Polymerase chain reaction in the diagnosis of T-cell lymphoma in paraffin-embedded bone marrow biopsies: a comparative study.

AIMS: In routine histological analysis of bone marrow biopsies, the distinction between reactive T-cell infiltrates and T-cell lymphoma can be difficult, even with the use of extensive immunohistochemistry. The aim of this study was to evaluate the diagnostic contribution of TCR-gamma gene rearrangement analysed by PCR. METHODS AND RESULTS: The samples studied consisted of 46 paraffin-embedded bone marrow biopsies (diagnosis, staging and follow-up) from 26 patients with T-cell lymphoma. The bone marrow biopsies were categorized into three groups according to the morphological and immunohistochemical results. Group 1, positive for T-cell lymphoma (24 bone marrow biopsies), group 2, suspicion of T-cell lymphoma (15 bone marrow biopsies) and group 3, negative for T-cell lymphoma (seven bone marrow biopsies). DNA could be amplified in 45/46 bone marrow biopsies (98%). Clonal rearrangement was detected in 30/45 bone marrow biopsies tested (67%) including 15/24 bone marrow biopsies (62.5%) of group 1, 11/14 (78.5%) of group 2 and 4/7 (57%) of group 3. In total, PCR analysis supported a diagnosis of T-cell lymphoma in 15/45 bone marrow biopsies (33%), in which histological and/or immunohistochemical examination provided inconclusive evidence of malignancy. CONCLUSIONS: TCR-gamma PCR is a complementary tool for the assessment of T-cell lymphoma in bone marrow biopsies. Optimal evaluation of bone marrow biopsies requires an integrative approach of all available results from morphology, immunohistochemistry, molecular biology and clinical data.

CD34/QBEND10 immunostaining in bone marrow biopsies: an additional parameter for the diagnosis and classification of myelodysplastic syndromes.

CD34/QBEND10 immunostaining has been assessed in 150 bone marrow biopsies (BMB) including 91 myelodysplastic syndromes (MDS), 16 MDS-related AML, 25 reactive BMB, and 18 cases where RA could neither be established nor ruled out. All cases were reviewed and classified according to the clinical and morphological FAB criteria. The percentage of CD34-positive (CD34 +) hematopoietic cells and the number of clusters of CD34+ cells in 10 HPF were determined. In most cases the CD34+ cell count was similar to the blast percentage determined morphologically. In RA, however, not only typical blasts but also less immature hemopoietic cells lying morphologically between blasts and promyelocytes were stained with CD34. The CD34+ cell count and cluster values were significantly higher in RA than in BMB with reactive changes (p<0.0001 for both), in RAEB than in RA (p=0.0006 and p=0.0189, respectively), in RAEBt than in RAEB (p=0.0001 and p=0.0038), and in MDS-AML than in RAEBt (p<0.0001 and p=0.0007). Presence of CD34+ cell clusters in RA correlated with increased risk of progression of the disease. We conclude that CD34 immunostaining in BMB is a useful tool for distinguishing RA from other anemias, assessing blast percentage in MDS cases, classifying them according to FAB, and following their evolution.

Prognostic value of follicular dendritic cells in nodular sclerosing Hodgkin's disease.

AIMS: In nodular sclerosing Hodgkin's disease (NSHD), the prognostic relevance of the histopathological grading in two subtypes NSI (low-grade) and NSII (high-grade) remains controversial. Analysis of follicular dendritic cells (FDC) may provide new prognostic parameters. METHODS AND RESULTS: Tumours from 59 patients with NSHD were studied. Mean follow-up time was 8 years. Forty-one cases were classified as NSI and 18 as NSII. FDC were immunostained with the paraffin-resistant monoclonal antibodies CD21 and CNA.42. We distinguished three patterns in the neoplastic tissue: FDC1, the presence of well-defined follicle-like structures (n = 20); FDC2, the presence of largely destroyed FDC networks (n = 25); and FDC3, no or a few isolated FDC (n = 14). The three groups differed clearly regarding the frequency of relapse and the survival. The longest survival was seen in the FDC1 group, the shortest in the FDC3 group, the FDC2 group being intermediate (P = 0.0025). FDC status was a discriminating prognostic factor for all patients, and within various age and stage categories. Combining the FDC status and the NSI-NSII grading defined the best survival group as FDC1-NSI. CONCLUSIONS: Assessment of FDC pattern, associated with histological subtyping, brings valuable data for predicting survival and outcome in NSHD.

