The comparison of the efficacy of swine FMD vaccine emulsified with oil adjuvant of ISA 201 VG or ISA 206 VG

doi:10.4236/jbm.2013.13005

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Journal of Biosciences and Medicines, 2013, 1, 22-25 JBM

http://dx.doi.org/10.4236/jbm.2013.13005 Published Online December 2013 (http://www.scirp.org/journal/jbm/)

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The comparison of the efficacy of swine FMD vaccine

emulsified with oil adjuvant of ISA 201 VG or ISA 206 VG

Dong Li*, Chunxue Zhou, Daliang She, Pinghua Li, Pu Sun, Xingwen Bai, Yingli Chen, Baoxia Xie,

Zaixin Liu*

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, OIE/Na-

tional Foot and Mouth Disease reference laboratory of China, Lanzhou Veterinary Research Institute, Chinese Academy of Agricul-

tural Sciences, Lanzhou, China

Email: *[email protected], *[email protected]

Received September 2013

ABSTRACT

The Seppic Company developed a new adjuvant

Montanide ISA 201 VG, the upgraded version of

Montanide ISA 206 VG, which keep the advantage

and added some chemical components on the basis of

ISA 206 to improve the cellular responses. The aim of

the study is to compare the efficacy of swine FMD

(foot-a nd-mouth) vaccine emulsified with oil adjuvant

of ISA 201 or ISA 206 respectively. The pigs were

vaccinated with FMD vaccine emulsified with inactive

FMD type O antigen and adjuvant ISA 201 or ISA

206 respectively, according to 2.0 ml (1/1 dose), 0.67

ml (1/3 dose), 0.22 ml (1/9 dose) to calculate their

PD50. The sera were collected from the vaccination of

the day 0, 3, 7, 14, 21, 28 and the ELISA FMD type O

antibody were detected. Furthermore, the PD50 were

calculated after the pigs were challenged with viru-

lent FMDV type O on 28 days post vaccination. The

ELISA antibody titers of 201vaccine were signifi-

cantly higher than that of 206 (except the third time).

The fifty percent of protection dose (PD50) of 201

vaccine (PD50 = 15.59) was higher than that of 206

vaccine (PD50 = 10.05). The above data showed that

the efficacy of the FMD vaccine emulsified with ISA

201 was better than which with ISA 206.

Keywords: Swine FMD Vaccine; Adjuvant; ISA 201;

ISA 206; Immune Efficacy

1. INTRODUCTION

Foot-and-mouth disease (FMD is an acute, febrile, and

contagious vesicular disease affecting cloven-hoofed ani-

mals. The causal agent, FMD virus (FMDV), is a mem-

ber of the genus Aphthovirus in the family Picornaviri-

dae and occurs as seven distinct serotypes throughout th e

world: A, O, C, Asia1 and South African Terri torie s (SAT)

1-3 [1-3]. Vaccination is the most important control and

eradication strategy for FMD, esp ecially the oil-adjuvant

vaccine in developing countries. Significant advances in

recent years have led to the introduction of ‘‘ready-to-

formulate” oil adjuvants. Montanide ISA 206, the miner-

al based oil, which readily forms water-in-oil-in-water

emulsion. Its efficacy in eliciting immune response was

studied in FMD vaccines against FMDV [4-7].

The advantage of oil adjuvant was attributed to depot

formation at the site of injection, a vehicle for transport

of the antigen throughout the lymphatic system and slow

antigen release with the stimulation of antibody produc-

ing cells. Moreover, being oil emulsion, Montanide ISA

206 had various advantages, like low viscosity, easy ad-

ministration, longer term protection, greater stability and

production of smaller nodules at the site of injec tion [5],

compared to other oil adjuvants, making as an ideal ad-

juvant candidate for FMD vaccines. It was also sug-

gested that Montanide ISA 206 could prevent the loss of

potency wa s due to the proteolysis o f VP1 or possibl y the

physical breakdown of the virus followed adsorption to

the aluminum hydroxide gel [8] and agree with the usage

of Montanide ISA 206 ready to formulate oil adjuvant

can be sued in all target species is ideal for emergency

vaccination [9]. The usage of oil adjuvant (Montanide

ISA 206) improve, enhance cell mediated immunity and

give higher level and long lasting immunity [10,11].

