bcl-2 rearrangement in Hodgkin's disease. Results of polymerase chain reaction, flow cytometry, and sequencing on formalin-fixed, paraffin-embedded tissue.

We examined 81 cases of Hodgkin's disease for evidence of the t(14;18) translocation, using the polymerase chain reaction assay on lysates of formalin-fixed, paraffin-embedded tissue. Seven of 74 amplifiable cases (9%) were positive for the translocation, which involves the bcl-2 oncogene and the immunoglobulin heavy chain gene. Two of these cases were sequenced and the breakpoints had the same pattern found in follicular lymphoma. The nuclei from one of the cases were sorted into large and small subpopulations. The t(14;18) signal was more intense in the large nucleus subpopulation, which contained a greater proportion of Reed-Sternberg-like nuclei. These results are consistent with the hypothesis that Reed-Sternberg cells carry the translocation, but they do not exclude the possibility that the translocation is found in cells representing the reactive component of Hodgkin's disease. The results also demonstrate that routinely processed material is suitable for polymerase chain reaction-based analysis of translocations, although the sensitivity is reduced 10- to 100-fold, compared with fresh tissue.     

Sequential karyotypes in non-Hodgkin lymphoma: their nature and significance

The examination of sequential karyotypes in hematologic disorders has demonstrated that karyotypic changes are often associated with concurrent changes in clinical behavior. Acquired abnormalities that recur among different patients may also suggest genomic areas important to tumor progression. We therefore examined sequential karyotypes in 21 patients with non-Hodgkin lymphoma (NHL). Sixteen of the 21 karyotypes demonstrated changes, including the majority of 6 small lymphocytic, 11 follicular, and 4 intermediate and high-grade diffuse lymphomas. The t(14;18)(q32;q21) occurred in ten initial karyotypes was retained in all cases. The band most frequently affected by newly acquired abnormalities was 14q32 (n = 5); chromosomes 1 and 2 (n = 5, each), and the 17p arm (n = 4) were also commonly affected. The acquired deletion of all or part of 17p appeared to be associated with a poor prognosis. Histologic transformation and karyotypic change did not correlate. This study of sequential karyotypes in NHL 1) confirms the primary importance of the t(14;18), 2) suggests that the 14q32 band is involved frequently in both primary and secondary cytogenetic events, and 3) suggests other genomic regions of potential significance to progression.     

Rosette formation in malignant lymphoma.

Lymph node biopsies in a patient with follicular lymphoma showed rosette structures as seen in neuroepithelial neoplasms. These specimens were studied by histologic, immunoperoxidase (for immunoglobulins, intermediate filaments (IF), actin and neuron-specific enolase), immunofluorescence (for immunoglobulins, and with a panel of monoclonal antibodies), and electron-microscopic examination. The rosettes were formed by neoplastic lymphocytes arranged around eosinophilic fibrillary material. Ultrastructurally, this was composed of cytoplasmic processes, projecting from the lymphocytes and containing thin and intermediate filaments. Immunohistochemically, it stained for monoclonal IgM lambda, all other antigens present on the neoplastic cells, and weakly for vimentin and actin. Based on recent information about lymphocyte surface changes, it is speculated that the rosettes might represent an aggregation of neoplastic lymphocytes activated by a microenvironmental stimulus, perhaps antigen-antibody binding at the cell membrane. The practical implication of this hitherto unreported finding is that the presence of rosettes cannot be used to rule out a lymphoma.     

Eosinophil-rich CD30+ lymphoproliferative disorder of the oral mucosa. A form of "traumatic eosinophilic granuloma"

We describe 3 patients who had oral mucosal lesions with features of traumatic eosinophilic granuloma (TEG) and containing CD30+ atypical cells. In 1 patient, the oral lesion was followed by skin nodules. All lesions were evaluated histologically, by immunohistochemical analysis, and by polymerase chain reaction (PCR) analysis of the T-cell receptor (TCR) gamma chain gene. All oral lesions were characterized by a dense and deeply infiltrative lymphoproliferation, showing epitheliotropism and massive eosinophilia. They contained atypical large lymphoid cells, which expressed T-cell markers and CD30. PCR analysis showed a monoclonal rearrangement of the TCR gamma chain gene in all lesions and, in 1 patient, the same rearrangement in the oral and cutaneous specimens. The lesions in these patients seem to be the oral counterpart of the spectrum of primary cutaneous CD30+ lymphoproliferative disorders and should be recognized as such to avoid a diagnosis of large T-cell lymphoma and possible consequent overtreatment. However, they represent only a subset among several others within the complex and heterogeneous category of disorders referred to as TEG.     

