The Effects of New Nonspecific Immunostimulators in Pig
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1 J. Fd Hyg. Safety 21(2), (2006)» w x 2 Á û Á Á xáy Á Á½ Á Á xá xá y 2 Á 2 Á «2 Á ½ 2 Á½» 2 Á 1 Á y 1 Á Á w wœ w gg lv œ w w wp w The Effects of New Nonspecific Immunostimulators in Pig Ji-Youn Jung 2, Nam-Shik Ahn, Joon-Suk Park, Eun-Hye Jo, Jae-Woong Hwang, Jung-Ran Park, Sun-Jung Kim, Yong-Geon Lee, Yun-Hyeok Jeong, Ji-Hye Chung, Seung-Ho Lee 2, Young-Seok Park 2, Byung-Kwon Park 2, Byeong-Soo Kim 2, Sang-Ki Kim 2, Sang-Bum Lee 1, Ke-Ho Lee 1, Kyung-Sun Kang, and Yong-Soon Lee %FQBSUNFOUPG7FUFSJOBSZ1VCMJD)FBMUI$PMMFHFPG7FUFSJOBSZ.FEJDJOF4FPVM/BUJPOBM6OJWFSTJUZ4FPVM,PSFB,PLPFOUFSQSJTF$P-UE$IFPOHXPOHVO$IVOHCVL,PSFB %FQBSUNFOUPG$PNQBOJPOBOE-BCPSBUPSZ"OJNBM4DJFODF,POHKV/BUJPOBM6OJWFSTJUZ$IVOHOBN,PSFB 3FDFJWFE+VOF"DDFQUFE+VOF ABSTRACT New nonspecific immunostimulators (Koko enterprise Co., Ltd., Korea) were evaluated for its effectiveness as a nonspecific immunostimulator in pigs. The effects were determined by analysis of cytokines using ELISA and blood-chemistry. IFN-r which is one of the cell mediated immune cytokines significantly increased in DIR-vitamineral 0.2% group posttreatment 4 weeks and was significantly higher IMF 0.2%, DIR-vita 0.1% and DIR-vitamineral 0.2% groups than control group in 3 months. DIR-vitamineral 0.2% had a most strong effectiveness as a nonspecific immunostimulator in our treatment materials in pigs. IMF 0.2% and DIR-vita 0.1% were seen that there had effectiveness of a nonspecific immunostimulator in posttreatment 3 months. IgG, IgM and Total Ig which were humoral immune globulins, were not significantly changed in posttreatment 4 weeks and 3 months. In conclusion, this study has demonstrated that new nonspecific immunostimulators had an immunostimulatory effect on pigs through cell mediated immune response. Key words: nonspecific immunostimulator, cytokines, IFN-r, IgG, ELISA xk ³ y z ù ƒ q x e w» w e ƒwš. w 1) x w» w ƒ w v, w³ ƒ w,» w³ w w e w š, w³ ü ³ q w» w w³ w w» v w š w. 2) ù, w e w s Author to whom correspondence should be addressed. w p w»» w ù» e w ƒ. 3) w³ w e z ù š w» w, w û z w w³ v w š ü ³ w y ùe w»w w. w, ƒ ù e š w³ w {» w³ ü ³ x w š w³ ü w mw e w j š. 1,3), e w 113
2 114 CtX yc yrp fw e w w e w ƒ y š, w w š w w³, y, ³ w wš. p w k 4,5) z ƒ ù, e w w w w p š š z t š ù, ü t 6-8) w. w ü xy ü Immu-Forteƒ p ƒ y w, x y. x» 9,10) y s l DIR-vita, DIR-vitamineral w w w y wš s y w IFNr, TNF-a x w, y w IgG, IgM, Total Ig x w ww. Materials x gg lv 5ƒ n x n w, 5ƒ Immuforte-a, DIR-vita 0.1%, DIR-vita 0.2%, DIR-vitamineral 0.1%, DIR-vitamineral 0.2%. ƒƒ 3-4 e w. n» 4 3 n w. Animals x w YLD j,, 3, n 21 w 22 l ƒ v w x 21 l 4, 3 n w. Methods ƒ. Sandwitch ELISA w me d x w me IFN-r, TNF-a, IgG, IgM, Total Ig w. s y w IFN-r, TNF-a x w, y w IgG, IgM, Total Ig x w. v w j plate y qw z,, ƒ cytokine t, ƒƒ plate 100 ul w. z, 30 ew. ƒ plate 300 ul 5z, w. cytokinez w 100ul ƒ well w. 30 ew ƒ plate 300 ul 5z wš w. 200 ul ƒ well w z, 15 ew, 50 ul ƒ well w. ELISA reader» 450 nm Ÿ d w. ù. x x ƒ wš q w» w x w. Statistical analysis w s l q, DIR-vita, DIRvitamineral s³e w» w e w Microcal Origin 6.0(Microcal Software, Inc., USA) w ANOVA test w, x» x w Student's t-test w w. x x 4 3 (1-1, 1-2, 1-3) w x ƒw x w w», x ƒ w ù x y w. s w x a. IFN-r IFN-r w 4 3 d w 4 w ƒ ùkü DIR-vitamineral 0.2%, y ùkü w s l q, DIR-vita 0.1%, DIR-vitamineral 0.2% ƒƒ ùkù.
