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Analysis of CD20-dependent cellular cytotoxicity by G-CSF-stimulated neutrophils

Leukemia volume 16, pages 693–699 (2002)Cite this article

Abstract

Rituximab, a chimeric CD20 monoclonal antibody (mAb), is widely used in the treatment of patients with low-grade non-Hodgkin's lymphoma. Possible anti-tumour mechanisms involve complement-mediated lysis and/or antibody-dependent cellular cytotoxicity (ADCC). Because G-CSF greatly enhances the cytotoxicity of neutrophils (PMN) in ADCC, the clinical efficacy of rituximab might be enhanced by the addition of G-CSF. Therefore, we investigated the neutrophil-mediated CD20-dependent cellular cytotoxicity in B cell lines. In contrast to previous studies by others, we found that G-CSF-primed PMN are capable of functioning as effector cells in CD20-dependent cellular cytotoxicity. However, HLA class II mAbs were far more effective. The differences between HLA class II- and CD20-mediated PMN-ADCC were not due to: (1) the use of chimeric (hIgG1) mAbs vs mIgG2a mAbs; (2) HLA class II-induced apoptosis as an ‘ADCC-sensitising’ mechanism; (3) CD20-induced inhibition of ADCC; (4) inferior membrane mobility of CD20. Analysis of Fcγreceptor (FcγR) involvement showed that although CD20-induced ADCC was mediated mainly via FcγRI, for optimal lysis FcγRI and FcγRII were both required. In contrast, in HLA class II-dependent ADCC both FcγRI and II were capable of independently inducing maximum lysis. The mechanism underlying these differences in FcγR-binding and activation remains to be elucidated.

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References

  1. Coiffier B, Haioun C, Ketterer N, Engert A, Tilly H, Ma D, Johnson P, Lister A, Feuring-Buske M, Radford JA, Capdeville R, Diehl V, Reyes F . Rituximab (anti-CD20 monoclonal antibody) for the treatment of patients with relapsing or refractory aggressive lymphoma: a multicenter phase II study Blood 1998 92: 1927–1932

    CAS  PubMed  Google Scholar 

  2. McLaughlin P, Grillo-López A, Link BK, Levy R, Czuczman MS, Williams ME, Heyman MR, Bence-Bruckler I, White CA, Cabanillas F, Jain V, Ho AD, Lister J, Wey K, Shen D, Dallaire BK . Rituximab chimeric anti-CD20 monoclonal antibody therapy for relapsed indolent lymphoma: half of patients respond to a four-dose treatment program J Clin Oncol 1998 16: 2825–2833

    Article  CAS  PubMed  Google Scholar 

  3. Davis TA, White CA, Grillo-López A, Velasquez WS, Link B, Maloney DG, Dillman RO, Williams ME, Mohrbacher A, Weaver R, Dowden S, Levy R . Single-agent monoclonal antibody efficacy in bulky non-Hodgkin's lymphoma: results of a phase II trial of rituximab J Clin Oncol 1999 17: 1851–1857

    Article  CAS  PubMed  Google Scholar 

  4. Reff ME, Carner K, Chambers KS, Chinn PC, Leonard JE, Raab R, Newman RA, Hanna N, Anderson DR . Depletion of B cells in vivo by a chimeric mouse human monoclonal antibody to CD20 Blood 1994 83: 435–445

    CAS  PubMed  Google Scholar 

  5. Tedder TF, Engel P . CD20: a regulator of cell-cycle progression of B lymphocytes Immunol Today 1994 15: 450–454

    Article  CAS  PubMed  Google Scholar 

  6. Shan D, Ledbetter JA, Press OW . Signaling events involved in anti-CD20-induced apoptosis of malignant human B cells Cancer Immunol Immunother 2000 48: 673–683

    Article  CAS  PubMed  Google Scholar 

  7. Hofmeister JK, Cooney D, Coggeshall KM . Clustered CD20 induced apoptosis: Src-family kinase, the proximal regulator of tyrosine phosphorylation calcium influx and caspase 3-dependent apoptosis Blood Cells Mol Dis 2000 26: 133–143

    Article  CAS  PubMed  Google Scholar 

  8. Golay J, Zaffaroni L, Vaccari T, Lazzari M, Borleri GM, Bernasconi S, Tedesco F, Rambaldi A, Introna M . Biologic response of B lymphoma cells to anti-CD20 monoclonal antibody rituximab in vitro: CD55 and CD59 regulate complement-mediated cell lysis Blood 2000 95: 3900–3908

    CAS  PubMed  Google Scholar 

  9. Harjunpaa A, Junnikkala S, Meri S . Rituximab (anti-CD20) therapy of B cell lymphomas: direct complement killing is superior to cellular effector mechanisms Scand J Immunol 2000 51: 634–641

