, 38:1221 | Cite as

Intramuscular injection of antigens and adjuvant preferentially decreases 18∶2n−6 and 18∶3n−3 in pig neck muscle

  • Richard P. BazinetEmail author
  • Holly Douglas
  • Ewen G. McMillan
  • Bruce N. Wilkie
  • Stephen C. Cunnane


Linoleic (18∶2n−6) and α-linolenic acids (18∶3n−3) have many important physiological functions including immunomodulation. We tested how immunization influences the metabolism of 18∶2n−6 and 18∶3n−3 in the neck muscle of pigs. At 35 d old, pigs received either an intramuscular neck injection containing hen egg white lysozyme (HEWL), killed Mycobacterium tuberculosis, and Freund’s complete adjuvant (immunized) or PBS (control). At 49 d old, immunized pigs received a booster injection of HFWI and Freund’s incomplete adjuvant, and the control pigs received PBS into the neck. At 56 d old, all pigs received an intradermal injection of Mycobacterium bovis into the hind leg to induce a delayed-type hypersensitivity (DTH) reaction. At 57 d old, immunized pigs had a twofold increase in serum haptoglobin, a 10-fold increase in antibodies to HEWL, and the skinfold at the DTH reaction site was 10 times thicker than the controls. Both 18∶2n−6 and 18∶3n−3 (% composition) were approximately 25% lower in muscle IG, 40% lower in FFA, 50% lower in phospholipids, but not different in cholesteryl esters of the neck muscle of immunized pigs. The antigens in this model induce an increased response in the innate (haptoglobin), humoral (antibodies), and cellular (DTH) immune systems as well as a preferential decrease of 18∶2n−6 and 18∶3n−3 in the inflamed neck muscle. It appears that 18∶2n−6 and 18∶3n−3 are preferentially metabolized (possibly β-oxidized) in response to antigens.


Cholesteryl Ester Neck Muscle Purify Protein Derivative Serum Haptoglobin PUFA Metabolism 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



