Follicular lymphoma with abundant PAS-positive extracellular material. Immunohistochemical and ultrastructural observations.

A finding of large amounts of extracellular amorphous material is rare in malignant lymphomas. We report four such cases of follicular lymphoma that were investigated by electron microscopy and immunohistochemistry using monoclonal antibodies. This extracellular material was clearly different from the sclero-hyalinosis commonly found in malignant lymphomas. Our observations revealed that the material is ultrastructurally composed of membranous structures, membrane-bound vesicles, and electron-dense bodies. Immunohistochemistry confirmed its neoplastic cell origin and its identity with cell membrane antigens (Dako-LC+, anti-B antibodies positive). In one case investigated using frozen sections, it showed the same immunophenotype as the neoplastic cells: SB4+ (CD19), SB2+ (CD21), TO15+ (CD22), SB3+ (CD37), IgML+, and Calla+. The mechanism for accumulation of this extracellular amorphous material is probably a function of a deregulation of cell wall synthesis and an exocytosis of cell membranes.

Monoclonal antibodies in the diagnosis of Hodgkin's disease. The search for a rational panel.

A novel, comprehensive panel of monoclonal antibodies was tested in a large series of routinely processed lymph node biopsy specimens from patients with Hodgkin's disease (69 cases), with the object of developing either definitive or adjunctive diagnostic criteria. B- and T-cell lymphomas and reactive states that could mimic Hodgkin's disease were also assessed with the same monoclonal antibody panel. In addition to the popularly used anti-Leu-M1 (CD15), the panel included the recently produced Ber-H2 (CD30) antibody, which detects a formalin-resistant epitope of the Ki-1 antigen. The other monoclonal antibodies were directed against epithelial membrane antigen (Dako-EMA) and leukocyte common antigen (Dako-LC) (CD45), as well as B-cell (LN-1 and LN-2) and T-cell (MT1) associated antigens. The results showed clear phenotypic separation of nodular lymphocyte predominant subtype of Hodgkin's disease from other subtypes. The lymphocytic and histiocytic cells of nodular lymphocyte predominant Hodgkin's disease were reactive for LN-1 (all cases) and anti-EMA (most cases) but negative for anti-Leu-M1 and Ber-H2. Within the other subtypes--i.e. nodular sclerosis and mixed cellularity--nearly all Reed-Sternberg cells and Hodgkin's cells were positive for both anti-Leu-M1 and Ber-H2. Ber-H2 monoclonal antibody was observed to react more frequently with Reed-Sternberg cells and Hodgkin's cells in Bouin's- or formalin-fixed tissues. Pleomorphic T-cell lymphomas, which could mimic Hodgkin's disease on morphology, created the same problem on phenotypic analysis. However, MT1 identified a significant proportion of T-cell lymphomas with Reed-Sternberg-like cells, having proven negative for Reed-Sternberg cells and Hodgkin's cells in Hodgkin's disease. Thus, a combination of anti-Leu-M1, Ber-H2, anti-EMA, LN-1, and MT1 monoclonal antibodies appears at present to be the most useful panel for the diagnosis and the differential diagnosis of Hodgkin's disease.

[Immunochemical diagnosis of hepatic localizations in malignant lymphoid hematologic diseases. Study of 80 cases]. / Diagnostic immunohistochimique des localisations hépatiques des hémopathies lymphoïdes malignes. Etude de 80 cas.

