Skip to main content
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1996 Mar;103(3):422–428. doi: 10.1111/j.1365-2249.1996.tb08297.x

Selected Th1 and Th2 cytokine mRNA expression by CD4+ T cells isolated from inflamed human gingival tissues

K FUJIHASHI *, M YAMAMOTO *, T HIROI , T V BAMBERG *, J R MCGHEE , H KIYONO *,†,
PMCID: PMC2200368  PMID: 8608641

Abstract

Elevated numbers of plasma cells are associated with localized and chronically inflamed gingiva of patients with adult periodontitis. However, only limited information is currently available as to how cytokines produced by CD4+ T cells are involved in these increased B cell responses in affected gingival tissues. When gingival mononuclear cells (GMC) were isolated from inflamed tissues and examined by flow cytometry, ∼20–30% of lymphocytes were CD4+ T cells. For the analysis of Th1 and Th2 cytokine expression by these CD4+ T cells, RNA was extracted and reverse transcriptase-polymerase chain reaction (RT-PCR) was performed by using specific 5' and 3' primers for interferon-gamma (IFN-γ) and IL-2 (Th1), IL-4, IL-5, IL-6, IL-10 and IL-13 (Th2) and β-actin (internal control). Two distinct cytokine profiles were noted based on the expression of selected Th1 and Th2 cytokines, where one pattern was represented by expression of mRNA for IFN-γ, IL-6, IL-10 and IL-13, while the second consisted of mRNA for IFN-γ, IL-6 and IL-13. In most samples, mRNA for IL-2, IL-4 and IL-5 were not detected by cytokine-specific RT-PCR. When RNA was isolated from CD4+ T cells of concanavalin A-stimulated peripheral blood mononuclear celts (PBMC) of the same patients and examined by RT-PCR. mRNA for all Th1 and Th2 cytokines were detected. These findings suggest that although human CD4+ T cells are capable of producing an array of Th1- and Th2-type cytokines, the CD4+ T cells associated with periodontitis are limited to production of IFN-γ, IL-6, IL-13 and in some instances IL-10. CD4+ T cells from diseased periodontal tissues are divisible into two groups based upon whether or not IL-10 is produced, together with IFN-γ, IL-6 and IL-13.

Keywords: cytokines, gingival tissues, inflammation, Th1 and Th2

Full Text

The Full Text of this article is available as a PDF (1,019.8 KB).