Other researchers [12,13] detected that DOE vaccine

containing Montanide ISA 206 is highly efficient, fluidy

with low viscosity which is easily dispersed from the

place of injection and gave high Ab titers and longest

duration of immunity than alhdyragel vaccine and Daoud

reported that the duration of immunity elicited by gel

FMD vaccine was short lived and antibody concentra-

tions rapidly fall after administration, while oils adju-

ventated FMD vaccines gave a longer duration of im-

munity and suggested that the oil adjuvenated vaccines

*Corresponding a uthors.

D. Li et al. / Journal of Biosciences and Medicines 1 (2013) 22-25

had potential as an alternative to the conventional alu-

minum hydroxide FMD vaccine [14]. Moreover Patil

reported that the oil adjuvant elicited a better immune

response at any time than did aluminum hydroxide gel

FMD quadrivalent vaccines in goats, and the response

developed quicker. Local tissue reactions such as granu-

lomas and cysts to oil-adjuvants have been not detected

[15].

Now the Seppic developed a new adjuvant Montanide

ISA 201, the upgraded version of Montanide ISA 206,

which keep the advantage and added some chemical

components on the basis of ISA 206 to improve the cel-

lular responses. The aim of the study is to compare the

efficacy and safety of swine FMD vaccine emulsified

with oil ad juvant of ISA 201 or ISA 2 0 6 respecti ve l y.

2. MATERIALS & METHODS

2.1. The Animals

Fifty 2-month-old pigs, sero-negative for FMDV type O

antibodies (titre 1:4 by LPB-ELISA) were randomly di-

vided into four groups. 45 pigs in group 1-3 and 15 pigs

in each group. The vaccine with ISA 206 was the first

group. The vaccine with ISA 201 was the second group.

The vaccine with PBS as the adjuvant was the third

group. Three small groups (Five pigs in it) were divided

in each group for three vaccine dose 2.0 ml (1/1 full

dose), 0.67 ml (1/3 full dose), 0. 22 ml (1/9 full dose) to

determine the PD50 (protective dose 50%). The left five

pigs were in the 4th group as the control. All the pigs

were bred in the P3 animal laboratory respectively.

2.2. Vaccine Preparation

The FMDV type O antigen was inactivated and safety

checked by conventional method, and emulsified with

ISA 201 or ISA 206 separately according to the prospec-

tus guide . The vacc ine was stored at 4˚C.

2.3. Vaccination, Sera Collection and Virus

Challenge

Three types of FMD vaccines were intramuscular ino-

culated at the ear-root-neck area of 45 pigs with 2 ml,

0.67 ml, 0.22 ml inactivated vaccine respectively, and

five pigs were bred without vaccination as negative con-

trol. Blood and sera samples were collected at 0, 3, 7, 14,

21, 28 dpv (days post vaccination) to assay the FMDV

type O antibody by L PB -ELISA. To demonstrate vaccin e

efficacy, all 50 pigs were challenged intramuscularly

with 1000 PID 50/2 ml of FMDV type O suckling mice

passaged strain at the ear-root-neck area after 28 dpv and

FMD symptoms were monitored for 10 days, and the

PD50 was calculated with Karber method.

3. THE RESULTS

Define abbreviations and acronyms the first time they are

used in the text, even after they have been defined in the

abstract. Abbreviations such as IEEE, SI, MKS, CGS, sc,

dc, and rms do not have to be defined. Do not use ab-

breviations in the title or heads unless they are unavoida-

ble.

3.1. The ELISA Antibody Titer of FMDV Type O

after Pigs Vaccinated

The ELISA antibody titer of 201-vaccine (1/1 dose) was

186.12 on 21st dpv, 201.82 on 28th dpv, much higher

than which of 206-titer (p < 0.05), because the titer of

206-vaccine (1/1 dose) was 69.76 on 21st dpv, 57.28 on

28th dpv. The ELISA antibody titer of 201-vaccine (1/3

dose) was 133.48 on 21st dpv, 120.62 on 28th dpv, much

higher than which of 206-titer r (p < 0.05), because the

titer of 206-vaccine (1/3 dose) was 69.3 on 21st dpv,

71.96 on 28th dpv. The ELISA antibody titer of 201-

vaccine (1/9 dose) was 65.54 on 21st dpv, 84.36 on 28th

dpv, much higher than which of 206-titer r (p < 0.05),

because the titer of 206-vaccine (1/9 dose) was 30.9 on

21st dpv, 29.56 on 28th dpv (Figure 1). These data

showed that the effect of 201-vaccine is better than 206-

vaccine.