The diagnostic value of immunohistochemistry using monoclonal antibodies was assessed in 100 liver biopsy specimens. The majority of these cases were hepatic localizations of lymphoid malignancies. Ten normal and reactive inflammatory liver biopsies were used as controls. Some monoclonal antibodies directed against leukocyte antigens revealed unexpected reactivities with normal liver structures: biliary tract (anti-CD10, anti-B MB2) and hepatocytes (anti-B LN1). In 12/17 cases of hepatic involvement by large cell malignancy, immunohistochemistry allowed the diagnosis of non Hodgkin's lymphoma (NHL); the remaining 5 cases were metastatic undifferentiated carcinoma. It was difficult to differentiate small cell liver NHL from reactive inflammatory infiltration. New anti-B (MB1, MB2, 4KB5, LN1 and LN2) and anti-T (MT1 and UCHL1) monoclonal antibodies suitable for use on paraffin sections were of value to phenotype NHL when only fixed material was available. But, information was too limited to distinguish malignant from reactive infiltrates. Immunohistochemistry on frozen sections was often necessary to diagnose inflammatory infiltrates and to phenotype NHL. Most NHL were of B cell origin (11/13 cases) and showed monotypic surface immunoglobulins as well as B cell-associated antigens (CD22+). The expression of the T CD5 antigen by B-cell NHL may have some diagnostic value. When monotypic surface immunoglobulins could not be demonstrated (due to background staining) the expression of this antigen by B lymphocytes was considered to be highly indicative of their neoplastic nature. Hairy cell leukemia exhibited a pathognomonic phenotype on frozen sections (CD11c+, CD22+, CD25+). T NHL were rare (2 cases) and difficult to diagnose due to the lack of clonal markers. The diagnosis of Hodgkin's disease in liver (15/20 cases) was facilitated by using paraffin sections of both monoclonal antibodies anti-CD15 (Leu M1) and anti-CD30 (Ber-H2) which detect fixation-resistant antigens expressed by Sternberg cells.

[Value of immunoperoxidase with monoclonal antibodies after dismounting of previously stained sections]. / Intérêt de l'immunopéroxydase avec anticorps monoclonaux après démontage de coupes déjà colorées.

Immunoperoxidase using monoclonal antibodies may be applied to paraffin sections previously stained. The precise nature of cells found by routine staining may thus be investigated. It also enables, with small biopsies, immunohistochemical identification of a tissue zone lost by serial sectioning.

[Merkel cell carcinoma of the skin. Anatomoclinical, ultrastructural and immunohistochemical study of 14 cases]. / Carcinome à cellules de Merkel de la peau. Etude anatomo-clinique, ultrastructurale et immunohistochimique de 14 cas.

The clinical and pathological features of 14 cases of Merkel cell carcinoma are reported. They commonly arise in the skin of elderly patients, particularly on the face and pelvis. They have a loco-regional aggressivity (nodal metastases in 4 cases) but some patients die with disseminated metastases (2 cases). These tumors are composed of round cells with scanty cytoplasm, arranged in solid or trabecular sheets. Mitotic figures are usually numerous. The ultrastructural study reveal secretory granules and paranuclear collection of intermediate filaments. Immunohistochemical phenotype is highly characteristic: cytoplasmic diffuse positivity with an anti-neuron-specific enolase polyclonal antibody; polar and/or diffuse positivity with anti-cytokeratin, anti-epithelial membrane antigen and anti-S100 protein monoclonal antibodies; polar positivity with an anti-neurofilament monoclonal antibody. The negativity with an anti-common leucocyte antigen monoclonal antibody is helpful to differentiate Merkel cell carcinoma from cutaneous malignant lymphoma.

[Cells of the phenotype HNK-1 + (CD 57 +) in reactive adenopathies, lymphomas and Hodgkin's disease]. / Les cellules de phénotype HNK-1 + (CD 57 +) dans les adénopathies réactionnelles, lymphomateuses et Hodgkiniennes.

The number and distribution of Leu-7 + cells, a subset of Natural Killer cells clustered as CD 57 + cells, were studied with an immunoperoxidase technique on reactive lymph nodes (n = 13), malignant lymphomas (n = 60) and Hodgkin's disease (n = 22). Results of paraffin-section immunocytochemistry were compared with those obtained of frozen sections of the same tissues. CD 57 + cells are small lymphocytes mainly located in the germinal centers of reactive lymph nodes and in their malignant counterpart, i.e. the follicular lymphomas. The paragranuloma of Hodgkin's disease, type I nodular, contained high numbers of CD 57+ cells. Nine cases of CD 57+ lymphomas are reported, which were of high grade of malignancy. Their histological subtypes were anaplastic (3 cases) immunoblastic (2 cases), pleomorphic medium and large cell (3 cases), unclassifiable (1 case). Their diverse T-cell (4 cases) or B-cell (2 cases) origin and the various expression of epithelial membrane antigen (EMA) or Kil/Ber-H2 (CD 30) suggest an aberrant CD 57+ phenotype of a subset of large cell lymphomas.