References

  • 1.Mosmann TR, Cherwinski H, Bond MW, et al. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol. 1986;136:2348–57. [PubMed] [Google Scholar]
  • 2.Mosmann TR, Coffman RL. Th1 and Th2 cells: different patterns of lymphokine secretion lead to different functional properties. Annu Rev Immunol. 1989;7:145–73. doi: 10.1146/annurev.iy.07.040189.001045. [DOI] [PubMed] [Google Scholar]
  • 3.Romagnani S. Human Th1 and Th2 subsets: doubt no more. Immunol Today. 1991;12:256–7. doi: 10.1016/0167-5699(91)90120-I. [DOI] [PubMed] [Google Scholar]
  • 4.Romagnani S. Lymphokine production by human T cells in disease states. Annu Rev Immunol. 1994;12:227–57. doi: 10.1146/annurev.iy.12.040194.001303. [DOI] [PubMed] [Google Scholar]
  • 5.Wierenga EA, Snoek M, deGroot C, et al. Evidence for compartmentalization of functional subsets of CD2+ T lymphocytes in atopic patients. J Immunol. 1990;144:4651–6. [PubMed] [Google Scholar]
  • 6.Del Prete GF, DeCarli M, Mastromauro C, et al. Purified protein derivative of Mycobacterium tuberculosis and excretory-secretory antigen(s) of Toxocara canis expand in vitro human T cells with stable and opposite (type 1 T helper or type 2 T helper) profile of cytokine production. J Clin Invest. 1991;8:346–50. doi: 10.1172/JCI115300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Parronchi P, Macchia D, Piccinni MP, et al. Allergen- and bacterial antigen-specific T cell clones established from atopic donors show a different profile of cytokine production. Proc Natl Acad Sci USA. 1991;88:4538–42. doi: 10.1073/pnas.88.10.4538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Yamamura M, Uyemura K, Deans RJ, et al. Defining protective responses to pathogens: cytokine profiles in leprosy lesions. Science. 1991;254:277–9. doi: 10.1126/science.254.5029.277. [DOI] [PubMed] [Google Scholar]
  • 9.Yssel H, Shanafelt MC, Soderberg C, et al. Borrelia burgdorferi activates a T helper type 1 like T cell subset in Lyme arthritis. J Exp Med. 174:593–601. doi: 10.1084/jem.174.3.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Schlaak J, Hermann E, Ringhoffer M, et al. Predominance of Th1-type T cells in synovial fluid of patients with Yersinia-induced reactive arthritis. Eur J Immunol. 1992;22:2771–6. doi: 10.1002/eji.1830221103. [DOI] [PubMed] [Google Scholar]
  • 11.Lahesmaa R, Yssel H, Batsford S, et al. Yersinia enterocoliticia activates a T helper type 1 like T cell subset in reactive arthritis. J Immunol. 1992;148:3079–85. [PubMed] [Google Scholar]
  • 12.Socransky SS, Haffajee AD. The bacterial etiology of destructive periodontal disease: current concepts. J Periodontol. 1992;63:322–31. doi: 10.1902/jop.1992.63.4s.322. [DOI] [PubMed] [Google Scholar]
  • 13.Williams RC. Periodontal disease. New Eng J Med. 1990;322:373–82. doi: 10.1056/NEJM199002083220606. [DOI] [PubMed] [Google Scholar]
  • 14.Okada H, Kida T, Yamagami H. Identification and distribution of immunocompetent cells in inflamed gingiva of human chronic periodontitis. Infect Immun. 1983;41:365–74. doi: 10.1128/iai.41.1.365-374.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Taubman MA, Stoufi ED, Ebersole JL, et al. Phenotypic studies of cells from periodontal disease tissues. J Periodontal Res. 1984;19:587–90. doi: 10.1111/j.1600-0765.1984.tb01320.x. [DOI] [PubMed] [Google Scholar]
  • 16.Seymour GJ. Possible mechanisms involved in the immunoregulation of chronic inflammatory periodontal disease. J Dent Res. 1987;66:2–9. doi: 10.1177/00220345870660010401. [DOI] [PubMed] [Google Scholar]
  • 17.McGhee ML, Ogawa T, Pitts AM, et al. Cellular analysis of functional mononuclear cells from chronically inflamed gingival tissue. Reg Immunol. 1989;2:103–10. [PubMed] [Google Scholar]
  • 18.Genco RJ, Mashimo PA, Kreggier G, et al. Antibody-mediated effects on the periodontium. J Periodontol. 1977;45:330–7. doi: 10.1902/jop.1974.45.5.330. [DOI] [PubMed] [Google Scholar]
  • 19.Mackler BF, Frostad KB, Robertson PB, et al. Immunoglobulin bearing lymphocytes and plasma cells in human periodontal disease. J Periodontol Res. 1977;12:37–45. doi: 10.1111/j.1600-0765.1977.tb00107.x. [DOI] [PubMed] [Google Scholar]
  • 20.Ogawa T, Tarkowski A, McGhee ML, et al. Analysis of human IgG and IgA subclass antibody-secreting cells from localized chronic inflammatory tissue. J Immunol. 1989;142:1150–8. [PubMed] [Google Scholar]
