Expression of Vimentin and Epithelial Membrane Antigen in Human Malignant Lymphomas

Immunoreactivity with monoclonal antibodies against the intermediate filament protein, vimentin, and epithelial membrane antigen (EMA) was examined in 330 cases of lymphoma (317 non-Hodgkin's and 13 Hodgkin's lymphomas), 12 reactive lymph nodes and mononuclear cells of the peripheral blood using either indirect immunoperox-idase staining or the avidin-biotin immunoperoxidase complex technique. The cell origin of each tumor was established using a panel of monoclonal antibodies against lymphocyte differentiation antigens. There were 41 T cell, 247 B cell and 29 undetermined lymphomas, and 13 cases of Hodgkin's disease in the series. Vimentin was expressed in 24 T-cell lymphomas (58.5%) and 60 B cell lymphomas (24.2%). This difference in frequency was statistically significant. Vimentin expression in follicular lymphomas was less frequent than in diffuse B-cell lymphomas. In diffuse lymphomas, small and medium cell types were more reactive with anti-vimentin than large cell types. Reed-Sternberg cells (R-S cells) in Hodgkin's disease were positive for vimentin in 11 cases (84.6%). The frequency of EMA reactivity in lymphomas was low, particularly in T cell lymphomas. No positive cases were found among follicular lymphomas. In diffuse non Hodgkin's lymphomas, EMA was expressed only in mixed and large cell types, but never in smaller ones. In conclusion, monoclonal antibodies against vimentin and EMA appear to be of limited usefulness for the diagnosis of non Hodgkin's lymphomas, but anti vimentin antibody may be used as an adjunct to the diagnosis of R-S cells in Hodgkin's disease.     

Study of vimentin expression in non-Hodgkin's lymphoma using paraffin sections.

Human non-Hodgkin's lymphomas were studied by means of an avidin-biotin complex immunoperoxidase method using several monoclonal antibodies against the intermediate filament protein, vimentin. The study cases were 61 B-cell lymphomas (including 2 plasmacytomas) and 30 T-cell lymphomas (including 8 cases of mycosis fungoides). Twelve of the 61 B-cell lymphomas were positive for vimentin, and were composed of extrafollicular-center cells such as immunoblastic and plasmacytoid cells. On the other hand, lymphomas of follicular center cell origin were negative for vimentin. All cases of T-cell lymphoma except for 14 (all of 9 AILD-type lymphomas, all of 4 lymphoblastic lymphomas and one diffuse mixed small/large lymphoma) were positive for vimentin. Although vimentin expression appeared to be influenced by various conditions such as the proportion of T- and B-cell subsets, or B-cell proliferation rate, follicular center cells were constantly negative for vimentin.     

波纹蛋白在NHL及RH中的表达及其在鉴别诊断中的意义

采用S-P法对38例非何杰金氏病淋巴瘤(NHL),40例淋巴组织反应性增生(RH)及3例何杰金氏病(HD)进行了波纹蛋白(Vimentin)标记,结果11例T-NHL中6例(54.5％)和27例B-NHL中22例(44.4％)表达Vimentin。Vimentin对NHL的分型作用是有限的。其中滤泡型淋巴瘤表达率低于弥漫型淋巴瘤;分化差的T淋巴母细胞性淋巴瘤及B小无裂细胞性淋巴瘤表达率低于中大细胞型淋巴瘤。Vimentin标记可显示RH组成细胞的多样性,显示血管分布的优越性以及某些滤泡型淋巴瘤表达Vimentin,而所有RH的生发中心Vimentin均呈阴性等特点有助于NHL与RH的鉴别断诊。另外Vimentin的表达亦是HD Reed Sternberg(R-S)细胞的有效标记物。     

Value of monoclonal antibodies in the diagnosis of Hodgkin's disease in paraffin sections]

