© 1990 S Karger AG, Basel 0302-2838/90/0186-0010$2 75/0
Eur Urol 1990; 18(suppl 2): 10-12
Monoclonal Antibodies to Renal Cancer Antigens
1808098
Neil H. Bander Laboratory of Human Cancer Immunology, Memorial Sloan-Kettering Cancer Center, and The James Buchanan Brady Foundation, Department of Surgery (Urology). The New York Hospital-Cornell Medical Center, New York, N.Y., USA
PTc
PT, LH„ LH„, DT
CT
T138 OKIal ■ J143 (UR0-1) _ C5H —« 050 S4 (UR0-2) — AJ8/J5 wmm OU HL60-4 ■■■ F23 (UR0-3) HÌTB241 OU HL60-3 anti-SSEA-1 1 A6H S27 (URO-4) T43 (URO-10) F31 (UR0-8) 10.32 (Tamm-Horslall) EMA C26 T16 BA-1 — BA-2 M2/S8
Fig. 1. Antigenic map of the human nephron. The bars indicate the site of binding of each mAb as determined by immunohistochemistry on frozen kidney sections. G = Glomerulus; PTC = proxi mal tubule, convoluted portion; PTS = proximal tubule, straight por tion; LHti = loop of Henle, descending limb; LHati = loop of Henle, ascending limb; DT = distal tubule; CT = collecting tubule.
exclusive; there are no cells which express both antigens. However, the study of developing fetal kidneys demon strated that proximal tubular progenitor cells express both URO-8 and URO-10. As these cells mature and assume their position along the nephron, the ‘adult’ phe notype supercedes.
Downloaded by: University of Exeter 144.173.6.94 - 5/5/2020 11:43:25 AM
A large number of monoclonal antibodies (mAbs) to kidney-associated antigens have been characterized in several laboratories (table 1). Few of these antigens had been defined prior to the mAb era. The normal site of expression of these antigens has been determined by standard immunofluorescence or immunoperoxidase as says on normal adult kidney sections and an ‘antigenic map’ of the kidney can be constructed (fig. 1). Determin ing the presence or absence of these antigens on a given cell defines the ‘antigenic phenotype' of that cell. For instance, cells of the straight portion of the proximal tubule are URO-1/URO-2+/URO-3+/URO-4+/URO-5-/ URO-8+/URO-10~ and can be distinguished from cells of the convoluted portion of the proximal tubule which are URO-1 -/ URO-2+/ URO-3U URO- 4+/ URO-5 / URO-8-/ URO-10+ [1-6]. Likewise, cells of the distal tubule are URO-1 -/ URO-2-/ URO- 3-/ URO- 4T URO- 5+/ URO-8U URO-10-, The antigenic phenotypes of normal adult kidney cells serve as a key reference point for a wide variety of studies such as normal and abnormal fetal development, defining the varying cell types growing in tissue culture of normal and neoplastic kidney or in the study of renal diseases such as renal cancer. Our laboratory has phenotyped a large number of kid ney tumor specimens using these mAbs on frozen sec tions. We found that the antigenic phenotypes are most consistent with their derivation from proximal tubular cells [2], consistent with earlier immunological [7] and ultrastructural [8, 9] studies. We have, however, been able to further refine the derivation. In the normal adult kidney, cells of the convoluted portion are URO-10+/ URO-8-, those of the straight portion are URO-10-/ URO-8L That is, typically (in the adult), the expression of URO-8 and URO-10 are reciprocal and mutually
Monoclonal Antibodies to Renal Cancer Antigens
11
