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. 1989 Feb;134(2):327–336.

Cell proliferation in non-Hodgkin's lymphomas. Digital image analysis of Ki-67 antibody staining.

B R Schwartz 1, G Pinkus 1, S Bacus 1, M Toder 1, D S Weinberg 1
PMCID: PMC1879592  PMID: 2464940

Abstract

Ki-67 is a monoclonal antibody to a nuclear antigen present in cycling human cells but not in resting cells. The authors have performed immunoperoxidase on non-Hodgkin's lymphomas using Ki-67 antibody in order to correlate proliferation rates with tumor grade and type, and compare Ki-67 staining with S-phase content as determined by flow cytometry. Ki-67 staining of 109 sections was quantitated using a digital image analysis system (CAS 100). There was a significant difference among mean overall Ki-67 staining values in Working Formulation low (13.7%), intermediate (42.6%), and high grade tumors (57.9%, P less than 0.00001). The level of significance improved when a revised grading system was formulated based on proliferative activity, with the inclusion of diffuse large cell lymphomas in the high grade category. Within nodular and a few diffuse lymphomas, there were well-defined proliferation centers in which Ki-67 staining showed no correlation with grade. Flow cytometric DNA determination was performed on 74 specimens, and there was a positive correlation between Ki-67 positivity and S phase content (r = 0.66). It is concluded that Ki-67 staining of tissue sections is an alternative to flow cytometric quantitation of cell cycle activity in lymphomas, and provides the advantage of revealing histologic patterns of proliferation. By including G1 phase cells, Ki-67 staining allows a more complete determination of total cell cycle activity in lymphomas.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akerman M., Brandt L., Johnson A., Olsson H. Mitotic activity in non-Hodgkin's lymphoma. Relation to the Kiel classification and to prognosis. Br J Cancer. 1987 Feb;55(2):219–223. doi: 10.1038/bjc.1987.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bauer K. D., Merkel D. E., Winter J. N., Marder R. J., Hauck W. W., Wallemark C. B., Williams T. J., Variakojis D. Prognostic implications of ploidy and proliferative activity in diffuse large cell lymphomas. Cancer Res. 1986 Jun;46(6):3173–3178. [PubMed] [Google Scholar]
  3. Braylan R. C., Benson N. A., Nourse V., Kruth H. S. Correlated analysis of cellular DNA, membrane antigens and light scatter of human lymphoid cells. Cytometry. 1982 Mar;2(5):337–343. doi: 10.1002/cyto.990020511. [DOI] [PubMed] [Google Scholar]
  4. Braylan R. C., Diamond L. W., Powell M. L., Harty-Golder B. Percentage of cells in the S phase of the cell cycle in human lymphoma determined by flow cytometry. Cytometry. 1980 Nov;1(3):171–174. doi: 10.1002/cyto.990010302. [DOI] [PubMed] [Google Scholar]
  5. Braylan R. C., Fowlkes B. J., Jaffe E. S., Sanders S. K., Berard C. W., Herman C. J. Cell volumes and DNA distributions of normal and neoplastic human lymphoid cells. Cancer. 1978 Jan;41(1):201–209. doi: 10.1002/1097-0142(197801)41:1<201::aid-cncr2820410129>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
  6. Christensson B., Tribukait B., Linder I. L., Ullman B., Biberfeld P. Cell proliferation and DNA content in non-Hodgkin's lymphoma. Flow cytometry in relation to lymphoma classification. Cancer. 1986 Sep 15;58(6):1295–1304. doi: 10.1002/1097-0142(19860915)58:6<1295::aid-cncr2820580620>3.0.co;2-h. [DOI] [PubMed] [Google Scholar]
  7. Costa A., Bonadonna G., Villa E., Valagussa P., Silvestrini R. Labeling index as a prognostic marker in non-Hodgkin's lymphomas. J Natl Cancer Inst. 1981 Jan;66(1):1–5. [PubMed] [Google Scholar]
  8. Diamond L. W., Nathwani B. N., Rappaport H. Flow cytometry in the diagnosis and classification of malignant lymphoma and leukemia. Cancer. 1982 Sep 15;50(6):1122–1135. doi: 10.1002/1097-0142(19820915)50:6<1122::aid-cncr2820500616>3.0.co;2-q. [DOI] [PubMed] [Google Scholar]