A comprehensive panel of monoclonal antibodies that mark R-S/H cell, T- and B-cell, monocyte/histocyte was tested in paraffin sections of 107 cases of Hodgkin's disease (HD) including 21 cases of lymphocyte predominance (LP) and 86 cases of Non-LP HD. Thirty cases each of peripheral T-cell lymphoma and B-cell lymphoma were also tested for comparison. R-S/H cells were not stained with T-cell marker (UCHL-1) or monocyte/histocyte marker (Mac387) in all of these cases of HD. The results showed presence of certain difference in the phenotype of LP from non-LP. The H+L type of R-S/H cells of LP often reacted with B-cell markers including L26, LN2, LN1 and MB2 (93.3%-100%), LCA (83.3%) and EMA (92.3%), but rarely with LeuM1,T mü 9, or BerH2. On the contrary, most of the R-S/H cells of non-LP reacted with LeuM1 (80%), T mu 9(84%), BerH2(65%) but not with B-cell markers, LCA or EMA. Our study suggests a B-cell (probably the follicular center cell) derivation for L+H type of R-S/H cells in LP. The fact that 1 case of LP in this group transformed to a large cell B-cell lymphoma also supports this consideration. PNA is a sensitive marker of R-S/H cells but is not a specific one, since PNA stains 43.3% of the peripheral T-cell lymphoma and 20% of the B-cell lymphomas. Our findings indicate that using a panel of antibodies which mark R-S/H cells, T- and B-cells in paraffin sections will be helpful in the diagnosis and subtyping of Hodgkin's disease.     

Study of Vimentin Expression in Non-Hodgkin's Lymphoma Using Paraffin Sections

Human non-Hodgkin's lymphomas were studied by means of an avidin biotin complex immunoperoxidase method using several monoclonal antibodies against the intermediate filament protein, vimentin. The study cases were 61 B cell lymphomas (including 2 plasmacytomas) and 30 T cell lymphomas (including 8 cases of mycosis fungoides). Twelve of the 61 B cell lymphomas were positive for vimentin, and were composed of extrafollicular center cells such as immunoblastic and plasmacytoid cells. On the other hand, lymphomas of follicular center cell origin were negative for vimentin. All cases of T cell lymphoma except for 14 (all of 9 AlLD- type lymphomas, all of 4 lymphoblastic lymphomas and one diffuse mixed small/ large lymphoma) were positive for vimentin. Although vimentin expression appeared to be influenced by various conditions such as the proportion of T- and B cell subsets, or B cell proliferation rate, follicular center cells were constantly negative for vimentin.     

Cytokeratin expression and vimentin content in large cell anaplastic lymphomas and other non-Hodgkin''s lymphomas.

The immunophenotypes of 74 malignant lymphomas (9 Hodgkin's disease, 19 low-grade B-cell, 20 high-grade B-cell, 8 T-cell, and 18 large cell anaplastic lymphomas [LCAL]) have been characterized with antibodies against leucocyte differentiation antigens, keratin, and vimentin. All the non-LCAL were CD45 positive and keratin negative. The LCALs had a more varied immunophenotype, with CD45 present only in 11 of 18 cases and keratin present in 5 of 18 of these rare lymphomas. The lymphoid origin of these latter cases was proven by gene rearrangement studies. All LCALs were CD30+, and, where tested, vimentin positive. Of four different vimentin monoclonal antibodies tested, V9 and MVI stained the highest number of lymphomas. Positive staining of tumor cells was seen in 61 of 71 cases. Vimentin-negative cases included Burkitt's as well as some follicular lymphomas.     

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.     

Reciprocal/dichotomic expression of vimentin and B cell differentiation antigens in Reed-Sternberg's cells

Summary An immunohistochemical study of 63 cases of Hodgkin's disease was undertaken using formalin-fixed paraffin embedded tissue sections. The antibodies used were against L26, LN-1, LN-2, EMA (epithelial membrane antigen), Leu-M1, Vimentin, UCHL-1, S-100, and lysozyme. Hodgkin's disease could be divided into three groups: the first group was LN-1+/L26+/vimentin-, the second LN-1-/L26+/vimentin+, and the third LN-1-/L26-/vimentin+). Sixteen cases of follicular lymphomas were also examined and were all positive for LN-1 and L26 and negative for vimentin. Thus the vimentin negativity of the first group, including 7 nodular lymphocyte-predominant cases, gives further evidence of their germinal center B-cell origin. Since vimentin is expressed mainly in the immature stage of B-lymphocytes, the second group of Hodgkin's disease may represent immature B-cell Hodgkin's disease. In the third group, vimentin was present in Reed-Sternberg's (RS) and Hodgkin's (H) cells in 45 of the 48 cases (92.5%). In none of 48 cases were these cells positive for S-100 or lysozyme, but strong vimentin-positivity still suggested monocytic or histiocytic origin. The results of our study suggest, at least, divergent origin of RS's and H's cells.     

Use of monoclonal antibodies for analyzing the distribution of the intermediate filament protein vimentin in human non-Hodgkin''s lymphomas.