In a study of renal cancers, 50 unselected specimens of primary renal cell carcinoma were phenotyped with mAbs [2], Nine cases were URO-8UURO-10“, 15 cases were URO-8~/URO-10+, 25 cases co-expressed both an tigens and only 1 case failed to express either antigen. The expression of these phenotypes by renal cancers is consistent with their respective derivations from the proximal convoluted tubule [URO-8~/URO-10+ (30% of cases)], the proximal straight tubule [URO-8VURO10~ (18% of cases)] or proximal tubule progenitor cells [URO-8+/URO-10+ (50% of cases)]. Only 1 of the 50 cases failed to express both the URO-8 and the URO10 antigens. This renal cancer did, however, express
other proximal tubular antigens (URO-2, URO-3 and URO-4). It is also interesting and important to point out that many renal cancers do not express a full complement of proximal tubular antigens. It appears that some antigens may be lost as part of the transformation process. This variability in antigen expression generates a number of different molecular subtypes of renal cancer. The clinical relevance of these subtypes is under study. We have found that the expressions of the URO-2, URO-3, URO-4 and URO-7 antigens are independent and incoordinate [1]. This, therefore, generates a signifi cant number of molecular subtypes of renal cancer. The
Table 1. mAbs to kidney-associated antigens mAb (immuno globulin class)
Defined antigen
Site of expression
Laboratory
T138 (IgM) J143/URO-1 (IgGl) AJ8 (IgGl) S4/URO-2 (IgG2a) T43/URO-10 (IgGl) F23/URO-3 (IgG2a) S23/URO-4a (IgGl) S27/URO-4 (IgGl) F31/URO-8 (IgM) C26 (IgG2a) M2 (IgM) S8 (IgM) Tl6 (IgG2b) S22 (IgGl) 10.32 (IgGl) OKIal (IgG2) Anti-SSEA-1 (IgM) A6H (IgGl) D5D (IgGl) C5H (IgGl) Anti-Bl (IgG2a) BA-1 (IgM) BA-2 (IgG3) E6, B7, C8, D8 DU ALL-1 DU HL60-4 DU HL60-3 (IgM) G250 (IgGl)
gp25 gpl40, 120, 30 gplOO (CALLA) gpl60 gp85 gpl40 gpl20 (ADAbp) gp!20 (ADAbp) glycolipid gp40 A blood group B blood group gp48, 42 gp 115 gp90 (Tamm-Horsfall) HLA-DR glycolipid ND ND pi 15 ND ND p24 ND p24 pl60 pl 5 ND
vascular endothelium glomerulus, urothelium glomerulus, proximal tubule glomerulus, proximal tubule proximal tubule (pars convoluta only) proximal tubule proximal tubule, loop of Henle proximal tubule, loop of Henle proximal tubule (pars recta only), loop of Henle distal tubule, collecting tubule, urothelium collecting tubule, urothelium collecting tubule, urothelium distal tubule, collecting tubule, urothelium renal cancer only loop of Henle, distal tubule endothelium, mesangium proximal tubule proximal tubule Bowman’s capsule glomerulus fetal ureteral bud Bowman’s capsule, distal tubule, collecting tubule Bowman’s capsule, distal tubule, collecting tubule renal cancer only glomerulus (mesangial cells), distal tubule proximal tubule, Bowman’s capsule proximal tubule (pars recta + convoluta) renal cancer only
MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC Cedarlane Labs Ortho Pharmaceuticals Wistar Institute University of Minnesota University of Minnesota University of Minnesota Dana-Farber University of Minnesota University of Minnesota Mainz, FRG Duke University Duke University Duke University University of Leiden, the Netherlands
Downloaded by: University of Exeter 144.173.6.94 - 5/5/2020 11:43:25 AM
gp = Glycoprotein (numbers are molecular weight in kilodaltons); MSKCC = Memorial Sloan-Kettering Cancer Center; CALLA = common acute lymphoblastic leukemia antigen; ADAbp = adenosine-deaminase-binding protein; ND = not defined.
