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. 1992 Nov;34(6):361–369. doi: 10.1007/BF01741745

Comparative efficiencies of bispecific F(ab′γ)2 and chimeric mouse/human IgG antibodies in recruiting cellular effectors for cytotoxicity via Fcγ receptors

John Greenman 1,2, Nancy Hogg 1,2, Suzanne Nikoletti 1,2, Christopher Slade 1,2, George Stevenson 1,2, Martin Glennie 1,2,✉
PMCID: PMC11038724  PMID: 1532922

Abstract

The three forms of Fcγ receptor carried by monocytes (FcγRI, II) and natural killer (NK) cells (FcγRIII) are all capable of mediating cell lysis. Here we compare the use of F(ab′γ)2 bispecific antibodies, specifically targetting individual FcγR, and chimeric IgG mouse/human antibodies which are capable of targetting all FcγR, for their ability to mediate target cell destruction. The derivatives are prepared by linking hinge sulphydryl residues via tandem thioether bonds, using a bismaleimide crosslinker: Fab′ from an anti-FcγR mAb linked to Fab′ from a common anti-target mAb (BsAb), or Fab′ from the common anti-target mouse antibody linked to human Fcγ (FabFc or bisFabFc). All the derivatives targetting chick red blood cells gave efficient lysis, although different effector cell donors yielded differences in both the lytic levels achieved and the comparative efficiencies of derivatives. In contrast, significant lysis of the guinea pig lymphoblastic leukaemia, L2C, regularly resulted only via the anti-FcγRIII BsAb and the chimeric derivatives. These results suggest that the chimeric, Fc-containing derivatives mediate tumour cell lysis principally through FcγRIII on NK cells. This is in contrast to the situation with the chick red blood cells where the chimeric derivatives appear capable of lysing erythrocytes by utilizing either monocytes or NK cells, because significant (≈50%) lysis occurred with effector cell populations magnetically depleted through either FcγRII or FcγRIII. A major difference between these two types of antibody derivative was their ability to function in the presence of high concentrations of normal human Fcγ. The lysis mediated by BsAb reactive with FcγRI or II was unaffected by the presence of human Fcγ at 2.5 mg/ml (a concentration comparable with that yielded by IgG in plasma) whereas the BsAb recognizing FcγRIII and all the Fc-containing derivatives were completely inhibited.

Key words: Antibody derivatives, FcγR, Cellular cytotoxicity

Footnotes

This work has been supported by Tenovus, the Cancer Research Campaign, the Leukaemia Research Fund, Italfarmaco, Milano, Italy and the Imperial Cancer Research Fund