A series of human non-Hodgkin's lymphomas was examined for immunoreactivity with monoclonal antibodies to the intermediate filament protein vimentin with the use of an avidin-biotin immunoperoxidase method. The lymphoid cell nature of each tumor was established with the use of a panel of monoclonal antibodies to lymphoid cell differentiation antigens. There were 28 B-cell and 2 T-cell lymphomas in the series; of the 30 tumors, 11 (37%) were immunoreactive for vimentin. There was no correlation between vimentin immunoreactivity and the histopathologic type of lymphoma. In some tumors, there was nonspecific stromal immunoreactivity for vimentin, but the neoplastic lymphocytes were not immunoreactive. The selective expression of vimentin in non-Hodgkin's lymphomas may be due to masking of the appropriate epitopes or to selective expression of the vimentin gene in certain tumors. On the basis of these results, monoclonal antibodies to vimentin appear to be of limited usefulness in establishing the diagnosis of non-Hodgkin's lymphoma.     

Distribution of HNK-1+ cells in malignant lymphomas

HNK-1, a murine monoclonal antibody, is known to react with most of the natural killer (NK) and killer (K) cells in peripheral blood. Cells reacting with this antibody (HNK-1+ cells) were studied on tissue sections of ninety two cases of malignant lymphomas (MLs) by using immunoperoxidase technique, in an attempt to elucidate the role of this type of cells in MLs. Follicular lymphomas were found to be highly infiltrated with HNK-1+ cells. The mode of infiltration in follicular lymphomas is just like in normal germinal centers. Many cases of diffuse lymphomas with cleaved nuclei, indicative of diffuse B-cell lymphomas of follicular center cell origin, as well as diffuse ML with heavy fibrosis (sclerosis) or histiocytic reaction, were also found to be infiltrated with abundant HNK-1+ cells. Meanwhile, other types of B-cell ML and all types of T-cell ML, as well as Hodgkin's disease, were shown to be very poor in HNK-1+ cell reaction. From a prognostic viewpoint, the low grade malignancy group in the NCI Working Formulation or Kiel Classification was found to be infiltrated with significantly much more HNK-1+ cells as compared to the high grade malignancy group. The significance of these findings are discussed, with the stress on the possible suppressive function of HNK-1+ cells on proliferation and differentiation of follicular center cell type B-cell MLs.     

Genotypic analysis of large cell lymphomas which express the Ki-1 antigen

The monoclonal antibody Ki-1 reacts with Reed-Sternberg cells in Hodgkin's disease and with the tumour cells in a minority of large cell non-Hodgkin's lymphomas. This study describes the results of immunophenotypic and DNA analysis in 30 cases of non-Hodgkin's lymphoma, all of which expressed the Ki-1 antigen. The genotypic analysis has been undertaken using both immunoglobulin and T-cell receptor gene probes. Sixteen cases were shown by this method to be of monoclonal T-cell origin, six of B-cell origin, while in eight cases there was no evidence of either T- or B-cell lineage. This confirms previous immunohistological data indicating that non-Hodgkin's lymphomas which express the Ki-1 antigen may be of either T-cell or B-cell origin.     

Distribution of T-cell subsets in follicular and diffuse lymphomas of B-cell type.

The authors examined the number and distribution of cells reacting with monoclonal antibodies to T-cell subsets in frozen tissue sections of B-cell lymphomas (30 follicular and 17 diffuse lymphomas). In five diffuse lymphomas (two lymphocytic, three small cleaved cell) the neoplastic B-lymphocytes reacted with the monoclonal antibody anti-T1. In all other cases, the monoclonal antibodies to T-cell subsets reacted only with small lymphocytes concentrated between the follicles of follicular lymphomas and distributed randomly in diffuse lymphomas. The distribution of T cells and the T4+/T8+ ratio in follicular small cleaved and mixed lymphomas was similar, although not identical, to that seen in hyperplastic lymphoid follicles. Fewer T cells and a decrease in the T4+/T8+ ratio were seen in follicular large cell lymphoma and in diffuse large cell lymphomas. The number and distribution of T cells in follicular lymphomas is consistent with the hypothesis that there is a functional interaction between neoplastic B cells and benign T cells. No tumors were found in which the neoplastic B cells reacted with anti-T3, anti-T4, or anti-T8.     

