Skip to main content
SpringerLink
Log in

BLT-esterase activity followingin vitro andin vivo activation of human lymphocytes with interleukin-2

In vivoIL-2 induces BLT-esterase

  • Published:
Biotherapy

Abstract

BLT-esterase and cytolytic activity by humanin vitro andin vivo generated Lymphokine Activated Killer (LAK) cells were measured. Lysates made from peripheral blood lymphocytes (PBL) of both normal donors and cancer patients receiving IL-2 therapy were assayed for BLT-esterase activity in a spectro-photometric assay. Cytotoxicity of PBL was measured in a51Cr-release assay. Both BLT-esterase activity and cytotoxicity increased when normal-donor PBL were stimulatedin vitro with IL-2, with greater activities at higher IL-2 concentrations. The activities also increased over time, peaking at 6 days ofin vitro stimulation. Patient PBL had increased BLT-esterase and cytotoxic activities after 4 weeks ofin vivo IL-2 treatment. This association of BLT-esterase activity and cytotoxicity with IL-2 activation is consistent with the model that LAK cytotoxicity is mediated by secretion of BLT-esterase associated cytolytic granules. Lymphocytes obtained afterin vivo IL-2 treatment and cultured for 3-4 hours in IL-2 show markedly augmented cytotoxic activity but no increase in their BLT-esterase activity. These results indicate that the increased cytotoxicity observed after this brief pulse ofin vitro IL-2 followingin vivo IL-2 treatment must result from effects of IL-2 other than the production of more esterase-containing cytolytic granules.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BLT:

N-α-carbamazepine-L-lysine thiobenzyl

HS-RPMI:

medium RPMI 1640 supplemented with human serum and antibiotics

IL-2:

interleukin-2

LAK:

lymphokine activated killer activity

LU:

lytic units

NK:

natural killer

OD:

optical density

PBL:

peripheral blood lymphocytes

References

  1. Young JD, Liu CC, Persechini PM, Cohn ZA. Performdependent and -independent pathways of cytotoxicity mediated by lymphocytes. Immunol Rev 1988; 103: 161–202.

    Google Scholar 

  2. Young JD, Liu C. Multiple mechanisms of lymphocytemediated killing. Immunol Today 1988; 9: 140–4.

    Google Scholar 

  3. Lowrey D, Hameed A, Lichtenheld M, Podack E. Isolation and characterization of cytotoxic granules from human lymphokine (interleukin-2) activated killer cells. Cancer Res 1988; 48: 4681–8.

    Google Scholar 

  4. Pasternack MS, Eisen HN. A novel serine esterase expressed by cytotoxic T lymphocytes. Nature 1985; 314: 743–5.

    Google Scholar 

  5. Pasternack MS, Verret CR, Liu MA, Eisen HN. Serine esterase in cytolytic T lymphocytes. Nature 1986; 322: 740–3.

    Google Scholar 

  6. Podack ER, Lowrey DM, Lichtenheld M, Hamid A. Function of granule perform and esterases in T cell-mediated reaction. In: Battisto JR, Plate J, Shearer G, eds. Cytotoxic T Cells Biology and Relevance to Disease. New York: New York Academy of Sciences, 1988: 292–302 (Annals of the New York Academy of Sciences; Vol 532).

    Google Scholar 

  7. Masson D, Nabholz M, Estrade C, Tschopp J. Granules of cytolytic T lymphocytes contain two serine esterases. EMBO J 1986; 5: 1595–1600.

    Google Scholar 

  8. Takayama H, Trenn G, Humphrey W, Bluestone J, Henkart P, Sitkovsky M. Antigen receptor-triggered secretion of a trypsin-type esterase from cytotoxic T lymphocytes. J Immunol 1987; 138: 566–9.

    Google Scholar 

  9. Garcia-Sanz J, Plaetinck G, Velotti R, Masson D, Tschopp J, MacDonald HR, Nabholz M. Perforin is present only in normal activated Lyt2 + T lymphocytes and not in L3T42+ cells, but the serine protease granzyme A is made by both subsets. EMBO J 1987; 6: 933–8.

    Google Scholar 

  10. Dennert G, Podack ER. Cytolysis by H-2-specific T killer cells: assembly of tubular complexes on target membranes. J Exp Med 1983; 157: 1483–95.

    Google Scholar 

  11. Millard PJ, Henkart MP, Reynolds CW, Henkart PA. Purification and properties of cytoplasmic granules from cytotoxic rat LGL tumors. J Immunol 1984; 132: 3197–204.

    Google Scholar 

  12. Taplits M, Henkart P, Hodes R. T-helper cell cytoplasmic granules: exocytosis in response to activation via the T cell receptor. J Immunol 1988; 141: 1–9.