[Immunoperoxidase study of normal and pathologic lymphoid tissue. Value of monoclonal antibodies]. / Etude en immunopéroxydase du tissu lymphoïde normal et pathologique. Intérêt des anticorps monoclonaux.

Immunoperoxidase study can be performed either on fixed and paraffin embedded biopsy specimens or on frozen sections. Advantages and limits of these two methods, as well as the results obtained on normal and pathologic lymphoid tissue are presented. Immunoperoxidase on paraffin sections (PAP technic) is a simple method which allows a good morphologic analysis. However, most of the fixatives destroy proteic antigens particularly those linked to the cell membrane. Thus surface immunoglobulins (S.Ig) cannot be detected. In contrast cytoplasmic immunoglobulins remain antigenic enough to be demonstrated in routine paraffin embedded sections. In lymphomas synthesizing monotypic immunoglobulins, the percentage of labelled cells varies from 5 to 80%. Beside the background staining, which can be attenuated by trypsinisation, absorption of extracellular substances is often responsible for a false positive staining. Pathologists are mainly confronted with the passive uptake of extracellular immunoglobulins (IgG K and IgG L), as well as other serum proteins (lysozyme etc...). Immunoperoxidase on frozen sections allows the use of monoclonal antibodies. A large number of surface and cytoplasmic antigens can be detected. First, the localization of B and T lymphocytes, NK cells, interdigitating cells and dendritic reticulum cells within the normal lymph node is described. In the second part, the interest of monoclonal antibodies in differential diagnosis between lymphoma and pseudo-lymphoma, and in phenotyping of lymphomas is discussed. Now, it is possible to perform an in situ immunologic characterization of most lymphomas. B cell lymphomas have sIg associated with other antigens (Pan B+, HLA-DR+). Cells of chronic lymphoid leukaemia and centrocytic (cleaved-cell) lymphomas frequently express T65 (T 101+ or Leu 1+) antigen which is usually found on normal or neoplastic T lymphocytes. Monoclonal antibodies provide new evidence of the germinal centre origin of follicular lymphomas. Thus, monoclonal antibody directed against dentritic reticulum cells (CRD) revealed the same network of DRC in follicular lymphomas as in reactive germinal centres. This finding could account for the nodular pattern of these lymphomas, neoplastic cells being in some way, enclosed within the DRC network. On the other hand, neoplastic follicles are surrounded by a large amount of t lymphocytes. Some T lymphocytes are also found within the follicles where they are associated with NK cells. Lastly, as reactive benign follicles, neoplastic follicles are labelled by the anti-Calla antibody.(ABSTRACT TRUNCATED AT 400 WORDS)

[Giant uterine leiomyoma and pregnancy. Clinical, radiologic, unusual histopathologic aspects]. / Léiomyome utérin géant et grossesse. Aspects cliniques, radiologiques, histopathologiques inhabituels.

The authors report a case of uterine leiomyomas diagnosed during the first trimester of pregnancy. The most unusual presentation of this observation did not allow any diagnosis through clinical exam, either by sonographic and X ray technics (magnetic resonance imaging, computer tomography) or by the classical histological study. The intra abdominal tumors compatible with imagery are argued: ovarian cyst, gelatinous disease of the peritoneum, desmoid tumor, lymphoma, lymphangiomyoma. The right diagnosis has been established as a last resort to immunocytochemistry.

[Diagnosis of undifferentiated tumors by means of monoclonal antibodies on paraffin sections]. / Diagnostic des tumeurs indifférenciées à l'aide d'anticorps monoclonaux utilisables sur coupes en paraffine.