References

  • 1.Anderson CL. Structural and functional polymorphism of human Fc receptors for IgG. Chem Immunol. 1989;47:1. doi: 10.1159/000318901. [DOI] [PubMed] [Google Scholar]
  • 2.Anderson CL, Guyre PM, Whitin JC, Ryan DH, Looney RJ, Fanger MW. Monoclonal antibodies to Fc receptors for IgG on human mononuclear phagocytes. J Biol Chem. 1986;261:12 856. [PubMed] [Google Scholar]
  • 3.Buckle AM, Hogg N. The effect of IFN-gamma and colony stimulating factors on the expression of neutrophil cell membrane receptors. J Immunol. 1989;143:2295. [PubMed] [Google Scholar]
  • 4.Cohen L, Sharp S, Kulczycki A., Jr Human monocytes, B lymphocytes and non-B lymphocytes each have structurally unique Fc receptors. J Immunol. 1983;131:378. [PubMed] [Google Scholar]
  • 5.Dearman RJ, Stevenson FK, Wrightham M, Hamblin TJ, Glennie MJ, Stevenson GT. Lymphokine-activated killer cells from normal and lymphoma subjects are cytotoxic for cells coated with antibody derivatives displaying human Fcγ. Blood. 1988;72:1985. [PubMed] [Google Scholar]
  • 6.Dougherty GJ, Selvendran Y, Murdoch S, Palmer DG, Hogg N. The human mononuclear phagocyte high-affinity Fc receptor, FcRI, defined by a monoclonal antibody. 10.1. Eur J Immunol. 1987;17:1453. doi: 10.1002/eji.1830171011. [DOI] [PubMed] [Google Scholar]
  • 7.Elliott TJ, Glennie MJ, McBride HM, Stevenson GT. Analysis of the interaction of antibodies with immunoglobulin idiotypes on neoplastic B lymphocytes: implications for immunotherapy. J Immunol. 1987;138:981. [PubMed] [Google Scholar]
  • 8.Fanger MW, Shen L, Graziano RF, Guyre PM. Cytotoxicity mediated by human Fc receptors for IgG. Immunol Today. 1989;10:92. doi: 10.1016/0167-5699(89)90234-X. [DOI] [PubMed] [Google Scholar]
  • 9.Fleit HB, Wright SD, Unkeless JC. Human neutrophil Fc gamma receptor distribution and structure. Proc Natl Acad Sci USA. 1982;79:3275. doi: 10.1073/pnas.79.10.3275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Glennie MJ, McBride HM, Worth AT, Stevenson GT. Preparation and performance of bispecific F(ab′γ)2 antibody containing thioether-linked Fab′γ fragments. J Immunol. 1987;139:2367. [PubMed] [Google Scholar]
  • 11.Graziano RF, Fanger MW. FcγRI and FcγRII on monocytes and granulocytes are cytotoxic trigger molecules for tumour cells. J Immunol. 1987;139:3536. [PubMed] [Google Scholar]
  • 12.Greenman J, Tutt AL, George AJT, Pulford KAF, Stevenson GT, Glennie MJ. Characterization of a new monoclonal anti-FcγRII antibody, AT10, and its incorporation into a bispecific F(ab′)2 derivative for recruitment of cytotoxic effectors. Mol Immunol. 1991;28:1243. doi: 10.1016/0161-5890(91)90011-8. [DOI] [PubMed] [Google Scholar]
  • 13.Grimm EA, Wilson DJ. The human lymphokine-activated killer system. Cell Immunol. 1985;94:568. doi: 10.1016/0008-8749(85)90280-1. [DOI] [PubMed] [Google Scholar]
  • 14.Guyre PM, Graziano RF, Vance BA, Morganelli PM, Fanger MW. Monoclonal antibodies that bind to distinct epitopes on FcγRI are able to trigger receptor function. J Immunol. 1989;143:1650. [PubMed] [Google Scholar]
  • 15.Hogg N, Horton MA, et al. Myeloid antigens: new and previously defined clusters. In: McMichael AJ, et al., editors. Leucocyte typing: III. White cell differentiation antigens. Oxford: Oxford University Press; 1987. p. 576. [Google Scholar]
  • 16.Holborow EJ, Johnson GD. Immunofluorescence. In: Weir DM, editor. Handbook of experimental immunology. Oxford: Blackwell Scientific; 1967. p. 580. [Google Scholar]
  • 17.Huizinga TWJ, van Kemande F, Koenderman L, Dolman KM, von dem Borne AEGKr, Tetteroo PAT, Roos D. The 40-kDa Fcγ receptor (FcRII) on human neutrophils is essential for the IgG-induced respiratory burst and IgG-induced phagocytosis. J Immunol. 1989;142:2365. [PubMed] [Google Scholar]
  • 18.Jones JF, Titus JA, Segal D. Antibody-dependent cell-mediated cytolysis (ADCC) with antibody-coated effectors: rat and human effectors versus tumour targets. J Immunol. 1981;126:2457. [PubMed] [Google Scholar]