  • 21.Kono Y, Beagley KW, Fujihashi K, et al. Cytokine regulation of localized inflammation. Induction of activated B cells and IL-6 mediated polyclonal IgG and IgA synthesis in inflamed human gingiva. J Immunol. 1991;146:1812–21. [PubMed] [Google Scholar]
  • 22.Matsuki Y, Yamamoto T, Hara K. Detection of inflammatory cytokine messenger RNA (mRNA)-expressing cells in human inflamed gingiva by combined in situ hybridization and immunohistochemistry. Immunology. 1992;76:42–47. [PMC free article] [PubMed] [Google Scholar]
  • 23.Gemmell E, Seymour GJ. Interleukin-1, interleukin-6 and transforming growth factor-β production by human gingival mononuclear cells following stimulation with Porphyromonas gingivalis and Fusobacterium nucleatum. J Periodont Res. 1993;28:122–9. doi: 10.1111/j.1600-0765.1993.tb01059.x. [DOI] [PubMed] [Google Scholar]
  • 24.Fujihashi K, Beagley KW, Kono Y, et al. Gingival mononuclear cells from chronic inflammatory periodontal tissues produce interleukin (IL)-5 and IL-6 but not IL-2 and IL-4. Amer J Pathol. 1993;142:1239–50. [PMC free article] [PubMed] [Google Scholar]
  • 25.Lundqvist C, Baranov V, Teglund S, et al. Cytokine profile and ultrastructure of intraepithelial γδ T cells in chronically inflamed human gingiva suggest a cytotoxic effector function. J Immunol. 1994;153:2302–12. [PubMed] [Google Scholar]
  • 26.Brenner CA, Tam AW, Nelson PA, et al. Message amplification phenotyping (MAPPing): a technique to simultaneously measure multiple mRNAs from small numbers of cells. Biotechniques. 1989;7:1096–103. [PubMed] [Google Scholar]
  • 27.Yamamoto M, Fujihashi K, Beagley KW, et al. Cytokine synthesis by intestinal intraepithelial lymphocytes. Both γ/δ T cell receptor-positive and α/b T cell receptor-positive T cells in the G1 phase of cell cycle produce IFN-γ and IL-5. J Immunol. 1993;150:106–14. [PubMed] [Google Scholar]
  • 28.Chomczynski P, Sacchi N. Single-step method of RNA isolation of acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–9. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  • 29.Stoufi ED, Taubman MA, Ebersole JL, et al. Phenotype analyses of mononuclear cells recovered from healthy and diseased human periodontal tissues. J Clin Immunol. 1987;7:235–45. doi: 10.1007/BF00915729. [DOI] [PubMed] [Google Scholar]
  • 30.Lundqvist C, Hammarström ML. T-cell receptor γδ-expressing intraepithelial lymphocytes are present in normal and chronically inflamed human gingiva. Immunology. 1993;79:38–45. [PMC free article] [PubMed] [Google Scholar]
  • 31.Becker S. Interferons as modulators of human monocyte-macrophage differentiation. I. Interferon-gamma increases HLA-DR expression and inhibits phagocytosis of zymosan. J Immunol. 1984;132:1249–54. [PubMed] [Google Scholar]
  • 32.Mangan DF, Robertson B, Wahl SW. IL-4 enhances programmed cell death (apoptosis) in stimulated human monocytes. J Immunol. 1992;148:1812–6. [PubMed] [Google Scholar]
  • 33.Zurawski G, deVries JE. Interleukin-13, an interleukin-4-like cytokine that acts on monocytes and B cells, but not on T cells. Immunol Today. 1994;15:19–26. doi: 10.1016/0167-5699(94)90021-3. [DOI] [PubMed] [Google Scholar]
  • 34.Punnonen J, Aversa G, Cocks BG, et al. Interleukin 13 induces interleukin 4-independent IgG4 and IgE synthesis and CD23 expression by human B cells. Proc Natl Acad Sci USA. 1993;90:3730–4. doi: 10.1073/pnas.90.8.3730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Minty A, Chalon P, Derocq JM, et al. Interleukin-13 is a new human lymphokine regulating inflammatory and immune responses. Nature. 1993;362:248–50. doi: 10.1038/362248a0. [DOI] [PubMed] [Google Scholar]
  • 36.Moore KW, O'Garra A, de Waal Malefyt R, et al. Interleukin 10. Annu Rev Immunol. 1993;11:165–90. doi: 10.1146/annurev.iy.11.040193.001121. [DOI] [PubMed] [Google Scholar]
  • 37.Taga K, Tosato G. IL-10 inhibits human T cell proliferation and IL-2 production. J Immunol. 1992;148:1143–8. [PubMed] [Google Scholar]
  • 38.Del Prete GF, De Carli M, Almerigogna F, et al. Human IL-10 is produced by both type 1 helper (Th1) and type 2 helper (Th2) T cell clones and inhibits their antigen-specific proliferation and cytokine production. J Immunol. 1993;150:353–60. [PubMed] [Google Scholar]
  • 39.de Waal Malefyt R, Abrams J, Bennett B, et al. Interleukin 10 (IL-10) inhibits cytokine synthesis by human monocytes: an autoregulatory role of IL-10 produced by monocytes. J Exp Med. 1991;174:1209–20. doi: 10.1084/jem.174.5.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES