Detection of 14;18 chromosomal translocation in paraffin-embedded lymphoma tissue.
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Chromosome Mapping↗
Biomedical subjects
D Y Mason
Publications and source records attributed to D Y Mason.
The utility of Ki67 immunostaining, nuclear organizer region counting, and morphology in the assessment of follicular lymphomas.
Three methods, the enumeration of large non-cleaved cells (LNCC), the enumeration of nucleolar organizer regions (AgNORs), and the estimation of the percentage of Ki67-positive cells, have been compared on a series of 36 follicular lymphomas for their utility in subclassification. All three methods produced similar subdivisions to those obtained using the histological classification of the Working Formulation for clinical usage. Twenty cases have been followed clinically to assess any relationship between these parameters and short-term (less than 4 years) survival. Each technique showed a trend towards identifying patients with a poor short-term prognosis with no one method being superior to any other. Unless larger series with longer follow-up suggest otherwise, there is no indication to use either the AgNOR or Ki67 methods which offer no advantage over the simple and inexpensive Working Formulation or Berard's LNCC counting.
Antibodies, Monoclonal↗
Expression of macrophage-associated antigens in tissues involved by Langerhans' cell histiocytosis (histiocytosis X).
The expression of macrophage antigens KP1, Mac, lysozyme, and alpha-1-antichymotrypsin was investigated on routine paraffin sections from 17 cases of Langerhans' cell histiocytosis (LCH). All the major clinical forms were represented, including single lesions and monosystemic and multisystemic disease. In all the cases, a variable fraction (3-35%) of LCH cells was immunoreactive with KP1 and anti-Mac; the staining pattern was quite typical because the immunoreaction product was often confined to the perinuclear space and the Golgi area. LCH cells containing lysozyme and AACT were detected less frequently; however, in positive cases the percentage of LCH cells immunoreactive for lysozyme and AACT was in the same range as that of KP1-positive cells. On immunostained cytosmears (one case), about 10% of the CD1a-positive cell population was reactive for the macrophage antigens CD14 and PAM-1. No association was noted between the number of KP1-positive cells and the clinical form and/or anatomic site of the lesion. Phagocytic macrophages were significantly and diffusely immunoreactive with KP1 and anti-Mac and for AACT and lysozyme. Multinucleated giant cells with irregular nuclei were frequently observed; these cells were rarely S-100 positive, were consistently stained by KP1 and AACT, and were occasionally anti-Mac positive. The authors' findings suggest that antimacrophage monoclonals, in conjunction with S-100 protein, may represent a useful tool to establish the diagnosis of LCH in paraffin-embedded material.
Antibodies, Monoclonal↗
Clonal silencing of self-reactive B lymphocytes in a transgenic mouse model.
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Animals↗
A translocation involving a specific breakpoint (q35) on chromosome 5 is characteristic of anaplastic large cell lymphoma ('Ki-1 lymphoma').
Two cases of non-Hodgkin's lymphoma are reported in which a chromosomal translocation was observed involving the same site (q35) on the long arm of chromosome 5. The other breakpoint involved in the translocation was chromosome 2 (p23) in one case and chromosome 3 (q12) in the other. Both cases were large cell lymphomas expressing CD30 antigen ('Ki-1 lymphoma'). One was clearly of T lymphoid origin, the other probably B cell derived. One other case of a Ki-1 lymphoma with 2;5 translocation (involving the same breakpoint on chromosome 5) has been reported previously, and it is suggested that this cytogenetic abnormality may be specifically associated with Ki-1 lymphoma. The literature contains a further eight cases of lymphoid neoplasms with a translocation involving a breakpoint at q35 on chromosome 5. They have all been described as cases of 'malignant histiocytosis', but the present findings make it likely that these cases were in reality also examples of Ki-1 lymphoma. The breakpoint at the q35 region on chromosome 5 is close to the position of the fms proto-oncogene, suggesting that an abnormality affecting this gene might possibly play a causal role in 'Ki-1 lymphoma'. However, DNA restriction fragment analysis of the present cases showed no evidence that the breakpoint on chromosome 5 involves the fms gene or its immediate vicinity.
Adolescent↗
The expression of the CD3 antigen in Hodgkin's disease.
