Distribution of the carcinoembryonic antigen CEA in gastric lesions. Immunohistochemical testing of three novel monoclonal antibodies.

Three mouse monoclonal antibodies MAB (CEA 12-140-1, -2 and -4) raised against different CEA epitopes were tested in 32 gastric adenocarcinomas (18 intestinal type and 14 diffuse type) and 34 gastric lesions with severe and moderate dysplasia. The MAB stained 13, 11 and 13 out of the 14 diffuse carcinomas and 11, 13 and 13 out of the 18 intestinal carcinomas. The dysplastic lesions were positive in 9, 9 and 6 out of 34 cases. Less than half of the cases with metaplastic epithelium adjacent to the carcinomas were also positive for MAB. All MAB showed the same pattern of reactivity without cross-reactivity. Their cumulative staining rate corresponded closely to that of polyclonal CEA antiserum, but the MAB stained more cells. The reactivity was confined to intracytoplasmic vacuoles in diffuse carcinomas and appeared diffusely in the cytoplasm or limited to the cell membrane in intestinal type of carcinomas. Our findings do not indicate CEA to be a reliable marker for malignant transformation in gastric mucosa.

Ki67 and 4F2 antigen expression as well as DNA synthesis predict survival at relapse/tumour progression in low-grade B-cell lymphoma.

Previous work has shown that parameters of cell activation studied on lymphoma biopsies can be used to discriminate between low-grade and high-grade non-Hodgkin's lymphomas and to predict prognosis in the low-grade malignancy group alone. We have now examined expression of several activation antigens and indicators of DNA synthesis in 29 patients with low-grade malignant B-cell lymphomas at the time of primary diagnosis and later at relapse and/or tumour progression. At both times, the level of 4F2 antigen expression examined by flow cytometry on cells in suspension as well as the number of Ki67 antigen-positive cells examined by immunohistochemistry were predictive of patient survival. DNA synthesis estimated by (3H-TdR) thymidine incorporation was of prognostic value at the second biopsy only. These parameters were more sensitive than histological demonstration of morphological transformation in secondary high-grade lymphomas in identifying high-risk patients at repeated biopsy. We propose that Ki67 or 4F2 expression or a marker of DNA synthesis (such as 3H-TdR incorporation or labelling index) should be evaluated when repeated biopsies are performed, in order to select patients for whom aggressive chemotherapy may be considered.

Low-grade intestinal lymphoma of intraepithelial T lymphocytes with concomitant enteropathy-associated T cell lymphoma: case report suggesting a possible histogenetic relationship.

In a low-grade small cell intestinal lymphoma found in a 41-year-old woman with celiac disease, the neoplastic intraepithelial lymphocytes (IELs) showed cytoplasmic granules, expressed the alpha/beta T cell antigen receptor (TCR), and stained positively for the IEL antibody, HML-1. The tumor cells carried the CD3 and CD7 T cell antigens, but were double negative for CD4 and CD8 and also lacked other T cell antigens (CD1, CD2, and CD5). Tumor cell monoclonality was proven by the demonstration of a rearranged TCR-beta chain gene. The special marker profile of the lymphoma may be explained by partial antigenic deletion of the phenotype which characterizes the majority of normal IELs (TCR alpha/beta+, CD3+, CD8+, or CD4+). Alternately, the tumorous cell clone might originate from an as yet unrecognized TCR alpha/beta+, CD3+, CD7+, CD4-, CD8- normal subset of IEL. In the presented case, a concomitant high-grade large cell lymphoma supports the assumption that enteropathy-associated T cell lymphomas (EATCLs) derive from the intestinal IEL population and suggests that EATCLs may be preceded by a low-grade IEL lymphoma.

[Aspiration cytology in palpable breast changes of the breast. Too many specimens without cell material to examine]. / Aspirasjonscytologi ved palpable forandringer i mamma. For mange prøver uten diagnostiserbart cellemateriale.

During the first half of 1988, 927 fine needle aspiration cytology samples from palpable non-cystic breast lesions were examined in our laboratory. Among 582 aspirations performed by members of the laboratory medical staff the frequency of technically non-diagnostic specimens was 4.3 per cent, as against 34.2 per cent among 345 referred specimens. In the latter group there was a clear relationship between the frequency of inadequate specimens and experience in aspiration technique as judged from the number of referred specimens from the individual clinician. It is concluded that education and training in aspiration technique is needed to improve the quality of this diagnostic method.