    Article  CAS  PubMed  Google Scholar 

  10. Wurflein D, Dechant M, Stockmeyer B, Tutt AL, Hu P, Repp R, Kalden JR, van de Winkel JG, Epstein AL, Valerius T, Glennie M, Gramatzki M . Evaluating antibodies for their capacity to induce cell-mediated lysis of malignant B cells Cancer Res 1998 58: 3051–3058

    CAS  PubMed  Google Scholar 

  11. Stockmeyer B, Valerius T, Repp R, Heijnen IA, Buhring HJ, Deo YM, Kalden JR, Gramatzki M, van de Winkel JG . Preclinical studies with Fc(gamma)R bispecific antibodies and granulocyte colony-stimulating factor-primed neutrophils as effector cells against HER-2/neu overexpressing breast cancer Cancer Res 1997 57: 696–701

    CAS  PubMed  Google Scholar 

  12. Elsasser D, Valerius T, Repp R, Weiner GJ, Deo Y, Kalden JR, van de Winkel JG, Stevenson GT, Glennie MJ, Gramatzki M . HLA class II as potential target antigen on malignant B cells for therapy with bispecific antibodies in combination with granulocyte colony-stimulating factor Blood 1996 87: 3803–3812

    CAS  PubMed  Google Scholar 

  13. Valerius T, Repp R, de Wit T, Berthold S, Platzer E, Kalden JR, Gramatzki M, van de Winkel J . Involvement of the high-affinity receptor for IgG (Fc gamma RI; CD64) in enhanced tumor cell cytotoxicity of neutrophils during granulocyte colony-stimulating factor therapy Blood 1993 82: 931–939

    CAS  PubMed  Google Scholar 

  14. Ottonello L, Morone P, Dapino P, Dallegri F . Monoclonal Lym-1 antibody-dependent lysis of B-lymphoblastoid tumor targets by human complement and cytokinine-exposed mononuclear and neutrophilic polymorphonuclear leukocytes Blood 1996 87: 5171–5178

    CAS  PubMed  Google Scholar 

  15. Cemerlic D, Dadey B, Han T, Vaickus L . Cytokine influence on killing of fresh chronic lymphocytic leukemia cells by human leukocytes Blood 1991 77: 2707–2715

    CAS  PubMed  Google Scholar 

  16. Kerst J, van de Winkel J, Evans A, de Haas M, Slaper-Cortenbach I, de Wit T, von dem Borne A, van der Schoot C, van Oers R . Granulocyte colony-stimulating factor induces hFc gamma RI (CD64 antigen)-positive neutrophils via an effect on myeloid precursor cells Blood 1993 81: 1457–1464

    CAS  PubMed  Google Scholar 

  17. Kerst J, de Haas M, van der Schoot C, Slaper-Cortenbach I, Kleijer M, von dem Borne A, van Oers R . Recombinant granulocyte colony-stimulating factor administration to healthy volunteers: induction of immunophenotypically and functionally altered neutrophils via an effect on myeloid progenitor cells Blood 1993 82: 3265–3272

    CAS  PubMed  Google Scholar 

  18. Hooijberg E, Sein JJ, van den Berk PC, Hekman A . Characterization of a series of isotype switch variants of a new CD20 monoclonal antibody Hybridoma 1996 15: 23–31

    Article  CAS  PubMed  Google Scholar 

  19. Parham P . On the fragmentation of monoclonal IgG1 IgG2a and IgG2b from BALB/c mice J Immunol 1983 131: 2895–2902

    CAS  PubMed  Google Scholar 

  20. Rosenfeld SI, Looney RJ, Leddy JP, Phipps DC, Abraham GN, Anderson CL . Human platelet Fc receptor for immunoglobulin G. Identification as a 40 000-molecular-weight membrane protein shared by monocytes J Clin Invest 1985 76: 2317–2322

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. van der Kolk LE, Grillo-López A, Gerritsen W, Baars JW, van Oers MHJ . Chimeric anti-CD20 monoclonal antibody (rituximab) plus G-CSF in relapsed B cell lymphoma: a phase I/II clinical trial Blood 1998 92: 4037a (Abstr.)

