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Enhancing Effect of Ginseng Stem-leaf Saponins on the Immune Responses in Vaccinated Calves with FMD Bivalent Vaccine



Sonia A. Rizk, Eman M. El-garf and Abeer A. Talaat
 
ABSTRACT

A comprehensive sero-immunological studies were conduced to reveal the adjuvant's effect of Ginseng Stem-leaf Saponins (GSLS) on the immune response of gel adjuvanted Bivalent Foot and Mouth Disease (FMD) vaccinated calves. These study conducted in two calve groups; group (A) vaccinated subcutaneously with bivalent Alhydragel adjuvanted (30)% FMD vaccine, while group (B) vaccinated subcutaneously with bivalent FMD vaccine adjuvanted with both Alhydragel and GSLS (10 mg/dose). The humeral and cellular immunoresponses were monitored in different tested groups that received the gel adjuvanted vaccine and the Alhydragel-GSLS adjuvanted vaccine. Results indicated that the higher immune responses were found in calves vaccinated with Alhydragel-GSLS adjuvanted vaccine up to 24 week while with Alhydragel alone was only up to 18 week.

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  How to cite this article:

Sonia A. Rizk, Eman M. El-garf and Abeer A. Talaat, 2011. Enhancing Effect of Ginseng Stem-leaf Saponins on the Immune Responses in Vaccinated Calves with FMD Bivalent Vaccine. International Journal of Virology, 7: 167-175.

DOI: 10.3923/ijv.2011.167.175

URL: https://scialert.net/abstract/?doi=ijv.2011.167.175
 
Received: June 20, 2011; Accepted: July 28, 2011; Published: October 22, 2011

INTRODUCTION

Foot and Mouth Disease (FMD) is an acute contagious viral disease of cloven footed animals (Radostits et al., 1995; Orsel et al., 2007). The causative agent is a single stranded positive-sense RNA virus that belongs to the genus Aphthovirus in the family Picornaviridae. There are seven immunologically distinct serotype of FMD virus, namely, O, A, C, Asia1, Sat1, Sat2 and Sat3 (Belsham, 1993). In Egypt, the disease is enzootic and outbreaks have been reported since 1950 (Mousa et al., 1974). Type O was the most prevalent since 1960 and onwards (Zahran, 1960; Daoud et al., 1988; Farag et al., 2005). Since 1950, 1953 and 1956 serotype A didn't recorded in Egypt (Zahran, 1960 recently serotype A FMD virus introduced to Egypt through live animals importation and the sever clinical signs occurred among cattle and buffaloes (Abed El-Rahman et al., 2006). The control of FMD in animals was considered to be important to effectively contain the disease in endemic areas, so that vaccination of animals is effective in limiting the spread of FMD (Nair and Sen, 1992) Most foot-and-mouth disease vaccines are made of BEI (binary Ethylenimine) inactivated virus that is adjuvanted with either aluminum hydroxide-saponin (AS) or oil adjuvant. Adjuvants, also can prolong the immune response and stimulate specific components of the immune response either humoral or cell mediated immunity (Dalsgarrd et al., 1990; Barnett et al., 2003; Pluimers, 2004; Lombard et al., 2007). Saponins extracted from Ginseng Stems and Leaves (GSLS) has an adjuvant effects on the immune responses of buffalo to vaccination against Foot-and-Mouth Disease Virus (FMDV) (Xie et al., 2004). This study was carried out as an attempt to detect the adjuvant effects of Saponin extracted from ginseng stems and leaves (GSLS) on the immune responses of calves to vaccination against foot-and-mouth disease virus (FMDV) to improve local inactivated FMD vaccine.

MATERIALS AND METHODS

Animals: Nine calves (local breed) were clinically healthy and free from antibodies against FMD virus as proved by using SNT and ELISA were used in this study.

FMD viruses: FMD viruses O1/3/93-Egypt Strain and A1/Egypt/2006 are locally isolated strains of cattle origin. The viruses were typed at Veterinary Serum and Vaccine Research Institute, Abbasia, Cairo and confirmed by Pirbright, International Reference Laboratories, United Kingdom.

FMD vaccines: Inactivated bivalent FMD vaccines were prepared using the local strains O1/3/93 Egypt and A1/Egypt/2006, propagated in BHK-21 cell line. The viruses had a titer of 108 TCID50 /mL for both and inactivated by Binary Ethylenemine (BEI), FMD vaccines with different adjuvant are formulized as follow:

Alhydragel: The inactivated FMD viruses suspension was mixed with 30% Alhydragel solution as adjuvant (Mousa et al., 1976).

