Abstract
In this study the problems encountered in staining immunoglobulin (Ig) in sections of paraffin-embedded human lymphoma samples have been investigated. It was found that the "masking' of cytoplasmic Ig, which occurs when tissues are fixed in formol saline (the fixative employed in most previous studies), can be avoided by the use of mercury-based fixatives. When non-Hodgkin's lymphoma samples fixed in this way were studied it was found that cytoplasmic Ig labelling of both lymphoid and histiocytic cells is often attributable to non-specific uptake of serum proteins. This phenomenon probably accounts for a number of published anomalous immunoperoxidase staining results in human lymphoma (e.g. the presence of both kappa and lambda chains in the same neoplastic cell). Double immunoenzymatic labelling (using alkaline phosphatase and peroxidase) proved valuable in the elucidation of this phenomenon. When staining due to absorbed Ig was discounted it was possible to demonstrate monoclonal Ig labelling in seven out of sixteen cases of non-Hodgkin's lymphoma. In each case IgM was found in association with a single light chain type and these results were in agreement with those obtained by direct immunofluorescent labelling of cryostat sections. In a further case u chains without associated light chains were demonstrated by immunoperoxidase staining. Seven cases of Hodgkin's disease were studied by immunoenzymatic techniques. Although IgG was frequently found in Reed-Sternberg and Hodgkin's cells its presence was not attributable to non-specific uptake of serum protein since albumin was absent or only present in small amounts. These findings are in support of the macrophage origin of these cells.
Full text
PDF![235](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/087bcef77106/clinexpimmunol00194-0015.png)
![236](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/774d93f3cf5e/clinexpimmunol00194-0016.png)
![237](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/d03a52c2d75e/clinexpimmunol00194-0017.png)
![238](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/25ed5597bd05/clinexpimmunol00194-0018.png)
![239](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/eace8f4b5515/clinexpimmunol00194-0019.png)
![240](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/e5d4719cd999/clinexpimmunol00194-0020.png)
![241](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/25119397d467/clinexpimmunol00194-0021.png)
![242](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/3957b604cf2a/clinexpimmunol00194-0022.png)
![243](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/cadcabb179e6/clinexpimmunol00194-0023.png)
![244](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/3430bf267985/clinexpimmunol00194-0024.png)
![245](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/e926f023b3cf/clinexpimmunol00194-0025.png)
![246](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/09d233a548f5/clinexpimmunol00194-0026.png)
![247](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/6d1c9637aabe/clinexpimmunol00194-0027.png)
![248](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1b/1536967/2e9f26ace642/clinexpimmunol00194-0028.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alexanian R. Monoclonal gammopathy in lymphoma. Arch Intern Med. 1975 Jan;135(1):62–66. [PubMed] [Google Scholar]
- Bosman F. T., Lindeman J., Kuiper G., van der Wal A., Kreunig J. The influence of fixation on immunoperoxidase staining of plasmacells in paraffin sections of intestinal biopsy specimens. Histochemistry. 1977 Jul 18;53(1):57–62. doi: 10.1007/BF00511210. [DOI] [PubMed] [Google Scholar]
- Brandtzaeg P. Mucosal and glandular distribution of immunoglobulin components. Immunohistochemistry with a cold ethanol-fixation technique. Immunology. 1974 Jun;26(6):1101–1114. [PMC free article] [PubMed] [Google Scholar]