Systemic mastocytosis following a malignant ovarian germ cell tumour.

Cases of mediastinal germ cell tumours associated with haematological disorders (two cases of systemic mastocytosis included) have been reported previously. This combination is more frequent than would be expected by chance alone. We report the case of a 30-year-old woman, who presented with a systemic mastocytosis following a malignant ovarian germ cell tumour which was treated by chemo- and radiotherapy. The patient predominantly complained of skeletal pains, which led to an erroneous radiological diagnosis of fibrous dysplasia for years. An aggressive variant of systemic mastocytosis was diagnosed on bone marrow examination. Systemic mastocytosis was confirmed by splenectomy, liver biopsy and finally autopsy. The present case is unique because of the ovarian location of the germ cell tumour. We suggest our observation could be related to the broad group of haematological malignancies associated with germ cell tumours.

Follicular dendritic cells in bone marrow lymphoproliferative diseases: an immunohistochemical study including a new paraffin-resistant monoclonal antibody, DR53.

Two-hundred and twenty-one bone marrow biopsies with lymphoid infiltrates were studied histologically and immunohistochemically, to assess the incidence and the pattern of follicular dendritic cells. Three monoclonal antibodies selective for follicular dendritic cells were used: CD21, CD35 and DR53, all reactive on paraffin-embedded material. Follicular dendritic cells were present in two of 38 benign lymphoid aggregates, 92 of 134 low grade B-cell lymphomas (45 of 62 lymphocytic, 16 of 27 lymphoplasmacytoid, 0 of six hairy cell leukaemias, five of six centrocytic, 19 of 21 centroblastic-centrocytic, seven of 12 low grade NOS), one of 23 high grade B-cell lymphomas, 0 of 10 T-cell lymphomas, 0 of three Hodgkin's disease and four of 13 suspicious infiltrates. Follicular dendritic cells were found in lymphomatous involvement with nodular, patchy and massive growth pattern, but not in interstitial ones. They formed follicle-like networks, whose number and size were directly correlated to the tumour mass. The origin and frequency distribution of follicular dendritic cells in bone marrow biopsy lymphomas is discussed and the diagnostic relevance of follicular dendritic cell immunostaining in routine bone marrow biopsy lymphoid infiltrates is assessed.

[Immunophenotyping of B lymphoma in bone marrow biopsies. Contribution of 4 monoclonal antibodies used on paraffin-embedded tissues:DDB.42,DNA.7,DNA.44 and DND.53]. / Immunophénotype des lymphomes B dans les biopsies médullaires. Apport de 4 anticorps monoclonaux utilisables en paraffine: DBB.42, DNA.7, DBA.44 et DND.53.

An immunohistochemical study was performed on 132 routinely processed trephine biopsies of the bone marrow (40 reactive lymphocytosis, 80 B-cell lymphomas, 7 T-cell lymphomas, 5 Hodgkin's diseases), using 4 monoclonal antibodies, DBB.42, DNA.7, DBA.44, DND.53, which are directed against B-lymphocyte associated antigens, not destroyed by fixation. DBB.42, DNA.7 and DBA.44 are formalin and Bouin resistant. They do not stain myeloid, erythroblastic, histiomonocytic and endothelial cells, and are particularly suitable for bone marrow biopsies. DBB.42 and DNA.7 in association, identify all B-cell lymphomas and no T-cell lymphoma. DBB.42 recognizes respectively 65/65 and 13/15 low and high grade malignancy B-cell lymphomas, and DNA.7, 54/65 and 15/15. In our hands, these 2 antibodies are more reliable than L26 and MB2 on bone marrow biopsies. DBA.44 identifies 15/15 hairy-cell leukemias, and interestingly stains only 10% of other B-cell lymphomas, and no T-cell lymphoma. DND.53 is less specific, but reveals Reed-Sternberg cells. Combined immunostaining with anti T-lymphocyte antibody CD3, let us appreciate the contribution and the limits of immunohistochemistry in the diagnosis of bone marrow lymphoproliferative diseases, especially for interstitial and nodular lymphoid infiltrates of undetermined significance. These results point out the value of these 4 monoclonal antibodies in routinely processed bone marrow biopsies, particularly of DBB.42 and DBA.44.

Sinus histiocytosis with massive lymphadenopathy (Rosai Dorfman disease) in an HIV-positive patient.