Immunoperoxidase study of lymphomas. Comparison of a one-step frozen section technic with indirect methods on paraffin sections

Fifty cases of non-Hodgkin's lymphoma (15 nodular and 35 diffuse) were studied to determine the sensitivity, specificity, and ease of several different immunoperoxidase methods. The methods included a rapid, simple one-step immunoperoxidase procedure on frozen sections compared with indirect immunoperoxidase technics on paraffin sections. The frozen-section immunoperoxidase technic stained 15 of 15 nodular lymphomas and 24 of 35 diffuse lymphomas for monoclonal light chain. The majority of the diffuse lymphomas that did not stain for light chains were morphologically and immunohistochemically consistent with T-cell lymphomas. The indirect method on B-5 and formalin-fixed tissues only rarely displayed monoclonal staining for nonplasmacytoid small cell lymphomas but did stain some large cell lymphomas and a majority of plasmacytoid lymphomas for monoclonal light chain. The frozen section technic presented in this report is sufficiently sensitive and reliable to detect immunoglobulins in any morphologic subtype of B-cell lymphoma, whereas paraffin-embedded tissues have only limited application.     

DISTRIBUTION OF HNK-1+ CELLS IN MALIGNANT LYMPHOMAS

HNK-1, a murine monoclonal antibody, is known to react with most of the natural killer (NK) and killer (K) cells in peripheral blood. Cells reacting with this antibody (HNK-1⁺ cells) were studied on tissue sections of ninety two cases of malignant lymphomas (MLs) by using immunoperoxidase technique, in an attempt to elucidate the role of this type of cells in MLs. Follicular lymphomas were found to be highly infiltrated with HNK-1⁺ cells. The mode of infiltration in follicular lymphomas is just like in normal germinal centers. Many cases of diffuse lymphomas with cleaved nuclei, indicative of diffuse B-cell lymphomas of follicular center cell origin, as well as diffuse ML with heavy fibrosis (sclerosis) or histiocytic reaction, were also found to be infiltrated with abundant HNK-1+ cells. Meanwhile, other types of B-cell ML and all types of T-cell ML, as well as Hodgkin's disease, were shown to be very poor in HNK-1⁺ cell reaction. From a prognostic viewpoint, the low grade malignancy group in the NCI Working Formulation or Kiel Classification was found to be infiltrated with significantly much more HNK-1⁺ cells as compared to the high grade malignancy group. The significance of these findings are discussed, with the stress on the possible suppressive function of HNK-1+ cells on proliferation and differentiation of follicular center cell type B-cell MLs. ACTA PATHOL. JPN. 35: 339–350, 1985.     

Dendritic cells in the bursal follicles and germinal centers of the chicken's caecal tonsil express vimentin but not desmin

Background: Immunohistochemical studies with anti-vimentin and anti-desmin monoclonal antibodies were designed to determine the origin of bursal secretory dendritic cells (SDC) and follicular dendritic cells. Methods: The binding sites of anti-vimentin, anti-desmin, and antichicken-IgG specific monoclonal antibodies were visualized with a biotinylated anti-mouse-IgG, ABC Elite kit, and 4-chloronaphthol. Cells were double stained (anti-vimentin and rabbit anti-chicken-IgG Fc) to determine if the vimentin positive cells possessed surface IgG. Results: Vimentin positive cells were observed in the cortex and medulla of the bursa and germinal center and lymphoepithelial compartment of the caecal tonsil. The mesenchymal reticular cell, the basic supporting cell of the germinal center, was stained prominently by anti-vimentin and anti-desmin. Both antibodies stained the bursal cortex but only anti-vimentin bound the bursal secretory dendritic cell of the medulla. In addition to being vimentin positive and desmin negative, the bursal secretory dendritic cell possessed and the follicular dendritic cell appeared to possess IgG on their surfaces. In all the observations, B-cells were vimentin negative. Conclusion: These studies suggest that follicular dendritic cells and mesenchymal reticular cells in the caecal tonsil's germinal centers may be functionally different cell populations while the bursal secretory dendritic cell and follicular dendritic cell of the caecal tonsil may have a common origin. © 1995 Wiley-Liss, Inc.     

Immunohistochemical determination of CD23 expression in Hodgkin's disease using paraffin sections

The leukocyte antigen CD23 is expressed during B-cell development, and functions as an IgE receptor and a lymphocyte growth factor. We studied the expression of CD23 in paraffin sections of lymphoid tissue using the monoclonal antibody BU38. Fifteen cases of Hodgkin's disease, ten reactive lymph nodes, eight B-cell, and seven T-cell non-Hodgkin's lymphomas were analysed immunohistologically. CD23 positivity was seen on follicular dendritic cells and a small number of lymphocytes in reactive nodes. Thirteen of the 15 cases of Hodgkin's disease showed CD23 expression in both neoplastic cells and reactive lymphocytic infiltrate. The antigen was demonstrated in four of the B-cell and one of the T-cell tumours. CD23 may be important in mediating the mixed cellular infiltrate characteristic of Hodgkin's lymphoma.     

