scholarly journals Isolation of Reed-Sternberg cells from lymph nodes of Hodgkin's disease patients

Blood ◽  
1989 ◽  
Vol 73 (1) ◽  
pp. 222-229
Author(s):  
G Sitar ◽  
E Brusamolino ◽  
C Bernasconi ◽  
E Ascari
Keyword(s):  
Lymph Nodes ◽  
Hodgkin's Disease ◽  
Mononuclear Cells ◽  
Density Gradient Centrifugation ◽  
Hodgkin’S Disease ◽  
Gradient Centrifugation ◽  
Cell Types ◽  
Velocity Sedimentation ◽  
Single Cell Suspension ◽  
Final Yield

This study describes a simple and relatively rapid method of purifying Reed-Sternberg (R-S) cells and their morphologic variants from the lymph nodes of patients affected by Hodgkin's disease. Our initial studies defined the optimal procedure for a quantitative disaggregation of Hodgkin's lymph nodes and the densities of R-S cells in several donors. These preliminary steps were helpful in the development of strategies for selectively concentrating R-S cells by density gradient centrifugation. We layered a single-cell suspension over Percoll of appropriate density, centrifuged these samples for 15 minutes, and collected a fraction enriched in R-S cells. Most of the R-S cells were distributed between densities of 1.060 and 1.072, with a peak at approximately 1.066 g/mL. R-S cells are denser than many mononuclear cells present in the lymph nodes of Hodgkin's patients and lighter than reactive cells such as eosinophils, mast cells, and neutrophils. However, the ranges of densities of these cell types overlap, making purification of R-S cells by isopyknic centrifugation impossible. Nevertheless, when this enriched fraction is further processed by velocity sedimentation in order to take advantage of the larger size of R-S cells as compared with all other cells, a substantial purification is achieved. We used three different velocity-sedimentation chambers to find the optimal conditions for obtaining the highest purity with a high final yield. The cells isolated by this method are viable, appear to be morphologically normal, and have been further characterized biologically.

scholarly journals Isolation of Reed-Sternberg cells from lymph nodes of Hodgkin's disease patients

Blood ◽  
1989 ◽  
Vol 73 (1) ◽  
pp. 222-229 ◽  
Author(s):  
G Sitar ◽  
E Brusamolino ◽  
C Bernasconi ◽  
E Ascari
Keyword(s):  
Lymph Nodes ◽  
Hodgkin's Disease ◽  
Mononuclear Cells ◽  
Density Gradient Centrifugation ◽  
Hodgkin’S Disease ◽  
Gradient Centrifugation ◽  
Cell Types ◽  
Velocity Sedimentation ◽  
Single Cell Suspension ◽  
Final Yield

Abstract This study describes a simple and relatively rapid method of purifying Reed-Sternberg (R-S) cells and their morphologic variants from the lymph nodes of patients affected by Hodgkin's disease. Our initial studies defined the optimal procedure for a quantitative disaggregation of Hodgkin's lymph nodes and the densities of R-S cells in several donors. These preliminary steps were helpful in the development of strategies for selectively concentrating R-S cells by density gradient centrifugation. We layered a single-cell suspension over Percoll of appropriate density, centrifuged these samples for 15 minutes, and collected a fraction enriched in R-S cells. Most of the R-S cells were distributed between densities of 1.060 and 1.072, with a peak at approximately 1.066 g/mL. R-S cells are denser than many mononuclear cells present in the lymph nodes of Hodgkin's patients and lighter than reactive cells such as eosinophils, mast cells, and neutrophils. However, the ranges of densities of these cell types overlap, making purification of R-S cells by isopyknic centrifugation impossible. Nevertheless, when this enriched fraction is further processed by velocity sedimentation in order to take advantage of the larger size of R-S cells as compared with all other cells, a substantial purification is achieved. We used three different velocity-sedimentation chambers to find the optimal conditions for obtaining the highest purity with a high final yield. The cells isolated by this method are viable, appear to be morphologically normal, and have been further characterized biologically.

scholarly journals Atypical hodgkin and reed-sternberg cells in peripheral blood of a patient with advanced stage of Hodgkin's disease - a case report