Ginseng Stems and leaves (GSLS) Saponin: The inactivated FMD viruses suspension was mixed with 30% Alhydragel solution with adding 10 mg/dose of Ginseng stems and leaves saponin (Song and Hu, 2009).

Experimental design: Two groups each group contain 3 calves, were vaccinated with the tested vaccines beside unvaccinated group (3 calves). Serum samples were collected weekly post vaccination for one month then every 2 weeks post-vaccination till the end of experiment. The immune response was evaluated through the estimation of cellular and humoral immune level using Lymphocyte blastogenesis assay, SNT and ELISA.

Serum neutralization tet (SNT): It was performed using the technique as described by Ferreira (1976).

Enzyme linked immunosrobent assay (ELISA): It was carried out according to the method described by Voller et al. (1976).

Indirect solid phase ELISA was applied, patently prepared at department of FMD, Veterinary Serum and Vaccine Research Institute, Abbasia, Egypt.

Evaluation of cell-mediated immunity in vitro using lymphocyte Proliferation (3-(4,5-Dimethylthiazol-2-yl)-2,5-(MTT) Assay: It was applied according to Lucy (1984) following by modification adopted by El-Watany et al. (1999) and Abeer (2001).

RESULTS

Humoral immune response of calves vaccinated with FMD vaccines: Results of humoral immune response revealed that serum antibody protective titer evaluated by mean of SNT and ELISA were as follow:

1st group: Started at 2nd week post vaccination with the titers of 1.3 log10 by SNT and 1.5 by ELISA for O1 was with titer of 1.4 log10 by SNT and 1.5 by ELISA for A1. The highest level of antibody titers were at the 6th week post vaccination as 2.1 log10 by SNT and 2.4 by ELISA for O1, as 2.2 log10 by SNT and 2.4 by ELISA for A1 and the immunity duration lasted for 18 weeks post vaccination as 1.2 log10 by SNT and 1.5 by ELISA for O1, as 1.2 log10 by SNT and 1.5 by ELISA for A1
2nd group: Started at 2nd week post vaccination with the titers of 1.5 log10 by SNT and 1.6 by ELIS for O1 and was with titer of 1.6 log10 by SNT and 1.6 by ELISA for A1. The highest level of antibody titers were at the 8th week post vaccination as 2.4 log10 by SNT and 2.6 by ELISA for O1, as 2.4 log10 by SNT and 2.6 by ELISA for A1 and the immunity duration lasted for 24 weeks post vaccination as 1.2 log10 by SNT and 1.5 by ELISA for O1, as 1.2 log10 by SNT and 1.5 by ELISA for A1 Table 1-4

Evaluation of cell-mediated immunity in vitro using lymphocyte Proliferation (MTT) Assay: Obtained results of cell mediated immune response using lymphocyte proliferation test for all animal groups expressed by ΔOD (Delta Optical Density) were as follow:

1st group: Delta Optical Density was (0.152-0.11-0.128) by using phytohaemagglutinin (PHA), Pokeweed (pok) mitogens and FMD virus at 3 day post vaccination and still rise reached its highest level (0.28-0.30-0.36) at 21 day post vaccination, then declined to (6 weeks)
2nd group: Delta Optical Density was (0.232-0.191-0.309) by using (PHA) (pok) and FMD virus at 3 day post vaccination and still rise reached its highest level (0.413-0.442-0.524) at 21st day post vaccination, then declined after (9 weeks) (Table 1-4).

Table 1: Immune status (SNT titer) of calves vaccinated with Alhydragel and modified Ginseng FMD vaccines against O1/3/93-Egypt virus
A1, A2 and A3: vaccinated calves no. SNT; serum neutralization test. *: The results of SNT expressed as log10 TCID50/mL. N.B.: The permissible protective level is 1.2 SNT titer

Table 2: Immune status (ELISA titer) of calves vaccinated with Alhydragel and modified ginseng FMD vaccines against O1/3/93-Egypt virus
A1, A2 and A3: vaccinated calves no. *: The results of ELISA expressed as log10 TCID50/mL. N.B.: The permissible protective level is 1.5 ELISA titer