3 Ssp8qqph qgpˆg y phtqthbxx y tx wf ƒ tyitr 115 Table 1. The changes of blood-chemistry in pig posttreatment Group Albumin (g/dl) T. Bililubin T. Protein (g/dl) Creatinine AST ALT T. CHOL ALP Potassium (mmol/l) 3 month Control IMF 0.2% DIR vita 0.1% DIR vita 0.2% DIR vitamine 0.1% DIR vitamine 0.2% weeks IMF 0.2% DIR vita 0.2% DIR vitamine 0.1% DIR vita 0.1% DIR vitamine 0.2% Control b. TNF-a TNF-a e ƒƒ n z d w 4, 3 x ƒ ùkü ùkù ù 4 DIRvitamineral 0.1%, DIR-vitamineral 0.2% w ƒw w ùkü. w x IgG 4 w 3 n w w ùkþ. ù, w IgG w n m w ƒƒ. wr, IgM total Ig 4 3 yƒ ùkù š ùkû. š» macrophage w Table 2. The changes of IFN-r in the blood of pigs posttreatment Control IMF 0.2% * DIR-vita 0.1% * DIR-vita 0.2% DIR-vitamineral 0.1% DIR-vitamineral 0.2% 54 * 128 * s T q w s B q w w. l ù 11,12) w ù, p T q ƒ w ù w y y k s wš p š w. w T q memory cell p x marker CD29 wì ùkùš v ùkùš ¾ Table 3. The changes of TNF-a in the blood of pigs posttreatment Control IMF 0.2% DIR-vita 0.1% DIR-vita 0.2% DIR-vitamineral 0.1% DIR-vitamineral 0.2% Table 4. The changes of IgG in the blood of pigs posttreatment Control IMF 0.2% DIR-vita 0.1% DIR-vita 0.2% DIR-vitamineral 0.1% DIR-vitamineral 0.2%
4 116 CtX yc yrp fw Table 5. The changes of IgM in the blood of pigs posttreatment Control IMF 0.2% DIR-vita 0.1% DIR-vita 0.2% DIR-vitamineral 0.1% DIR-vitamineral 0.2% Table 6. The changes of Total Ig in the blood of pigs posttreatment Control IMF 0.2% DIR-vita 0.1% DIR-vita 0.2% DIR-vitamineral 0.1% DIR-vitamineral 0.2% wš. 13) w w q w w t T B q ù. T q T s (T cell receptor, TCR) ƒ, TCR ab TCR gd TCR ƒ ù B q st (cell surface immunoglobulin) w ƒ š. 14) ù ù ³ û T q ƒ w w ù ƒ ƒwš w w ƒw ùkû. 15) w» w w w T q B q y y d w» w, me IFN-r, TNF-a, IgG, IgM, Total Ig w. s y w IFN-r, TNF-a x w, y w IgG, IgM, Total Ig x w. w,» ( s l q, DIR-vita, DIR-vitamineral) w» 4 3 w ü y y d w, DIR-vita DIRvitamineral ƒƒ 0.1% 0.2% ù y y d w. T q y y w w e IFN-r n» d w, 4 w ƒ ùkü DIRvitamineral 0.2%, y ùkü w s l q, DIR-vita 0.1%, DIR-vitamineral 0.2% ƒƒ ùkù. wr, TNF-a 4, 3 x ƒ ùkü ùkù ù 4 DIR-vitamineral 0.1%, DIR-vitamineral 0.2% w ƒw w ùkü. d w IgG, IgM, Total Ig w y ùkü. w l x w» ü» s T q w y y, x DIR-vitamineral 0.2% ƒ z ü» y y ùkü q. x 5ƒ ü» y y z w, ƒ z DIR-vitamineral 0.2% x, DIR-vitamineral 0.1%, s l q, DIR-vita 0.1% z q. ww, x w» z y w, w wì s z» x w. gg lv 2006 w w l (KRF-005-E00076). x n z 4 3 x d w» w w w w q w» w x w. w