Predictability of immunologic phenotype of malignant lymphomas by conventional morphology: a study of 60 cases.

In order to test the immunologic validity of the Lukes-Collins (L-C) classification of the non-Hodgkin's lymphomas (ML), 60 ML in adult patients were studied and their B- or T-cell nature was predicted on a morphologic basis, without knowledge of the clinical history or the results of surface marker (SM) studies (SIg, C', Fc and E-rosettes), which were performed on cell suspensions and cryostat sections from the same specimens used for histology. There was a good correlation between morphologic types and SM (97% for the nodular ML, 81% for the diffuse). The predictions were not confirmed in 6 instances: one nodular ML typed as T; one convoluted lymphocytic ML typed as histiocytic; and four diffuse ML, predicted to be of B-type, bore no detectable SM ("Null" ML). It is concluded that the L-C morphologic criteria do allow in most cases the recognition of the B- or T-cell nature of ML, but cannot detect variations of the SM pattern which seem to affect the clinical behavior of these neoplasias. Thus, while the L-C classification seems immunologically sound, its clinical relevance still needs to be demonstrated.     

Follicular (nodular) lymphoma in childhood: a rare clinical-pathological entity. Report of eight cases from four cancer centers.

Eight cases of follicular lymphoma (FL) were found amount 318 children with non-Hodgkin's lymphoma seen in two decades in four large institutions (overall incidence: 2.5%). The children's ages ranged from 3 to 13 years (median 8). Four patients presented with localized peripheral lymphadenopathy, two with tumors of the digestive tract, two with disseminated disease. Five were tentatively classified as stage I or II and 3 as stage IV. The existence of other diseases responsible for lymphadenopathy could satisfactorily be exclued. All patients are alive after follow-up periods of 1 to 14 years from diagnosis (median 4). Morphologically, 5 lymphomas were mixed and 3 histiocytic. The growth pattern was "expansile" in the younger patients (3 to 9 years), and "infiltrative," as in the adult disease, in the three older children (11 to 13 years). The histiocytic cytology correlated with stage IV disease. FL in children appears to be different from both its adult counterpart and the diffuse childhood lymphomas. Differences with the former include the absence of tumors of poorly differentiated lymphocytic type, the higher frequency of stage I-II disease and the better prognosis. This last feature, as well as the higher frequency of peripheral node involvement and the absence of leukemic conversion or CNS disease, differentiates the follicular from the diffuse childhood lymphomas.     

Incidence and patterns of bone marrow and blood involvement by lymphoma in relationship to the Lukes-Collins classification.

Pretreatment lymph nodes, bone marrow, and blood were examined in 176 cases of non-Hodgkin's lymphoma. By the criteria of the Lukes and Collins functional--morphological classification, 158 (90%) were B-cell lymphomas and 17 (10%) were T-cell lymphomas. Bone marrow involvement was present in 53% of cases: 51% of B-cell types and 65% of T-cell types. Marrow involvement was most frequent in small lymphocyte (B) (89%), convoluted lymphocyte (60%), and small cleaved follicular center cell (FCC) lymphomas (55%). The pattern of bone marrow involvement was most frequently focal paratrabecular in B-cell lymphomas and diffuse in T-cell lymphomas. Blood involvement was present in 50% of cases with bone marrow lymphoma and generally reflected extensive bone marrow disease. There was a higher incidence of both bone marrow and blood involvement in pediatric patients than in adults.     

Tumoral non-amyloidotic monoclonal immunoglobulin light chain deposits ('aggregoma'): presenting feature of B-cell dyscrasia in three cases with immunohistochemical and biochemical analyses

Tumoral monoclonal immunoglobulin (Ig) light chain non-fibrillar deposits ('aggregomas'), which can be considered analogous to solitary light chain amyloidomas, are a rare presenting feature of B-cell dyscrasias. It is not certain if they are truly localized or if in reality they represent an initial expression of a silent systemic non-amyloid light chain deposition disease (LCDD). This report describes three patients, two of whom presented with cervical masses and the third with a solitary lung nodule, each comprising granular aggregates of monoclonal kappa light chain. Extracted deposits from the lymph node of one patient were shown by N-terminal amino acid sequence analysis to belong to the variable-region kappa I (Vkappa I) light chain subgroup, the first reported kappa-LCDD protein encoded by the L9 gene and the first report of an expressed protein related to this gene. Extracted deposits from the lung nodule of the second patient belonged to the Vkappa IV light chain subgroup encoded by the B3 germ line gene. The N-terminal amino acid sequences of the light chains from the aggregomas were compared with the related germ line sequences and to the N-terminal amino acid sequences of the nine other known kappa-LCDD light chains reported thus far from patients with systemic LCDD.