Bander
12
References 1
Bander, N.H.: Renal cancer: a model system for the study of human neoplasia; in Riethmüller, von Kleist. Koprowski, Munk, Genes and antigenes in cancer cells: the monoclonal antibody approach. Contr. Oncol., voi. 19 (Karger, Basel 1984). 2 Bander, N.H.; Finstad, C.; Cordon-Cardo, C; Whitmore, W.F.; Vaughan, E.D.. Jr.; Oettgen, H.F.; Melamed, M.R.; Old, L.J.: Analysis of a mouse monoclonal antibody which reacts with a specific region of the human proximal tubule and subsets of renal cell carcinomas. Cancer Res. 49■ 6774-6780 (1989). 3 Ueda, R.; Ogata, S.I.; Morrissey, D.: Finstad, C.L.; Szkudlarek, J.; Whitmore, W.F.; Oettgen, H.F.; Lloyd, K.O.; Old, L.J.: Cell surface antigens of human renal cancer defined by mouse mono clonal antibodies: identification of tissue specific kidney glyco proteins. Proc. natn. Acad. Sci. USA 78. 5122-5126 (1981). 4 Finstad. C.L.; Cordon-Cardo, C.; Bander, N.H.; Whitmore, W.F.; Oettgen, H.F.; Melamed, M.R.; Old, L.J.: Specificity anal ysis of mouse monoclonal antibodies detecting cell surface anti gens of human renal cancer. Proc. natn. Acad. Sci. USA 82. 2955-2959 (1985). 5 Cordon-Cardo, C.; Bander, N.H.; Fradet, Y.; Finstad. C.; Whit more, W.F.; Oettgen, H.F.; Melamed, M.R.; Old, L.J.: Immunoanatomic dissection of the human urinary system with mouse monoclonal antibodies. J. Flistochem. Cytochem. 32: 1035 (1984). 6 Bander, N.H.; Cordon-Cardo, C.; Finstad, C.; Whitmore, W.F.; Vaughan, E.D., Jr.; Oettgen, H.F.; Melamed, M.R.; Old, L.J.: Immunohistologic dissection of the human kidney using mono clonal antibodies. J. Urol. 133: 502-505 (1985). 7 Wallace, A.C.; Nairn, R.C.: Renal tubular antigens in kidney tumors. Cancer 29. 977 (1972). 8 Seljelid. R.; Ericsson, L.E.: Electron microscopic observations of the cell surface in renal clear cell carcinoma. Lab. Invest. 14■ 435 (1965). 9 Tannenbaum, M.: Ultrastructural pathology of human renal cell tumors. Pathol. A. 6: 249 (1971). 10 Nanus, D.M.; Pfeffer, L.M.; Bander, N.H.; Bahn, S.; Albino. A.P.: Anti-proliferative and anti-tumor effect of alpha-interferon in renal carcinoma: Correlation to the expression of a kidney associated differentiation glycoprotein. Cancer Res. (in press).
Neil H. Bander. MD The James Buchanan Brady Foundation Department of Surgery (Urology) The New York Hospital-Cornell Medical Center 525 East 68th Street New York, NY 10021 (USA)
Downloaded by: University of Exeter 144.173.6.94 - 5/5/2020 11:43:25 AM
issue is whether these molecular subtypes have any prac tical clinical relevance. There clearly appears to be a direct correlation between the degree of differentiation and the number of proximal tubular antigens expressed. Perhaps immunopathologie typing will provide an objec tive means of grading renal cancers. Preliminary evi dence in our laboratory also indicates a correlation between antigen expression and prognosis. This observa tion needs to be extended to a large number of patients with adequate follow-up to determine if this apparent correlation is independent of stage and/or grade. Finally, it is possible that the definition of molecular subtypes will allow an enhanced ability to select treat ment for individual patients. For instance, it is known that about 15-20% of patients with renal cancer respond to interferon-a (IFN-a) or that 17-30% respond to interleukin-2-based protocols. It would certainly represent a significant advance to be able to predict, prior to treat ment, who would respond and who would not. At this time we have evidence using cell growth in vitro as well as renal cancer xenograft growth in nude mice that we can predict IFN-a sensitivity [10]. On the basis of these findings a prospective clinical treatment pilot study has been submitted for approval. Current clinical trials of mouse mAbs are at a very early stage. These studies will define the inherent ability of an mAb to reach and bind its respective antigen, the inherent cytotoxicity of the antibody alone and the tox icity of the antibody. These data will provide the ground work on which to build more effective treatments includ ing antibody combinations, antibody-cytotoxic conju gates, and combinations of antibody with chemotherapy and/or biological response modifiers. In conclusion, mAb technology, little more than 10 years old, has already had a profound impact on the study of basic cell biology. The technology has now reached the stage of clinical application where it may be anticipated to have a similar impact in the area of diag nostic testing. On the horizon, mAb technology, proba bly in conjunction with cellular immune techniques and recombinant DNA methods, may well allow us to reach the goal of effectively treating cancer.