The diagnostic value of 3 monoclonal antibodies applied on to routinely processed surgical biopsies was assessed. These antibodies were directed against keratin polypeptide (KL1), epithelial membrane antigen (DAKO-EMA) and leucocyte common antigen (DAKO-LC). First, using a three-step immunoperoxidase procedure, we determined the phenotype of well differentiated carcinomas (21 cases), non-Hodgkin's malignant lymphomas (44 cases), malignant histiocytoses (3 cases), melanomas (5 cases), sarcomas (6 cases) and miscellaneous tumors (16 cases). Nineteen out of the 21 carcinomas reacted with KL1 and DAKO-EMA antibodies but not with DAKO-LC. Forty out of the 44 non-Hodgkin's malignant lymphomas reacted with DAKO-LC. All these tumors were negative with KL1 antibodies but three of them, as well as 3 cases of malignant histiocytosis, expressed the epithelial membrane antigen. The value of these 3 antibodies was then assessed in the differential diagnosis of 30 undifferentiated tumors. A definite diagnosis was made in 28 cases: there were 11 undifferentiated carcinomas and 11 large cell malignant lymphomas. The phenotype of 6 tumors was highly suggestive of malignant histiocytosis, the peculiarity of which is to express both leucocyte common (DAKO-LC+) and epithelial membrane antigens (DAKO-EMA+). Only two tumors did not react with these 3 antibodies. We conclude that it is now possible to determine the nature of nearly all undifferentiated tumors on paraffin-embedded biopsy specimens.

[Coccidioidomycosis presenting as a pulmonary nodule after pneumonia]. / Opacité tumorale apparue au decours d'une pneumonie fébrile révélant une coccidioïdomycose thoracique.

Coccidioidomycosis is a fungal infection endemic in the south west of the United States. Sixty percent of infected individuals remain asymptomatic. Symptomatic disease manifests itself with variable signs such as pneumonia, pleural effusion, empyema or acute respiratory distress syndrome. Residual disease includes pulmonary nodules and fibrosis. We report a case of a woman, returning from a trip to Arizona, presenting with an acute respiratory infection associated with erythema nodosum and arthralgia. She was successfully treated with routine antibiotics. The acute pneumonia resolved and the radiological infiltrate contract into a solitary pulmonary nodule. We suspected a malignant nodule in a previous smoker. The diagnosis of pulmonary Coccidioidomycosis was made after surgical resection. One year later, the patient is asymptomatic and well. This review focuses on the most common clinical manifestations, the diagnostic strategy and the treatment and management of pulmonary Coccidioidomycosis.

Sinus histiocytosis with massive lymphadenopathy (Destombes-Rosai-Dorfman syndrome) occurring as a single enlarged submandibular lymph node: a light and immunohistochemical study with review of the literature.

Sinus histiocytosis with massive lymphadenopathy (SHML) is primarily a disease of children and young adults in which there is a pronounced and persistent cervical lymph node enlargement that usually is bilateral and is accompanied by fever. The histology, which varies according to the stage of the disease, is characterized by an exuberant intrasinusoidal histiocytic proliferation. The present case involves a 4-year-old girl who had several episodes of upper respiratory infection and otitis media; subsequently, a walnut-sized enlargement developed in the left anterior portion of the neck. Results of a physical examination were essentially normal. A laboratory work-up was noncontributory. Serologic tests for toxoplasmosis, infectious mononucleosis, and cat-scratch disease were negative. Immunoelectrophoresis disclosed normal values for IgG, IgM, IgA, and IgE. The histopathology was characteristic of SHML. The lymph node demonstrated pericapsular and capsular fibrosis and widely dilated subcapsular, trabecular, and medullary sinuses packed with histiocytes and plasma cells. "Lymphophagocytosis" and large atypical histiocytes resembling Reed-Sternberg cells were noted. Immunohistochemistry demonstrated a polyclonal population of plasma cells mostly stained with rabbit anti-human igG. The cytoplasm of the histiocytes, having ingested lymphocytes, was positively stained for IgG. Other groups of lymph nodes were affected during the next several months. The patient's condition has now been followed for 2 years, and the lymphadenopathy has almost completely regressed. The site distribution of the head and neck extranodal manifestations of SHML was analyzed in 54 cases.