Tissue from 86 cases of Hodgkin's disease, fixed in formalin and embedded in paraffin, was immunostained for the T-cell marker CD3. Of these cases, 20 were selected on the basis of previous reactivity of Reed-Sternberg cells for T-cell associated antigens in frozen sections whilst the remaining 66 were retrieved from the routine pathology files. Five of the 20 selected cases and 22 of the retrieved cases showed predominantly cytoplasmic positivity in a subpopulation of Hodgkin and Reed-Sternberg cells. CD3 positive cells were present in all subtypes of Hodgkin's disease including three of nine lymphocyte predominance cases. It therefore appears that some Hodgkin and Reed-Sternberg cells can express the major T-cell antigen CD3. Although these findings are open to other interpretations, they are consistent with the hypothesis that at least some cases of Hodgkin's disease arise from activated T-cells.
Antigens, Differentiation, T-Lymphocyte↗
Langerhans cells in human lung tumours: an immunohistological study.
In an immunocytochemical study of 41 human lung tumours we have shown that Langerhans cells can be reliably identified using the anti-CD1 monoclonal antibody NA1/34. Langerhans cells are present in all the main varieties of human lung tumour although they are infrequent in both small cell carcinoma and carcinoid tumour. There is considerable variation in numbers of Langerhans cells in both adenocarcinomas and squamous cell carcinomas. In this study tumours were divided into those with high numbers of Langerhans cells (greater than 2 per high power field) and those with low numbers (less than 2 per high power field). Analysing these results against patient survival showed a markedly worse survival in those tumours with a high number of Langerhans cells for all the tumours as a single group and for squamous cell carcinoma as a single entity.
Adenocarcinoma↗
The prognostic significance of immunophenotype in high-grade non-Hodgkin's lymphoma.
The prognostic value of immunophenotyping lymphomas was assessed by studying 51 cases of high-grade non-Hodgkin's lymphoma for which long term clinical follow-up (14-28 years) was available. Using antibodies which identify T- and B-cell-related antigens in formalin-fixed, paraffin-embedded material, 43 were shown to be of B-cell and eight of T-cell phenotype. In terms of survival, cases of high-grade T-cell lymphoma fared significantly worse (P less than 0.05) than cases of high-grade B-cell subtype. These findings support the belief that T-cell lymphomas have a more aggressive clinical course than their B-cell counterparts.
Antibodies, Monoclonal↗
Diagnosis of acute myeloid leukaemia with the use of monoclonal anti-neutrophil elastase (NP57) reactive with routinely processed biopsy samples.
The reactivity of a monoclonal anti-neutrophil elastase antibody (NP57) with routinely processed biopsy samples from various acute leukaemias has been examined and compared with that of chloroacetate esterase and CD15 (hapten X), two other myeloid cell-associated markers detectable in paraffin sections. No staining was seen with these markers in 14 cases of acute lymphoblastic leukaemia. In contrast the neoplastic cells in 27 of 37 acute myeloid leukaemias were NP57 positive. Twenty of these were also positive for chloroacetate esterase, whereas CD15 was expressed in only six cases. These results indicate that detection of elastase with monoclonal NP57 forms a useful supplement to traditional methods for the histopathological diagnosis of acute myeloid leukaemias.
Antibodies, Monoclonal↗
Automation of APAAP immunocytochemical technique.
A tissue processing instrument (the Histokinette) was modified by the addition of an electronic timing device which allows an immunocytochemical staining technique (the APAAP method) to be performed as a semiautomated procedure. After incubation with primary monoclonal antibodies (applied by hand) slides (up to 72 in a batch) are placed in racks and cycled through tanks of reagents, comprising anti-mouse Ig followed by APAAP complexes with intervening timed draining and washing stages. This semiautomated process gave consistent staining results and offered considerable savings in time compared with conventional methods. The same reagent baths were used over four months on an almost daily basis without deterioration in staining intensity, and consequently the calculated overall cost of the staining procedure was less than if the reagents had been applied by hand and then discarded. The machine is now into its eleventh month of operation; the reagents have been changed twice. It is suggested that this approach, because of savings in time and increased consistency, may be an attractive technique for the routine immunocytochemical staining of slides, and that the nature of the APAAP method is particularly suitable for automation as the necessary reagents can be produced at low cost.
Humans↗
Detection of T cells in paraffin wax embedded tissue using antibodies against a peptide sequence from the CD3 antigen.