Eur Urol 1990; 18(suppl 2): 10-12
Monoclonal Antibodies to Renal Cancer Antigens
1808098
Neil H. Bander Laboratory of Human Cancer Immunology, Memorial Sloan-Kettering Cancer Center, and The James Buchanan Brady Foundation, Department of Surgery (Urology). The New York Hospital-Cornell Medical Center, New York, N.Y., USA
PTc
PT, LH„ LH„, DT
CT
T138 OKIal ■ J143 (UR0-1) _ C5H —« 050 S4 (UR0-2) — AJ8/J5 wmm OU HL60-4 ■■■ F23 (UR0-3) HÌTB241 OU HL60-3 anti-SSEA-1 1 A6H S27 (URO-4) T43 (URO-10) F31 (UR0-8) 10.32 (Tamm-Horslall) EMA C26 T16 BA-1 — BA-2 M2/S8
Fig. 1. Antigenic map of the human nephron. The bars indicate the site of binding of each mAb as determined by immunohistochemistry on frozen kidney sections. G = Glomerulus; PTC = proxi mal tubule, convoluted portion; PTS = proximal tubule, straight por tion; LHti = loop of Henle, descending limb; LHati = loop of Henle, ascending limb; DT = distal tubule; CT = collecting tubule.
exclusive; there are no cells which express both antigens. However, the study of developing fetal kidneys demon strated that proximal tubular progenitor cells express both URO-8 and URO-10. As these cells mature and assume their position along the nephron, the ‘adult’ phe notype supercedes.
Downloaded by: University of Exeter 144.173.6.94 - 5/5/2020 11:43:25 AM
A large number of monoclonal antibodies (mAbs) to kidney-associated antigens have been characterized in several laboratories (table 1). Few of these antigens had been defined prior to the mAb era. The normal site of expression of these antigens has been determined by standard immunofluorescence or immunoperoxidase as says on normal adult kidney sections and an ‘antigenic map’ of the kidney can be constructed (fig. 1). Determin ing the presence or absence of these antigens on a given cell defines the ‘antigenic phenotype' of that cell. For instance, cells of the straight portion of the proximal tubule are URO-1/URO-2+/URO-3+/URO-4+/URO-5-/ URO-8+/URO-10~ and can be distinguished from cells of the convoluted portion of the proximal tubule which are URO-1 -/ URO-2+/ URO-3U URO- 4+/ URO-5 / URO-8-/ URO-10+ [1-6]. Likewise, cells of the distal tubule are URO-1 -/ URO-2-/ URO- 3-/ URO- 4T URO- 5+/ URO-8U URO-10-, The antigenic phenotypes of normal adult kidney cells serve as a key reference point for a wide variety of studies such as normal and abnormal fetal development, defining the varying cell types growing in tissue culture of normal and neoplastic kidney or in the study of renal diseases such as renal cancer. Our laboratory has phenotyped a large number of kid ney tumor specimens using these mAbs on frozen sec tions. We found that the antigenic phenotypes are most consistent with their derivation from proximal tubular cells [2], consistent with earlier immunological [7] and ultrastructural [8, 9] studies. We have, however, been able to further refine the derivation. In the normal adult kidney, cells of the convoluted portion are URO-10+/ URO-8-, those of the straight portion are URO-10-/ URO-8L That is, typically (in the adult), the expression of URO-8 and URO-10 are reciprocal and mutually
Monoclonal Antibodies to Renal Cancer Antigens
11
