CD30-positive, anaplastic large-cell lymphomas that express CD15 but lack CD45. A possible diagnostic pitfall.

We report the immunohistochemical and clinical features of two cases of morphologically typical anaplastic large-cell lymphoma. One patient had lymph node and focal visceral involvement, and the other patient had multiple-organ involvement by lymphoma. In both cases, the lymphoma cells were CD30 (Ber-H2) and CD15 (Leu-M1) positive and CD45 (common leukocyte antigen) negative--a phenotype that is commonly seen in Hodgkin's disease. This unusual phenotype in large-cell anaplastic lymphoma led to an initial misinterpretation in one of the cases. Large-cell anaplastic lymphomas are highly variable, both in immunophenotype and clinical presentation. Because of this variability, a broad immunophenotypic panel, in conjunction with morphological features, should be used to establish the correct diagnosis.     

Fine-needle aspiration biopsy of Ki-1-positive anaplastic large-cell lymphoma.

Five cases of Ki-1-positive anaplastic large-cell lymphoma diagnosed by fine-needle aspiration biopsy are reviewed, and cytologic, histologic, and ultrastructural findings in these cases are correlated. In all cases, the diagnosis of anaplastic large-cell lymphoma was suggested on the basis of the morphological appearance in aspiration smears. This diagnosis was confirmed by immunohistochemistry, which revealed strong positivity of most of the cells by Ki-1 antibody. Two of the lymphomas were T-cell type, one was B-cell type, and the remaining 2 were composed of null cells. In 2 cases, intracytoplasmic inclusions were seen in some of the tumor cells in aspiration smears. These were ultrastructurally correlated with large lysosomal bodies of variable morphology. Fine-needle aspiration combined with immunohistochemistry may be an effective technique for diagnosing this neoplasm.     

Detection of Epstein-Barr virus by polymerase chain reaction.

The polymerase chain reaction (PCR) was used to study DNA extracted from the blood of 25 transplant patients, 5 patients with infectious mononucleosis, and 13 healthy subjects and autopsy or biopsy tissue from 29 patients with lymphoproliferative disorders. Primers were directed to conserved regions of the Epstein-Barr virus (EBV) genome encoding capsid protein gp220 and Epstein-Barr nuclear antigen 1. Specific EBV amplification was found in the blood of 11 of 25 transplant patients and all patients with infectious mononucleosis. All patients with lymphoproliferative disorders occurring in the presence of immunosuppression (eight organ transplant patients and two patients with acquired immunodeficiency syndrome) had biopsies positive for EBV by PCR. Only 1 of 19 samples from lymphomas or leukemias unrelated to immunosuppression contained EBV. PCR confirmed the very close association of EBV and lymphoproliferative disorders occurring in the presence of immunosuppression. The significance of detecting EBV sequences in the blood of transplant patients, particularly in relationship to lymphoproliferation, requires further study.     

A case of primary gastric CD30-positive anaplastic large-cell lymphoma

Gastric CD30-positive anaplastic large-cell lymphoma is a very rare disease. It is sometimes difficult to distinguish it from undifferentiated carcinoma, sarcoma and so on. We report here on a case of primary gastric anaplastic large-cell lymphoma. A 50-yr-old woman complained of epigastric pain and severe chest pain for 1 week. The gastroendoscopic examination revealed geographic mucosal irregularities with shallow ulceration at the antrum. She underwent a total gastrectomy. The gross finding of the resected stomach was an 8 x 4.5 cm sized ulceroinfiltrative lesion at the pyloric antrum along the lesser curvature. The microscopic examination revealed diffuse and solid proliferations of large atypical cells with pleomorphic nuclei. Immunohistochemically, the tumor cells were positive for CD30, vimentin and CD3, and this was a finding compatible with anaplastic large-cell lymphoma. To the best of our knowledge, this is the first such reported case in Korea.     

Detection of Epstein-Barr virus sequences in Hodgkin''s disease by the polymerase chain reaction.

The authors examined paraffin-embedded lymph node biopsies from 65 cases of Hodgkin's disease for the presence of Epstein-Barr virus (EBV) DNA, using the highly sensitive polymerase chain reaction technique. Overall 40% of the cases were positive for EBV DNA; there were no statistically significant differences in the frequency of EBV positivity among the different subtypes of Hodgkin's disease. These results are in agreement with those of previous studies that employed less sensitive detection techniques and suggest that EBV either is present in pathologic tissues only in some phases of the evolution of Hodgkin's disease or is a pathogenetic factor involved in only a portion of cases.     

Primary and secondary cutaneous Ki-1+ (CD30+) anaplastic large cell lymphomas. Morphologic, immunohistologic, and clinical-characteristics.

Skin biopsies were collected from 15 patients with cutaneous tumors morphologically similar to Ki-1+ anaplastic large cell (ALC) lymphoma of the lymph nodes, including Ki-1+ ALC lymphoma of childhood. The histology and immunophenotype of these cutaneous tumors are reported and follow-up data on the patients are given. The tumorous infiltrates were composed of large, sometimes very bizarre cells with one nucleolus or multiple nucleoli. All tumor cells expressed the lymphoid cell activation antigen Ki-1 (CD30) in conjunction with CD25 and the beta-chain of the T cell receptor. In 11 patients, Ki-1+ cutaneous tumors developed primarily in the skin. In nine patients, these were restricted to the skin in follow-up periods ranging from 3 months to 6 years. Two patients developed lymph node involvement after 2 months and 2 years, indicating the spreading potential of these cutaneous tumors. Morphologic and immunophenotypical identity of the atypical cells found in primary cutaneous ALC lymphoma, in regressing atypical histiocytosis (RAH), and in lymphomatoid papulosis (LyP) of type A, together with the protracted clinical course in all three conditions, suggests that primary cutaneous ALC lymphoma, RAH, and LyP of type A represent clinical variants of the same lymphoma entity. Secondary development of Ki-1+ ALC skin tumors was observed in two patients with cutaneous T cell lymphomas of mycosis fungoides type. These secondary Ki-1+ ALC lymphomas of the skin showed rapid systemic progression similar to the primary lymphonodal Ki-1+ ALC lymphomas. Concomitant or subsequent occurrence of Ki-1+ ALC tumors in cutaneous T cell lymphomas thus may be a bad prognostic sign.     

The ultrastructural and immunohistochemical heterogeneity of CD-30-positive neoplasms: so-called anaplastic large cell Ki-1 lymphomas

Anaplastic large cell lymphoma (ALCL), also referred to as Ki-1 lymphomas, was first recognized as an entity with characteristic light microscopic appearance in 1985. This tumor is composed of variably cohesive cells, often with large, markedly atypical, and multinucleated cellular forms. The recognition of ALCL resulted from the development of a monoclonal antibody in Kiel, Germany, named Ki-1, which was initially believed to be a putative marker for Reed-Sternberg cells. This antibody was later found to be specific against the epitope CD-30. Attempts to create strict criteria to preserve this neoplasm as a specific entity have undergone evolution. However, it is now clear that included in this group are a variety of pleomorphic neoplasms with CD-30 immunoreactivity. Some of these neoplasms are nonlymphoid and show marked heterogeneity in their immunohistochemical and ultrastructural profiles. This article aims to highlight the ultrastructural spectrum of neoplasms exhibiting CD-30 positivity that are within the spectrum of ALCL. It remains to be determined if there are subgroups of these CD-30-positive neoplasms that can be segregated on the basis of ultrastructural and immunohistochemical criteria with corresponding clinical correlates that may impact on their management, treatment, and prognosis. We review here the heterogeneity of CD-30-positive neoplasms (so-called anaplastic large cell Ki-1 lymphomas).     

Detection and characterisation of bluetongue virus using the polymerase chain reaction.

Pairs of oligodeoxynucleotide primers whose sequences were based on those of RNA segment 3 that encodes the bluetongue virus serogroup-reactive protein VP3, were synthesized for three BTVs from different geographic regions of the world and for seven Australian orbiviruses. Each pair of primers was then tested for the synthesis of cDNA and in subsequent polymerase chain reactions (PCR) with all ten virus groups. All primers were serogroup-specific at low or high stringency. One pair of primers was specifically designed for its ability to serogroup a BTV isolate irrespective of its geographic origin. At either high or low stringency, this primer-pair resulted in a common and specific PCR product for each of the BTVs tested but not for the other orbiviruses. Eight pairs of primers based on RNA2 sequences (the gene segment encoding the serotype-specific protein VP2) were also synthesized for the eight Australian serotypes of BTV. Each primer-pair was serotype-specific at low or high stringency except for the BTV16A pair, which cross-reacted with BTV3A and also gave a non-specific product that differed in Mr from the authentic PCR product. Using the PCR and BTV1A RNA3-based primers, BTV1A was detected in blood samples from two sheep at 9 days post inoculation. Virus was found in the platelet, buffy-coat and packed red blood cell fractions, but not in whole blood.     

Variable expression of leucocyte-common (CD45) antigen in CD30 (Ki1)-positive anaplastic large-cell lymphoma: implications for the differential diagnosis between lymphoid and nonlymphoid malignancies.

Monoclonal antibodies (mAbs) directed against the leucocyte common (CD45) antigen have been proposed as a useful tool for the differential diagnosis between malignant lymphomas (CD45+) and poorly differentiated nonhemopoietic tumors (CD45-). Thanks to the availability of mAbs directed against fixative-resistant epitopes of the CD45 molecule, this distinction can now easily be made even in routinely processed tissues. However, a small percentage of morphologically poorly defined neoplasms are difficult to diagnose even with the help of immunohistochemistry. The investigators report that 63 out of 165 anaplastic large-cell (ALC) lymphomas did not show any reactivity for the CD45 antigen in paraffin sections. In routine biopsies, the lymphomatous nature of these cases, most of which had been sent for consultation, could be always unequivocally established by demonstrating negativity for cytokeratins (mAb KL1) and clear dot-like and/or surface reactivity with the Ber-H2 mAb, which is directed against a fixative-resistant epitope of the lymphoid cell activation antigen CD30. Strikingly, 54% of the CD45-cases reacted with mAbs directed against fixative-resistant epitopes of the T cell-restricted CD45RO antigen (mAb UCHL1) or the B-restricted molecules CD45RA (mAb 4KB5) and L26 (unclustered). In order to avoid confusion of ALC lymphomas with anaplastic nonlymphoid tumors, pathologists must be aware of the existence of CD30+/CD45- ALC lymphomas, as they can mimic the above-mentioned malignancies both morphologically (due to the sinusoidal growth pattern) and phenotypically (due to the expression of EMA). The investigators conclude that the combined use of mAbs directed against fixative-resistant epitopes of the CD30, CD45RO, CD45RA, and L26 antigens and cytokeratins is essential for the correct diagnosis and treatment of these equivocal cases.     

Detection of reticuloendotheliosis virus infection using the polymerase chain reaction

The polymerase chain reaction (PCR) was shown to be a sensitive and useful method for detection of infection by members of the group of avian reticuloendotheliosis virus (REV). Genomic DNA extracted from chick embryo fibroblasts (CEFs), blood and tumours of chickens experimentally infected with the spleen necrosis virus (SNV) strain of REV was used as the target for chain elongation. Deoxyoligonucle-otide primers that encompass a portion of the unique 3', repeat and unique 5' region of the long terminal repeat (LTR) served to prime chain elongation. Products characteristic of the SNV LTR were also produced from DNA extracted from CEF infected with other strains of REV, namely chick syncytial virus, duck infectious anaemia virus, and the T-strain of REV. SNV-LTR sequences were amplified from DNA of SNV-experimentally infected chicks between 5 days and 19 weeks after infection. SNV-LTR sequences were also amplified from DNA from tumours and brains of SNV infected chickens, but not from DNA from avian leukosis virus- or Marek's disease virus-induced tumours. Results indicate that PCR is a sensitive and specific method for detection of REV infection and tumours.     

间变性大细胞性T细胞性淋巴瘤中EB病毒检测及CD56的表达

目的：研究间变性大细胞性Ｔ细胞性淋巴瘤ＥＢ病毒表达情况及其与ＣＤ５６阳性表达间的关系。方法：采用免疫组织化学ＬＳＡＢ法检测１５例ＡＬＴＣＬ中Ｋｉ－１及ＣＤ５６的表达,并用原位杂交法检测其ＥＢＥＲｓ。结果：１５例ＡＬＴＣＬ中Ｋｉ－１均阳性（１００％）,５例ＣＤ５６阳性（３３．３％）,９例ＥＢＥＲＳ阳性。其中,３例ＡＬＴＣＬ中ＥＢＥＲＳ和ＣＤ５６共同阳性。结论：ＡＬＴＣＬ的发生同ＥＢ病毒感染有一定关系     

Measurement by the polymerase chain reaction of the Epstein-Barr virus load in infectious mononucleosis and AIDS-related non-Hodgkin's lymphomas

A polymerase chain reaction (PCR) assay for the detection of Epstein-Barr virus (EBV) sequences in various clinical samples, especially peripheral blood leukocytes (PBL) and serum, was carried out and the results obtained were compared with specific EBV serology. One hundred seventy patients were enrolled in the study: 89 healthy blood donors, 22 asymptomatic patients, 36 individuals with primary EBV infection (including 19 patients with infectious mononucleosis [IM]), 22 HIV-infected subjects (including 4 with hairy oral leukoplakia, 3 with central nervous disorders, and 15 with non-Hodgkin's lymphoma). All the serum samples from the healthy blood donors were negative, In patients with IM and in AIDS-non Hodgkin's lymphoma (ARNHL), PCR was strongly positive in leukocytes (>2, 000 genome equivalents/l 0⁴ cells), which was correlated with detectable amounts of EBV DNA in serum. The overall positivity rate of PCR in serum was 58.8%, 68%, and 73% of cases for non-IM primary EBV infections, IM, and ARNHL, respectively. In two cases of EBV primary infection, the viral DNA was detected in serum, respectively 1 month and 2 months before IgM positivity and IgG rise. In one case of ARNHL followed up for several months, PCR (viral load of 2, 000 genome equivalents/10⁴ cells) became positive concurrently with appearance of lymphoma. In immunocompromised individuals, PCR EBV, if carried out in larger prospective studies, could be considered as a tumor marker, useful for predicting EBV-driven lymphoma and follow-up therapy. © 1995 Wiley-Liss, Inc.     

Morphological manifestations of large cell anaplastic Ki1(CD30)-positive lymphoma in children

Classic morphological and lymphohistiocytic variant were found in 46 and 2 cases, respectively, of 49 LCAL cases studied pathohistologically. One patient had a variant with a predominance of small tumor cells. Ultrastructurally, cells with feature of histiocytic and lymphoid differentiation and undifferentiated cells in various proportions were observed.     

Large-cell anaplastic (Ki-1-positive) lymphoma complicated by disseminated intravascular coagulation.

Large-cell anaplastic lymphoma (Ki-1-positive lymphoma) was first described as a type of large-cell lymphoma that has morphologic, enzymatic, and immunologic similarities to malignant histiocytosis. To date, an association between Ki-1-positive lymphoma and disseminated intravascular coagulation has not been described, but cases of malignant histiocytosis with disseminated intravascular coagulation have been reported. We report a case of anaplastic, Ki-1-positive, large-cell lymphoma complicated by clinical and laboratory evidence of disseminated intravascular coagulation with histologic evidence of vascular invasion and fibrin thrombi.     

Detection of influenza B virus in throat swabs using the polymerase chain reaction.

An assay protocol based on exploiting the polymerase chain reaction (PCR) for the direct detection of influenza B virus in throat swabs is described. By the use of PCR with nested primers, it was possible to detect the virus in throat swabs. Dilution experiments showed that as little as 1 plaque forming unit of virus was sufficient for detecting the HA gene by the PCR. All throat swab samples from which influenza B virus had been isolated by conventional methods were also positive by the PCR method.     

Analysis of adhesion molecules in Ki-1 anaplastic large-cell lymphoma

We analysed the expression of adhesion molecules on lymphoma cells in 13 patients with Ki-1 (CD30)-positive anaplastic large-cell lymphoma (Ki-1 ALCL; lymph nodes in 6, extranodal tumours in 6, and both lymph nodes and bone in 1). Very late activation antigen (VLA)-4 (CD49d) and Hermes lymph node homing receptor (CD44) were constantly expressed in all specimens, and intercellular adhesion molecule-1 (ICAM-1; CD54) was frequently expressed in 10 of the 14 specimens. The expressions of lymphocyte function-associated antigen-1 (LFA-1; CD11a) and VLA-5 (CD49e) occurred in 5 of 14 and 4 of 14 specimens, respectively. The expressions of VLA-2 (CD49b), endothelial leukocyte adhesion molecule-1, neural cell adhesion molecule (CD56) and E cadherin were always lacking. VLA-6 (CD49f) was absent in all but one specimen. The expression of VLA-5 on Ki-1 ALCL was high in subcutis-cutis but absent in lymph nodes. Furthermore, in one case, LFA-1 was detected in the primary lymph node, but was absent in a metastatic bone lesion. These results suggest that the expression of ICAM-1 is partially responsible for aleukemic behaviour in Ki-1 ALCL and, moreover, that the Ki-1 ALCL cells modify their expression of adhesion molecules at each of the involved organs.