Neoplastic Cells In Peripheral Blood Research Articles

Identification of peripheral blood involvement in dogs with large B-cell lymphoma: Comparison of different methods

Stage V lymphoma is defined as the presence of neoplastic cells in peripheral blood (PB), bone marrow, or any other non-lymphoid tissue. Still, official guidelines do not specify which technique should be used to assess infiltration.We assessed the agreement among flow cytometry (FC), blood smear evaluation, and ADVIA120 (LUC and BASO) to quantify PB infiltration in 100 dogs with large B-cell lymphoma (LBCL).Significant errors were found for all methods compared to FC. A moderate agreement was present between FC and blood smear evaluation, whereas LUC and BASO had excellent specificity but unsatisfactory sensitivity in detecting FC infiltrated PB samples.The different techniques should not be used alternatively. We support the use of LUC/BASO as a speedy preliminary test to detect infiltrated samples, and the joined use of blood smear evaluation and FC to quantify definitively the infiltration. Our results are valid only within canine LBCL staging workup, once the diagnosis has been confirmed.

PCM1-JAK2 Fusion Product Selectively Activates MAP-Kinase Pathway and Fails to Phosphorylate JAK2 and STAT5: Implications for Therapeutic Strategies.

AbstractAbstract 2858Janus-activated kinase 2 (JAK2) gene mutations and translocations are involved in the pathogenesis of a variety of hematologic malignancies. Among different translocation partners, pericentriolar material 1 (PCM1)-JAK2 fusion products have been described in rare cases of both lymphoid and myeloid neoplasms characterized by morphological (myeloproliferaton, eosinophilia, myelofibrosis) and clinical (striking male predominance, aggressive course) similarities.We recently identified a new case of the rare translocation PCM1-JAK2 in a 29-year-old man presenting with atypical chronic myeloid leukemia (aCML) and peculiar aspects of diserythropoiesis in the bone marrow (BM): abundant paratrabecular clusters of proerythroblasts associated with marked reduction of mature erythroid compartment (Sammarelli et al., S.I.E.S. 12th Meeting, 2012).For the first time we describe here the erythroid differentiation capacity of ex-vivo expanded CD34+ cells from this PCM1-JAK2 fusion case, as well as the signaling pathways activated in peripheral blood neoplastic cells (PBNC) harboring the translocation.Presence of the PCM1-JAK2 fusion transcript in PBNC was confirmed by nested RT-PCR using primers derived from PCM1 exon 25 and JAK2 exon 9 described in Reiter et al. Cancer Res. 2005 (Figure 1).CD34+ cells were isolated from the peripheral blood (PB) of the patient and cultured in serum-free medium supplemented with erythropoietin (EPO), interleukin-3 (IL-3) and stem cell factor (SCF) to induce erythroid differentiation; erythroid cell output [evaluated in terms of fold increase (FI) and glycophorin-A (GlyA) expression at day 14 of culture] was compared to the one obtained from PB CD34+ cells from a polycythemia vera patient (PV), in which JAK2 is constitutively activated by V617F point mutation, and to CD34+ cells from a G-CSF-mobilized donor (M).As shown in Figure 2, FI and GlyA expression were significantly lower in our patient compared to M and PV (FI: 0.63, 6.47 and 7.92 respectively; GlyApos cells: 4.2%, 51.6% and 64.2%, respectively) consistently with the diserythropoietic picture in the BM.We then investigated the activation of the 3 main signaling pathways associated to Receptor tyrosine kinases and most commonly turned on in cancer: Mitogen-activated protein (MAP) kinase pathway, JAK/Signal transducer and activator of transcription (STAT) pathway and phosphatidylinositol 3-kinase (PI3K)/AKT pathway, evaluating, by Western Blot analysis, levels of phosphorylation of Extracellular signal-Regulated Kinase (ERK1/2), JAK2, STAT5 and AKT in PBNC from our PCM1-JAK2 case and in PB mononuclear cells (PBMC) from 5 healthy control subjects (C1-C5).Although these signaling cascades are deeply interconnected, we surprisingly found a selective activation of the sole MAP-kinase pathway in PBNC (Figure 3).These data suggest that, while presence of JAK2V617F mutation leads to ligand-independent activation of STAT5, AKT and ERK1/2 (Laubach et al. Exp. Hematol. 2009), PCM1-JAK2 fusion product fails to activate JAK/STAT and PI3K/AKT axis. Specifically, reduced STAT5 activation might explain impaired erythroid differentiation of CD34+ cells in vitro as well as the marked aspects of diserythropoiesis in the BM.The signaling signature of PMC1-JAK2 neoplastic cells described here has also relevant implications on the treatment strategy for these patients. In particular, given the lack of activation of JAK2 and its down-stream partner STAT5, JAK-inhibitor therapy does not seem the ideal candidate in this specific setting. [Display omitted] [Display omitted] [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Carcinocythaemia (carcinoma cell leukaemia) in a dog: an acute leukaemia‐like picture due to metastatic carcinoma