[Giant cell anaplastic anemia]. / Storcellet anaplastisk lymfom.

The Ki-1-antibody was raised against a cell-line of Hodgkin-cells, and reacts with most Hodgkin- and Reed-Sternberg-cells. By use of this antibody a new entity of non-Hodgkin's lymphomas has been identified; the anaplastic large cell lymphoma. We report our experience from studying 16 such tumours diagnosed during the period October 1986 to February 1988. In three cases the anaplastic large cell lymphoma appeared in patients who presented Hodgkin's disease as well. Of 15 cases studied using frozen section immunohistochemistry, ten showed T-, one B-, and one monocyte/macrophagephenotype, whereas three cases presented a mixed T-, B- and monocyte/macrophagephenotype. All cases showed a high proliferative activity as evaluated by presence of activation antigens. In paraffin sections six of 16 cases expressed epithelial membrane antigen (EMA). In 13 cases EMA, leucocyte common antigen (LCA) and Ber-H2 (a Ki-1-antibody) reactivity was compared in paraffin and frozen sections. The EMA reactivity was of nearly equal intensity in paraffin and frozen sections, while the reactivity of LCA and Ber-H2 was significantly reduced in paraffin sections, implying only partial preservation of the antigens. Clinically, most cases were in an advanced (III and IV) stage by the time of diagnosis. Of 14 actively treated patients, nine went into complete and two into partial remission, while three patients did not respond. Six patients are dead, five after less than one year. The coexistence of anaplastic large cell lymphoma and Hodgkin's disease and the common Ki-1 (CD30) antigen expression in these tumours suggest a possible histogenetic relationship.

[Multicentric Castleman's disease]. / Multisentrisk Castlemans sykdom.

We describe a case of histologically proven multicentric Castleman's disease in a 68 year old man. The clinical picture was dominated by severe hemolytic anemia. The outcome was fatal despite active treatment. We discuss the main pathological and clinical characteristics distinguishing multicentric Castleman's disease from the localized variant. The different age distribution, localized versus multicentric disease, different responses to treatment, and outcome, indicate that the two types of Castleman's disease represent different entities presenting common histological features in the lymphoid lesions.

Carcinoma of the nasopharynx. Histopathological examination with supplementary immunohistochemistry.

One hundred and one nasopharyngeal malignancies, clinically accepted and treated as carcinomas, were histologically reviewed. Originally, all of them had been given the histopathological diagnosis of carcinoma or possible carcinoma. A wide variety of diagnostic formulations had been used, some of them inconclusive. The review was based on strict morphological WHO criteria, and a definite diagnosis was attained in most cases. Three of the neoplasms, however, did not fulfil the criteria of carcinoma, and were given the diagnosis of malignant tumour, probable lymphoma. Immunohistochemistry with routinely processed tissue was performed on 69 of the poorly differentiated non-keratinizing neoplasms, including the three possible non-Hodgkin's malignant lymphomas. The neoplasms were positive for cytokeratin PKK1 with four exceptions: the three possible lymphomas and a large cell tumour with epithelial growth and prominent nucleoli which was found to be positive only for neurone-specific enolase. Two of three possible lymphomas were verified as such by being positive for leucocyte common antigen. This study showed that the WHO classification is quite useful when strictly applied. The histopathological diagnosis of this category of neoplasms can easily be confirmed by immunohistochemistry on routinely processed material and this adjunct can usually resolve questionable cases.

The activation-associated antigen 4F2 predicts patient survival in low-grade B-cell lymphomas.

Expression of the activation-associated 4F2 antigen, transferrin receptor and interleukin-2 (IL-2) receptor on suspended cells from 75 biopsied low-grade non-Hodgkin lymphomas (L-NHL) of B-cell origin was correlated to patient survival, clinical prognostic parameters and estimated DNA synthesis. 4F2 antigen expression correlated significantly with poor patient survival, high DNA synthesis and transferrin receptor expression. Transferrin receptor expression was associated with high DNA synthesis and treatment response, but not with patient survival. On the other hand, IL-2 receptor was correlated neither to patient survival nor to other studied markers for cell activation, but seemed to be expressed on certain subsets of lymphomas. We suggest that monoclonal antibody (MAb) against the activation-associated 4F2 antigen could be used to select patients with L-NHL for aggressive chemotherapy.

Single cell studies on the immunological marker profile of plasmacytoid T-zone cells.

