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.     

Autocrine growth regulation of CD30 ligand in CD30-expressing Reed-Sternberg cells: distinction between Hodgkin's disease and anaplastic large cell lymphoma

Persistent expression of high levels of CD30 in Hodgkin's Reed-Sternberg (H-RS) cells and anaplastic large-cell lymphoma (ALCL) cells, but not in most T- or B-cell lymphomas, suggests the presence of an underlying mechanism leading to the abnormality and possible involvement of CD30 in the growth and survival of these two unique types of tumors. In this study, we examined the effect of CD30 ligand (CD30L) on CD30-positive H-RS and ALCL cells in long-term cultures or in primary cultures. CD30L induced various degrees of proliferation in three long-term cultured H-RS cell lines (L428, HDLM-2, and KM-H2) as well as in primary cultures of H-RS cells obtained directly from patients with Hodgkin's disease. In contrast, significant inhibition was observed in one of the ALCL cell lines (SU-DHL-1), but no growth inhibition or promotion in responding to exogenous CD30L was detected in three others (PB-1, JB-6, and McG-2), nor in primary cultures of ALCL cells. Expression of CD30L was determined by polymerase chain reaction in long-term cultured cells and by an immunohistochemical method in H-RS or ALCL cells de novo in tissue sections. None of the H-RS and ALCL cell lines was positive for CD30L. In tissue sections, we noticed that ALCL cells were generally CD30L-negative. In contrast, the anti-CD30L antibody reacted with a majority of H-RS cells with diffuse cytoplasmic staining. Most H-RS cells were CD30-positive, indicating co-expression of CD30 and CD30L in the majority of lymphoma cells. The persistent high levels of CD30 and CD30L expression in H-RS cells suggest that the autocrine CD30L-CD30 cytokine-receptor loop, in addition to having the paracrine activity previously thought to exist, could play important roles in the proliferation of H-RS cells. In contrast, the CD30L-mediated cytotoxicity may participate in the regression or slow progression of ALCL during the early phase of the disease in selected patients. However, when the disease progresses, the ALCL cells are likely to become resistant to exogenous CD30L.     

Differential regulation of leucocyte L-selectin (CD62L) expression in normal lymphoid and inflamed extralymphoid tissues.

AIMS: To study tissue expression of L-selectin, a leucocyte cell surface molecule that is considered to be involved in adhesion to certain endothelia, particularly in peripheral lymph nodes and during inflammation, and is shed upon leucocyte activation. METHODS: Leucocytes were examined by immunohistochemistry and double immunofluorescence staining in various lymphoid sites and normal and inflamed extralymphoid tissues. RESULTS: L-selectin was present on mantle zone B lymphocytes in different lymphoid sites, including in intestinal lymphoid tissue, but was absent on germinal centre B cells. Splenic white pulp B cells also expressed L-selectin. The proportion of T lymphocytes expressing L-selectin depended on the site under study, being greatest in peripheral lymph nodes (mean 48% of T cells positive), and lower in mucosal lymphoid sites and spleen (9 and 11% positive, respectively). Non-lymphocytic L-selectin staining was observed on follicular dendritic cells in tonsils and on macrophages in thymus. L-selectin positive leucocytes were rare in normal extralymphoid tissues, and relatively few were seen in most inflammatory settings. However, in rejecting renal transplants, a higher proportion (30%) of leucocytes expressed L-selectin. CONCLUSIONS: Overall, the results indicate how the degree of L-selectin expression by leucocytes in particular tissues may reflect a requirement for L-selectin expression for entry into those tissues and the activation state of leucocytes once localised there.     

Phenotypic expression of T lymphocytes in thymus and peripheral lymphoid tissues.