5 Ssp8qqph qgpˆg y phtqthbxx y tx wf ƒ tyitr 117 ƒ, l x e ƒ w ƒ q w ƒ. j s ù s t me IFN-r d w, 4 w ƒ ùkü DIR-vitamineral 0.2%, y ùkü w s l q, DIR-vita 0.1%, DIR-vitamineral 0.2% ƒƒ ùkù. l DIR-vitamineral 0.2% 4 3 š y w, s l q, DIR-vita 0.1% 3 y ùkü w ùkü. w, DIR-vita 0.2% 3 z y ƒ ùkù DIR-vita 0.1% z q. wr, d w IgG, IgM, Total Ig w y ùkü. w l x w» ü» s T q w y y, x DIR-vitamineral 0.2% ƒ z ü» y y ùkü q. x 5ƒ ü» y y z w, ƒ z DIR-vitamineral 0.2% x, DIR-vitamineral 0.1%, s l q, DIR-vita 0.1% z q. ww, x w» z y w, w wì s z» x w. š x 1. Anadon, A., Martinez-Larranaga, M.R.: Residuesof antimicrobial drugs and feed additives in animal products; regulatory aspects. Livest. Prod. Sci., 59, (1999) 2. Pedersen, K.B., Aarestrup, F.M., Jensen, N.E., Bager, F., Jensen, L.B., Jorsal, S.E., Nielsen, T.K., Hansen, H.C., Meyling, A., Wegner, H.C.: The need for a veterinary antibiotic policy. Vet. Record. 145, (1999) 3. Bonneau, M. and Laarveld, B.: Biotechnology in animal nutrition, physiology and health. Livest.Prod. Sci. 59, (1999) 4. Berg, R.D.: Probiotics, prebiotics or conbiotics. Trends Microbiol. 6, (1998) 5. Terance, H., Alan, L., David, W.: New drug targets in inflammation and immunomodulation. DDT. 3, (1998) 6. Dunier, M., Vergnet, C., Siwicki, A.K., Verlhac, V.: Effect of lindane exposure on rainbow trout immunity. IV. Prevention of nonspecific and specific immunosuppression by dietary vitamin C. Ecotoxicol. Environ. Saf. 30, (1995) 7. Siwicki, A.K., Anderson, D.P., Dixon, O.W.: Comparisons of nonspecific and specific immunomodulation by oxolinic acid, oxytetracycline and levamisole in salmonids. Vet. Immunol. Immunopathol. 23, (1989) 8. Siwicki, A.K., Klein, P., Morand, M., Kiczka, W., Studnicka, M.: Immunostimulatory effects of dimerized lysozyme (KLP- 602) on the nonspecific defense mechanisms and protection against furunculosis in salmonids. Vet. Immunol. Immunopathol. 61, (1998) 9. Jung JY, Ahn NS, Park JS, Jo EH, Hwang JW, Lee SH, Park JR, Kim SJ, Lee YG, Jeong YH, Chung JH, Lee SJ, Lee SB, Goo YS, Kang KS, and Lee YS: Immunostimulatory Effects of Immu-Forte at 3 Months Post-Treatment in Mice. J. Fd Hyg. Safety. 20, (2005) 10. Jung JY: Changes of immunostimulatory effects by Immu- Forte on mice. Korean J. Vet. Res. 45, (2005) 11. Dutton, R.W., Bradley, L.M., Swain, S.L.: T cell memory. Annu. Rev. Immunol. 16, (1998) 12. Gonser, S., Weber, E., Folkers, G.: Peptides and polypeptides as modulators of the immune response: thymopentin-an example with unknown mode of action. Pharmaceutica Acta Helvetiae. 73, (1999) 13. Zuckermann, F.A. and Husmann, R.J.: Functional and Phenotypic analysis of porcine peripheral blood CD4/CD8 double-positive T cells. Immunology. 87, (1996) 14. Lunn, D.P.: A comparative review of human and equine leukocyte differentiation antigens. Br. Vet. J. 149, (1993) 15. Pescovitz, M.D., Sakopoulos, A.G., Gaddy, J.A., Husmann, R.J., Zuckermann, F.A. : Porcine peripheral blood CD4/CD8 dual expressing T-cells. Vet. Immunol. Immunopathol. 43, (1994)
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