Abstract
LL2 is a murine monoclonal antibody IgG2a reactive with B cells and non-Hodgkin's B-cell lymphoma, which, in a radioiodinated form, induces responses in lymphoma patients [Goldenberg et al. (1991) J Clin Oncol 9:548–564]. In this report we identify LL2 as a member of the CD22 cluster. The molecular size of the antigen, its expression profile, and competitive blocking studies were used to establish this identification. By Western blot analysis and immunoprecipitation studies using the Raji Burkitt's lymphoma cell line metabolically labelled with [3H]leucine, the LL2 antigen was determined to correspond to a molecular mass of 140 kDa. The molecular mass of the LL2 antigen, and the B-cell-restricted reactivity of the LL2 antibody, were consistent with both the CD21 and CD22 clusters. To assess additional similarities and differences between LL2 and anti-CD22 and anti-CD21, the binding of these mAb to cultured cell lines. Nalm-6 and Molt-4, was compared by flow cytometry. The binding profile of LL2 on these cell lines was consistent with anti-CD22, but not anti-CD21. Sequential immunoprecipitation and cross-blocking studies with anti-CD22 monoclonal antibodies recognizing established CD22 epitopes were performed to confirm that LL2 reacts with CD22 and to determine which epitope LL2 recognizes. Binding of131I-LL2 to Raji cells is inhibited over 90% by prior incubation of the target cells with unlabelled RFB4, indicating that LL2 belongs to the same epitope group as RFB4, i.e., epitope B.
Similar content being viewed by others
References
Aruffo A, Kanner SB, Sgroi D, Ledbetter JA, Stamenkovic I (1992) CD22-mediated stimulation of T cells regulates T-cell receptor/CD3-induced signaling. Proc Natl Acad Sci USA 89: 10242
Becker W, Lsuer U, Marienhagen J, Goldenberg DM, Wolf F (1992) Comparison of Tc-99m-antigranulocyte (NCA-95) Fab′-fragments and scintigraphy with In-111-oxin/Tc-99m-HMPAO-labelled leukocytes in infectious diseases. J Nucl Med 33: 903
Buchsbaum DJ, Wahl RL, Normolle DP, Kaminski MS (1992) Therapy with unlabeled and131I-labeled pan-B-cell monoclonal antibodies in nude mice bearing Raji Burkitt's lymphoma xenografts. Cancer Res 52:6476
DeNardo SJ, DeNardo GL, O'Grady LF, Hu E, Sytsma VM, Mills SL, Levy NB, Macey DJ, Miller CH, Epstein AL (1988) Treatment of B cell malignancies with131I Lym-1 monoclonal antibodies. Int J Cancer [Suppl] 3: 96
Dorken B, Moldenhauer G, Pezzutto A, Schwartz R, Feller A, Kiesel S, Nadler LM (1986) HD39 (B3), a B lineage-restricted antigen whose cell surface expression is limited to resting and activated human B lymphocytes. J Immunol 136: 4470
Dorken B, Moller P, Pezzutto A, Schwartz-Albiez R, Moldenhauer G (1989) B-cell antigens: CD22. In: Knapp W, Dorken B, Gilks WR, Rieber EP, Schmidt RE, Stein H, von dem Borne AEGKr (eds) Leukocyte typing, IV. Oxford University Press, Oxford, p 63
Ghetie M-A, May RD, Till M, Uhr JW, Ghetie V, Knowles PP, Relf M, Brown A, Wallace PM, Janossy G, Amlot P, Vitetta ES, Thorpe PE (1988) Evaluation of ricin A chain-containing immunotoxins directed against CD19 and CD22 antigens on normal and malignant human B-cells as potential reagents for in vivo therapy. Cancer Res 48: 2610
Ghetie M-A, Richardson J, Tucker T, Jones D, Uhr JW, Vitetta ES (1991) Antitumor activity of Fab' and IgG-anti-CD22 immunotoxins in disseminated human B lymphoma grown in mice with severe combined immunodeficiency disease: effect on tumor cells in extranodal sites. Cancer Res 51: 5876
Goldenberg DM (1989) Future role of radiolabeled monoclonal antibodies in oncological diagnosis and therapy. Semin Nucl Med 19: 332
Goldenberg DM, Horowitz JA, Sharkey RM, Hall TC, Murthy S, Goldenberg H, Lee RE, Stein R, Siegel JA, Izon DO, Burger K, Swayne L, Belisle E, Hansen HJ, Pinsky CM (1991) Targeting, dosimetry and radioimmunotherapy of B-cell lymphomas with iodine-131-labeled LL2 monoclonal antibody. J Clin Oncol 9: 548
Hurwitz R, Hozier J, LeBien T, Minowada J, Gajl-Peczalska K, Kubonishi I, Kersey J (1979) Characterization of a leukemic cell line of the pre-B phenotype. Int J Cancer 23: 174