Plasmacytoid T cells (PTC) are known to home to thymic (T) zones in human lymph nodes and are characterized by their abundant, concentrically layered, rough endoplasmic reticulum. These cells have been found in reactive and neoplastic conditions. Three cases of PTC lymphomas have so far been reported. All of them were complicated by a myelomonocytic leukemia leading to the assumption of a functional relationship between PTC and the myeloid system. The immunologic phenotype of PTC, as revealed on frozen tumor tissue sections, comprised the expression of CD5 (T1), CD4 (T4), and HLA-DR, but not CD8 (T8) and CD2 (T11) and suggested an affiliation to the T cell system. Extending our previous report on one of these cases we here present the first study on the immunological marker profile of suspended PTC. The employment of unfractionated or PTC-enriched tumor cell suspensions rendered possible the application of a panel of monoclonal antibodies (moAbs) on both fixed and unfixed cells and enabled us to allocate various markers either to the intracytoplasmic or surface domain of this cell type. Our results suggest that PTC from our case rest in the G0/G1 phase of the cell cycle. They express the transferrin receptor, but not the Il-2 receptor (CD25) or the nuclear antigen Ki-67. No T cell antigen was demonstrated on the surface of unfixed suspended PTC. Under these conditions only HLA-DR and a predominantly monocytic antigen (CD36/moAb 5F1) were identified. Fixed cells, however, showed a weak cytoplasmic reactivity for CD5 and two myelomonocytic antigens (CD15/moAb 1G10 and CD14/moAb My4). Our findings do not sustain positive evidence for a T cell nature of PTC. Whether their phenotypical pattern indicates terminal differentiation with concomitant loss of T cell antigens or points to a cytogenetic relationship of PTC to the myeloid system, remains speculative. Until the cytogenesis of PTC is clarified we propose the noncommitted term "plasmacytoid T-zone cells" for this elusive cell type.

T-zone lymphoma with predominance of 'plasmacytoid T-cells' associated with myelomonocytic leukaemia--a distinct clinicopathological entity.

In this paper we present a further case of a new clinicopathological entity combining a rare type of non-Hodgkin lymphoma with a myelomonocytic leukaemia. The characteristic feature of the lymphoma is massive infiltration of the T-zones of lymph nodes by plasmacytoid cells originally described by Lennert in non-specific lymphadenitis. Two lymphoma cases of this type have recently been published, by Müller-Hermelink et al. who named the cells 'plasmacytoid T-cells' (PTC), and by Prasthofer et al. These three cases have similar clinical and pathological features and appear to form a distinct clinicopathological entity. In contrast to the two previously published cases the present lymphoma also contained irregular lymphoid cells accompanying the PTC in the lymph node lesion and focally infiltrating the bone marrow. An accumulation of polytypic IgG positive plasma cells was observed in the remaining lymph node follicles. Immunohistological analysis with a range of monoclonal antibodies showed the PTC of our case to be CD5(T1)+, CD4(T4)+, CD3(T3)-, CD8(T8)-, CD2(T11)-, and CD25(TAC)-, but HLA-DR+ and transferrin receptor positive. The nature of this peculiar lymphoid lesion and its relationship to myelomonocytic leukaemia are discussed.

Sarcoid reaction of hilar and paratracheal lymph nodes in patients treated for testicular cancer.

Bilateral hilar lymph node enlargement developed in four patients 1 to 8 years after successful radiotherapy for testicular cancer Stage I was performed. Two had additional paratracheal involvement. Biopsy showed sarcoid reaction in all four cases. An increased frequency of sarcoid reaction is assumed in patients treated for testicular cancer. The pathogenesis remains unknown. The clinical and radiologic findings should not be misinterpreted as showing recurrent malignant disease. In an otherwise disease-free patient in whom mediastinal lymph node enlargement develops, a biopsy should always be performed.

Immunologic subsets in B cell lymphomas defined by surface immunoglobulin isotype and complement receptor--their relationship to survival.

Cell surface marker profiles were studied on cell suspensions from monoclonal B cell lymphomas. Surface immunoglobulin (sIg) was examined in 178 cases, whereas combined data with sIg and receptors for complement (C3) were available in 99/178 cases. The results showed that B cell lymphomas can be divided into distinct immunological subsets according to surface marker expression. Whereas some histologic subgroups (diffuse centroblastic/centrocytic, centrocytic, immunocytoma) (Kiel classification) consisted of few immunological subtypes, others were more heterogeneous (follicular centroblastic/centrocytic, centroblastic, lymphocytic). By combining immunological and histological subgroups, more than 40 phenotypes could be identified; this diversity most likely reflects, the heterogeneity of the B cell compartment. As part of the same study the prognostic significance of cell marker phenotypes was examined. Survival analysis undertaken on 149/178 patients did not uncover any significant relationship between type of heavy or light chain expression, C3 receptor expression and clinical outcome. Our data do not confirm recent findings that the type of immunological phenotype may be of prognostic significance.