linoleic acid


α-linolenic acid


arachidonic acid




cell-mediated immune/immunity


delayed-type hypersensitivity


Freund’s complete adjuvant


hen egg white lysozyme


hormone-sensitive lipase






purified protein derivative


  1. 1.
    Aaes-Jørgensen, E. (1961) Essential Fatty Acids, Physiol. Rev. 41, 1–51.PubMedGoogle Scholar
  2. 2.
    Burr, G.O., and Burr, M.M. (1929) A New Deficiency Disease Produced by the Rigid Exclusion of Fat from the Diet, J. Biol. Chem. 82, 345–367.Google Scholar
  3. 3.
    Salem, N.J. (1989) New Protective Roles of Selected Nutrients, in Human Nutrition (Spiller, G., and Scale, J., eds.), pp. 109–228, Alan R. Liss, New York.Google Scholar
  4. 4.
    Neuringer, M., Connor, W.E., Lin, D.S., Barstad, L., and Luck, S. (1986) Biochemical and Functional Effects of Prenatal and Postnatal Omega-3 Fatty Acid Deficiency on Retina and Brain in Rhesus Monkeys, Proc. Natl. Acad. Sci. USA 83, 4021–4025.PubMedCrossRefGoogle Scholar
  5. 5.
    Serhan, C.N., and Oliw, E. (2001) Unorthodox Routes to Prostanoid Formation: New Twists in Cyclooxygenase-Initiated Pathways, J. Clin. Invest. 107, 1481–1489.PubMedCrossRefGoogle Scholar
  6. 6.
    Kinsella, J.E., Lokesh, B., Broughton, S., and Whelan, J. (1990) Dietary Polyunsaturated Fatty Acids and Eicosanoids: Potential Effects on the Modulation of Inflammatory and Immune Cells: An Overview, Nutrition 6, 24–44; discussion 59–62.PubMedGoogle Scholar
  7. 7.
    Hwang, D. (2000) Fatty Acids and Immune Responses—A New Perspective in Searching for Clues and Mechanism, Ann. Rev. Nutr. 20, 431–456.CrossRefGoogle Scholar
  8. 8.
    Devchand, P.R., Keller, H., Peters, J.M., Vazquez, M., Gonzalez, F.J., and Wahli, W. (1996) The PPARα-Leukotriene B4 Pathway to Inflammation Control, Nature 384, 39–43.PubMedCrossRefGoogle Scholar
  9. 9.
    Calder, P.C., and Grimble, R.F. (2002) Polyunsaturated Fatty Acids, Inflammation and Immunity, Eur. J. Clin. Nutr. 56 (Suppl.) 3, S14-S19.PubMedCrossRefGoogle Scholar
  10. 10.
    Calder, P.C. (2001) Polyunsaturated Fatty Acids, Inflammation, and Immunity, Lipids 36, 1007–1024.PubMedGoogle Scholar
  11. 11.
    Frankenfield, D.C., Wiles, C.E., 3rd, Bagley, S., and Siegel, J.H. (1994) Relationships Between Resting and Total Energy Expenditure in Injured and Septic Patients, Crit. Care Med. 22, 1796–1804.PubMedCrossRefGoogle Scholar
  12. 12.
    Hwang, T.L., Huang, S.L., and Chen, M.F. (1993) The Use of Indirect Calorimetry in Critically Ill Patients—The Relationship of Measured Energy Expenditure to Injury Severity Score, Septic Severity Score, and APACHE II Score, J. Trauma 34, 247–251.PubMedCrossRefGoogle Scholar
  13. 13.
    Nanni, G., Siegel, J.H., Coleman, B., Fader, P., and Castiglione, R. (1984) Increased Lipid Fuel Dependence in the Critically Ill Septic Patient, J. Trauma 24, 14–30.PubMedGoogle Scholar
  14. 14.
    Stoner, H.B., Little, R.A., Frayn, K.N., Elebute, A.E., Tresadern, J., and Gross, E. (1983) The Effect of Sepsis on the Oxidation of Carbohydrate and Fat, Br. J. Surg. 70, 32–35.PubMedGoogle Scholar
  15. 15.
    Iriyama, K., Kusaka, N., Nishiwaki, H., Teranishi, T., Mori, H., and Suzuki, H. (1986) Metabolism of Non-protein Energy-Substrates in Septic Rats Receiving Parenteral Nutrition, Int. Surg. 71, 5–8.PubMedGoogle Scholar
  16. 16.
    Raina, N., Matsui, J., Cunnane, S.C., and Jeejeebhoy, K.N. (1995) Effect of Tumor Necrosis Factor-α on Triglyceride and Phospholipid Content and Fatty Acid Composition of Liver and Carcass in Rats, Lipids 30, 713–718.PubMedGoogle Scholar
  17. 17.
    Bazinet, R.P., McMillan, E.G., Seebaransingh, R., Hayes, A.M., and Cunnane, S.C. (2003) Whole-Body β-Oxidation of 18∶2ω6 and 18∶3ω3 in the Pig Varies Markedly with Weaning Strategy and Dietary 18∶3ω3, J. Lipid Res. 44, 314–319.PubMedCrossRefGoogle Scholar
  18. 18.
    Mallard, B.A., Wilkie, B.N., Kennedy, B.W., and Quinton, M. (1992) Use of Estimated Breeding Values in a Selection Index to Breed Yorkshire Pigs for High and Low Immune and Innate Resistance Factors, Anim. Biotechnol. 3, 257–280.CrossRefGoogle Scholar
  19. 19.
    Pond, C.M. (2002) Adipose Tissue, the Immune System and Exercise Fatigue: How Activated Lymphocytes Compete for Lipids, Biochem. Soc. Trans. 30, 270–275.PubMedCrossRefGoogle Scholar
  20. 20.
    Bazinet, R.P., Douglas, H., and Cunnane, S.C. (2003) Whole-Body Utilization of n−3 PUFA in n−6 PUFA-Deficient Rats, Lipids 38, 187–189.PubMedGoogle Scholar
  21. 21.