Kaminski MS, Fig LM, Zasadny KR, Koral KF, DelRosario RB, Francis IR, Hanson CA, Normolle DP, Mudgett E, Liu CP, Moon S, Scott P, Miller RA, Wahl RL (1992) Imaging, dosimetry, and radioimmunotherapy with iodine 131-labeled anti-CD37 antibody in B-cell lymphoma. J Clin Oncol 10: 1696
Kaminski MS, Zasadny KR, Moon S, Roesner J, Milik AW, Butchko G, Glenn S, Wahl RL (1992) Initial clinical radioimmunotherapy results with 131-I-anti-B1 (anti-CD20) in refractory B-cell lymphoma. Antibody Immunoconjugate Radiopharmacol 5: 345
Kreitman RJ, Hansen HJ, Jones AL, FitzGerald DJP, Goldenberg DM, Pastan I (1993)Pseudomonas exotoxin-based immunotoxins containing the antibody LL2 or LL2-Fab' induce regression of subcutaneous human B-cell lymphoma in mice. Cancer Res 53: 819
Li J-L, Shen G-L, Ghetie M-A, May RD, Till M, Ghetie V, Uhr JW, Janossy G, Thorpe PE, Amlot P, Vitetta ES (1989) The epitope specificity and tissue reactivity of four murine monoclonal anti-CD22 antibodies. Cellular Immunol 118: 85
Lu HH, Shih LB, Xuan H, Goldenberg DM (1992) Internalization and cellular distribution of an anti-B-cell lymphoma antibody, LL2. Proc Am Assoc Cancer Res 33: 342
McConahey PJ, Dixon FJ (1966) A method of trace iodination of proteins for immunologic studies. Int Arch Allergy 29: 185
Minowada J, Ohnuma T, Moore GE (1972) Rosette-forming human lymphoid cell lines 1. Establishment and evidence for origin of thymus-derived lymphocytes. J Natl Cancer Inst 49: 891
Pawlak-Byczkowska EJ, Hansen HJ, Dion AS, Goldenberg DM (1989) Two new monoclonal antibodies, EPB-1 and EPB-2, reactive with human lymphoma. Cancer Res 49: 4568
Pezzutto A, Dorken B, Moldenhauer G, Clark EA (1987) Amplification of human B cell activation by a monoclonal antibody to the B cell-specific antigen CD22, Bp130/140. J Immunol 138: 98
Pezzutto A, Rabinovitch PS, Dorken B, Moldenhauer G, Clark EA (1988) Role of the CD22 human B cell antigen in B cell triggering by anti-immunoglobulin. J Immunol 140: 1791
Press OW, Eary JF, Badger CC, Martin PJ, Appelbaum FR, Levy R, Miller R, Brown S, Nelp WB, Krohn K, Fisher D, DeSantes K, Porter B, Kidd P, Thomas ED, Bernstein ID (1989) Treatment of refractory non-Hodgkin's lymphoma with radiolabeled MB-1 (anti-CD37) antibody. J Clin Oncol 7: 1027
Pulvertaft RJV (1964) Cytology of Burkitt's lymphoma. Lancet 1: 238
Scheinberg DA, Straus DJ, Yeh SD, Divgi C, Garin-Chesa P, Graham M, Pentlow K, Coit D, Oettgen HF, Old LJ (1990) A phase I toxicity, pharmacology, and dosimetry trial of monoclonal antibody OKB7 in patients with non-Hodgkin's lymphoma: effects of tumor burden and antigen expression. J Clin Oncol 8: 792
Schwartz-Albiez R, Dorken B, Moldenhauer G (1989) The carbohydrate moiety of the CD22 antigen can be modulated by inhibitors of the glycosylation pathway. In: Knapp W, Dorken B, Gilks WR, Rieber EP, Schmidt RE, Stein H, von dem Borne AEGKr (eds) Leukocyte typing. IV. Oxford University Press, Oxford, p 65
Stamenkovic I, Sgroi D, Aruffo A, Sy MS, Anderson T (1991) The B lymphocyte adhesion molecule CD22 interacts with leukocyte common antigen CD45RO on T cells and α2-6 sialyltransferase, CD75, on B cells. Cell 66: 1133
Stein R, Basu A, Chen S, Shih LB, Goldenberg DM (1993) Specificity and properties of MAb RS7-3G11 and the antigen defined by this pancarcinoma momoclonal antibody. Int J Cancer (in press)
Stein R, Chen S, Grossman W, Goldenberg DM (1989) Human lung carcinoma monoclonal antibody specific for the Thomsen-Friedenreich antigen. Cancer Res 49: 32
Vitetta ES, Stone M, Amlot P, Fay J, May R, Till M, Newman J, Clark P, Collins R, Cunningham D, Ghetie C, Uhr JW, Thorpe PE (1991) Phase I immunotoxin trial in patients with B-cell lymphoma. Cancer Res 51: 4052
Author information
Authors and Affiliations
Additional information
This work was supported in part by USPHS grant CA39841 from the NIH
Rights and permissions
About this article
Cite this article
Stein, R., Belisle, E., Hansen, H.J. et al. Epitope specificity of the anti-(B cell lymphoma) monoclonal antibody, LL2. Cancer Immunol Immunother 37, 293–298 (1993). https://doi.org/10.1007/BF01518451
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01518451