3.2. The FMD V irule nt Virus Challenge to

Immunized Pigs on 28dpv

The challenge result was presented in Table 1. Fifty pig s

were all protected in 201-vaccine group (PD50-201 =

15.59). Thirteen pigs were protected in 206-vaccine

group (2 pigs had clinic symptom with 1/9 dose, PD50-

206 = 10.05). Four pigs were protected in PBS-vaccine

group (5 pigs had clinic symptom with 1/9 dose, 3 pigs

had clinic symptom with 1/3 dose, 3 pigs had clinic

symptom with 1/1 dose, PD50-PBS = 1.39). Generally,

the FMD vaccine is qualified when PD50 comes up to

6.0. The above data showed that the PD50 of 201 emul-

sified FMD vaccine was higher than which of 206 emul-

sified vaccine.

Figure 1. Comparisons of the ELISA antibody titers among

201-vaccine, 206-vaccine, PBS-vaccine.

D. Li et al. / Journal of Biosciences and Medicines 1 (2013) 22-25

Table 1. The FMD clinical incidence after challenge.

Groups Pig No & clinical incidence

206 (1/1) 3051 (−) 3056 (−) 3057 (−) 3076 (−) 3098 (−)

206 (1/3) 4441 (−) 3074 (−) 3047 (−) 3075 (−) 3077 (−)

206 (1/9) 3072 (−) 3060 (+) 3063 (−) 3065 (+) 3083 (−)

201 (1/1) 3058 (−) 3059 (−) 4477 (−) 3079 (−) 3086 (−)

201 (1/3) 3097 (−) 3073 (−) 3094 (−) 3082 (−) 3043 (−)

201 (1/9) 3064 (−) 3092 (−) 3081 (−) 3070 (−) 3069 (−)

PBS (1/1) 3055 (−) 3067 (−) 3093 (+) 3084 (+) 3089 (+)

PBS (1/3) 3054 (−) 3061 (+) 3095 (+) 3068 (−) 3088 (+)

PBS (1/9) 3044 (+) 3090 (+) 3087 (+) 3085 (+) 3096 (+)

Control 3099 (+) 1040 (+) 1439 (+) 3080 (+) 3071 (+)

Note: (+) means FMD clinic sighs appeared.

4. DISCUSS

The ISA206 adjuvant has applied in FMD vaccine pro-

duction and other vaccines for 20 years, because it has

lower side-reactions and viscosity, easier to emulsify and

injection, and higher stability. Recen tly, T he ISA 201 has

developed on the base of ISA 206, which inherited ad-

vantages of ISA 206, and added some chemical sub-

stances with the hope of enhanci ng animals’ cellular im-

mune reactions [16]. In this report, the comparisons be-

tween ISA 201 and ISA 206 were done as the FMD vac-

cine respectively.

First the ELISA antibodies against FMDV type O

were compared. The antibody titer induced by 201-vac-

cine were higher than which of 206-vaccine on 3dpv,

7dpv, 14dp v, 21 dpv , 2 8dpv. This means that the effect of

201-vaccine in inducing antibody is better than which of

206-vaccine.

The virus challenge trial to vaccinated animals is a

very important index for evaluating the potency of FMD

vaccine, which is indicated with PD50 (50 percent pro-

tection dose) [17,18]. Here, fifty pigs were all protected

immunized with 201 vaccine, but thirty pigs were pro-

tected with 206 vaccine, which means that the potency of

201 vacci ne is better than that of 206 va c c i ne.

5. CONCLUSION

From above, the efficacy of the FMD vaccine emulsified

with ISA 201 is su pe rior to ISA 206.

6. ACKNOWLEDGEMENTS

This work has been supported by “Seppic, France”.

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