The pattern of expression of cell surface antigens and their relationship to the sequence of T-cell differentiation were delineated by the application of a series of monoclonal antibodies against T cells on tissue sections. The morphologic features and location of T-cell differentiation can be categorized into four stages: 1) subcapsular thymocytes, 2) cortical thymocytes, 3) medullary thymocytes, and 4) peripheral T cells. Phenotypically, on the basis of reactivity with monoclonal antibodies, T cells could be separated into two groups: immature and mature T cells. All stages of T cells expressed T200, Lyt 3, Leu 1, OKT3, Leu 9, and TA1, although staining intensities varied between thymic cortex and medulla. Mature T cells (medullary thymocytes and peripheral T cells) stained more intensely than immature T cells for some antigens, such as Leu 1, OKT3, and Leu 9, and, therefore, probably have a greater antigen density. Immature T cells, exemplified by subcapsular and cortical thymocytes, were characterized by the expression of Tdt, Leu M3, OKT6, OKT9, J5, BA-2, OKT10, and usually coexpressed both helper (T4/Leu 3a) and suppressor (T8/Leu 2a) antigens. With further maturation, medullary thymocytes and peripheral T cells lost reactivity for the above-mentioned markers, acquired A1G3 and Leu 8, and segregated into either T4+/Leu 3a+ or T8+/Leu 2a+ cells. A subpopulation of T cells in germinal centers differs from the majority of peripheral T cells by virtue of an absence of expression of Leu 8/A1G3. This histologically localized subset of T cells may be responsible for the mediation of helper function within the germinal center.     

Distribution of porcine CD4/CD8 double-positive T lymphocytes in mucosa-associated lymphoid tissues.

The present study describes the distribution of CD4/CD8 double-positive (DP) T cells in lymph nodes and mucosa-associated lymphoid tissues of pigs at 6-7 months of age. Proportions of lymphocytes isolated from peripheral, bronchial and mesenteric lymph nodes expressing CD4 and/or CD8 molecules were similar and ranged from 10-13% CD4/CD8 DP cells, 25-27% CD4 single-positive (SP) cells, and 27-32% CD8 SP cells. Mucosa-associated lymphoid tissues had significantly smaller proportions of CD4+ and/or CD8+ T cells than lymph nodes and CD4/CD8 DP cells accounted for a larger proportion of the total CD4+ lymphocytes than in lymph nodes. Compared to the lymph nodes, the predominant CD4+ and/or CD8+ T-cell subset in tonsils was the CD4/CD8 DP population (18%), because of both a higher proportion of these cells and a lower proportion of CD4 SP (12%) and CD8 SP (14%) lymphocyte subsets. Jejunal Peyer's patches were comprised of 12% CD4 SP, 28% CD8 SP and 10% CD4/CD8 DP lymphocytes. In contrast, the mid-section of the continuous Peyer's patch in the ileum contained 7% CD4 SP, 8% CD8 SP and 4% CD4/CD8 DP lymphocytes. The distribution of T lymphocytes in porcine mucosal lymphoid aggregates agrees with the reported correlation between high and low rates of lymphocyte entry into these tissues and the abundance or scarcity of T cells, respectively. Defining the role of CD4/CD8 DP lymphocytes and their unique distribution in mucosa-associated lymphoid tissues in the pig may reveal novel T-cell-mediated regulatory pathways.     

CD30 distribution. Immunohistochemical study on formaldehyde-fixed, paraffin-embedded Hodgkin's and non-Hodgkin's lymphomas.

Two hundred Hodgkin's and non-Hodgkin's lymphomas were immunohistochemically studied for the presence of the CD30 (Ki-1) activation antigen using a monoclonal antibody BerH2 on paraffin-embedded, formaldehyde-fixed tissue. Immunohistochemistry was performed by using the avidin-biotin complex technique and was preceded by enzymatic digestion with pepsin (0.05% for 20 minutes). Ninety percent (56/64) of cases of Hodgkin's disease, other than lymphocyte predominance type, showed positive tumor cells, although the positivity was often focal. In contrast, lymphocyte predominance type showed CD30 in only two of nine cases. CD30 was commonly seen in non-Hodgkin's lymphomas. Five of 37 large-cell lymphomas showed extensive CD30 positivity and morphologically represented large-cell anaplastic lymphomas ("Ki-1 lymphomas"). Apart from this, occasional CD30-positive cells were seen in nine of 32 large-cell non-Hodgkin's lymphomas. About half of the nodular small cleaved-cell lymphomas contained CD30-positive cells, two of them showing large numbers of positive cells both within and outside the nodules. Lymphocytic lymphoma sometimes (6/17) showed a few CD30-positive cells. Peripheral T-cell lymphomas showed positive cells in three of eight cases. The positive cases were one lymphoma with small groups of epithelioid cells (Lennert's lymphoma) and two immunoblastic lymphadenopathylike peripheral T-cell lymphomas. The results show that CD30 is more widespread than originally thought in non-Hodgkin's lymphomas and that especially nodular small cleaved-cell lymphomas often contain positive cells. These findings have to be considered in the immunohistochemical differential diagnosis of lymphomas. Obviously, CD30 alone cannot be used to differentiate between Hodgkin's and non-Hodgkin's lymphomas. The CD30-positive cells in non-Hodgkin's lymphoma may represent a link between Hodgkin's and non-Hodgkin's lymphomas.     

