed	Mice shedding	C. trachomatis IFU shed	Mice shedding	C. trachomatis IFU shed
MOMP	CpG + Montanide	1/20 (5)^c	0 (0-960)^d	2/20 (10)^c	0 (0-12)^d	0/20 (0)	0	0/20 (0)	0	0/20 (0)	0	3/20 (15)^c
Ovalbumin	CpG + Montanide	12/20 (60)	2 (0-2,368,060)	13/20 (65)	14 (0-534,586)	5/20 (25)	0 (0-134,720)	1/20 (5)	0 (0-6)	0/20 (0)	0	17/20 (85)
MOMP	CpG + alum	7/20 (35)^c	0 (0-36,868)^d	10/20 (50)^c	13 (0-2,272)^d	4/20 (20)	0 (0-1,160)	0/20 (0)	0	0/20 (0)	0	15/20 (75)
Ovalbumin	CpG + alum	13/20 (65)	18 (0-891,922)	17/20 (85)	391 (0-302,034)	5/20 (25)	0 (0-309,124)	1/20 (5)	0 (0-7,820)	0/20 (0)	0	19/20 (95)
C. trachomatis		3/21 (14)^c	0 (0-18)^d	0/21 (0)^c	0^d	0/21 (0)	0	0/21 (0)	0	0/21 (0)	0	3/21 (14)^c
MEM-0		11/20 (55)	2 (0-8,500)	10/20 (50)	327 (0-372,934)	1/20 (5)	0 (0-2,380)	1/20 (5)	0 (0-867)	0/20 (0)	0	15/20 (75)

↵ a No. of mice that shed/total no. of mice per group (% positive).
↵ b Median no. of C. trachomatis IFU shed per group (range).
↵ c P < 0.05 by the Fisher exact test, compared with the corresponding ovalbumin- or MEM-0-immunized control group.
↵ d P < 0.05 by the Mann-Whitney U test, compared with the corresponding ovalbumin- or MEM-0-immunized control group.

TABLE 4.

Results of fertility studies

Antigen	Adjuvant	No. of mice that had embryos in uterine horns/total no. of mice/group (%)			Mean no. of embryos in uterine horns ± 1 SD
Antigen	Adjuvant	Both	Right	Left^a	Both	Right	Left^a
MOMP	CpG + Montanide	15/20 (75)^c^,^e	20/20 (100)	15/20 (75)^c^,^e	5.7 ± 3.0^d^,^f	3.1 ± 1.4	2.6 ± 2.4^d^,^f
Ovalbumin	CpG + Montanide	2/20 (10)	14/20 (70)	2/20 (10)	3.0 ± 2.5	2.9 ± 2.4	0.2 ± 0.5
MOMP	CpG + alum	7/20 (35)	17/20 (85)	8/20 (40)	4.2 ± 2.2^d^,^f	3.3 ± 1.9	0.9 ± 1.3
Ovalbumin	CpG + alum	2/20 (10)	12/20 (60)	2/20 (10)	2.3 ± 2.4	2.0 ± 2.1	0.3 ± 0.8
C. trachomatis		17/21 (81)^c^,^e	20/21 (95)	18/21 (86)^c^,^e	5.7 ± 2.6^d^,^f	3.1 ± 1.7	2.6 ± 1.5^d^,^f
MEM-0		6/20 (30)	16/20 (80)	7/20 (35)	3.7 ± 2.1	3.0 ± 2.0	0.8 ± 1.2
Fertility control^b		20/24 (83)	23/24 (96)	21/24 (88)	5.8 ± 2.5	3.2 ± 1.6	2.7 ± 1.8

↵ a Ovarian bursae inoculated with C. trachomatis MoPn.
↵ b Fertility control: mice were neither immunized nor challenged.
↵ c P < 0.05 by the Fisher exact test, compared to the corresponding ovalbumin- or MEM-0-immunized control group.
↵ d P < 0.05 by the Mann-Whitney U test, compared to the corresponding ovalbumin- or MEM-0-immunized control group.
↵ e P > 0.05 by the Fisher exact test, compared to the fertility control group.
↵ f P > 0.05 by the Mann-Whitney U test, compared to the fertility control group.