- Cawley J. C., Smith J., Goldstone A. H., Emmines J., Hamblin J., Hough L. IgA and IgM cytoplastic inclusions in a series of cases of chronic lymphocytic leukaemia. Clin Exp Immunol. 1976 Jan;23(1):78–82. [PMC free article] [PubMed] [Google Scholar]
- Curran R. C., Gregory J. The unmasking of antigens in paraffin sections of tissue by trypsin. Experientia. 1977 Oct 15;33(10):1400–1401. doi: 10.1007/BF01920206. [DOI] [PubMed] [Google Scholar]
- Curran R. C., Jones E. L. Hodgkin's disease: an immunohistochemical and histological study. J Pathol. 1978 May;125(1):39–51. doi: 10.1002/path.1711250107. [DOI] [PubMed] [Google Scholar]
- Curran R. C., Jones E. L. Immunoglobulin in Reed-Sternberg and Hodgkin cells. J Pathol. 1978 Sep;126(1):35–37. doi: 10.1002/path.1711260105. [DOI] [PubMed] [Google Scholar]
- Curran R. C., Jones E. L. Immunoglobulin-containing cells in human tonsils as demonstrated by immunohistochemistry. Clin Exp Immunol. 1977 Apr;28(1):103–115. [PMC free article] [PubMed] [Google Scholar]
- Curran R. C., Jones E. L. The lymphoid follicles of the human palatine tonsil. Clin Exp Immunol. 1978 Feb;31(2):251–259. [PMC free article] [PubMed] [Google Scholar]
- Davey F. R., Halliday D., Marucci A. A., Gottlieb A. J. Detection of intracellular and cell surface immunoglobulin in non-Hodgkin's lymphoma. Hum Pathol. 1978 May;9(3):285–294. doi: 10.1016/s0046-8177(78)80086-0. [DOI] [PubMed] [Google Scholar]
- Feremans W. W., Neve P., Caudron M. IgM lambda cytoplasmic crystals in three cases of immunocytoma: a clinical, cytochemical, and ultrastructural study. J Clin Pathol. 1978 Mar;31(3):250–258. doi: 10.1136/jcp.31.3.250. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garvin A. J., Spicer S. S., McKeever P. E. The cytochemical demonstration of intracellular immunoglobulin. In neoplasms of lymphoreticular tissue. Am J Pathol. 1976 Mar;82(3):457–478. [PMC free article] [PubMed] [Google Scholar]
- Garvin A. J., Spicer S. S., Parmley R. T., Munster A. M. Immunohistochemical demonstration of IgG in Reed-Sternberg and other cells in Hodgkin's disease. J Exp Med. 1974 May 1;139(5):1077–1083. doi: 10.1084/jem.139.5.1077. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guillouzo A., Belanger L., Beaumont C., Valet J. P., Briggs R., Chiu J. F. Cellular and subcellular immunolocalization of alpha1-fetoprotein and albumin in rat liver. Reevaluation of various experimental conditions. J Histochem Cytochem. 1978 Nov;26(11):948–959. doi: 10.1177/26.11.82574. [DOI] [PubMed] [Google Scholar]
- Halliday D., Davey F. R., Marucci A. A. Detection of intracellular immunoglobulin in nodular lymphomas. Am J Clin Pathol. 1978 Jun;69(6):587–593. doi: 10.1093/ajcp/69.6.587. [DOI] [PubMed] [Google Scholar]
- Hartmann L., Liacopoulos P. Lymphoid cells of the normal man intestinal mucosa possessing both kappa and lambda light chain specifities. Biomedicine. 1977 Apr;27(3):123–127. [PubMed] [Google Scholar]
- Isaacson P., Wright D. H. Intestinal lymphoma associated with malabsorption. Lancet. 1978 Jan 14;1(8055):67–70. doi: 10.1016/s0140-6736(78)90004-1. [DOI] [PubMed] [Google Scholar]
- Kadin M. E., Stites D. P., Levy R., Warnke R. Exogenous immunoglobulin and the macrophage origin of Reed-Sternberg cells in Hodgkin's disease. N Engl J Med. 1978 Nov 30;299(22):1208–1214. doi: 10.1056/NEJM197811302992203. [DOI] [PubMed] [Google Scholar]
- Landaas T. O., Godal T., Halvorsen T. B. Characterization of immunoglobulins in Hodgkin cells. Int J Cancer. 1977 Nov 15;20(5):717–722. doi: 10.1002/ijc.2910200511. [DOI] [PubMed] [Google Scholar]
- Li C. Y., Harrison G., Jr Histochemical and immunohistochemical study of diffuse large-cell lymphomas. Am J Clin Pathol. 1978 Nov;70(5):721–732. doi: 10.1093/ajcp/70.5.721. [DOI] [PubMed] [Google Scholar]