In a study of renal cancers, 50 unselected specimens of primary renal cell carcinoma were phenotyped with mAbs [2], Nine cases were URO-8UURO-10“, 15 cases were URO-8~/URO-10+, 25 cases co-expressed both an tigens and only 1 case failed to express either antigen. The expression of these phenotypes by renal cancers is consistent with their respective derivations from the proximal convoluted tubule [URO-8~/URO-10+ (30% of cases)], the proximal straight tubule [URO-8VURO10~ (18% of cases)] or proximal tubule progenitor cells [URO-8+/URO-10+ (50% of cases)]. Only 1 of the 50 cases failed to express both the URO-8 and the URO10 antigens. This renal cancer did, however, express
other proximal tubular antigens (URO-2, URO-3 and URO-4). It is also interesting and important to point out that many renal cancers do not express a full complement of proximal tubular antigens. It appears that some antigens may be lost as part of the transformation process. This variability in antigen expression generates a number of different molecular subtypes of renal cancer. The clinical relevance of these subtypes is under study. We have found that the expressions of the URO-2, URO-3, URO-4 and URO-7 antigens are independent and incoordinate [1]. This, therefore, generates a signifi cant number of molecular subtypes of renal cancer. The
Table 1. mAbs to kidney-associated antigens mAb (immuno globulin class)
Defined antigen
Site of expression
Laboratory
T138 (IgM) J143/URO-1 (IgGl) AJ8 (IgGl) S4/URO-2 (IgG2a) T43/URO-10 (IgGl) F23/URO-3 (IgG2a) S23/URO-4a (IgGl) S27/URO-4 (IgGl) F31/URO-8 (IgM) C26 (IgG2a) M2 (IgM) S8 (IgM) Tl6 (IgG2b) S22 (IgGl) 10.32 (IgGl) OKIal (IgG2) Anti-SSEA-1 (IgM) A6H (IgGl) D5D (IgGl) C5H (IgGl) Anti-Bl (IgG2a) BA-1 (IgM) BA-2 (IgG3) E6, B7, C8, D8 DU ALL-1 DU HL60-4 DU HL60-3 (IgM) G250 (IgGl)
gp25 gpl40, 120, 30 gplOO (CALLA) gpl60 gp85 gpl40 gpl20 (ADAbp) gp!20 (ADAbp) glycolipid gp40 A blood group B blood group gp48, 42 gp 115 gp90 (Tamm-Horsfall) HLA-DR glycolipid ND ND pi 15 ND ND p24 ND p24 pl60 pl 5 ND
vascular endothelium glomerulus, urothelium glomerulus, proximal tubule glomerulus, proximal tubule proximal tubule (pars convoluta only) proximal tubule proximal tubule, loop of Henle proximal tubule, loop of Henle proximal tubule (pars recta only), loop of Henle distal tubule, collecting tubule, urothelium collecting tubule, urothelium collecting tubule, urothelium distal tubule, collecting tubule, urothelium renal cancer only loop of Henle, distal tubule endothelium, mesangium proximal tubule proximal tubule Bowman’s capsule glomerulus fetal ureteral bud Bowman’s capsule, distal tubule, collecting tubule Bowman’s capsule, distal tubule, collecting tubule renal cancer only glomerulus (mesangial cells), distal tubule proximal tubule, Bowman’s capsule proximal tubule (pars recta + convoluta) renal cancer only
MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC MSKCC Cedarlane Labs Ortho Pharmaceuticals Wistar Institute University of Minnesota University of Minnesota University of Minnesota Dana-Farber University of Minnesota University of Minnesota Mainz, FRG Duke University Duke University Duke University University of Leiden, the Netherlands
Downloaded by: University of Exeter 144.173.6.94 - 5/5/2020 11:43:25 AM
gp = Glycoprotein (numbers are molecular weight in kilodaltons); MSKCC = Memorial Sloan-Kettering Cancer Center; CALLA = common acute lymphoblastic leukemia antigen; ADAbp = adenosine-deaminase-binding protein; ND = not defined.