  • 19.Karpovsky B, Titus JA, Stephany DA, Segal DM. Production of target-specific effector cells using hetero-cross-linked aggregates containing anti-target cell and anti-Fcγ receptor antibodies. J Exp Med. 1984;160:1686. doi: 10.1084/jem.160.6.1686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Klaus GGB, Bijsterbosch MK, O'Garra A, Harnett MM, Rigley KP. Receptor signalling and crosstalk in B lymphocytes. Immunol Rev. 1987;99:19. doi: 10.1111/j.1600-065x.1987.tb01170.x. [DOI] [PubMed] [Google Scholar]
  • 21.Lamoyi E, Nisonoff A. Preparation of F(ab′γ)2 fragments from mouse IgG of various subclasses. J Immunol Methods. 1983;56:235. doi: 10.1016/0022-1759(83)90415-5. [DOI] [PubMed] [Google Scholar]
  • 22.Lanier LL, Kipps TJ, Phillips JH. Functional properties of a unique set of cytotoxic CD3+ T lymphocytes that express Fc receptors for IgG (CD16/Leu-11 antigen) J Exp Med. 1985;162:2089. doi: 10.1084/jem.162.6.2089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Lanier LL, Ruitenberg JJ, Phillips JH. Functional and biochemical analysis of CD16 antigen on natural killer cells and granulocytes. J Immunol. 1988;141:3478. [PubMed] [Google Scholar]
  • 24.Lovchik JC, Hong R. Antibody-dependent cell-mediated cytolysis (ADCC): analyses and projections. Prog Allergy. 1977;22:1. [PubMed] [Google Scholar]
  • 25.Pulford K, Rafkiaer E, MacDonald SM, Erber WN, Falini B, Gatter KC, Mason DY. A new monoclonal antibody (KB61) recognising a novel antigen which is selectively expressed on a subpopulation of human B lymphocytes. Immunology. 1986;57:71. [PMC free article] [PubMed] [Google Scholar]
  • 26.Rosenberg SA, Lotze MT, Muul LM, Chang AE, Avis FP, Leitman S, Lineham WM, Robertson CN, Lee RE, Rubin JT, Seipp CA, Simpson CG, White DE. A progress report on the treatment of 157 patients with advanced cancer using lymphokine-activated killer cells and interleukin-2 or high dose interleukin-2 alone. N Engl J Med. 1987;316:889. doi: 10.1056/NEJM198704093161501. [DOI] [PubMed] [Google Scholar]
  • 27.Ruegg J, Jungi TW. Antibody-mediated erythrolysis and erythrophagocytosis by human monocytes, macrophages and activated macrophages. Evidence for distinction between involvement of high-affinity and low-affinity receptors for IgG by using different erythroid target cells. Immunology. 1988;63:513. [PMC free article] [PubMed] [Google Scholar]
  • 28.Simmons D, Seed B. The Fcγ receptor of natural killer cells is a phospholipid-linked membrane protein. Nature. 1988;333:568. doi: 10.1038/333568a0. [DOI] [PubMed] [Google Scholar]
  • 29.Stevenson GT, Pindar A, Slade CJ. A chimeric antibody with dual Fc regions (bisFabFc) prepared by manipulations at the IgG hinge. Anti-Cancer Drug Design. 1989;3:219. [PubMed] [Google Scholar]
  • 30.Tutt A, Greenman J, Stevenson GT, Glennie MJ. Bispecific F(ab′γ)3 antibody derivatives for redirecting unprimed cytotoxic T cells. Eur J Immunol. 1991;21:1351. doi: 10.1002/eji.1830210604. [DOI] [PubMed] [Google Scholar]
  • 31.Unkeless JC. Function and heterogeneity of human Fc receptors for immunoglobulin G. J Clin Invest. 1989;83:355. doi: 10.1172/JCI113891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Unkeless JC, Scigliano E, Freedman VH. Structure and function of human and murine receptors for IgG. Annu Rev Immunol. 1988;6:251. doi: 10.1146/annurev.iy.06.040188.001343. [DOI] [PubMed] [Google Scholar]
  • 33.Vadas MA, David JR, Butterworth A, Pisani NT, Siongok TA. A new method for the purification of human eosinophils and neutrophils, and a comparison of the ability of these cells to damage schistosomula ofSchistosoma mansoni . J Immunol. 1979;122:1228. [PubMed] [Google Scholar]
  • 34.Van de Winkel JGJ, Boonen GJJC, Janssen PLW, Vlug A, Hogg N, Tax WJM. Activity of two types of Fc receptors, FcγRI and FcγRII, in human monocyte cytotoxicity to sensitized erythrocytes. Scand J Immunol. 1989;29:23. doi: 10.1111/j.1365-3083.1989.tb01095.x. [DOI] [PubMed] [Google Scholar]

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