Hodgkin''s disease, lymphocyte predominance type, nodular--further evidence for a B cell derivation. L & H variants of Reed-Sternberg cells express L26, a pan B cell marker.

Immunoreactivity for L26, a highly effective pan B cell marker that can be detected in paraffin sections, was evaluated in 72 cases of Hodgkin's disease of various histologic types. In all cases of nodular lymphocyte predominance type of Hodgkin's disease, L & H variants of Reed-Sternberg cells uniformly exhibited strong immunoreactivity for L26. Other variants of Reed-Sternberg cells, eg, lacunar, mononuclear, and diagnostic forms, present in nodular sclerosis, mixed cellularity, and lymphocyte depletion types of Hodgkin's disease, infrequently expressed L26 reactivity. In 55 of 63 cases (87%) of these combined types, less than 5% of Reed-Sternberg cells or variants were L26 positive. In the remaining cases, a larger proportion of these cells expressed L26. Topographic patterns of immunoreactivity for small lymphocytes in these different types of Hodgkin's disease also varied. In nodular lymphocyte predominance type, L26 positive lymphocytes (presumptive B cells) were mainly localized to nodular areas of the proliferation. In other types of Hodgkin's disease, L26 positive cells occurred in small or large aggregates and generally represented a minor proportion of the population of lymphoid cells. These studies further support a B cell derivation for L & H variants of Reed-Sternberg cells and provide additional evidence that nodular lymphocyte predominance type Hodgkin's disease may represent a distinct entity, possibly an unusual low grade B cell lymphoma. These data also suggest that some Reed-Sternberg cells and variants present in other histologic types of Hodgkin's disease may be of B cell derivation, and precludes the use of L26 as a diagnostic discriminant in cases in which the distinction between Hodgkin's disease and non-Hodgkin's lymphoma is unclear.     

Granulocyte and HLA-D region specific monoclonal antibodies in the diagnosis of Hodgkin''s disease.

Tissue sections embedded in paraffin and fixed in formalin from 32 patients with Hodgkin's disease, representing the major histological subtypes, were studied using two granulocyte specific monoclonal antibodies (Leu-M1 and 3C4) and an HLA-D region specific monoclonal antibody (TAL-IB5). Reed-Sternberg cells were stained with one or other of the antigranulocyte antibodies in the nodular sclerosing and lymphocyte depleted subtypes. Reed-Sternberg cells in all but three cases of mixed cellularity Hodgkin's disease were positive with both Leu-M1 and 3C4. One case stained with only Leu-M1, and two cases were consistently negative with both antibodies. HLA-DR was widely expressed in the Reed-Sternberg cells of all three subtypes. In the four cases of lymphocyte predominant Hodgkin's disease the multinucleated Reed-Sternberg cells did not stain with either antigranulocyte antibody but were strongly positive with anti-HLA-DR. Twenty five cases of non-Hodgkin's lymphoma, in which there were multinucleated giant cells resembling Reed-Sternberg cells, were studied in a similar way. These cases included pleomorphic T cell and B cell lymphomas, histiocytic lymphomas, and malignant histiocytosis of the intestine. In none of these did the multinucleated cells stain with either antigranulocyte antibody, but in most cases the multinucleated cells stained with anti-HLA-DR. In two cases of the tumour stage of mycosis fungoides dot like intracytoplasmic staining was shown in the tumour cells with both antigranulocyte markers. The monoclonal antigranulocyte antibodies Leu-M1 and 3C4 are of considerable value in both the diagnosis and the differential diagnosis of Hodgkin's disease and are particularly valuable in that they can be applied to tissue fixed in formalin and embedded in paraffin. Antibody to HLA-DR, while useful, is of less value.     

Monoclonal antibodies reactive in routinely processed tissue sections of malignant lymphoma, with emphasis on T-cell lymphomas