Cytokeratin and CD30 expression in dysgerminoma

Dysgerminoma is a malignant germ cell tumor of the ovary that shares morphological, immunophenotypic, and genetic features with its testicular counterpart, seminoma. Recent evidence supports the hypothesis that seminoma can differentiate into non-seminomatous germ cell tumor types. The progression of these tumors can be measured by their acquisition of the potential to express cytokeratin intermediate filaments, a characteristic specific to epithelial differentiation. Although testicular seminomas have been widely investigated, little is known about cytokeratin or E-cadherin expression in dysgerminomas. We investigated 26 formalin-fixed, paraffin-embedded ovarian dysgerminomas with immunohistochemical stains for CAM5.2, AE1/AE3, epithelial membrane antigen, cytokeratin 7, cytokeratin 20, high-molecular-weight keratin, and E-cadherin. In addition, we investigated the CD30 and vimentin immunoreactivity of these tumors. Immunoreactivity for CAM5.2 and for AE1/AE3 was present in more than 10% of neoplastic cells in 5 (19.2%) of 26 cases and in 2 (7.7%) of 26 cases, respectively. Cytokeratin 7 showed only focal positivity and never showed positive staining in greater than 10% of dysgerminoma cells. E-cadherin staining was positive in 2 cases showing weak membranous immunostaining in more than 10% of cells. Vimentin immunoreactivity was observed in only 2 dysgerminomas, both of which had less than 10% of the neoplastic cells staining. Cytokeratin 20, epithelial membrane antigen, high-molecular-weight keratin, and CD30 were consistently negative in all cases. Our study demonstrates that cytokeratin expression in dysgerminomas is not unusual and is consistent with the hypothesis that dysgerminomas have the capacity to differentiate along epithelial lines. Furthermore, the immunohistochemical staining patterns for cytokeratins, E-cadherin, and CD30 in dysgerminomas need to be considered when assessing differential diagnoses in difficult cases of primary ovarian tumors.     

Recombinant CD30 ligand and CD40 ligand share common biological activities on Hodgkin and Reed-Sternberg cells

The CD30 ligand (CD30L) and CD40L are members of the tumor necrosis factor (TNF) protein superfamily. CD30L and CD40L are mainly expressed as membrane-bound proteins by activated T cells. CD30L and CD40L are costimulatory for T cell proliferation and activation. Further, CD40L is a critical signal for T cell-dependent activation of B cells. Primary and cultured Hodgkin and Reed-Sternberg (H-RS) cells, the neoplastic component of Hodgkin's disease (HD), express high levels of the counterreceptors CD30 and CD40. We have found that both the recombinant CD30L and CD40L enhanced interleukin (IL)-6, TNF and lymphotoxin (LT)-α release from cultured H-RS cells. In addition, CD40L, but not CD30L, induced IL-8 secretion. CD30L and CD40L seem to share some redundant biological activities involved in the deregulated secretion of cytokines known to play a central role in the clinical presentation and pathology of HD. Further, CD30L enhanced surface expression of intercellular adhesion molecule-1 (ICAM-1/CD54) on cultured H-RS cells, which is frequently overexpressed on primary H-RS cells. CD30L- and CD40L-enhanced CD54 surface expression is followed by elevated shedding of CD54, as shown by detection of elevated 82-kDa soluble (s) CD54 levels in culture supernatants after stimulation with both ligands. CD30L and CD40L share common pleiotropic biological activities on CD30⁺/CD40⁺ H-RS cells and are elements of the cytokine and cell contact-dependent activation network typical for HD, a tumor of cytokine producing cells.     