Immunophenotyping of mesothelial cells and carcinoma cells with monoclonal antibodies to cytokeratins, vimentin, CEA and EMA improves the cytodiagnosis of serous effusions.

This paper presents an immunocytochemical study performed on cytocentrifuged deposits from 109 peritoneal and pleural effusions including 20 transudates, 43 malignant metastatic effusions and 46 effusions containing atypical cells, unidentifiable as reactive mesothelial or malignant epithelial cells on the classical morphological criteria. A panel of four monoclonal antibodies (MAb) was used, including KL1 directed to cytokeratins (KER), V9 to vimentin (VIM), NEO 723 to carcinoembryonic antigen (CEA) and E29 to epithelial membrane antigen (EMA). In most transudates the reactive mesothelial cells coexpressed VIM and KER with a ring-like pattern for the latter proteins. In contrast, they were unreactive to anti-CEA and weakly and inconsistently reactive to anti-EMA. In malignant effusions, most carcinoma cells coexpressed EMA, CEA and KER with a predominant diffuse cytoplasmic pattern for the latter. Only a few malignant epithelial cells from five metastatic adenocarcinomas weakly expressed VIM. When used on the 46 effusions with unidentifiable cells, the panel of MAb allowed reactive mesothelial cells and malignant epithelial cells to be distinguished from each other in 39 of 46 cases (85%).

Hodgkin's lymphoma of the oropharyngeal region: report of four cases and diagnostic value of monoclonal antibodies in detecting antigens associated with Reed-Sternberg cells.

In a review of more than 500 cases from the Lymphoma Registry, Department of Anatomic Pathology, University Hospital Purpan, Université Paul Sabatier, Toulouse, France, four cases of primary lesions in the oropharyngeal region were found. There were two lesions in the palatine tonsil, one in the nasopharynx, and one in the parotid gland. The average age of the patients was 54.5 years (range, 37 to 70 years), and all patients were men. The histologic types were lymphocyte predominance (one case), nodular-sclerosis (one case), and mixed cellularity (two cases). The patients were respectively staged as IAa, IIAa, and IIIA. They were treated with radiation, chemotherapy, or both. All four patients are now free of tumor and have been followed from 18 months to more than 6 years after definitive diagnosis and appropriate therapy. Immunohistochemistry significantly contributes to the differential diagnosis of atypical Hodgkin's disease from non-Hodgkin's lymphomas, especially in extranodal sites. Dako-EMA and Leu-M1 monoclonal antibodies are reactive, respectively, with L and H variants (Dako-EMA+, Leu-M1-) in the lymphocyte predominance type (Type 1) and with Reed-Sternberg cells (Dako-EMA-, Leu-M1+) in the nodular-sclerosis (Type 2), mixed cellularity (Type 3), and lymphocyte depletion (Type 4) types.

Reactivity of Leu 1 and T101 monoclonal antibodies with B cell lymphomas (correlations with other immunological markers).

The reactivity of Leu 1/T101 monoclonal antibodies (MoAb) was studied in a series of 69 lymphomas with B cell differentiation and was correlated with other cell markers. A three step immunoperoxidase technique on frozen sections was used to test a panel of 20 MoAb: anti-human Ig (heavy and light chains), To 15 (Pan B cells), Leu 1, T101, Leu 4, Leu 3a, Leu 5, OKT 8, OKT 6, Leu 7, anti-CALLA (IOT 5), Leu 10, anti-HLA-DR, OKM 1 and anti-dendritic reticulum cells (R 4/23). T101/Leu 1 antigen was detected in 24 cases: CLL (11 of 11), diffuse centrocytic lymphomas (four of 11), follicular lymphomas (none of 12), follicular and diffuse lymphomas (seven of 10) and one unclassified low grade lymphoma. This antigen was observed in only one high grade malignant lymphoma. In follicular lymphomas, two results deserve attention: (1) T101+ lymphomas showed most frequently IgM+, IgD+ surface Ig. Inversely, T101 unreactive lymphomas displayed IgM+, IgD+ phenotype. (2) Tp67 antigen (T101, Leu 1) and CALLA (GP 100) were found to be mutually exclusive in these lymphomas. These results suggest that follicular lymphomas could be derived from two distinct germinal center cell populations: IgM+ Ig'D-, Calla+, Leu 1-/T101- and IgM+, IgD+, CALLA-, Leu+/T101+.