- Mason D. Y. Measurement of kappa:lambda light chain ratios by haemagglutination techniques. J Immunol Methods. 1975 Jan;6(3):273–282. doi: 10.1016/0022-1759(75)90070-8. [DOI] [PubMed] [Google Scholar]
- Mason D. Y., Sammons R. E. The labeled antigen method of immunoenzymatic staining. J Histochem Cytochem. 1979 Apr;27(4):832–840. doi: 10.1177/27.4.109496. [DOI] [PubMed] [Google Scholar]
- Mason D. Y., Sammons R. Alkaline phosphatase and peroxidase for double immunoenzymatic labelling of cellular constituents. J Clin Pathol. 1978 May;31(5):454–460. doi: 10.1136/jcp.31.5.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mepham B. L., Frater W., Mitchell B. S. The use of proteolytic enzymes to improve immunoglobulin staining by the PAP technique. Histochem J. 1979 May;11(3):345–357. doi: 10.1007/BF01005033. [DOI] [PubMed] [Google Scholar]
- Papadimitriou C. S., Stein H., Lennert K. The complexity of immunohistochemical staining pattern of Hodgkin and Sternberg-reed cells--demonstration of immunoglobulin, albumin, alpha1-antichymotrypsin and lysozyme. Int J Cancer. 1978 May 15;21(5):531–541. doi: 10.1002/ijc.2910210502. [DOI] [PubMed] [Google Scholar]
- Poppema S., Elema J. D., Halie M. R. The significance of intracytoplasmic proteins in Reed-Sternberg cells. Cancer. 1978 Oct;42(4):1793–1803. doi: 10.1002/1097-0142(197810)42:4<1793::aid-cncr2820420418>3.0.co;2-l. [DOI] [PubMed] [Google Scholar]
- Sternberger L. A., Hardy P. H., Jr, Cuculis J. J., Meyer H. G. The unlabeled antibody enzyme method of immunohistochemistry: preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem. 1970 May;18(5):315–333. doi: 10.1177/18.5.315. [DOI] [PubMed] [Google Scholar]
- Taylor C. R. An immunohistological study of follicular lymphoma, reticulum cell sarcoma and Hodgkin's disease. Eur J Cancer. 1976 Jan;12(1):61–75. doi: 10.1016/0014-2964(76)90125-0. [DOI] [PubMed] [Google Scholar]
- Taylor C. R., Burns J. The demonstration of plasma cells and other immunoglobulin-containing cells in formalin-fixed, paraffin-embedded tissues using peroxidase-labelled antibody. J Clin Pathol. 1974 Jan;27(1):14–20. doi: 10.1136/jcp.27.1.14. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor C. R. Immunocytochemical methods in the study of lymphoma and related conditions. J Histochem Cytochem. 1978 Jul;26(7):496–512. doi: 10.1177/26.7.357639. [DOI] [PubMed] [Google Scholar]
- Taylor C. R., Mason D. Y. The immunohistological detection of intracellular immunoglobulin in formalin-paraffin sections from multiple myeloma and related conditions using the immunoperoxidase technique. Clin Exp Immunol. 1974 Nov;18(3):417–429. [PMC free article] [PubMed] [Google Scholar]
- Taylor C. R., Russell R., Chandor S. An immunohistologic study of multiple myeloma and related conditions, using an immunoperoxidase method. Am J Clin Pathol. 1978 Oct;70(4):612–622. doi: 10.1093/ajcp/70.4.612. [DOI] [PubMed] [Google Scholar]
- Taylor C. R. The nature of Reed-Sternberg cells and other malignant "reticulum" cells. Lancet. 1974 Oct 5;2(7884):802–807. doi: 10.1016/s0140-6736(74)91071-x. [DOI] [PubMed] [Google Scholar]
- Warnke R., Levy R. Immunopathology of follicular lymphomas. A model of B-lymphocyte homing. N Engl J Med. 1978 Mar 2;298(9):481–486. doi: 10.1056/NEJM197803022980903. [DOI] [PubMed] [Google Scholar]
- Zulman J., Jaffe R., Talal N. Evidence that the malignant lymphoma of Sjögren's syndrome is a monoclonal B-cell neoplasm. N Engl J Med. 1978 Nov 30;299(22):1215–1220. doi: 10.1056/NEJM197811302992204. [DOI] [PubMed] [Google Scholar]
- van den Tweel J. G., Taylor C. R., Parker J. W., Lukes R. J. Immunoglobulin inclusions in non-Hodgkin's lymphomas. Am J Clin Pathol. 1978 Mar;69(3):306–313. doi: 10.1093/ajcp/69.1.306. [DOI] [PubMed] [Google Scholar]