An eight-year-old entire female boxer was presented with a two-week history of anorexia and lethargy and two-day history of unilateral left epistaxis. Clinical findings and laboratory test results suggested disseminated intravascular coagulation. On blood smear evaluation, occasional large epithelioid-like unclassified cells were detected. Occasionally these cells were organised in small clusters. Bone marrow examination revealed a marked infiltration by a malignant population of the same epithelioid-like cells. The dog was euthanased because of the guarded prognosis. Following histology and immunohistochemistry, a widespread undifferentiated carcinoma of unknown primary origin was diagnosed. To the authors' knowledge, this is the first case of carcinoma cell leukaemia reported in a dog. Carcinoma cell leukaemia is a rare oncological condition previously described in humans, characterised by non-haematopoietic neoplastic cells in peripheral blood.

Disseminated histiocytic sarcoma with peripheral blood involvement in a Bernese Mountain dog

A 6-year-old Bernese Mountain dog was presented with a history of lethargy and weight loss of 2 weeks duration. On physical examination the dog had pale mucous membranes and tachypnea. Ultrasound examination revealed hepatomegaly, splenomegaly, and mesenteric lymphadenomegaly. Results of a CBC included marked normocytic normochromic nonregenerative anemia, marked thrombocytopenia, and moderate leukocytosis with mild neutrophilia and a large population of unclassified round cells (6.2 x 10(3)/microL). The unclassified cells occasionally were bi- or multinucleated and had variably abundant pale basophilic cytoplasm that contained multiple irregular clear vacuoles and occasionally erythrocytes. Fine needle aspirate specimens of the mesenteric lymph nodes and spleen were composed of a population of round pleomorphic cells with the same features as the circulating cells. On flow cytometric analysis of peripheral blood, the unclassified cells expressed CD18, CD45, CD11c, CD1c, and CD14; immunocytochemical analysis of blood smears also indicated the cells were positive for CD1c, CD1a, and CD11c. The dog died a few hours after referral. The histologic interpretation of samples collected from spleen, liver, and lymph nodes was malignant neoplasia of histiocytic origin. Immunohistochemical staining yielded negative results for CD11d, a marker of red-pulp macrophages, ruling out hemophagocytic histiocytic sarcoma. Based on clinical and pathologic findings, the final diagnosis was disseminated histiocytic sarcoma (DHS) with peripheral blood involvement. To our knowledge, DHS in a dog with evidence and immunophenotyping of neoplastic cells in peripheral blood has been reported only rarely.

Agressive Lymphoma without Lymphadenopathy

Hepatosplenic T cell lymphomas are among a rare breed of aggressive non Hodgkin tumors that usually comprise of immature cytotoxic T cells. They most commonly involve young adults who generally present with constitutional symptoms of fever, weight loss and night sweats. Diagnosis can be challenging in the absence of lymphadenopathy and paucity of bone marrow findings. The discovery of neoplastic cells in peripheral blood is often a finding late in the clinical course. Demonstation of sinusoidal infiltration of T cells on either liver or splenic biopsy is ususally diagnostic. These cells are positive for clonal rearrangement of the γ gene of the T cell receptor which is a hallmark of this disease. Prognosis remains poor and most patients die within two years of diagnosis. We report a case of a patient who presented with unexplained fever and hepatosplenomegaly and after extensive workup was diagnosed with hepatosplenic T cell lymphoma.A 42 year old male with no significant past medical history presented with complains of abdominal pain, fever, malaise and 30lb weight loss over a period of six weeks. Pain was described as intermittent episodes of bilateral flank fullness associated with nausea and vomiting. He had traveled to Mexico 8 weeks earlier and according to him his symptoms began soon after his trip. He denied any headaches, cough, shortness of breath,. No dysuria, and no diarrhea. No smoking alcoholism or illicit drug use was reported. No sick or new sexual contacts. Family history was noncontributory. Vital signs were significant for temperature of 101 F. Physical examination revealed presence of marked hepatosplenomegaly. There was no lympadenopathy as well as no murmurs or rubs. Labs showed pancytopenia with blood counts of 2.7, 9.8 and 101 for WBC, Hb and platelets respectively. Additionally, alkaline phosphatase was elevated while rest of the liver functions was normal. Preliminary diagnosis of infectious versus malignant etiology was made. Peripheral smear did not show any malignant cells. Infectious workup returned negative for HIV, hepatitis, histoplasma, brucella, leptospira, coccidioidomycosis, syphilis, leishmaniasis as well as malaria and dengue fever. A bone marrow biopsy was performed for further evaluation and showed moderately hypercellular bone marrow with erythroid and megakaryocytic hyperplasia but no evidence of malignancy or granulomatous process. Meanwhile patient remained persistently febrile without any growth on blood cultues. An echo was normal with no evidence of endocarditis. Tuberculin test was non reactive. Given the results of the bone marrow biopsy and hepatosplenomegaly without any lymphadenopathy it was thought that a localized infiltrative process of he liver or spleen was likely. Patient thus underwent a liver biopsy disclosing intra sinusoidal collections of atypical CD3, CD7, TIA-1 and CD56 positive lymphocytes also bearing T cell receptor clonal rearrangement consistent with a diagnosis of hepatosplenic T-cell lymphoma which was further classified as gammadelta type.. He was subsequently started on ECHOP regimen and is currently in remission on a three month followup Hepatosplenic lymphoma, though is a rare tumor, should be considered in patients with unexplained hepatosplenomegaly without lymphadenopathy and normal peripheral smear. Liver biopsy is diagnostic most of the time.