    Google Scholar 

  22. Verhoeven AJ, van SM Roos D, Weening RS . Detection of carriers of the autosomal form of chronic granulomatous disease Blood 1988 71: 505–507

    CAS  PubMed  Google Scholar 

  23. De Boer M, Reijneke R, Van de Griend RJ, Loos JA, Roos D . Large-scale purification and cryopreservation of human monocytes J Immunol Meth 1981 43: 225–239

    Article  CAS  Google Scholar 

  24. Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, van Oers M . Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis Blood 1994 84: 1415–1420

    CAS  PubMed  Google Scholar 

  25. Newell MK, Van der Wall J, Beard KS, Freed JH . Ligation of major histocompatibility complex class II molecules mediates apoptotic cell death in resting B lymphocytes Proc Natl Acad Sci USA 1993 90: 10459–10463

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. van de Winkel J, Capel PJ . Human IgG Fc receptor heterogeneity: molecular aspects and clinical implications Immunol Today 1993 14: 215–221

    Article  CAS  PubMed  Google Scholar 

  27. Clynes RA, Towers TL, Presta LG, Ravetch JV . Inhibitory Fc receptors modulate in vivo cytoxicity against tumor targets Nat Med 2000 6: 443–446

    Article  CAS  PubMed  Google Scholar 

  28. Cassel DL, Keller MA, Surrey S, Schwartz E, Schreiber AD, Rappaport EF, McKenzie SE . Differential expression of Fc gamma RIIA Fc gamma RIIB and Fc gamma RIIC in hematopoietic cells: analysis of transcripts Mol Immunol 1993 30: 451–460

    Article  CAS  PubMed  Google Scholar 

  29. Brooks DG, Qiu WQ, Luster AD, Ravetch JV . Structure and expression of human IgG FcRII(CD32). Functional heterogeneity is encoded by the alternatively spliced products of multiple genes J Exp Med 1989 170: 1369–1385

    Article  CAS  PubMed  Google Scholar 

  30. Hooijberg E, Sein JJ, van den Berk PC, Hart AA, van der Valk MA, Kast WM, Melief CJ, Hekman A . Eradication of large human B cell tumors in nude mice with unconjugated CD20 monoclonal antibodies and interleukin 2 Cancer Res 1995 55: 2627–2634

    CAS  PubMed  Google Scholar 

  31. Davis TA, Maloney DG, Grillo-Lopez AJ, White CA, Williams ME, Weiner GJ, Dowden S, Levy R . Combination immunotherapy of relapsed or refractory low-grade or follicular non-Hodgkin's lymphoma with rituximab and interferon-alpha-2a Clin Cancer Res 2000 6: 2644–2652

    CAS  PubMed  Google Scholar 

  32. Locker GJ, Kapiotis S, Veitl M, Mader RM, Stoiser B, Kofler J, Sieder AE, Rainer H, Steger GG, Mannhalter C, Wagner OF . Activation of endothelium by immunotherapy with interleukin-2 in patients with malignant disorders Br J Haematol 1999 105: 912–919

    Article  CAS  PubMed  Google Scholar 

  33. Margolin KA . Interleukin-2 in the treatment of renal cancer Semin Oncol 2000 27: 194–203

    CAS  PubMed  Google Scholar 

  34. Weiss K . Safety profile of interferon-alpha therapy Semin Oncol 1998 25: 9–13

    CAS  PubMed  Google Scholar 

  35. Trask PC, Esper P, Riba M, Redman B . Psychiatric side effects of interferon therapy: prevalence proposed mechanisms and future directions J Clin Oncol 2000 18: 2316–2326

    Article  CAS  PubMed  Google Scholar 

  36. Sacchi S, Federico M, Vitolo U, Boccomini C, Vallisa G, Baldini L, Petrini M, Rupoli S, Di Raimondo F, Merli F, Liso V, Tabilio A, Saglio G, Di Renzo N, Gobbi P, Pitini G, Quarta G, Brugiatelli M, Vinci G, Dastoli G . Phase II study of rituximab after priming with IFN in patients with relapsed LG/F B cell lymphoma Blood 1999 94: 399a (Abstr.)

    Google Scholar 

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Authors and Affiliations

  1. Department of Haematology, Academic Medical Center, Amsterdam, The Netherlands

    LE van der Kolk & MHJ van Oers

  2. CLB Sanquin Blood Supply Foundation, Amsterdam, The Netherlands

    M de Haas

  3. IDEC Pharmaceuticals, San Diego, CA, USA

    AJ Grillo-López

  4. Antoni van Leeuwenhoek Hospital/The Netherlands Cancer Institute, Amsterdam, The Netherlands

    JW Baars

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  1. LE van der Kolk
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  4. JW Baars
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  5. MHJ van Oers
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van der Kolk, L., de Haas, M., Grillo-López, A. et al. Analysis of CD20-dependent cellular cytotoxicity by G-CSF-stimulated neutrophils. Leukemia 16, 693–699 (2002). https://doi.org/10.1038/sj.leu.2402424

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