    Google Scholar 

  13. Ferguson WS, Verret CR, Reilly EB, Iannini MJ, Eisen HN. Serine esterase and hemolytic activity in human cloned cytotoxic T lymphocytes. J Exp Med 1988; 167: 528–40.

    Google Scholar 

  14. Berke G, Rosen D. Highly lytic in vivo primed cytolytic T lymphocytes devoid of lytic granules and BLT-esterase activity acquire these constituents in the presence of T cell growth factors upon blast transformationin vitro. J Immunol 1988; 141: 1429–36.

    Google Scholar 

  15. Sondel PM, Hank JA., Rosenthal N, Moore KH, Hong R, Storer B, Kohler PC. Clinical testing of IL-2:in vivo administration of IL-2 induces IL-2 dependent non-MHC restricted cytotoxicity (NRC). In: Truitt RL, Gale RP, Bortin MM, eds. Cellular Immunotherapy of Cancer. New York: Alan R. Liss, Inc., 1987: 161–72.

    Google Scholar 

  16. Hank JA, Kohler PC, Weil-Hillman G et al.In vivo induction of the lymphokine-activated killer phenomenon: interleukin-2-dependent human non-major histocompatibility complex-restricted cytotoxicity generatedin vivo during administration of human recombinant interleukin-2. Cancer Res 1988; 48: 1965–71.

    Google Scholar 

  17. Takayama H, Trenn G, Sitkovsky MV. A novel cytotoxic T lymphocyte activation assay: optimized conditions for antigen receptor triggered granule enzyme secretion. J Immunol Methods 1987; 104: 183–90.

    Google Scholar 

  18. Pross HF, Maroun JA. The standardization of NK cell assays for use in studies of biological response modifiers. J Immunol Methods 1984; 68: 235–49.

    Google Scholar 

  19. Sosman JA, Kohler PC, Hank JA et al. Repetitive weekly cycles of recombinant human interleukin-2 (IL-2): responses of renal carcinoma with acceptable toxicity. J Nat Cancer Inst 1988; 80: 60–3.

    Google Scholar 

  20. Sosman JA, Kohler PC, Hank JA et al. Repetitive weekly cycles of interleukin-2 (IL-2): II. Clinical and immunologic effects of dose, schedule and indomethacin. J Nat Cancer Inst 1988; 80: 1451–60.

    Google Scholar 

  21. osenthal NS, Hank JA, Kohler PC et al. Thein vitro function of lymphocytes from 25 cancer patients receiving four to seven consecutive days of recombinant IL-2. J Biol Resp Mod 1988; 7: 123–39.

    Google Scholar 

  22. Grimm EA, Maxumder A, Zhang HZ, Rosenberg SA. The lymphokine activated killer cell phenomenon: lysis of NK resistant fresh solid tumor cells by IL-2 activated autologous human peripheral blood lymphocytes. J Exp Med 1982; 155: 1823.

    Google Scholar 

  23. Berke G. Lymphocyte mediated cytolysis: the mechanisms whereby early membrane derangements results in target cell death. In: Battisto JR, Plate J, Shearer G, eds. Cytotoxic T cells; biology and relevance to disease. NY Academy of Science, 1988: 314–35.

  24. Roberts K, Lotze M. Interleukin-2 promotes conjugate formation by purified LAK precursors and T lymphocytes: evaluation of conjugates using flow cytometric techniques. J Biol Resp Mod 1988; 7: 249–66.

    Google Scholar 

  25. Yanelli JR, Thurman GB, Mrowca-Bastin A, Pennington CS, West WH, Oldham RK. Enhancement of human lymphokine activated killer cell cytolysis and a method for increasing lymphokine activated killer cell yields to cancer patients. Cancer Res 1988; 48: 5696–700.

    Google Scholar 

  26. Ortaldo JR, Longo DL. Human natural lymphocyte effector cells: definition, analysis of activity, and clinical effectiveness. J Nat Cancer Inst 1988; 80: 999–1010.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Human Oncology, University of Wisconsin, Madison, Wisconsin, USA

    Adria Cannon, Jacquelyn A. Hank & Paul M. Sondel M.D., Ph.D.

  2. Department of Pediatrics, University of Wisconsin, Madison, Wisconsin, USA

    Paul M. Sondel M.D., Ph.D.

  3. Department of Genetics, University of Wisconsin, Madison, Wisconsin, USA

    Paul M. Sondel M.D., Ph.D.

Authors
  1. Adria Cannon
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jacquelyn A. Hank
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Paul M. Sondel M.D., Ph.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cannon, A., Hank, J.A. & Sondel, P.M. BLT-esterase activity followingin vitro andin vivo activation of human lymphocytes with interleukin-2. Biotherapy 3, 253–260 (1991). https://doi.org/10.1007/BF02171689

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02171689

Key words

Search

Navigation