We report a case of a 31-year-old HIV infected black female, who presented with asymptomatic generalized lymphadenopathy. Three particularly enlarged lymph nodes were biopsied (2 cervical and 1 axillary). The histological picture was consistent with a diagnosis of sinus histiocytosis with massive lymphadenopathy (SHML) or Rosai-Dorfman disease. Large histiocytes, positive for a variety of macrophage markers and for the S-100 protein, were observed in the distended sinuses. A few hyperplastic follicles, such as usually seen in HIV-infection-associated lymphadenopathy, were present at the periphery of one lymph node. No infections agent besides HIV could be detected by histological or microbiological analysis or by in situ hybridization. This is the first reported case of SHML associated with HIV infection. The possible relationship between the two diseases is discussed.

[Bone mastocytosis and osteomalacia. Apropos of a case with histodynamic analysis on non-decalcified bone]. / Mastocytose osseuse et ostéomalacie. A propos d'un cas avec étude histodynamique sur os non décalcifié.

A patient presented with cutaneous mastocytosis, bone pains, biologic disorders typical of osteomalacia, and intestinal malabsorption. Bone biopsy with histomorphometric analysis and measure of calcification rate on undecalcified bone, confirmed the diagnosis of bone mastocytosis and osteomalacia. There are only 2 other reports of osteomalacia caused by malabsorption. The latter is possibly bound to a mast-cell infiltrate of the intestinal mucosa. Sometimes only villosity atrophy or oedema of the mucosa are observed.

[Osteomalacia. Histologic definition and significance of the histodynamic analysis on undecalcified bone]. / L'ostéomalacie. Définition histologique et intérêts de l'étude histodynamique sur os non décalcifié.

Bone histomorphometry has a major significance in metabolic bone diseases, and especially in osteomalacia. This technique needs a strict methodology with quantitative analysis on undecalcified bone and measure of calcification rate. Histologic definition of osteomalacia includes hyperosteoidosis, characterized by thickened osteoid edges, and decrease of calcification rate. Histodynamic analysis on undecalcified bone provides interest in diagnosis, prognosis and supervision of this affection.

[Axillary lymph nodes in breast cancer. Comparison of standard histological analytical technics and macroscopic serial sections]. / Ganglions axillaires dans les cancers du sein. Comparaison des techniques d'analyse histologique standard et en coupes macroscopiquement sériées.

Lymph nodes in breast cancer have been examined by two histological techniques in 324 patients who underwent axillary node dissection. A routine pathologic examination consisted in the examination of only one pathological section while in the other, the lymph nodes were macroscopically serially sectioned. With the latter, an increase of 18.8 per cent in the number of detected metastases was noted. However this increase can be considered of prognostic and therapeutic importance only in 8.6 per cent of cases: half corresponding to those cases in which more than the initial threshold number of three lymph nodes were found, the other half corresponding to those cases in which one lymph node was found while the initial examination was negative. In the latter case, the increase of lymph node involvement was mainly due to "clandestine" metastases, i.e. lymphatic embolisation of micrometastases.

[Contribution of drill-biopsies to pre-treatment investigation of breast adenocarcinomas (author's transl)]. / Apport de la drill-biopsie dans le bilan préthérapeutique des adénocarcinomes mammaires.

The sensitivity of drill-biopsies (Rousseau's technique) was studied before using them routinely in pre-treatment investigation of breast adenocarcinomas. Results of histological examination, and estrogen (ER) and progesterone (PR) receptors levels obtained preoperatively by this technique, were compared with those from the corresponding mastectomy or tumourectomy specimens in 85 cases. Cytological examination of the various specimens, performed on prints, was used to compare their cellular density. Measurement of receptors included the determination of cytosolic and nuclear sites, and was performed by the single point dextran charcoal method. The limits of positivity established for ER and PR were 10 and 15 fM/mg protein respectively. The results showed sensitivity of 91.8 p. cent for pathology examinations; furthermore, the Scarff-Bloom-Richardson histoprognosis grading could be performed satisfactorily on drill-biopsy specimens. No significant difference was observed between the percentage of positivity for ER and PR from surgical and drill-biopsy specimens. Good correlation existed between values obtained from drill-biopsy and from surgery both for ER (r = 0.86, p less than 10(-5)) and PR (r = 0.86, p less than 10(-5)).