The immunoreactivity of eight monoclonal antibodies was evaluated on 45 routinely processed lymphomas (22 T-cell lymphomas, 11 B-cell lymphomas, and 12 cases of Hodgkin's disease). Two antibodies reactive with leukocyte common (T200) antigens (PD7/26 and 2B11) stained most of the B- and T-cell lymphomas but did not stain the Reed-Sternberg cells and variants in Hodgkin's disease. Two antibodies known to stain B cells (LN-1 and LN-2) reacted with some of the B-cell lymphomas, but LN-2 also reacted with the neoplastic cells in six of 22 T-cell lymphomas and with the Reed-Sternberg variants in eight of 12 cases of Hodgkin's disease. The granulocyte antibody anti-Leu M1 reacted with most cases of Hodgkin's disease but also reacted with two of 11 B-cell non-Hodgkin's lymphomas. An antibody to epithelial membrane antigen (anti-EMA) stained some cases of T-cell lymphoma, B-cell lymphoma, and Hodgkin's disease. Leu 7 was expressed in one T-cell lymphoma and in one case of Hodgkin's disease. A novel antibody reactive with T cells (L60) stained all cases of T-cell lymphoma but also stained some cases of B-cell lymphoma and one case of Hodgkin's disease. We conclude that none of these antibodies, when used alone on routinely fixed paraffin-embedded material, is completely sensitive and specific for T-cell lymphoma, B-cell lymphoma, or Hodgkin's disease. However, a panel of antibodies is useful in distinguishing Hodgkin's disease from non-Hodgkin's lymphoma and in suggesting the B- or T-cell phenotype of non-Hodgkin's lymphomas.     

Childhood lymphoma. A clinicopathological and immunohistological study of 58 cases

Fifty-eight cases diagnosed as malignant lymphoma in patients younger than 15 years between 1976 and 1986 in the Kanto area were reviewed and reclassified as follow: 48 non-Hodgkin's lymphomas, 9 Hodgkin's disease and one malignant histiocytosis. Lymphoblastic type consisted of 26 cases or 54.2%; large cell type, 11 cases or 22.9%; Burkitt's type, 7 cases or 14.5%; medium-sized cell type and mixed cell type consisted of 4 cases. There was no follicular lymphoma case. A rare sclerosing mediastinal lymphoblastic lymphoma and diffuse large cell lymphomas with T-zone involvement as well as primary epidural Burkitt's lymphomas were found. Immunohistochemical studies using paraffin sections were performed in 43 non-Hodgkin's lymphomas and phenotypes of 37 cases were determined as follows; T cell origin in 24, B cell origin in 11 and non-T non-B in 2 cases. Of 25 lymphoblastic lymphomas, LCA was positive only in 11 cases. Reed-Sternberg cells and their variants of Hodgkin's disease reacted with anti-Leu M1 antibody in 3 of 8 examined but not with EMA antibody. This study revealed that the survival was related to sites of the primary lesion, regardless of histological type and immunologic phenotypes.     

CHILDHOOD LYMPHOMA. A Clinicopathological and Immunohistological Study of 58 Cases

Fifty-eight cases diagnosed as malignant lymphoma in patients younger than 15 years between 1976 and 1986 in the Kanto area were reviewed and reclassified as follow: 48 non-Hodgkin's lymphomas, 9 Hodgkin's disease and one malignant histiocytosis. Lymphoblastic type consisted of 26 cases or 54.2%; large cell type, 11 cases or 22.9%; Burkitt's type, 7 cases or 14.5%; medium-sized cell type and mixed cell type consisted of 4 cases. There was no follicular lymphoma case. A rare sclerosing mediastinal lymphoblastic lymphoma and diffuse large cell lymphomas with T-zone involvement as well as primary epidural Burkitt's lymphomas were found. Immunohistochemical studies using paraffin sections were performed in 43 non-Hodgkin's lymphomas and phenotypes of 37 cases were determined as follows; T cell origin in 24, B cell origin in 11 and non-T non-B in 2 cases. Of 25 lymphoblastic lymphomas, LCA was positive only in 11 cases. Reed-Sternberg cells and their variants of Hodgkin's disease reacted with anti-Leu Ml antibody in 3 of 8 examined but not with EMA antibody. This study revealed that the survival was related to sites of the primary lesion, regardless of histological type and immunologic phenotypes. ACTA PATHOL JPN 38: 1149∼1166, 1988.     

Monoclonal antibody (UCHL1) that recognises normal and neoplastic T cells in routinely fixed tissues.