Composite thymoma and chronic lymphocytic leukemia/small lymphocytic lymphoma involving the anterior mediastinum.

c Simultaneous involvement of the anterior mediastinum by thymoma and B-cell chronic lymphocytic leukemia/ small lymphocytic lymphoma (CLL/SLL), to our knowledge, has not been previously reported. We describe a composite tumor composed of thymoma and CLL/SLL incidentally discovered in a 62-year-old man who had no history of malignant diseases or immunologic disorders. The preoperative peripheral blood specimen showed a normal complete blood cell count and differential count. The diagnosis was established by histologic examination and immunophenotypic studies of the surgically excised anterior mediastinal mass. Postoperatively, bone marrow aspiration and biopsy specimens showed morphologic evidence of CLL/SLL, and the presence of neoplastic cells in peripheral blood and bone marrow was confirmed by flow cytometry immunophenotypic analysis. (Arch Pathol Lab Med. 2003;127:e76‐e79)

Increased risk of tumor seeding after percutaneous radiofrequency ablation for single hepatocellular carcinoma.

Radiofrequency (RF) ablation is an alternative to percutaneous ethanol injection (PEI) for single nonsurgical hepatocellular carcinoma (HCC) and is currently used as adjuvant therapy before liver transplantation. This phase II study assesses the treatment-related complications and response rate of RF for the treatment of single HCC < or = 5 cm. Percutaneous RF was performed under conscious sedation and ultrasound (US) guidance with an electrical generator connected to a single cooled-tip electrode. Neoplastic cells in peripheral blood (reverse transcription-polymerase chain reaction for alpha fetoprotein [AFP] messenger RNA) were analyzed before and after RF. Treatment response was assessed by spiral computed tomography (CT) at 1 month and every 3 months by US or spiral CT thereafter. Thirty-two patients (20 men; age 67 +/- 4 years; 78% hepatitis C virus; 24 Child-Pugh A) with a mean tumor size of 2.8 cm (25 patients < or = 3 cm) were treated by RF (1.25 sessions; mean time, 22.1 +/- 2 minutes). Adjuvant PEI was performed in 9 cases. Complete response was achieved in 21 patients (65%), being significantly higher for HCC < or = 3 cm (76% vs. 29%, P = .03). After a median follow-up of 10 months, 8 patients showed treatment-related morbidity. Four of them (12.5%) showed biopsy-proven needle-track seeding detected between 4 to 18 months. Neoplastic seeding was related to subcapsular location (P = .009), poor differentiation degree (P = .02), and baseline AFP levels (P = .02). Thus, RF ablation with cooled-tip needle for HCC is associated with a high risk of neoplastic seeding. Iatrogenic dissemination was related to subcapsular location or an invasive tumoral pattern, and has to be considered when selecting curative treatments for HCC or adjuvant therapies before liver transplantation.

Changes in the distribution of cells expressing tumour-associated antigen in lymph nodes during the progression of enzootic bovine leukosis

A monoclonal antibody (MAb), c143, that recognizes a tumour-associated antigen that is "upregulated" on neoplastic B cells in cattle with enzootic bovine leukosis (EBL), was used as a marker to study disease progression. An immunohistochemical examination of neoplastic tissue and superficial cervical lymph nodes from 14 animals with EBL revealed three morphologically definable stages of change in the structure of lymph nodes, associated with the distribution of c143-positive cells: (1) the presence of c143-positive cells at the marginal sinus with no apparent changes in lymph node structure; (2) the presence of positive cells extending into and distorting the architecture of the lymph node, with clear evidence of proliferation before overt changes (enlargement of lymph nodes) were evident; and (3) the presence of positive cells throughout the lymph node with total disruption of lymph node structure when clinical signs of lymph node enlargement were evident. The results indicated that the bovine leukaemia virus-transformed lymphocytes or neoplastic cells in peripheral blood accumulate in the marginal sinus area at the earliest stages, and subsequently proliferate and infiltrate into follicles, leading to the development of clinical signs of lymphosarcoma.

Surface Markers of Neoplastic Cells in Peripheral Blood from Leukemic Lymphoma

Surface Markers of Neoplastic Cells in Peripheral Blood from Leukemic Lymphoma