Immunohistochemical profile of cutaneous B-cell lymphoma on cryostat and paraffin sections.

Thirty cases of primary (23 cases) and secondary (seven cases) cutaneous B-cell lymphoma (CBCL) were studied by immunohistochemistry using a selected monoclonal antibody (MoAb) panel on both cryostat and paraffin sections. On cryostat sections all CBCL so tested were positive for surface membrane immunoglobulins (IgMk most often) and B-cell antigens (CD22+, CD37+) with a variable T-cell-reactive component identified by MoAbs against T-cell antigens (CD2, CD3, CD4, CD5, CD8). CD4-positive stromal T-cells were usually more numerous than CD8-positive cells. A strong (50-75% of total cells) stromal T-cell (CD2+, CD3+) reaction was found in centroblastic-centrocytic lymphoma. Small numbers of CD1+ Langerhans cells were found in most cases, but they were present in large numbers in follicular lymphoma. On paraffin sections, a combination of MoAbs against B-associated antigens (LN-1, MB2) identified B-cell lineage in virtually all cases of CBCL. CBCL was negative for MoAbs against T-associated antigens (MT1, UCHL1) with rare exceptions (two cases). However, MT1 and UCHL1 combined identified the T-cell nature of all cases of nonepidermotropic, nonmycosis T-cell lymphoma, which were initially predictive of B-lineage by histologic pattern.

Stage IE nonHodgkin's lymphoma of the testis: a need for a brief aggressive chemotherapy.

Primary nonHodgkin's lymphoma of the testis is a localized disease in 50 per cent of the cases. Clinical records and pathological material from 9 stage IE cancer patients treated at our institutions were reviewed. All but 1 patient had B cell type lymphomas of intermediate (6) or high (3) grade according to the Working Formulation. Mean survival was 49 months and actuarial survival was 74 per cent at 5 years. Chemotherapy differed with time and frequently was associated with subdiaphragmatic involved field and prophylactic contralateral testis radiotherapy. In view of the good prognosis of patients receiving doxorubicin-based chemotherapy and recent reports on low stage nonHodgkin's lymphoma we recommend an aggressive brief therapy for stage IE lymphoma of the testis after orchiectomy.

Diagnostic features of primary malignant lymphomas of the thyroid with monoclonal antibodies.

Primary malignant lymphomas (ML) of the thyroid are rare and their conclusive morphologic diagnosis is not always possible. The authors report diagnostic features of 11 cases of ML and one case of plasmacytoma of thyroid compared with chronic lymphocytic thyroiditis and undifferentiated carcinomas of thyroid in an immunohistochemical study using monoclonal antibodies (MoAb). The lymphoid nature of tumors could be identified in all cases with three MoAb on paraffin sections. In ML, tumor cells expressed leucocyte common antigen (Dako-LC+) with negativity for epithelial membrane antigen (Dako-EMA-) and cytokeratin (KL1-). Newer MoAb identifying B-cell (LN-1, LN-2, MB2) and T-cell-associated antigens (MT1, UCHL1) not denatured by fixation, revealed B-cell nature of tumor cells in all cases of ML. Among anti-B MoAb, LN-1 and MB2 were most consistent in their reactivity. In cryostat sections of three ML cases, the tumor cells expressed one or more B-cell-associated antigens. Plasmacytoma was negative for Dako-LC and KL1 but positive for Dako-EMA and monotypic cytoplasmic Ig.