Rabbit polyclonal antibodies were raised against a proline rich, peptide sequence, comprising 13 amino acids, in the cytoplasmic domain of the CD3 epsilon chain. Immunoprecipitation experiments showed that this antibody preparation recognised the CD3 antigen on human T lymphoblasts. The antibody stained normal T cells strongly in tissue sections which had been fixed in formalin or Bouin's solution and embedded in paraffin wax. Its reactivity with T cell lymphoma, when evaluated on a series of 96 previously phenotyped cases, closely agreed with the results obtained on cryostat sections. These results indicate that the specific detection of T cells in routinely processed tissue biopsy specimens is now technically feasible on a wide scale in diagnostic laboratories using CD3 peptide antibodies, and they also suggest that in future the use of anti-peptide antibodies may detect other lineage specific antigenic markers in paraffin wax sections.
Amino Acid Sequence↗
KP1: a new monoclonal antibody that detects a monocyte/macrophage associated antigen in routinely processed tissue sections.
A new monoclonal antibody, KP1, raised against a lysosomal fraction of human lung macrophages, recognises a fixation-resistant epitope in a wide variety of tissue macrophages (such as Kupffer cells germinal centre, splenic, and lamina propria macrophages), and in granulocyte precursors. Its broad reactivity with cells of the mononuclear phagocytic lineage was established by testing on routinely processed samples of normal and reactive lymphoid tissues. Interdigitating reticulum cells were unstained or showed limited cytoplasmic staining while Langerhans' cells and follicular dendritic reticulum cells were unreactive. KP1 recognises a molecule of about 110 kilodaltons in macrophage-rich human tissue when tested by either immunoprecipitation or Western blotting (although the latter procedure also shows two additional components with molecular weights of 70 and 40 kilodaltons). KP1 should be of considerable value for studying disorders of the monocyte/macrophage system, including both reactive and neoplastic states (such as true histiocytic proliferations).
Antibodies, Monoclonal↗
Molecule detected in formalin fixed tissue by antibodies MT1, DF-T1, and L60 (Leu-22) corresponds to CD43 antigen.
Three monoclonal antibodies MT1, L60 (Leu-22), and DF-T1, were reported independently as recognising human T cells in routinely processed, paraffin wax embedded tissue. The present study was performed to compare these three reagents in terms of their immunocytochemical reactions and target molecule(s). On Western blotting of white cell extracts the three antibodies reacted with antigens of the same molecular weight (range 110-160 kilodaltons). Furthermore, their immunocytochemical reactivity with normal human cells, as analysed by two-colour flow cytometry, was essentially identical (labelling of monocytes, most T lymphocytes, and weak reactions with some B cells), and the antibodies gave closely similar reactions on 54 white cell derived neoplasms. To identify the target antigen for these three reagents, antibodies from the Third International Workshop on Leucocyte Antigens were reviewed and it was shown that the Western blotting and immunocytochemical reactions of MT1, L60 (Leu-22), and DF-T1 were identical with those of the reagents which defined the CD43 antigen (also known as leucosialin or sialophorin). Furthermore, all these antibodies reacted with cells transfected with a cDNA clone encoding CD43. It is concluded that antibodies MT1, L60 (Leu-22), and DF-T1 all recognise the heavily glycosylated myeloid/lymphoid associated CD43 antigen.
Antibodies, Monoclonal↗
Double immunocytochemical labeling of cell and tissue samples with monoclonal anti-bromodeoxyuridine.
We describe a new monoclonal antibody (designated Bu20a) against bromodeoxyuridine (BrdU). This antibody was selected by screening against human tissues using the APAAP technique, and shows no crossreactivity with normal nuclei. It stains BrdU incorporated into the nuclei of a wide range of cell types, including human tonsil lymphoid cells, normal mouse tissues, and human tumors growing in nude mice. A double-labeling technique is described using this antibody in which cell smears or tissue sections are first labeled by an immunoperoxidase procedure for a cellular antigen (e.g., mouse or human histocompatibility class II antigen, T-lymphocyte antigen, keratin) and BrdU is then detected by indirect immunofluorescence. This procedure, which was applied to a variety of human and animal cells and tissues, is of wide potential value in analyzing the phenotype of S-phase cells and in co-localizing antigen expression and BrdU incorporation in tissue sections.
Animals↗
Isolation and characterization of human fetal macrophages from placenta.