Table 3: Immune status (SNT titer) of calves vaccinated with Alhydragel and modified Ginseng FMD vaccines against A1/Egypt/2006 virus
A1, A2 and A3: vaccinated calves No. SNT: serum neutralization test. *: The results of SNT expressed as log10 TCID50/mL. N.B.: The permissible protective level is 1.2 SNT titer

Table 4: Immune status (ELISA titer) of calves vaccinated with Alhydragel and modified Ginseng FMD vaccines against A1/Egypt/2006 virus
A1, A2 and A3: vaccinated calves No. *: The results of ELISA expressed as log10 TCID50/mL N.B.: The permissible protective level is 1.5 ELISA titer

DISCUSSION

The control of FMD in animals was considered to be important to effectively contain the disease in endemic areas, so that vaccination of animals is effective in limiting the spread of FMD.

So, this study is to improvement of inactivated FMD Alhydragel vaccine with adding Ginseng stems and leaves saponin as an adjuvant.

From Table 1 and 2 the results revealed that SNT and ELISA titers for Alhydragel FMD vaccines, go in hand with the results obtained are consistent with the statement of Hamblin et al. (1986) who explained that the SNT measures those antibodies which neutralize the infectivity of FMD virion, while ELISA probably measure all classes of antibodies even those produced against incomplete and non-infectious virus.

From Table 3 and 4 the results revealed that SNT and ELISA titers for Alhydragel and Ginseng FMD vaccine agreed with Rivera et al. (2003), Hu et al. (2003), Sun et al. (2005), Yan et al. (2007) and Song and Hu (2009). who showed that adjuvant properties of Ginseng Extract as potent adjuvant induced higher antibody titers than the vaccine adjuvanted with Al(OH)3 and improved the potency of adjuvants. Results supported also by Scaglione et al. (1996), Rivera et al. (2003) and Wang et al. (2009) who found that ginseng might help the vaccine work more effectively, increasing antibody production.

Table 5: Cell-mediated Immune response of calves Vaccinated with Alhydragel and modified Ginseng FMD vaccines
*: Δ Optical densities, **: Type of mitogen, PHA: Phytohaemagglutinin, V: FMD Virus, POK: Pokeweed, N.B.: The permissible protective level is 0.250 delta optical density

From Table 5, the results of evaluation of cell mediated immune response using lymphocyte proliferation test for all animal groups expressed by ΔOD (Delta Optical Density). Supported by Knudsen et al. (1979) and Sharma (1981) who reported that cell mediated immune response was a constitute of immune response against FMD virus. And in agreement in some points with Garcia-Valcarcel et al. (1996) El-Watany et al. (1999) and Abeer (2001). Mansour (2001) and Samir (2002) that FMD vaccine stimulated the cellular immune response and lymphocyte stimulation by FMDV was greater than by mitogens (PHA) and (POK) and appeared increased in 1st and 2nd weeks post vaccination. While disagreed with El-Watany et al. (1999) and Mansour (2001) in that cell mediated immune response reach its highest level on the 14th day.

The obtained results were in agreement with Song et al. (2002), Chen et al. (2008) and Sun et al. (2009) who stated that Ginseng extract act as an activator of the TH1 response. The Th1 type is characterized by the production of antigen-specific IgG2a a Th1 nd the secretion of gamma interferon, interleukins which favor cellular immunity.

Our results also were supported by (Xie et al., 2004) and Wang et al. (2007) who mentioned that Ginseng extract enhanced interleukins which enhance cell mediated immune response.

Finally, it can conclude that: The usage of Saponin extracted from ginseng stems and leaves in inactivated FMD vaccine gave long lasting immunity than that which with Alhydragel adjuvant alone GSLS and improve both cellular and humoral immunity and gave earlier and more long lasting immunity.

REFERENCES
Abed El-Rahman, A.O., M.A. Farag, S. El-Kilany, M.A. Eman, M.A. El-Yazed and S. Zeidan, 2006. Isolation and identification of foot and mouth disease virus during an outbreak of 2006 in Egypt. Kafr El-Sheikh Vet. Med. J., 4: 451-464.
Direct Link  |  

Abeer, E.M., 2001. Studies on the effect of mycotoxins in ration on the immune responce of FMD vaccinated farm animals (cattle and sheep). Ph.D. Thesis, Faculty of Veterinary Medicine, Cairo University.