2003 ◽  
Vol 131 (9-10) ◽  
pp. 400-402 ◽  
Author(s):  
Rajko Milosevic ◽  
Milica Colovic ◽  
Vesna Cemerikic-Martinovic ◽  
Natasa Colovic ◽  
Marina Bogunovic
Keyword(s):  
Bone Marrow ◽  
Lymph Nodes ◽  
Peripheral Blood ◽  
Hodgkin's Disease ◽  
Mononuclear Cells ◽  
Hodgkin’S Disease ◽  
Lymphoid Cells ◽  
High Dose ◽  
Advanced Stage ◽  
Neck Lymph Nodes

The occurrence of abnormal Hodgkin's and Reed-Sternberg cells in the peripheral blood in a patient suffering from Hodgkin's disease has been noticed exceptionally rare in a previous period, and especially rare in last ten years primarily due to successfull treatment of this disease. The presence of atypical mononuclear cells in peripheral blood which cytomorphologically resembled Reed-Sternberg cells was registered in 8 patients till 1966. During the last decade, the presence of atypical mononuclear cells in the peripheral blood was used for their isolation cultivation, and detailed immunophenotypic and genetic analysis. The analysis of mononuclear cells in rare patients with Hodgkin's disease was established that they belong to the B-lymphoid cells with expression of CD30 and CD15 antigens. The examination of presence of Hodgkin's cells in the peripheral blood of patients with Hodgkin's disease is important for patients with advanced stage of the disease in which autologous stem cell transplantation and high dose chmeotherapy is planned. The authors present a 33-year-old patient, who noticed enlarged neck lymph nodes in September 2000, high temperature and loss in weight. On physical examination enlarged neck lymph nodes 5x8 cm and hepatosplenomegaly were found. There was anemia and thrombo-cytopenia, and normal WBC count with 24% of lymphoid elements in differential formula. On histologic examination of lymph nodes Hodgkin?s disease, type nodular sclerosis with mixed cellularity was found. Histology of bone marrow showed nodal lymphomatous infiltration. Immunohistochemistry with monoclonal antibodies of concentrate of peripheral blood cells showed expression of CD30+ and CD15+, immunophenotypically and morphologically matching Reed-Sternberg cells. Cytogentic analysis of mononuclear cells of the bone marrow showed normal karyotype. The patient was in clinical stage IV/V of the disease and chemotherapy with 9 cycles of ABVD+Mp protocol was applied. He is still in remission.

EXTRAVASAL FIBRIN STABILIZATION BY FACTOR XIII IN LYMPH NODES WITH HODGKIN’S DISEASE

1987 ◽  
Author(s):  
R Adày ◽  
A Szegedi ◽  
Z Nemes ◽  
L Muszbek
Keyword(s):  
Lymph Nodes ◽  
Hodgkin's Disease ◽  
Factor Xiii ◽  
Mononuclear Cells ◽  
Hodgkin’S Disease ◽  
Tumor Stroma ◽  
Urea Solution ◽  
Immunoperoxidase Technique ◽  
Long Time ◽  
Covalently Linked

The formation of extravasal fibrin deposits in various tumors has been recognized a long time ago and it has been implicated in various aspects of tumor growth. However, no adequate information is available on the nature of intratumoral fibrin. In this study we attempted to find out if fibrin deposit in human lymph nodes with Hodgkin’s disease is stabilized and made resistant to fibrinolysis by factor XIII /FXIII/ of blood coagulation. The two main tasks for FXIII in fibrin stabilization is to attach a^antiplas-min the main phyiological inhibitor of fibrinolysis to fibrin strands and crosslink fibrin chains. By double immunofluorescent labeling for fibrin and α2-antiplasmin a complete colocalization of the two antigens could be observed. A part of fibrin strands also stained for α2-antiplasmin-plasmin-complex-neoantigen revealing that α2-antiplasmin covalently linked to fibrin inhibited intratumoral fibrinolysis. The finding that immunolabeling for fibrin was preserved following the treatment of sections by concentrated urea solution clearly demonstrates that fibrin chains became crosslinked by FXIII. These results were further supported by SDS PAGE analysis of intratumoral fibrin deposits. There are two theoretical possibilities for the appearance of FXIII in the interstitial space: 1/ plasmatic FXIII can get acrossthe vessel wall when increased permeability is induced 2/ FXIII can be produced and released by certain cells of the tumorous tissue. We explored the secondpossibility by various immunomorphological and enzymcytochemical techniques. Alarge number of FXIII positive cells were detected by immunoperoxidase technique in the follicular and interfollicular region of malignant, but not in normal lymph nodes. These relatively large, multipolar, mononuclear cells possesseda macrophage-like appearance and showedANAE-positivity,i.e.,they belong to thegroup of tumor associated macrophages. FXIII containing cells were labeled by monoclonal anti-Leu M3 (a monocyte/macrophage marker), but not by anti-HLA-DR.They were often found in the immediate vicinity of malignant Hodgkin’s cells and also showed an intimate relationshipwith extravasal fibrin formation.lt is suggested that FXIII secreted byintactor released from damaged macrophages might be involved in the stabilization offibrin in the tumor stroma or around tumor cells.