UCHL1 is a murine monoclonal antibody that recognises a 180-185 kD determinant on CD4 (72%) and CD8 (36%) positive T cells. This antibody is effective in formalin fixed and paraffin embedded tissues, using the immunoperoxidase method. One hundred and forty three cases of malignant lymphoma were examined. Neoplastic cells in 100% of cases of Mycosis fungoides (n = 10), 83% of cases of peripheral T cell lymphoma (n = 25), and 78% of cases of (T-ALL) T acute lymphoblastic lymphoma (n = 9) were stained by this antibody. In addition, staining was seen in 100% of cases of malignant histiocytosis of the intestine (n = 13), a condition now thought to be a T cell lymphoma. Two cases of true histiocytic lymphoma were also positive. This antibody stained neither the neoplastic cells in a wide range of B cell lymphomas (n = 62) nor Reed-Sternberg cells in 16 cases of Hodgkin's disease. UCHL1 also stained neoplastic cells in four cases of granulocytic sarcoma. A panel of normal tissues was similarly studied. Staining was seen in normal T cells and mucosal intraepithelial lymphocytes, macrophages, mature myeloid cells, and endometrial stromal granulocytes. UCHL1 is a monoclonal antibody that identifies T cells in formalin fixed paraffin embedded tissues, and should prove useful for diagnosing T cell lymphomas, especially when only formalin fixed tissue is available for diagnosis.     

Immunohistochemical analysis of CDw52 antigen expression in non-Hodgkin's lymphomas.

AIM--To determine the antigen expression of CDw52 using Campath-1 antibodies in a series of non-Hodgkin's lymphomas (NHLs). METHODS--Tissue sections of lymphoma were stained immunohistochemically using rat Campath-1G and humanised Campath-1H with avidin-biotin-peroxidase complex techniques. Fifty-two fresh frozen lymphomas and a further 26 paraffin wax embedded sections were studied. RESULTS--Thirty-seven out of 41 B cell lymphomas were positive with Campath-1H in frozen sections (low grade, 24 of 24; high grade, 13 of 17) as were three out of five T cell lymphomas. Reed-Sternberg cells in six cases of Hodgkin's disease did not react. Eleven out of 16 high grade B cell lymphomas also stained positively with Campath-1G in paraffin wax sections as did five out of 10 T cell lymphomas. CONCLUSIONS--The Campath-1 antibodies showed that CDw52 antigen expression was present in all cases of low grade B cell NHL examined. Immunohistochemical staining in high grade B cell NHL and in T cell NHL was variable. These findings may be relevant to patient selection when considering treatment with Campath-1 antibodies.     

Transforming growth factor beta 1 messenger RNA in Reed-Sternberg cells in nodular sclerosing Hodgkin's disease.

AIMS--To determine the cellular origin of the most potent cytokine present in Hodgkin's disease, transforming growth factor (TGF) beta, the polycellular population of Hodgkin's tissue was studied using in situ hybridisation. METHODS--A biotin labelled oligo-complementary DNA (cDNA) was constructed according to the previously determined sequence for TGF beta 1 cDNA. Forty three frozen and paraffin wax embedded tissue samples replaced by Hodgkin's disease or non-Hodgkin's lymphoma, three Reed-Sternberg cell lines, one Ki1 positive lymphoma cell line, and an epithelial cell line were studied for expression of TGF beta 1 messenger RNA (mRNA) as well as secretion of the TGF beta 1 protein and expression of the CD30 epitope. RESULTS--The results obtained with the 24 frozen tissue samples confirmed that the TGF beta antigen is found predominantly in the nodular sclerosing Hodgkin's disease (NSHD) subtype. Nineteen paraffin wax embedded tissue samples were used to measure the simultaneous expression of CD30 and TGF beta 1 mRNA. The latter was found in eight of eight NSHD samples, two of six mixed cellularity samples, and two of five non-Hodgkin's lymphoma samples. No evidence of fibroblast expression of TGF beta 1 mRNA was noted. CONCLUSIONS--Activated lymphocytes in NSHD express TGF beta 1 mRNA, but binucleate Reed-Sternberg cells and mononuclear Hodgkin's cells are the primary sources of activated TGF beta in Hodgkin's disease.     

Leu M1 and peanut agglutinin stain the neoplastic cells of Hodgkin's disease

It has been suggested that the malignant cells of Hodgkin's disease (HD), Reed-Sternberg cells, and their mononuclear variants may be related to cells of the monocyte-histiocyte system. To test this hypothesis, 20 cases of HD were tested with nine antibodies, monoclonal or polyclonal, that normally react with cells of monocytic/histiocytic/granulocytic lineages, as well as PNA, which binds to histiocytes directly. Only two reagents, Leu M1 and PNA bound to the neoplastic cells in 20/22 and 13/22 cases tested, respectively. Leu M1 was the most sensitive reagent and was negative in only two cases of lymphocyte predominant HD. Leu M1 also could be employed in routinely fixed and processed formalin or B5-fixed tissue. This antibody, which was negative in 27 cases of non-Hodgkin's lymphoma, including 12 peripheral T-cell lymphomas, should prove to be of value in differential diagnosis of HD and morphologically similar reactive and neoplastic lymphoid lesions.     