Barnett, P.V., R.J. Statham, W. Vosloo and D.T. Haydon, 2003. Foot-and-mouth disease vaccine potency testing: Determination and statistical validation of a model using a serological approach. Vaccine, 21: 3240-3248.
PubMed  |  Direct Link  |  

Belsham, G.J., 1993. Distinctive features of FMDV, a member of the Picorna virus family, aspects of virus protein synthesis, protein processing and structure. Prog. Biophys. Mol. Biol., 60: 241-260.

Chen, C.F., W.F. Chiou and J.T. Zhang, 2008. Comparison of the pharmacological effects of Panax ginseng and Panax quinquefolium. Acta Pharmacol. Sin., 29: 1103-1108.
PubMed  |  Direct Link  |  

Dalsgarrd, K., L. Hilgers and G. Trouve, 1990. Classical and new approaches to adjuvant use in domestic food animals. Adv. Vet. Sci. Comp. Med., 35: 121-160.
PubMed  |  Direct Link  |  

Daoud, A., A. Omar, M. El Bakry, N. Metwally, M. El Mekkawi and S. El Kilany, 1988. Strains of FMD virus recovered from 1987 outbreak in Egypt. J. Egypt. Vet. Med. Ass., 48: 63-71.

El-Watany, H., M. Shawky, O.M. Roshdy and S. El- Kelany, 1999. Relationship between cellular and humoral immunity responses in animal vaccinated with FMD vaccine. Zag. Vet. J., 27: 137-143.
Direct Link  |  

Farag, M.A., M.A. Aggour and A.M. Daoud, 2005. ELISA as a rapid method for detecting the correlation between the field isolates of FMD and the current used vaccine strain in Egypt. Vet. Med. J. Giza, 53: 949-955.

Ferreira, M.E.V., 1976. Prubade microneutralization poraestudies de anticueropos de la fibrea fsta. BItn Centro Panamericano Fiebre Aftosa, 21-22: 17-24.

Garcia-Valcarcel, M., T. Doel, T. Collen, M. Ryan and R.M. Parkhouse, 1996. Recognition of foot-and-mouth disease virus and its capsid protein VP1 by bovine peripheral T lymphocytes. J. Gen. Virol., 77: 727-735.
PubMed  |  Direct Link  |  

Hamblin, C., I.T. Barnett and R.S. Hedger, 1986. A new Enzyme-Linked Immunosorbent Assay (ELISA) for the detection of antibodies against foot-and-mouth disease virus. I. Development and method of ELISA. J. Immunol. Methods, 93: 115-121.
PubMed  |  

Hu, S., C. Concha, F. Lin and K.P. Waller, 2003. Adjuvant effect of ginseng extracts on the immune responses to immunization against Staphylococcus aureus in dairy cattle. Vet. Immunol. Immunopathol., 91: 29-37.
PubMed  |  Direct Link  |  

Knudsen, R.C., C.M. Groocock and A.A. Aaderson, 1979. Immunity to foot-and-mouth disease virus in guinea pigs: Clinical and immune responses. Infect. Immun., 24: 787-792.
Direct Link  |  

Lombard, M., P.P. Pastoret and A.M. Moulin, 2007. A brief history of vaccines and vaccination. Rev. Sci. Tech., 26: 29-48.
Direct Link  |  

Lucy, F.L., 1984. Proliferative response of chicken B and T lymphocytes to mitogens. chemical regulation of enhancing effect of Gsls on vaccinated calves with FMD 167. Immun. Vet. Med., 15: 44-52.

Mansour, A., 2001. Some studies on the effect of mycotoxins on immune response of FMD vaccinated animals. Ph.D. Thesis, Faculty of Veterinary Medicine, Cairo University.

Mousa, A.A., S.M. Boulaus, F.S. Elsayed and H.O. Bohm, 1974. Typing and subtyping of a strain of FMD isolated from sharquia province, 1970. J. Egy. Assuit. Vet. Med., 34: 413-419.

Mousa, A.A.M., M.H. Ibrahim and K. Hussein, 1976. Preliminary study on antibody response of cattle after experimental infection with FMDV. Proceedings of the 13th Arab Veternary Congeress, February 13-18, 1976, Cairo, Egypt -.