Primary gastric lymphomas: Morphologic, immunohistochemical and immunogenetic analyses

Morphologic, lmmunohistochemical and lmmunogenetic studies were performed on 28 cases of primary gastric lymphoma from fresh frozen tissue. Eight cases were diagnosed as diffuse large B-cell lymphoma, four as follicular center lymphoma (follicular), five as mucosa-associated lymphoid tissue (MALT) lymphoma, three as plasmacytoma, and three as T-cell lymphoma, two as mantle cell lymphoma, one as follicular center lymphoma (diffuse, predominantly small cell), and one as lymphoplasmacytoid lymphoma, and one as Hodgkin's disease.
From lmmunohistochemical studies, four types of morphologically similar low-grade lymphomas can be differentiated by a combination of various monoclonal antibodies. Cases of diffuse large B-cell lymphoma may have a germinal center origin. We observed lympho-epithelial lesions in cases of non-MALT lymphomas. We therefore consider that the current diagnostic criterion for MALT lymphoma may not always be valid.
Except for cases of T-cell lymphoma and Hodgkin's disease, 17 out of 22 cases revealed clonal rearrangement bands of the JH gene. In situ hybridization (ISH) and polymerase chain reaction (PCR) studies revealed the presence of Epstein-Barr (EB) virus genomes in two and three cases, respectively. Epstein-Barr virus may play a role in lympho-magenesis, although on relatively rare occasions.     

Clusterin expression in malignant lymphomas: a survey of 266 cases.

Clusterin expression has been reported to be characteristic of systemic anaplastic large cell lymphoma and usually negative in cutaneous anaplastic large cell lymphoma as well as other lymphoma types. We surveyed clusterin expression using immunohistochemical methods in 266 cases of non-Hodgkin's lymphoma and Hodgkin's disease to further assess the diagnostic utility of this marker. Clusterin immunostaining was observed in 40 of 49 (82%) systemic anaplastic large cell lymphomas and 12 of 29 (41%) cutaneous anaplastic large cell lymphomas. Clusterin also was expressed in 5 of 43 (12%) diffuse large B-cell lymphomas (4 of 5 CD30+), 1 of 14 (7%) peripheral T-cell lymphomas, 1 of 32 (3%) cases of nodular sclerosis Hodgkin's disease, and 1 case of mycosis fungoides in large cell transformation. Clusterin was negative in all other neoplasms assessed including follicular lymphoma of all grades (n = 24), mantle cell lymphoma (n = 13), marginal zone B-cell lymphoma (n = 12), precursor T-cell or B-cell lymphoblastic leukemia/lymphoma (n = 10), mixed cellularity Hodgkin's disease (n = 8), chronic lymphocytic leukemia/small lymphocytic lymphoma (n = 7), Burkitt lymphoma (n = 7), mycosis fungoides (n = 4), nodular lymphocyte predominant Hodgkin's disease (n = 3), lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (n = 2), and plasmacytoma (n = 2). We conclude that clusterin is a marker of anaplastic large cell lymphoma and that addition of clusterin to antibody panels designed to distinguish systemic anaplastic large cell lymphoma from classical Hodgkin's disease is useful. However, clusterin is also positive in a substantial subset of cutaneous anaplastic large cell lymphomas, a smaller subset of diffuse large B-cell lymphomas, and rarely in cases of peripheral T-cell lymphoma and nodular sclerosis Hodgkin's disease.     

Primary mediastinal non-Hodgkin's lymphomas: a morphologic and immunologic study of 19 cases

Nineteen, primary, non-lymphoblastic, non-Hodgkin's lymphomas were investigated by conventional morphologic studies as well as immunologic studies using the application of a battery of monoclonal antibodies to frozen tissue sections. Seventeen of the lymphomas were diffuse large cell; one was large cell immunoblastic and one was a follicular and diffuse lymphoma of intermediate differentiation. Thirty-seven percent of the lymphomas showed prominent sclerosis, sometimes associated with the superior vena cava syndrome. Six of the cases showed evidence of immunoglobulin production with light chain restriction. Twelve additional cases were shown to be of B-cell lineage by B1/T015 expression but did not show evidence of immunoglobulin production. One case was a T-cell lymphoma of helper phenotype. Ia expression was found in 14 of 18 cases studied.