CD40 expression in Hodgkin's disease.

CD40 is a transmembrane protein that belongs to a superfamily of proteins related to nerve growth factor receptor. CD40 is expressed on B cells and some B cell malignancies. It appears to be involved in B cell proliferation and the prevention of apoptosis in germinal center cells, which is accompanied by expression of bcl-2. Its expression is up-regulated by the EBV protein latent membrane protein-1 and cytokines interleukin-4 and interferon-gamma. The expression of CD40 in 37 cases of Hodgkin's disease and 23 cases of non-Hodgkin's lymphoma (11 T cell lymphomas and 12 B cell lymphomas) was examined by paraffin section immunohistochemistry using the BB-20 monoclonal antibody. In 26 of 37 cases of Hodgkin's disease the Reed-Sternberg cells showed strong membrane or cytoplasmic expression of CD40. Only 3 of 23 non-Hodgkin's lymphomas showed any expression of CD40 and then only weakly. There was no correlation between expression of bcl-2 or latent membrane protein-1 with CD40 expression. These results show that there is probable hyperexpression of CD40 in Hodgkin's disease and suggest that dysregulation of CD40 expression may play a role in the pathogenesis of Hodgkin's disease.     

An immunocytochemical study of T-cell lymphomas using monoclonal and polyclonal antibodies effective in routinely fixed wax embedded tissues

Formalin fixed and paraffin wax embedded tissue from 24 cases of T-cell lymphoma diagnosed using immunocytochemistry on cryostat sections was examined using a panel of eight monoclonal and three polyclonal antisera. The monoclonal antibodies UCHL1 and MT1 proved to be comparable and reliable markers of neoplastic cells in T-cell lymphomas. The B-cell specific marker, MB1, strongly stained all cells in two cases of pleomorphic large cell T-cell lymphoma, large cells in two cases of pleomorphic mixed medium and large cell lymphoma, and isolated clusters of blast cells in four cases of T-zone and angioimmunoblastic lymphadenopathy-like T-cell lymphoma. The cells stained by MB1 expressed T suppressor/cytotoxic surface markers on frozen section. Epithelial membrane antigen, as detected by a polyclonal anti-EMA and the monoclonal antibody HMFG2, was expressed in 36% of tumours especially those of monomorphic large cell and pleomorphic large cell phenotype. Single granules or finely dispersed cytoplasmic granularity was seen in four tumours using the anti-granulocyte reagent Leu M1. Tumour cells in one case stained in a pattern identical to Reed-Sternberg cells in Hodgkin's disease. Granular alpha-1-antitrypsin staining was found in 10 cases of pleomorphic large cell and monomorphic large cell lymphoma. No staining was observed using anti-lysozyme or the monoclonal macrophage specific marker Mac411. Monomorphic and pleomorphic large cell lymphomas tended to show a common immunophenotype with the majority of cells co-expressing alpha-1-antitrypsin HLA-DR and epithelial membrane antigen. Scattered large transformed blast cells in cases of angioimmunoblastic lymphadenopathy-like T-cell lymphomas and T-zone lymphomas shared a similar immunophenotype with the large cell lymphomas. Using a panel of monoclonal antibodies effective in paraffin embedded tissue, diagnostically useful staining profiles which correlate with the morphological phenotype can be established in T-cell lymphomas.     