  9. Donhuijsen K. Mitoses in non-Hodgkin's lymphomas. Frequency and prognostic relevance. Pathol Res Pract. 1987 Jun;182(3):352–357. doi: 10.1016/S0344-0338(87)80071-7. [DOI] [PubMed] [Google Scholar]
  10. Donhuijsen K. Mitosis counts: reproducibility and significance in grading of malignancy. Hum Pathol. 1986 Nov;17(11):1122–1125. doi: 10.1016/s0046-8177(86)80417-8. [DOI] [PubMed] [Google Scholar]
  11. Ellison D. J., Nathwani B. N., Metter G. E., Turner R. R., Burke J. S., Miller T. P., Mann R. B., Barcos M., Kjeldsberg C. R., Winberg C. D. Mitotic counts in follicular lymphomas. Hum Pathol. 1987 May;18(5):502–505. doi: 10.1016/s0046-8177(87)80035-7. [DOI] [PubMed] [Google Scholar]
  12. Evans H. L., Butler J. J., Youness E. L. Malignant lymphoma, small lymphocytic type: a clinicopathologic study of 84 cases with suggested criteria for intermediate lymphocytic lymphoma. Cancer. 1978 Apr;41(4):1440–1455. doi: 10.1002/1097-0142(197804)41:4<1440::aid-cncr2820410432>3.0.co;2-g. [DOI] [PubMed] [Google Scholar]
  13. Gerdes J., Dallenbach F., Lennert K., Lemke H., Stein H. Growth fractions in malignant non-Hodgkin's lymphomas (NHL) as determined in situ with the monoclonal antibody Ki-67. Hematol Oncol. 1984 Oct-Dec;2(4):365–371. doi: 10.1002/hon.2900020406. [DOI] [PubMed] [Google Scholar]
  14. Gerdes J., Lemke H., Baisch H., Wacker H. H., Schwab U., Stein H. Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol. 1984 Oct;133(4):1710–1715. [PubMed] [Google Scholar]
  15. Gerdes J., Schwab U., Lemke H., Stein H. Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer. 1983 Jan 15;31(1):13–20. doi: 10.1002/ijc.2910310104. [DOI] [PubMed] [Google Scholar]
  16. Gerdes J., Stein H., Pileri S., Rivano M. T., Gobbi M., Ralfkiaer E., Nielsen K. M., Pallesen G., Bartels H., Palestro G. Prognostic relevance of tumour-cell growth fraction in malignant non-Hodgkin's lymphomas. Lancet. 1987 Aug 22;2(8556):448–449. doi: 10.1016/s0140-6736(87)90977-9. [DOI] [PubMed] [Google Scholar]
  17. Grogan T. M., Lippman S. M., Spier C. M., Slymen D. J., Rybski J. A., Rangel C. S., Richter L. C., Miller T. P. Independent prognostic significance of a nuclear proliferation antigen in diffuse large cell lymphomas as determined by the monoclonal antibody Ki-67. Blood. 1988 Apr;71(4):1157–1160. [PubMed] [Google Scholar]
  18. Hall P. A., Richards M. A., Gregory W. M., d'Ardenne A. J., Lister T. A., Stansfeld A. G. The prognostic value of Ki67 immunostaining in non-Hodgkin's lymphoma. J Pathol. 1988 Mar;154(3):223–235. doi: 10.1002/path.1711540305. [DOI] [PubMed] [Google Scholar]
  19. Juneja S. K., Cooper I. A., Hodgson G. S., Wolf M. M., Ding J. C., Ironside P. N., Thomas R. J., Parkin J. D. DNA ploidy patterns and cytokinetics of non-Hodgkin's lymphoma. J Clin Pathol. 1986 Sep;39(9):987–992. doi: 10.1136/jcp.39.9.987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kvaløy S., Godal T., Marton P. F., Steen H., Brenhovd I. O., Abrahamsen A. F. Spontaneous (H)-thymidine uptake in histological subgroups of human B-cell lymphomas. Scand J Haematol. 1981 Mar;26(3):221–234. doi: 10.1111/j.1600-0609.1981.tb01650.x. [DOI] [PubMed] [Google Scholar]
  21. Lage J. M., Driscoll S. G., Yavner D. L., Olivier A. P., Mark S. D., Weinberg D. S. Hydatidiform moles. Application of flow cytometry in diagnosis. Am J Clin Pathol. 1988 May;89(5):596–600. doi: 10.1093/ajcp/89.5.596. [DOI] [PubMed] [Google Scholar]
  22. Lang W., Kienzle S., Diehl V. Proliferation kinetics of malignant non-Hodgkin's lymphomas related to histopathology of lymph node biopsies. Virchows Arch A Pathol Anat Histol. 1980;389(3):397–407. doi: 10.1007/BF00430662. [DOI] [PubMed] [Google Scholar]
  23. Macartney J. C., Camplejohn R. S., Alder J., Stone M. G., Powell G. Prognostic importance of DNA flow cytometry in non-Hodgkin's lymphomas. J Clin Pathol. 1986 May;39(5):542–546. doi: 10.1136/jcp.39.5.542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Meyer J. S., Higa E. S-phase fractions of cells in lymph nodes and malignant lymphomas. Arch Pathol Lab Med. 1979 Feb;103(2):93–97. [PubMed] [Google Scholar]