Granulocytic sarcoma with expression of CD30.

A case of a young man with a spinal epidural tumour, initially diagnosed as large cell anaplastic malignant lymphoma, is reported. The tumour consisted of poorly differentiated cells showing immunoreactivity with antibodies directed against CD30 and CD45. Ten months later the patient developed acute myeloid leukaemia. The histological slides of the epidural tumour were reviewed, including additional enzymochemical and immunochemical stains. As the tumour showed immunoreactivity for myeloperoxidase and chloroacetate esterase, it was reclassified as a granulocytic sarcoma.     

CD30 expression in non-Hodgkin''s lymphoma

The CD30 antigen has been reported as the immunophenotypic hallmark of a recently described category of non-Hodgkin's lymphoma, termed anaplastic large cell lymphoma. From a series of approximately 500 lymphomas, 17 cases showing typical anaplastic features have been identified. They were strongly labelled by monoclonal antibodies recognizing CD30 (Ki-1 or BerH2). However, 36 other lymphomas, mainly high-grade, of non-anaplastic cytology also expressed CD30, either diffusely or focally, with a staining pattern identical to that seen in anaplastic large cell lymphomas. This clearly suggests that such lymphomas cannot be identified solely on the basis of being high-grade non-Hodgkin's lymphomas showing CD30 positivity. From the present results, the distinction between the anaplastic and non-anaplastic types would be better made with antibodies to epithelial membrane antigen than to CD30. Clinical data, available for 48 of the patients (16 with anaplastic large cell lymphomas and 32 with non-anaplastic) revealed no significant differences with regard to age at presentation, sex or clinical signs. A short-term follow-up study of 25 patients revealed that for the first 2 years after diagnosis there were no significant differences in patient survival between anaplastic large cell lymphoma, other CD30+ high-grade lymphomas and all high-grade non-Hodgkin's lymphomas considered together. These findings, which must be confirmed by larger studies, suggest that in a general lymphoma clinic there is probably little justification for differentiating anaplastic large cell lymphomas or CD30+ lymphomas from other high-grade non-Hodgkin's lymphomas.     

CD 30 expression in follicular lymphoma

The CD30 antigen is a characteristic phenotypic feature of Sternberg-Reed and Hodgkin cells and is also found in a subset of large cell non-Hodgkin's lymphomas. The finding of CD30 positive cells in some centroblastic/centrocytic (cb/cc) follicular lymphomas prompted us to characterize the presence and distribution of CD30 positive cells in this type of lymphoma, using the monoclonal antibody BerH2. CD30 positive cells were present in 17/19 of the cases studied, located mainly at the edge of the neoplastic follicles, but also in some cases in perinodular or T-cell areas. This distribution resembles that found in reactive tonsils and lymph nodes. The majority of these CD30 positive cells in cb/cc lymphoma seem to be B-cells, as suggested by their reactivity with B-cell markers demonstrated by double immunostaining. The nature of these CD30 positive cells is unclear, but they should be taken into consideration in the differential diagnosis of cb/cc lymphoma with lymphocyte predominance Hodgkin's disease.     

CD30 expression in follicular lymphoma

CONTEXT: CD30(+) anaplastic large cell lymphomas were originally described as being of T-cell, null cell, and B-cell origin. CD30, however, is not a specific marker of anaplastic large cell lymphoma and has been found to be expressed in reactive as well as neoplastic populations as a probable activation marker. In addition, CD30(+) cells have also been described in both diffuse large B-cell and follicular lymphomas (FLs), resembling the pattern seen in reactive tonsils and lymph nodes. OBJECTIVE: We report an index case of FL with CD30 expression, which on initial touch preparations and flow cytometric immunophenotyping revealed a prominent population of CD30(+) cells with marked cellular pleomorphism (anaplasia) in a background of typical FL. Immunohistochemistry of the paraffin section for CD30 in our index case confirmed unequivocal CD30(+) pleomorphic cells in the malignant nodules in occasional clusters. This case prompted a study of additional cases of FL for pattern of immunoreactivity with CD30 on paraffin sections. DESIGN: Twenty-two additional cases of FL (grades 1-3) were retrieved for CD30 immunoperoxidase staining as in the index case. RESULTS: This study demonstrated 32% of the additional cases of FL had definitive CD30(+), large, pleomorphic malignant cells by paraffin immunohistochemistry. In 2 cases (9%), the pattern of immunoreactivity with CD30 showed clustering and variable staining of large cells, as our index case. CONCLUSION: This study underscores the morphologic and immunophenotypic spectrum of FL that includes CD30 staining and cellular pleomorphism.     