Paraffin section immunohistochemistry. II. Hodgkin''s disease and large cell anaplastic (Ki1) lymphoma

A panel of antibodies that recognize antigens that survive fixation and conventional processing have been applied to 43 cases of Hodgkin's disease and five cases of large cell anaplastic lymphoma. Reed-Sternberg cells in all five cases of nodular lymphocyte predominance Hodgkin's disease were positive with leucocyte common (CD45) and B-cell antibodies, and negative with LeuM1 (CD15) and BerH2 (CD30) antibodies. In other types of Hodgkin's disease, Reed-Sternberg cells were positive with BerH2 in all cases, positive with LeuM1 in 63% of cases (with enzymic predigestion), positive with at least one B-cell antibody in 29% of cases and positive for CD45 in 8% of cases. In 19% of all cases, Reed-Sternberg cells were positive for epithelial membrane antigen and in 93% they were positive with TAL1B5 (anti-class II MHC). No case showed immunoreactivity with anti-T-cell antibodies. The patterns of immunoreactivity of large cell anaplastic lymphoma were similar, except that none was positive with B-cell antibodies and three were positive with T-cell antibodies. All five were positive with BerH2 (CD30) and TAL1B5. Comparison of the results with those seen in other cases of non-Hodgkin's lymphoma indicates that, with the currently available reagents, this immunohistological profile cannot be used as the sole diagnostic discriminant of these conditions; this must still be based upon careful morphological assessment.     

Presence of 3-fucosyl-N-acetyllactosamine shown by monoclonal antibody AGF 4.48 in Reed-Sternberg cells.

A series of 50 specimens of Hodgkin's disease and 10 of reactive follicular hyperplasia were examined by means of indirect immunoperoxidase staining with a monoclonal antibody AGF 4.48: this is known to bind to 3-fucosyl-N-acetyllactosamine, which, in particular, is expressed by granulocyte series cells. Most Reed-Sternberg and many Hodgkin's cells were labelled by the antibody after pretreatment with neuraminidase. Routinely processed paraffin wax embedded sections proved suitable for staining. The findings were comparable with those reported by others with monoclonal antibodies to various other granulocyte markers. This technique is of potential diagnostic value.     

A new mdr-1 encoded P-170 specific monoclonal antibody: (6/1C) on paraffin wax embedded tissue without pretreatment of sections.

AIMS: The generation and characterisation of a monoclonal antibody that specifically recognises the mdr-1 encoded protein, P-glycoprotein (P-170), on routinely processed formalin fixed, paraffin wax embedded tissue sections. METHODS: The monoclonal antibody, designated 6/1C, was produced following a combination of in vivo and in vitro immunisation regimens in Balb/c mice with a synthetic 12 amino acid peptide that corresponds to amino acids 21-32 (believed to be intracellularly located) of P-170 and has insignificant homology with the mdr-3 encoded P-170. Antibody 6/1C was characterised by western blotting and immunocytochemistry on cytospins of paired multidrug resistant or sensitive cell lines, including mdr-1 and mdr-3 transfected cells, and by immunohistochemistry on normal and malignant formalin fixed paraffin wax embedded tissue sections. RESULTS: Antibody 6/1C showed a single band at 170 kDa on western blots of multidrug resistant cell lysates and mdr-1 transfected cell lysates that was absent on similar preparations of drug sensitive cells and mdr-3 transfected cells. Immunocytochemical studies on cytospins of multidrug resistant cells and mdr-1 transfected cells revealed strong inner plasma membrane/cytoplasmic staining. Staining was negligible on drug sensitive cells and cells transfected with the mdr-3 gene. Immunohistochemical studies on formalin fixed, paraffin wax embedded normal adult kidney, liver, and breast tissue and a range of fetal tissues exhibited staining patterns of a variety of secretory surfaces consistent with documented mdr-1 specific staining. Specific staining of malignant cells in similarly treated sections of breast tumours was seen also with antibody 6/1C. Staining on paraffin wax embedded tissue with this antibody did not require any pretreatment of tissue sections. CONCLUSIONS: This new monoclonal antibody, chosen for its specificity with the mdr-1 encoded P-170 and its reactivity on routinely fixed paraffin wax embedded tissue samples without pretreatment, appears to be useful for the investigation of P-170 in archival material. It is especially useful for retrospective studies on pretreatment and post-treatment tissue sections, and could help establish when and how rapidly mdr-1 associated drug resistance develops during chemotherapeutic regimens. Immunohistochemical assessment of P-170 expression in many cancers has potential for diagnostic purposes and may influence the choice of chemotherapeutic drugs used in the treatment of refractory tumours.     

