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Journal of Clinical Immunology

, Volume 5, Issue 3, pp 172–179 | Cite as

Suppression ofin vitro lymphocyte and neutrophil responses by a low molecular weight suppressor active peptide from burn-patient sera

  • A. Nuri Ozkan
  • John L. Ninnemann
Original Articles

Abstract

Thermal injury produces profound pathophysiological changes in the severely burned patient. Primary among these is the modulation of immunity, leading to episodes of immunosuppression and thus increasing the risk of sepsis and possible death. We herein report the isolation of a low molecular weight suppressor active peptide (SAP) which appears to be responsible for many of the observed immunologic changes in burned patients. SAP suppressed T-lymphocyte blastogenesis in the mixed lymphocyte reaction (MLR) and inhibited neutrophil chemotaxis (CTX)in vitro. Characterization of SAP revealed a complex structure comprised of (1) a peptide component rich in glycine, serine, and alanine; (2) a carbohydrate component containing sialic acid; and (3) a fatty acid component, tentatively identified as prostaglandin E. The immunosuppressive activity of SAP is dependent upon the presence of all three structural components. The molecular weight of SAP was estimated to be 3654 as determined by Amicon cell ultrafiltration and amino acid analysis. The isoelectric point of SAP was estimated by chromatofocusing and ion-exchange chromatography to be between 3.2 and 3.6. We hypothesize that the suppressor active peptide may be comprised of cellular or tissue components released into the circulation at the time of injury.

Key words

Immunosuppression thermal injury lymphocyte neutrophil prostaglandin 

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References

  1. 1.
    Davis JM, Dineen P, Gallins JI: Neutrophil degranulation and abnormal chemotaxis after thermal injury. J Immunol 124:1467–1471, 1980Google Scholar
  2. 2.
    Gelfand J, Donelan M, Hawiger A, Burke J: Alternative complement pathway activation increases mortality in a model of burn injury in mice. J Clin Invest 70:1170–1176, 1982Google Scholar
  3. 3.
    Hansbrough J, Peterson V, Kortz E, Diacentine J: Postburn immunosuppression in an animal model: Monocyte dysfunction induced by burned tissue. Surgery 93:415–423, 1982Google Scholar
  4. 4.
    Ninnemann JL: Immunologic defenses against infection: Alterations following thermal injuries. J Burn Care Rehab 3:355–366, 1982Google Scholar
  5. 5.
    Ninnemann JL, Stein MD, Condie JT: The nature of impaired lymphocyte response following thermal injury. J Burn Care Rehab 2:196–199, 1982Google Scholar
  6. 6.
    Hakim AA: An immunosuppressive factor from serum of thermally traumatized patients. J Trauma 17:908–919, 1977Google Scholar
  7. 7.
    Garner WD, Prager MD, Baxter CR: Multiple inhibitors of lymphocyte transformation in serum from burn patients. J Burn Care Rehab 2:97–105, 1981Google Scholar
  8. 8.
    Goodwin JS, Ceuppens J: Regulation of the immune response by prostaglandins. J Clin Immunol 3:295–315, 1983Google Scholar
  9. 9.
    Winkelstein A, Kelley VE: The pharmacologic effect of PGE1 on murine lymphocytes. Blood 55:437–443, 1980Google Scholar
  10. 10.
    Hansbrough J, Peterson V, Zapata-Sirvent R, Claman HN: Postburn immunosuppression in an animal model. II. Restoration of cell-mediated immunity by immunomodulatory drugs. Surgery 95:290–295, 1984Google Scholar
  11. 11.
    Ninnemann JL, Stockland AE: Participation of prostaglandin E in immunosuppression following thermal injury. J Trauma 24:201–207, 1984Google Scholar
  12. 12.
    Richey J: A comprehensive separation technique for biopolymers. Am Lab 10:1–17, 1982Google Scholar
  13. 13.
    Udenfriend S, Stein S, Boehlen P, Dairman W, Leimgruber W, Weigele M: Fluorescamine: a reagent for assay of amino acids, peptides, proteins and primary amines in the picomole range. Science 178:871–872, 1972Google Scholar
  14. 14.
    Harveth L, Falk WR, Leonard EJ: Rapid quantitation of neutrophil chemotaxis: Use of a polyvinylpyrrolidone-free polycarbonate membrane in a multiwell assembly. J Immunol Methods 37:39–45, 1980Google Scholar
  15. 15.
    Falk WR, Goodwin RH, Leonard EJ: A 48-well micro chemotaxis assembly for rapid and accurate measurement of leukocyte migration. J Immunol Methods 33:239–247, 1980Google Scholar
  16. 16.
    Ninnemann JL, Stockland AE, Condie JT: Induction of prostaglandin synthesis dependent suppressor cells with endotoxin: Occurrence in patients with thermal injuries. J Clin Immunol 3:142–150, 1983Google Scholar
  17. 17.
    Chen RF: Removal of fatty acids from serum albumin by charcoal treatment. J Biol Chem 212:173–181, 1967Google Scholar
  18. 18.
    Matsudaira PT, Burgess DR: SDS microslab linear gradient polyacrylamide gel electrophoresis. Anal Biochem 87:386–396, 1978Google Scholar
  19. 19.
    Laemmli UK, Favre M: Maturation of the head of bacteriophage T4. J Mol Biol 80:575–599, 1973Google Scholar
  20. 20.
    Raz A: Interaction of prostaglandins with blood plasma proteins: Binding of prostaglandin E2 in vitro and in vivo. Biochim Biophys Acta 280:602–613, 1972Google Scholar
  21. 21.
    Koop PR, Morgan ET, Tarr GE, Coon MJ: Purification and characterization of a unique isozyme of a cytochrome P-450 from liver microsomes of ethanol-treated rabbits. J Biol Chem 257:8472–8480, 1982Google Scholar
  22. 22.
    Walker RI, Porvaznik M: Association of bacteria and endotoxin with posttrauma events. In Traumatic Injury: Infection and Other Immunologic Sequelae, JL Ninnemann (ed). Baltimore, University Park Press, 1983, pp 1–15Google Scholar
  23. 23.
    Ninnemann JL, Condie JT, Davis SE, Crockett RA: Isolation of immunosuppressive serum components following thermal injury. J Trauma 22:837–844, 1982Google Scholar

Copyright information

© Plenum Publishing Corporation 1985

Authors and Affiliations

  • A. Nuri Ozkan
    • 1
  • John L. Ninnemann
    • 1
  1. 1.Divisions of Plastic Surgery and TraumaUniversity of California, San Diego, School of MedicineLa Jolla

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