Diagnostic Cell Pathology in Lymph Nodes Diseases

1972 ◽  
Vol 58 (5) ◽  
pp. 289-310 ◽  
Author(s):  
Silvana Pilotti ◽  
Franco Rilke
Keyword(s):  
Lymph Nodes ◽  
Hodgkin's Disease ◽  
Hodgkin’S Disease ◽  
Cell Types ◽  
Malignant Lymphomas ◽  
Histological Sections ◽  
Histiocytic Lymphoma ◽  
Poorly Differentiated ◽  
Nuclear Structures ◽  
Metastatic Malignant Tumor

The investigation is based on the cytological findings in imprints, stained by the modified Papanicolaou E.A. 50 method, of 190 lymph nodes, of which 10 were normal 60 with benign diseases, 58 with malignant lymphomas and 62 with metastases. The cytological data were controlled by and correlated with histological sections. Touch preparations of lymph nodes do not present technical difficulties, are rapidly prepared and are always satisfactory. Cytological details are much better preserved and more recognizable than in histological sections but little information on lymph node organization is yielded. Imprints do not replace tissue diagnoses or even frozen sections, but under certain circumstances they may supply sufficient data for immediate diagnostic purposes, such as findings of metastatic malignant tumor cells or of Reed-Sternberg cells and their variants in lymph nodes during laparatomy for the staging of Hodgkin's disease. In addition to their supplementary diagnostic as well as teaching value, touch preparations allow minute caryological observations in malignant lymphomas. Peculiar basic nuclear structures in both well and poorly differentiated lymphocytic lymphomas and two different nuclear types in histiocytic lymphoma are described. The variations and frequency of several cell types (reticulum cells, lymphocytes, histiocytes, Reed-Sternberg and lacunar cells) in Hodgkin's disease are described and correlated with the histologic type.

Chemotherapy in hodgkin’s disease (lymphadenoma)

1973 ◽  
Vol 11 (15) ◽  
pp. 57-59
Keyword(s):  
Lymph Nodes ◽  
Young Women ◽  
Hodgkin's Disease ◽  
Important Determinant ◽  
Hodgkin’S Disease ◽  
X Rays ◽  
Single Factor ◽  
Node Biopsy ◽  
Choice Of Treatment ◽  
Choice Of Therapy

The extent of Hodgkin’s disease at the time of diagnosis is the most important single factor affecting prognosis and the choice of treatment.1 2 Thorough investigation is needed to determine the stage of the disease,3 and it is best to refer patients to a centre with special experience of the condition. The staging process requires haematological, biochemical and radiographic studies, including abdominal lymphography.4 Some patients may also need laparotomy with splenectomy and hepatic and node biopsy to localise any intra-abdominal spread, particularly into the liver which is ominous and an important determinant in the choice of therapy.5 6 Laparotomy is less usually performed in patients over 60. In children under 6 splenectomy should not be carried out because it makes them more susceptible to infection.7 In young women the ovaries may be sewn to the abdominal wall in the mid-line so that they can be shielded from x-rays directed at iliac lymph nodes.

Tissue T-lymphocytes in untreated Hodgkin's disease morphologic and functional correlations in spleens and lymph nodes

Cancer ◽  
1982 ◽  
Vol 50 (2) ◽  
pp. 259-268 ◽  
Author(s):  
Carlo D. Baroni ◽  
Luigi Ruco ◽  
Stefania Uccini ◽  
Antonio Foschi ◽  
Massimo Occhionero ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Lymph Nodes ◽  
Hodgkin's Disease ◽  
Hodgkin’S Disease

Pediatric Hodgkin's disease in India.