    Magnusson, U., Wilkie, B.N., Artursson, K., and Mallard, B.A. (1999) Interferon-α and Haptoglobin in Pigs Selectively Bred for High and Low Immune Response and Infected with Mycoplasma hyorhinis, Vet. Immunol. Immunopathol. 68, 131–137.PubMedCrossRefGoogle Scholar
  22. 22.
    Yaqoob, P., Newsholme, E.A., and Calder, P.C. (1994) Fatty Acid Oxidation by Lymphocytes, Biochem. Soc. Trans. 22, 116S.Google Scholar
  23. 23.
    Lengle, E.E., Gustin, N.C., Gonzalez, F., Menahan, L.A., and Kemp, R.G. (1978) Energy Metabolism in Thymic Lymphocytes of Normal and Leukemia AKR Mice, Cancer Res. 38, 1113–1119.PubMedGoogle Scholar
  24. 24.
    Calder, P.C., Yaqoob, P., and Newsholme, E.A. (1994) Triacylglycerol Metabolism by Lymphocytes and the Effect of Triacylglycerols on Lymphocyte Proliferation, Biochem. J. 298, Pt. 3, 605–611.PubMedGoogle Scholar
  25. 25.
    Bergman, R. (1980) Cell-Mediated Immune Response in Pigs Persistently Infected with a Mycobacterium avium Strain, Res. Vet. Sci. 28, 315–320.PubMedGoogle Scholar
  26. 26.
    Binns, R.M., Whyte, A., Licence, S.T., Harrison, A.A., Tsang, Y.T., Haskard, D.O., and Robinson, M.K. (1996) The Role of E-Selectin in Lymphocyte and Polymorphonuclear Cell Recruitment into Cutaneous Delayed Hypersensitivity Reactions in Sensitized Pigs, J. Immunol. 157, 4094–4099.PubMedGoogle Scholar
  27. 27.
    Binns, R.M., Licence, S.T., and Whyte, A. (1996) Transfer of T-Cell-Mediated, Antigen-Specific Delayed Type Hypersensitivity Reactions to Naive Recipient Inbred Pigs, Res. Vet. Sci. 60, 24–28.PubMedCrossRefGoogle Scholar
  28. 28.
    Hiramatsu, K., and Arimori, S. (1982) Rapid Determination of Lipids in Healthy Human Lymphocytes, J. Chromatogr. 227, 423–431.PubMedGoogle Scholar
  29. 29.
    Marinetti, G.V., and Cattieu, K. (1982) Composition and Metabolism of Phospholipids of Human Leukocytes, Chem. Phys. Lipids 31, 169–177.PubMedCrossRefGoogle Scholar
  30. 30.
    Field, C.J. (2000) Use of T Cell Function to Determine the Effect of Physiologically Active Food Components, Am. J. Clin. Nutr. 71, 1720S-1725S; discussion 1726S–1727S.PubMedGoogle Scholar
  31. 31.
    Chapel, H.M., and August, P.J. (1976) Report of Nine Cases of Accidental Injury to Man with Freund’s Complete Adjuvant, Clin. Exp. Immunol. 24, 538–541.PubMedGoogle Scholar
  32. 32.
    Black, C.A. (1999) Delayed Type Hypersensitivity: Current Theories with an Historic Perspective, Dermatol. Online J. 5, 5–27.Google Scholar
  33. 33.
    Mattacks, C.A., and Pond, C.M. (1997) The Effects of Feeding Suet-Enriched Chow on Site-Specific Differences in the Composition of Triacylglycerol Fatty Acids in Adipose Tissue and Its Interactions in vitro with Lymphoid Cells, Br. J. Nutr. 77, 621–643.PubMedCrossRefGoogle Scholar
  34. 34.
    Pond, C.M. (1999) Physiological Specialisation of Adipose Tissue, Prog. Lipid Res. 38, 225–248.PubMedCrossRefGoogle Scholar
  35. 35.
    Holm, C., Kirchgessner, T.G., Svenson, K.L., Fredrikson, G., Nilsson, S., Miller, C.G., Shively, J.E., Heinzmann, C., Sparkes, R.S., Mohandas, T., et al. (1988) Hormone-Sensitive Lipase: Sequence, Expression, and Chromosomal Localization to 19 cent-q13.3, Science 241, 1503–1506.PubMedCrossRefGoogle Scholar
  36. 36.
    Halliwell, K.J., Fielding, B.A., Samra, J.S., Humphreys, S.M., and Frayn, K.N. (1996) Release of Individual Fatty Acids from Human Adipose Tissue in vivo After an Overnight Fast, J. Lipid Res. 37, 1842–1848.PubMedGoogle Scholar
  37. 37.
    Gavino, V.C., and Gavino, G.R. (1992) Adipose Hormone-Sensitive Lipase Preferentially Releases Polyunsaturated Fatty Acids from Triglycerides, Lipids 27, 950–954.PubMedGoogle Scholar
  38. 38.
    Haemmerle, G., Zimmermann, R., Hayn, M., Theussl, C., Waeg, G., Wagner, E., Sattler, W., Magin, T.M., Wagner, E.F., and Zechner, R. (2002) Hormone-Sensitive Lipase Deficiency in Mice Causes Diglyceride Accumulation in Adipose Tissue, Muscle, and Testis, J. Biol. Chem. 277, 4806–4815.PubMedCrossRefGoogle Scholar
  39. 39.
    Gavino, G.R., and Gavino, V.C. (1991) Rat Liver Outer Mitochondrial Carnitine Palmitoyltransferase Activity Towards Long-Chain Polyunsaturated Fatty Acids and Their CoA Esters, Lipids 26, 266–270.PubMedGoogle Scholar

Copyright information

© AOCS Press 2003

Authors and Affiliations

  • Richard P. Bazinet
    • 1
    Email author
  • Holly Douglas
    • 1
  • Ewen G. McMillan
    • 2
  • Bruce N. Wilkie
    • 3
  • Stephen C. Cunnane
    • 1
  1. 1.Department of Nutritional Sciences, Faculty of MedicineUniversity of TorontoTorontoCanada
  2. 2.Maple Leaf Foods AgresearchGuelphCanada
  3. 3.Department of PathobiologyUniversity of GuelphGuelphCanada

Personalised recommendations