Bander
12
References 1
Bander, N.H.: Renal cancer: a model system for the study of human neoplasia; in Riethmüller, von Kleist. Koprowski, Munk, Genes and antigenes in cancer cells: the monoclonal antibody approach. Contr. Oncol., voi. 19 (Karger, Basel 1984). 2 Bander, N.H.; Finstad, C.; Cordon-Cardo, C; Whitmore, W.F.; Vaughan, E.D.. Jr.; Oettgen, H.F.; Melamed, M.R.; Old, L.J.: Analysis of a mouse monoclonal antibody which reacts with a specific region of the human proximal tubule and subsets of renal cell carcinomas. Cancer Res. 49■ 6774-6780 (1989). 3 Ueda, R.; Ogata, S.I.; Morrissey, D.: Finstad, C.L.; Szkudlarek, J.; Whitmore, W.F.; Oettgen, H.F.; Lloyd, K.O.; Old, L.J.: Cell surface antigens of human renal cancer defined by mouse mono clonal antibodies: identification of tissue specific kidney glyco proteins. Proc. natn. Acad. Sci. USA 78. 5122-5126 (1981). 4 Finstad. C.L.; Cordon-Cardo, C.; Bander, N.H.; Whitmore, W.F.; Oettgen, H.F.; Melamed, M.R.; Old, L.J.: Specificity anal ysis of mouse monoclonal antibodies detecting cell surface anti gens of human renal cancer. Proc. natn. Acad. Sci. USA 82. 2955-2959 (1985). 5 Cordon-Cardo, C.; Bander, N.H.; Fradet, Y.; Finstad. C.; Whit more, W.F.; Oettgen, H.F.; Melamed, M.R.; Old, L.J.: Immunoanatomic dissection of the human urinary system with mouse monoclonal antibodies. J. Flistochem. Cytochem. 32: 1035 (1984). 6 Bander, N.H.; Cordon-Cardo, C.; Finstad, C.; Whitmore, W.F.; Vaughan, E.D., Jr.; Oettgen, H.F.; Melamed, M.R.; Old, L.J.: Immunohistologic dissection of the human kidney using mono clonal antibodies. J. Urol. 133: 502-505 (1985). 7 Wallace, A.C.; Nairn, R.C.: Renal tubular antigens in kidney tumors. Cancer 29. 977 (1972). 8 Seljelid. R.; Ericsson, L.E.: Electron microscopic observations of the cell surface in renal clear cell carcinoma. Lab. Invest. 14■ 435 (1965). 9 Tannenbaum, M.: Ultrastructural pathology of human renal cell tumors. Pathol. A. 6: 249 (1971). 10 Nanus, D.M.; Pfeffer, L.M.; Bander, N.H.; Bahn, S.; Albino. A.P.: Anti-proliferative and anti-tumor effect of alpha-interferon in renal carcinoma: Correlation to the expression of a kidney associated differentiation glycoprotein. Cancer Res. (in press).
Neil H. Bander. MD The James Buchanan Brady Foundation Department of Surgery (Urology) The New York Hospital-Cornell Medical Center 525 East 68th Street New York, NY 10021 (USA)
Downloaded by: University of Exeter 144.173.6.94 - 5/5/2020 11:43:25 AM
issue is whether these molecular subtypes have any prac tical clinical relevance. There clearly appears to be a direct correlation between the degree of differentiation and the number of proximal tubular antigens expressed. Perhaps immunopathologie typing will provide an objec tive means of grading renal cancers. Preliminary evi dence in our laboratory also indicates a correlation between antigen expression and prognosis. This observa tion needs to be extended to a large number of patients with adequate follow-up to determine if this apparent correlation is independent of stage and/or grade. Finally, it is possible that the definition of molecular subtypes will allow an enhanced ability to select treat ment for individual patients. For instance, it is known that about 15-20% of patients with renal cancer respond to interferon-a (IFN-a) or that 17-30% respond to interleukin-2-based protocols. It would certainly represent a significant advance to be able to predict, prior to treat ment, who would respond and who would not. At this time we have evidence using cell growth in vitro as well as renal cancer xenograft growth in nude mice that we can predict IFN-a sensitivity [10]. On the basis of these findings a prospective clinical treatment pilot study has been submitted for approval. Current clinical trials of mouse mAbs are at a very early stage. These studies will define the inherent ability of an mAb to reach and bind its respective antigen, the inherent cytotoxicity of the antibody alone and the tox icity of the antibody. These data will provide the ground work on which to build more effective treatments includ ing antibody combinations, antibody-cytotoxic conju gates, and combinations of antibody with chemotherapy and/or biological response modifiers. In conclusion, mAb technology, little more than 10 years old, has already had a profound impact on the study of basic cell biology. The technology has now reached the stage of clinical application where it may be anticipated to have a similar impact in the area of diag nostic testing. On the horizon, mAb technology, proba bly in conjunction with cellular immune techniques and recombinant DNA methods, may well allow us to reach the goal of effectively treating cancer.