1985 ◽  
Vol 3 (12) ◽  
pp. 1605-1612 ◽  
Author(s):  
K Dinshaw ◽  
S Pande ◽  
S Advani ◽  
G Ramakrishnan ◽  
C Nair ◽  
...  
Keyword(s):  
Hodgkin's Disease ◽  
Hodgkin’S Disease ◽  
Initial Period ◽  
Age Groups ◽  
Cell Types ◽  
Clinical Staging ◽  
Mixed Cell ◽  
Adequate Treatment ◽  
Female Ratio ◽  
Tata Memorial Hospital

Twenty-one percent of all Hodgkin's disease in India was seen in the pediatric age groups at the Tata Memorial Hospital (Bombay, India). From 1975 to 1982, 151 cases of children were reviewed. The youngest presentation was at 3 years in three patients, with a marked male: female ratio of 5.5:1. Twenty-six patients were previously treated before referral while the remaining 125 cases were investigated and treated according to the prevalent protocols in 1975 to 1978 and 1979 to 1982. Clinical staging revealed 54% of patients in stages I and II with symptoms in 20%, and 46% of patients in stages III and IV with symptoms in 67%. Staging laparotomy was performed in 27 patients, with a total changes of staging in 17 children (63%). The mixed cell types (46%) and lymphocytic predominant types (31%) were the most common histologic presentations. Nine percent nodular sclerosis and 9% lymphocytic-depleted varieties were also observed. Five percent of all cases were not classifiable. Minimum adequate treatment was completed in 87 cases. Comparisons were made between the treatments administered to 40 patients during the initial period 1975 to 1978 when individualized treatment was administered, and the later 47 patients during the 1979 to 1982 period, when chemotherapy was the mainstay of treatment with involved field radiation.

Detection of Clonal Hodgkin and Reed-Sternberg Cells With Identical Somatically Mutated and Rearranged VH Genes in Different Biopsies in Relapsed Hodgkin’s Disease

Blood ◽  
1998 ◽  
Vol 92 (8) ◽  
pp. 2899-2907 ◽  
Author(s):  
Martina Vockerodt ◽  
Marta Soares ◽  
Holger Kanzler ◽  
Ralf Küppers ◽  
Dieter Kube ◽  
...  
Keyword(s):  
Single Cell ◽  
Hodgkin's Disease ◽  
Mononuclear Cells ◽  
Residual Disease ◽  
Hodgkin’S Disease ◽  
Clinical Importance ◽  
Variable Region ◽  
High Sensitivity ◽  
Gene Rearrangements ◽  
American Society

Abstract Hodgkin’s disease (HD) represents a malignant lymphoma in which the putative malignant Hodgkin and Reed-Sternberg (H-RS) cells are rare and surrounded by abundant reactive cells. Single-cell analyses showed that H-RS cells regularly bear clonal Ig gene rearrangements. However, there is little information on the clinical evolution of HD in a given patient. In this study, we used the single-cell polymerase chain reaction (PCR) to identify H-RS cells with clonal Ig gene rearrangements in biopsy specimens of patients with relapsed HD. The obtained clonal variable region heavy-chain (VH) gene rearrangements were used to construct tumor-clone-specific oligonucleotides spanning the complementarity determining region (CDR) III and somatically mutated areas in the rearranged VHgene. A number of biopsies were obtained during a period of 3 years from two HD patients. H-RS cells with identical VHrearrangements were detected in two separate infiltrated lymph nodes from one patient with nodular sclerosis HD. In a second patient with mixed cellularity HD subtype, clonal VH rearrangements with identical sequences were detected in infiltrated spleen and two lymph node biopsies. Despite the high sensitivity of the PCR method used (one clonal cell in 105 mononuclear cells), residual H-RS cells were not found in peripheral blood, leukapheresis material, purified CD34+ stem cells or bone marrow. The results show that different specimens from relapsed patients suffering from classical HD carry the same clonotypic IgH rearrangements with identical somatic mutations, demonstrating the persistence and the dissemination of a clonal tumor cell population. Thus, PCR assays with CDRIII-specific probes derived from clonal H-RS cells are of clinical importance in monitoring the dissemination of HD and tumor progression and could be useful for analysis of minimal residual disease after autologous stem cell transplantation. © 1998 by The American Society of Hematology.

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