The demonstration of B-cell, T-cell and myeloid antigens in paraffin sections

The monoclonal antibodies MB1 and MT1, which detect B cells and T cells respectively, have been applied to human lymphoid tissues. The distribution of staining within paraffin sections was compared with that observed using frozen sections and was found to be identical. The antibodies were then applied to paraffin sections of 19 B-cell lymphomas and 10 T-cell lesions in which full immunophenotyping had been performed. The B lymphomas all consisted of a large majority of MB1 positive cells with a variable infiltrate of small MT1 positive lymphocytes. The T cell lesions consisted of MT1 positive cells with few MB1 positive cells except in residual B cell areas of lymph nodes. In paraffin sections from cases of Hodgkin's disease anti-Leu M1 identified Reed-Sternberg cells and their variants and MB1 and MT1 showed a similar distribution of B cells and T cells to that demonstrated in previous studies using frozen sections. The results show that MB1 and MT1 are useful markers for B and T cells in routinely fixed paraffin embedded tissue. In conjunction with anti-Leu M1 they provide a valuable panel of antisera for the examination of lymph nodes and other biopsies when frozen tissue is not available.     

Identification of common germinal-center B-cell precursors in two patients with both Hodgkin's disease and non-Hodgkin's lymphoma

BACKGROUND: Hodgkin's disease and non-Hodgkin's B-cell lymphoma occasionally occur in the same patient. The identification of a common precursor of the two types of lymphoma would show definitively that Reed-Sternberg cells originate from B cells. METHODS: We studied lymphomas from two patients, one with a composite lymphoma (classic Hodgkin's disease and a follicular lymphoma in the same lymph node) and the other with a T-cell-rich B-cell lymphoma that was followed by classic Hodgkin's disease. Single Reed-Sternberg cells and non-Hodgkin's lymphoma cells from frozen sections were micromanipulated. The rearranged immunoglobulin variable-region genes (V genes) of the heavy and light chains were amplified by the polymerase chain reaction from genomic DNA and sequenced. RESULTS: In both patients, the Reed-Sternberg cells were related clonally to the non-Hodgkin's lymphoma B cells. The V genes carried somatic mutations (a hallmark of germinal-center B cells and their descendants). In both patients, some somatic mutations were shared by the Reed-Sternberg and non-Hodgkin's lymphoma cells, whereas other somatic mutations were found exclusively in one or the other cell type. CONCLUSIONS: In two patients with classic Hodgkin's disease and non-Hodgkin's B-cell lymphoma, we identified a common B-cell precursor, probably a germinal-center B-cell, for both lymphomas. This finding suggests that the two types of lymphoma underwent both shared and distinct transforming events and provides proof of the B-cell derivation of Reed-Sternberg cells in classic Hodgkin's disease.     

Immunohistological analysis of the immunoreactivity of normal lymphoid cells and lymphomas with the monoclonal antibody OPD4

OPD4 is a recently described monoclonal antibody that recognizes a fixation-resistant 200 kD antigen restricted to a subset of T cells. Immunolabelling with OPD4 in paraffin sections of normal lymphoid tissues and cases of malignant lymphoma was compared with that of other antibodies in common use, including the T-cell restricted antibodies MT1 and UCHL1 and the B-cell restricted antibodies MB1, F8-11-13, and L26. OPD4 showed similar immunoreactivity to UCHL1 in normal tissues. OPD4 did not stain Reed-Sternberg cells in Hodgkin's disease. In non-Hodgkin's lymphomas, OPD4, like UCHL1, reacted with only 2/22 B-cell lymphomas. OPD4 was, however, less useful as a marker of T-cell lymphomas, staining only 11/32 cases, while UCHL1 stained 22/32 cases. We conclude that OPD4 is not a useful antibody for the routine diagnosis of T-cell lymphoma.     

Expression of the matrix metalloproteinase 9 in Hodgkin's disease is independent of EBV status.