3H-thymidine uptake in B cell lymphomas--relationship to treatment response and survival.

Cell suspensions were obtained from biopsy tissue from 149 patients with B cell lymphomas and analysed with regard to DNA-synthesis as assessed by 3H-thymidine uptake, response to therapy and survival. The 3H-thymidine uptake was significantly increased in lymphomas of high versus low grade malignancy (p = 0.0001), in patients with stage I and II versus stage III and IV (p = 0.014), and in patients with general symptoms (p = 0.0025) as opposed to asymptomatic cases. The complete response rate was significantly higher in patients with increased thymidine uptake than in those with low uptake, 26/51 (51%) cases versus 24/83 (29%) cases, respectively (p = 0.014). 55 patients with increased 3H-thymidine uptake survived for significantly shorter times than (94 patients) with low uptake (p = 0.0056). Furthermore, a markedly larger group of high-risk patients was identified by the 3H-thymidine assay than by histopathology alone, 55 cases versus 23 cases, respectively. Among the patients (126 cases) with low grade tumours, those with increased 3H-thymidine uptake (40 cases) had poorer outcome than those with low uptake (86 cases) (p = 0.045). The data suggest that DNA-synthesis in this study, as assessed by 3H-thymidine uptake, is an independent indicator of survival in NHL. Furthermore, it may be a useful parameter in laying down guidelines for therapy in B cell neoplasms, especially in low grade tumours.

Induction of maturation in different types of B-cell lymphomas in vitro with TPA and antibodies to surface immunoglobulin.

Cells from eight selected cases of human non-Hodgkin lymphomas of various histological types (lymphocytic, centrocytic, centrocytic/centroblastic, and immunocytomas) were stimulated in vitro with 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and anti-immunoglobulins (anti-Ig) against the surface immunoglobulin (sIg) on the tumour cells. Six of these cases responded by intracellular Ig accumulation as measured by flow cytofluorometry and direct phenotypical change into immunoblasts/plasmablasts as detected by light microscopic immunocytochemistry. However, the response to TPA alone varied considerably from case to case. These findings suggest that many, if not all, B-cell subsets have the capacity to develop directly into Ig-synthesizing cells (immunoblasts and plasmablasts). However, conditions for eliciting such events may vary, depending on the phenotypical properties and differentiation stage.

Induction of maturation of human B-cell lymphomas in vitro. Morphologic changes in relation to immunoglobulin and DNA synthesis.

In vitro stimulation of cells from 8 non-Hodgkin's lymphomas comprising several histologic types with a tumor promotor (TPA) and with or without anti-immunoglobulins directed against the surface immunoglobulin of the tumor cells is reported. Morphologic transformation to immunoblastic and plasmablastic cells, but not to plasma cells, and induction of Ig and DNA synthesis were observed. A comparative analysis, including flow cytofluorometry, light microscopy combined with immunocytochemistry, and electron microscopy, suggests that the three events may not always be associated phenomena at the single-cell level even in monoclonal cell populations.

Hexagonal crystals in the bone marrow in patients with myeloproliferative disease and preleukaemia.

In epoxy resin-embedded bone marrow biopsies from 238 cases of myeloproliferative disease and preleukaemia, crystalline inclusions were found in the bone marrow cells of 24 (10.1%), most often in patients with acute myeloproliferative disease (23%). By light microscopy the crystals were easily recognized after Giemsa staining, lying within the cytoplasm of bone marrow macrophages. By electron microscopy the hexagonal crystals were encountered chiefly within secondary lysosomes of the macrophages, but also within immature myeloid cells in one of the two cases studied. In the latter a tight-fitting membrane around the crystals suggested that they were of lysosomal origin and represented a form of abnormal granulation. We suggest that the presence of crystals in myeloid cells represents a kind of abnormal granula formation in the neoplastic cells. In the macrophages the crystals lodge in secondary lysosomes, probably after phagocytosis of crystal-bearing non-viable myeloid cells. The accumulation of crystals in lysosomes indicates resistance to degradation.