  25. Nathwani B. N., Diamond L. W., Winberg C. D., Kim H., Bearman R. M., Glick J. H., Jones S. E., Gams R. A., Nissen N. I., Rappaport H. Lymphoblastic lymphoma: a clinicopathologic study of 95 patients. Cancer. 1981 Dec 1;48(11):2347–2357. doi: 10.1002/1097-0142(19811201)48:11<2347::aid-cncr2820481102>3.0.co;2-x. [DOI] [PubMed] [Google Scholar]
  26. Nathwani B. N., Winberg C. D., Diamond L. W., Bearman R. M., Kim H. Morphologic criteria for the differentiation of follicular lymphoma from florid reactive follicular hyperplasia: a study of 80 cases. Cancer. 1981 Oct 15;48(8):1794–1806. doi: 10.1002/1097-0142(19811015)48:8<1794::aid-cncr2820480817>3.0.co;2-m. [DOI] [PubMed] [Google Scholar]
  27. Payne C. M., Grogan T. M., Cromey D. W., Bjore C. G., Jr, Kerrigan D. P. An ultrastructural morphometric and immunophenotypic evaluation of Burkitt's and Burkitt's-like lymphomas. Lab Invest. 1987 Aug;57(2):200–218. [PubMed] [Google Scholar]
  28. Pinkus G. S., Said J. W. Characterization of non-Hodgkin's lymphomas using multiple cell markers. Immunologic, morphologic, and cytochemical studies of 72 cases. Am J Pathol. 1979 Feb;94(2):349–380. [PMC free article] [PubMed] [Google Scholar]
  29. Porwit-Ksiazek A., Christensson B., Lindemalm C., Mellstedt H., Tribukait B., Biberfeld G., Biberfeld P. Characterization of malignant and non-neoplastic cell phenotypes in highly malignant non-Hodgkin lymphomas. Int J Cancer. 1983 Dec 15;32(6):667–674. doi: 10.1002/ijc.2910320604. [DOI] [PubMed] [Google Scholar]
  30. Roos G., Dige U., Lenner P., Lindh J., Johansson H. Prognostic significance of DNA-analysis by flow cytometry in non-Hodgkin's lymphoma. Hematol Oncol. 1985 Oct-Dec;3(4):233–242. doi: 10.1002/hon.2900030403. [DOI] [PubMed] [Google Scholar]
  31. Scarffe J. H., Crowther D. The pre-treatment proliferative activity of non-Hodgkin's lymphoma cells. Eur J Cancer. 1981 Jan;17(1):99–108. doi: 10.1016/0014-2964(81)90218-8. [DOI] [PubMed] [Google Scholar]
  32. Schrape S., Jones D. B., Wright D. H. A comparison of three methods for the determination of the growth fraction in non-Hodgkin's lymphoma. Br J Cancer. 1987 Mar;55(3):283–286. doi: 10.1038/bjc.1987.54. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shackney S. E., Levine A. M., Fisher R. I., Nichols P., Jaffe E., Schuette W. H., Simon R., Smith C. A., Occhipinti S. J., Parker J. W. The biology of tumor growth in the non-Hodgkin's lymphomas. A dual parameter flow cytometry study of 220 cases. J Clin Invest. 1984 Apr;73(4):1201–1214. doi: 10.1172/JCI111306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Srigley J., Barlogie B., Butler J. J., Osborne B., Blick M., Johnston D., Kantarjian H., Reuben J., Batsakis J., Freireich E. J. Heterogeneity of non-Hodgkin's lymphoma probed by nucleic acid cytometry. Blood. 1985 May;65(5):1090–1096. [PubMed] [Google Scholar]
  35. Swerdlow S. H., Habeshaw J. A., Murray L. J., Dhaliwal H. S., Lister T. A., Stansfeld A. G. Centrocytic lymphoma: a distinct clinicopathologic and immunologic entity. A multiparameter study of 18 cases at diagnosis and relapse. Am J Pathol. 1983 Nov;113(2):181–197. [PMC free article] [PubMed] [Google Scholar]
  36. Weisenburger D. D., Linder J., Daley D. T., Armitage J. O. Intermediate lymphocytic lymphoma: an immunohistologic study with comparison to other lymphocytic lymphomas. Hum Pathol. 1987 Aug;18(8):781–790. doi: 10.1016/s0046-8177(87)80051-5. [DOI] [PubMed] [Google Scholar]
  37. Weisenburger D. D., Nathwani B. N., Diamond L. W., Winberg C. D., Rappaport H. Malignant lymphoma, intermediate lymphocytic type: a clinicopathologic study of 42 cases. Cancer. 1981 Sep 15;48(6):1415–1425. doi: 10.1002/1097-0142(19810915)48:6<1415::aid-cncr2820480625>3.0.co;2-n. [DOI] [PubMed] [Google Scholar]
  38. Weiss L. M., Strickler J. G., Medeiros L. J., Gerdes J., Stein H., Warnke R. A. Proliferative rates of non-Hodgkin's lymphomas as assessed by Ki-67 antibody. Hum Pathol. 1987 Nov;18(11):1155–1159. doi: 10.1016/s0046-8177(87)80384-2. [DOI] [PubMed] [Google Scholar]

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