Human fetal macrophages expressing class II major histocompatibility complex (MHC) antigens have been isolated from the stroma of the chorionic plate of term placentas, using enzymatic digestion procedures, and enriched by Percoll density centrifugation. These cells are adherent, phagocytic and express Fc receptors for IgG. By rosetting with bovine erythrocytes coated with IgG, they can be enriched to 77-95% purity. Placental macrophages isolated in this way stimulate the proliferation of lymphocytes from unrelated donors in mixed-cell cultures, and act as accessory cells in oxidative mitogenesis. In a family study, placental macrophages stimulated proliferation of maternal and paternal lymphocytes but there was no evidence for either priming to, or suppression by, the fetal cells when the responses of lymphocytes from the mother and her HLA identical twin were compared. The possibility that these cells can protect the fetus from infection and/or stimulate the production of maternal anti-fetal HLA-antibodies is discussed.
Cell Adhesion↗
Colocalization of elastase and myeloperoxidase in human blood and bone marrow neutrophils using a monoclonal antibody and immunogold.
The authors have localized elastase in human blood and bone marrow neutrophils by immunoelectron microscopy using a monoclonal anti-human elastase antibody (NP 57) and compared its distribution with myeloperoxidase (MPO) and lactoferrin (LF), which mark primary and secondary neutrophil granule, respectively. Human bone marrow and blood polymorphonuclear leukocytes (PMN), either unstimulated or after phagocytosis of latex microbeads, were fixed in 4% paraformaldehyde. Ultrathin frozen sections were immunolabeled with NP 57, followed by an immunogold probe. In bone marrow granulocyte precursors elastase appeared simultaneously in the immature first granules of myeloblasts with MPO. As these granules became denser with maturation, labeling for both enzymes became weaker and sometimes negative (possibly due to masking of immunoreactivity). The ellipsoidal primary granules were strongly labeled by NP57. LF positive granules appeared later, at the myelocyte stage, and contained neither MPO nor elastase. In mature neutrophils, immunolabeling for elastase was found together with MPO in the large electron-dense primary granules and in a different granule population from the LF-positive secondary granules. Double labeling with two different-sized gold particles was used to compare the kinetics of degranulation of secondary and primary granules. The observation and the analysis of single phagosome content was made possible by this new technique. In conclusion, immunoelectron microscopy was used to show elastase in the primary granules of neutrophils, where it appears simultaneously with MPO. This technique has also allowed comparison of the kinetics of degranulation of both types of granules, and could be applied to different experimental and pathologic conditions.
Antibodies, Monoclonal↗
Diagnosis of myelomonocytic and macrophage neoplasms in routinely processed tissue biopsies with monoclonal antibody KP1.
A new monoclonal antibody, KP1, against the CD68 antigen, which labels macrophages and other members of the mononuclear phagocyte lineage in routinely processed tissue sections, has been used to stain a range of lymphoid, histiocytic, and myelomonocytic proliferations. All 20 neoplasms of myeloid, myelomonocytic, and presumed macrophage derivation reacted with antibody KP1. None of the 22 cases of T cell neoplasia had positive reactions. Although 14 of 41 B lineage lymphomas and leukaemias were stained by antibody KP1, staining was usually confined to small dots of reactivity, in contrast to the strong and extensive cytoplasmic staining seen in the neoplasms of myeloid and macrophage/monocyte origin. Furthermore, positive B cell neoplasms were almost all small cell proliferations, which are unlikely to be confused with myelomonocytic malignancies. It was concluded that antibody KP1 is a valuable addition to a panel of monoclonal antibodies for phenotyping lymphomas, particularly in routinely fixed tissues. It should assist the pathologist in the recognition of extramedullary presentation of leukaemia, aid in the diagnosis of suspected cases of true histiocytic neoplasia, and allow for quantitation of macrophages infiltrating lymphomas and other solid tumors.
Antibodies, Monoclonal↗
Immunocytochemical analysis of cytocentrifuged fine needle aspirates. A study based on lung tumors aspirated in vitro.
The value of Cytospin preparations of fine needle aspiration (FNA) biopsy material for immunocytochemical analysis was investigated using aspirates obtained from 23 resected human lung tumors. The results were compared with those on cryostat sections from the same tumors. The Cytospin preparations of the FNA biopsies gave the best immunostaining reactions and enabled a comprehensive range of monoclonal antibodies (MAbs) to be utilized. The quality of the Cytospin immunostaining compared favorably with that on cryostat sections of the same tumors and generally yielded a similar immunophenotype. However, the Cytospin preparations were not suitable for staining with MAb Ki67, which detects an antigen associated with cellular proliferation. With Ki67, conventionally prepared smears were much superior and enabled an assessment of tumor growth fraction that concurred with the growth fraction calculated from cryostat sections in most cases.
Antibodies, Monoclonal↗