Constitutive expression of tenascin in T-dependent zones of human lymphoid tissues.

Tenascin is a major extracellular matrix glycoprotein that can interfere with the action of fibronectin by inhibiting cell adhesion and spreading. Although tenascin is able to exert important immunomodulatory activities on T and B cells and macrophages, little is known about its distribution in different lymphohemopoietic tissues. In this study we have analyzed tenascin immunoreactivity on cryostat and paraffin sections of normal and pathological lymphoid tissues using two different monoclonal antibodies. We demonstrated strong tenascin expression in all peripheral lymphoid tissues, whereas it was barely detectable in the thymus and in bone marrow. In reactive lymph nodes, tenascin was mainly found in T-dependent zones, forming a variably close-woven reticular network corresponding to fibroblastic reticulum cells and blood vessels basal laminae, showing a partial co-localization with fibronectin. In B-dependent zones, tenascin was restricted to blood vessels. Using double-marker analysis, we performed a thorough study comparing tenascin expression in different compartments of lymphoid microenvironments. Tenascin network appeared much thicker in chronically stimulated tissues, where CD4+ lymphocytes with "memory" phenotype (CD45RO+/CD45RA-) were predominant, and at sites of ongoing inflammation. In particular, a striking increase of tenascin was observed in sarcoid lymph node, as well as in myasthenic hyperplastic thymuses. In addition, tenascin can be abnormally synthesized in tissue involved by various types of lymphomas, including Hodgkin's disease and hairy cell leukemia.     

Simultaneous expression of activated lymphoid cell-associated and granulocytic cell-associated antigens on Hodgkin's and Reed-Sternberg cells in Hodgkin's disease

The surface antigens of Hodgkin cells and Reed-Sternberg cells (H- and R-S cells), including lacunar cells, were analyzed with a large panel of monoclonal and polyclonal antibodies by an immunohistochemical method and an immunoelectron microscopic technique. H- and R-S cells in each histologic subtype of nodular sclerosis, mixed cellularity and lymphocyte depletion were stained similarly with anti-Leu-M1, anti-Leu-11b, TG8, anti-HLA-DR, anti IL-2R, RSC-1 (Ki-1) and anti-alpha-1-antitrypsin, but not with other antibodies examined. These findings suggest the following: (1) H- and R-S cells of nodular sclerosis, mixed cellularity and lymphocyte depletion are not heterogeneous, at least in terms of surface antigen expression, and (2) H- and R-S cells may be lymphoid cells which simultaneously express activated lymphoid cell-associated antigens (e.g., HLA-DR, RSC-1 and IL-2R) and granulocytic cell-associated antigens (e.g., Leu-M1 and TG8).     

Immunoglobulin idiotype expression in reactive lymphoid tissues and B-cell lymphomas

Summary In an investigation of the immunoglobulin idiotypic expression of non-tumour and neoplastic B lymphocytes in situ, fresh-frozen specimens of reactive tonsils, lymph nodes and B-cell malignant lymphomas (B-MLs) from Japanese patients were studied immuno-histochemically with 39 different anti-idiotype antibodies. In reactive lymphoid tissues, while idiotype-bearing cells were largely distributed sparsely in follicles and perifollicular areas, some were heavily crowded in particular germinal centres (GCs), suggesting the presence of oligoclonal proliferations of B-cells in GCs. Forty-eight out of 100 B-MLs reacted with anti-idiotype antibodies. This proportion was significantly higher than those reported in Western cases (27–36%), indicating that Japanese B-MLs share public idiotypes much frequently than western cases. The idiotypes demonstrated in these lymphomas, in contrast to those not expressed in any B-ML, were found much commonly in non-tumour lymphocytes, suggesting that such public idiotypes as were common in B-MLs were frequently shared by normal B-lymphocytes.