Nair, S.P. and A.K. Sen, 1992. A comparative study on the immune response of sheep to foot and mouth disease virus vaccine type Asia-1 prepared with different inactivants and adjuvants. Comp. Immunol. Microbiol. Infect. Dis., 15: 117-124.
CrossRef  |  Direct Link  |  

Orsel, K., M.C. de Jong, A. Bouma, J.A. Stegeman and A. Dekker, 2007. Foot and mouth disease virus transmission among vaccinated pigs after exposure to virus shedding pigs. Vaccine, 25: 6381-6391.
PubMed  |  Direct Link  |  

Pluimers, F.H., 2004. Foot-and-mouth disease control using vaccination: The dutch experience in 2001. Dev. Biol. (Basel), 119: 41-49.
PubMed  |  Direct Link  |  

Radostits, O.M., D.C. Blood and C.C. Goy, 1995. Veterinary Medicine. In: Educational Low Priced Blooks Scheme, Shriver, D.F., P.W. Atkins and C.H. Langford (Eds.)., ELBS., UK., pp: 965-973.

Rivera, E., S. Hu and C. Concha, 2003. Ginseng and aluminium hydroxide act synergistically as vaccine adjuvants. Vaccine, 21: 1149-1157.
Direct Link  |  

Samir, M.A.A., 2002. Studies on preparation of newly oil adjuvanted FMD vaccine. Ph.D. Thesis, Cairo University, Cairo.

Scaglione, F., G. Cattaneo, M. Alessandria and R. Cogo, 1996. Efficacy and safety of the standardised ginseng extract G115 for potentiating vaccination against the influenza syndrome and protection against the common cold. Drugs Exp. Clin. Res., 22: 65-72.
PubMed  |  Direct Link  |  

Sharma, S.K., 1981. Foot and mouth disease in sheep and goats. Vet. Res. J., 4: 1-21.

Song, J.Y., S.K. Han, E.H. Son, S.N. Pyo, Y.S. Yun, and S.Y. Yi, 2002. Induction of secretory and tumoricidal activities in peritoneal macrophages by ginsan. Int. Immunopharmacol., 2: 857-865.
CrossRef  |  PubMed  |  Direct Link  |  

Song, X. and S. Hu, 2009. Adjuvant activities of saponins from traditional Chinese medicinal herbs. Vaccine, 27: 4883-4890.
CrossRef  |  Direct Link  |  

Sun, H.X., F. Qin and Y.P. Ye, 2005. Relationship between haemolytic and adjuvant activity and structure of protopanaxadiol-type saponins from the roots of Panax notoginseng. Vaccine, 23: 5533-5542.
PubMed  |  Direct Link  |  

Sun, H.X., Y. Xie and Y.P. Ye, 2009. Advances in saponin-based adjuvants. Vaccine, 27: 1787-1796.
CrossRef  |  PubMed  |  Direct Link  |  

Voller, A., D.E. Bidwell and A. Bartlett, 1976. Enzymeimmunoassay in diagnostic medicine. Bull. World Health Org., 53: 55-65.
Direct Link  |  

Wang, C.L., D.Z. Shi and H.J. Yin, 2007. Effect of panax quinquefolius saponin on angiogenesis and expressions of VEGF and bFGF in myocardium of rats with acute myocardial infarction. Zhongguo Zhong Xi Yi Jie He Za Zhi, 27: 331-334.
PubMed  |  

Wang, H., D. Peng and J. Xie, 2009. Ginseng leaf-stem: Bioactive constituents and pharmacological functions. Chin. Med., 4: 20-20.
PubMed  |  

Xie, J.T., S.R. Mehendale, A. Wang, A.H. Han, J.A. Wu, J. Osinski and C.S. Yuan, 2004. American enhancing effect of gsls on vaccinated calves with fmd 69 ginseng leaf: Ginsenoside analysis and hypoglycemic activity. Pharmacol. Res., 49: 113-117.

Yan, B., G.A.J. Wang, L. Xie and H. Hao et al., 2007. Construction of the fingerprints of ginseng stem and leaf saponin reference substances and spiked plasma sample by LC-ESI/MS and its application to analyzing the compounds absorbed into blood after oral administration of ginseng stem and leaf saponin in rat. Biol. Pharm. Bull., 30: 1657-1662.
PubMed  |  

Zahran, G.E.D., 1960. Foot and mouth disease in southern region of URA. Bull. Off. Int. Epiz., 13: 390-393.

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