BACKGROUND: In vitro the Epstein-Barr virus (EBV) encoded latent membrane protein 1 (LMP-1) has been shown to upregulate expression of matrix metalloproteinase 9 (MMP-9), a member of a family of zinc dependent endopeptidases that is believed to facilitate tumour invasion and metastasis by degradation of the extracellular matrix. AIM: To test whether the expression of MMP-9 in Hodgkin's disease correlates with EBV status and survival and to investigate whether LMP-1 expression affects MMP-9 concentrations in the Hodgkin's disease cell line, L428. METHODS: MMP-9 expression was measured by means of immunohistochemistry in a series of Hodgkin's disease tumours and this expression was correlated with EBV status and survival. The influence of LMP-1 on MMP-9 expression was also investigated in the Hodgkin's disease cell line, L428. RESULTS: MMP-9 expression was demonstrated in the malignant Hodgkin and Reed-Sternberg cells of all (n = 86) formalin fixed, paraffin wax embedded Hodgkin's disease tumours examined. Although the intensity of MMP-9 immunostaining varied between cases, there was no correlation between MMP-9 expression and EBV status or survival. MMP-9 expression was also detected in a variety of non-malignant cells, including fibroblasts. MMP-9 was detected by zymography in the L428 and KMH2 Hodgkin's disease cell lines, whereas low or undetectable amounts of MMP-9 were found in the L591 Hodgkin's disease cell line. Induction of LMP-1 expression in the Hodgkin's disease cell line L428 did not result in a detectable increase in the values of MMP-9 as measured by zymography. CONCLUSIONS: These results demonstrate that MMP-9 is consistently expressed by the Hodgkin and Reed-Sternberg cells of Hodgkin's disease tumours and by the Hodgkin's disease cell lines, L428 and KMH2. However, this expression does not appear to be related either to LMP-1 values or to survival.     

Localization of proliferating cell nuclear antigen (PCNA/Cyclin) in workshop cases of Hodgkin's disease and non-Hodgkin's lymphoma.

Proliferating cell nuclear antigen (PCNA/Cyclin) is a 36-kD protein that is present in cycling cells but not in resting cells, and therefore represents a marker of tumor proliferation. Application of anti-PCNA/Cyclin monoclonal antibodies has shown that this protein is localized to the nucleus of cycling cells, with the exception of cells in mitosis, which demonstrate faint cytoplasmic reactivity. Recently, Benjamin and Gown found that Reed-Sternberg cells and variants show nuclear and cytoplasmic staining with anti-PCNA/Cyclin antibody 19A2, and suggested that this feature may be useful in distinguishing Hodgkin's disease from other tumors. This report describes the reactivity of 42 workshop cases that were stained with anti-PCNA/Cyclin antibodies 19A2 and/or PC10. Thirty-three (79%) of the 42 cases showed adequate reactivity to allow for interpretation of staining localization. In the group of reactive cases, 26 (79%) showed nuclear and cytoplasmic staining. The localization of PCNA/Cyclin was compared with the consensus diagnosis in each case. Eighty percent of cases classified as Hodgkin's disease, 67% of cases classified as non-Hodgkin's lymphoma, and 100% of unresolved cases showed both nuclear and cytoplasmic staining. The incidence of cytoplasmic PCNA/Cyclin was not different between Hodgkin's disease and non-Hodgkin's lymphoma in this study.     

An evaluation of the utility of anti-granulocyte and anti-leukocyte monoclonal antibodies in the diagnosis of Hodgkin's disease.

The immunoreactivity of six different monoclonal antigranulocyte antibodies (Leu M1, TG1, 3C4, BY/87a, BY/37a, and 3CD1) has been evaluated in 23 cases of Hodgkin's disease (7 lymphocyte predominant, 12 nodular sclerosing, and 5 mixed cellularity); in a variety of non-Hodgkin's lymphomas and in a series of reactive and benign lesions of lymph nodes. Applying a monoclonal antibody (PD7/26) to leukocyte common antigen (T200), we have also investigated reports that the L&H variants in nodular lymphocyte predominant Hodgkin's disease are strongly immunoreactive for leukocyte common antigen in contrast to the lack of reactivity of Reed-Sternberg (RS) cells and variants thereof in other forms of Hodgkin's disease. All six monoclonal anti-granulocyte antibodies reacted against RS cells and "Hodgkin's cells" in the nodular sclerosing (NSHD) and mixed cellularity (MCHD) types, with strong cell membrane and juxtanuclear (Golgi) staining. In contrast an anti-leukocyte antibody PD7/26 was unreactive with RS cells and variants thereof in NSHD and MCHD. On the other hand, RS cells and L&H variants thereof in the nodular L&H form of Hodgkin's disease (nodular lymphocyte predominant type) showed reactivity with PD7/26 but not with the anti-granulocyte markers. Rare L&H cells in 2 cases of diffuse lymphocyte predominant type showed reactivity with some, but not all, of the anti-granulocyte antibodies. These findings provide further support for the concept that the nodular L&H type of Hodgkin's disease represents an entity distinct from other forms of this disorder. Our studies also demonstrate the usefulness of these immunoperoxidase techniques when applied to formalinfixed, paraffin-embedded tissues.