CD34 Antigen Expression Research Articles

Transcription factor ERG is a specific and sensitive diagnostic marker for hepatic angiosarcoma

To investigate the expression of ERG, CD34, CD31 (PECAM-1, platelet/endothelial cell adhesion molecule 1) and factor VIII-related antigen (FVIIIRAg) in the diagnosis of hepatic angiosarcoma patients. Patient samples were collected from January 1986 to December 2012 from the People's Liberation Army General Hospital in Beijing, China. We obtained twenty-four samples of hepatic angiosarcoma (HAS) that were confirmed by two pathologist. The samples were the result of three autopsy cases, eight biopsy cases and 13 patients who underwent surgical tumor removal. The HAS cases accounted for 2.23% (24/1075) of all hepatic vascular tumors at the hospital during the same time period. Patient histories including age, gender, clinical manifestations, medical treatments, laboratory tests, radiological images, histological observations and outcomes for each case were analyzed in detail. All samples were evaluated histologically with hematoxylin and eosin staining. Using immunohistochemistry, the expression and localization of ERG was examined in all HAS specimens and compared to the known endothelial markers CD34, CD31 and FVIIIRAg. The endothelial markers were also evaluated in a panel of non-HAS tumors. This cohort of 24 HAS cases is, to the best of our knowledge, currently the largest cohort in the world in the publicly available literature. Hepatic angiosarcoma tissue samples were obtained from 14 males and 10 females with a mean age of 50.6 years (range: 7-86 years). The patients presented with the following clinical manifestations: abdominal pain (16/24), back pain (3/24), heart palpitations (1/20), cough (1/24) or no clinical symptoms (3/24). Tumors were predominantly localized in the right hepatic lobe (15/24) or left hepatic lobe (6/24), or a diffuse growth on the right and left hepatic lobes (3/24). Eleven patients underwent surgical resection (45.8%), two patients received a liver transplant (8.3%), eight patients received interventional therapy (33.3%) and three patients received no treatment (lesions discovered at autopsy, 12.5%). Postoperative follow-up of patients revealed that 87.5% (21/24) of patients had died and three cases were not able to be tracked. In all cases, the mean survival time was 12.1 mo. While 100% of the HAS samples were positive for ERG expression, expression of the other markers was more variable. CD31 was expressed in 79.2% (19/24) of samples, CD34 was expressed in 87.5% (21/24) of samples and FVIIIRAg was expressed in 41.7% (10/24) of samples. ERG is a more sensitive and specific diagnostic marker for hepatic angiosarcoma in comparison to CD31, CD34 and FVIIIRAg.

Aberrant myeloid antigen co-expression is correlated with high percentages of CD34-positive cells among blasts of acute lymphoblastic leukemia patients: an Indian tertiary care center perspective.

BackgroundAberrant myeloid antigen (MA) co-expression and high expression of CD34 antigen on the blasts of acute lymphoblastic leukemia (ALL) patients are independently reported to have a role in pathogenesis and prognosis. This study was conducted to determine whether these two parameters are related.MethodsA total of 204 cases of ALL were included in an analysis of blast immunophenotypic data. CD34 expression was categorized as low when less than 50% of blasts were CD34-positive (CD34low) and as high when 50% or more were CD34-positive (CD34high).ResultsOf 204 cases of ALL, 163 and 41 were of B-cell origin (B-ALL) and T-cell origin (T-ALL), respectively. Of all cases, 132 (64.7%) showed co-expression of MA and among these, 101 (76.51%) were CD34high, while the remaining 31 (23.48%) were CD34low. Of 72 cases without MA co-expression, 25 (34.72%) were CD34high and 47 (67.25%) were CD34low. Furthermore, of 163 cases of B-ALL, 111 showed co-expression of MA and 84 of these were CD34high. Of 52 cases of B-ALL without MA expression, 22 were CD34high. Among 41 cases of T-ALL, 21 co-expressed MA, 17 of which were CD34high. Moreover, all 20 cases of T-ALL without co-expression of MA were CD34low. These differences were statistically significant.ConclusionWe observed a strong correlation between aberrant MA expression and CD34high expression on the blasts of ALL. We hypothesize that these different patient subsets may represent unique prognostic characteristics.

Histological and immmunohistochemical assessment of the immunological aspect of certain cells in human palatine tonsils

Background Tonsils contain four specialized lymphoid compartments that together are involved in immune functions. The capacity of tonsillar lymphocytes to counter infections may be altered during one’s lifetime. The classification of lymphocytes by CD antigen expression is now widely used in clinical medicine and experimental immunology. Purpose The present work was designed to study the distribution of CD4 and CD8 antigen expression in T lymphocytes in human tonsils at different periods of life. Materials and methods Sixty-two tonsillar specimens were obtained from still birth infants and from children aged 1–9 years. Paraffin sections were prepared and stained with H&E and with immunohistochemical stains to demonstrate CD4 and CD8 T lymphocytes. The distribution of these cells in the different components of the tonsils was evaluated with an image analyzer. The obtained data were statistically analyzed using SPSS. Results There was a significant increase in the distribution of stained CD4 and CD8 T lymphocytes in the interfollicular areas, mantle zones of lymphoid follicles, and partially in the germinal centers of the examined tonsils with the advancement of age. Conclusion Activated T lymphocytes differentiate into several subtypes, among which are CD4 and CD8 cells. These types of T lymphocytes express surface antigens, which can interact with different foreign pathogens.

Epigenetic Status Of CEBPA Promoter and Expression Of CEBPα Correlates With Molecular Genetic Subtype and CD2 Aberrant Expression In B Cell Precursor ALL

CCAAT/enhancer binding protein alpha (CEBPα) is one of the crucial transcription factors involved in hematopoietic differentiation and leukemogenesis. CEBPα promotes myeloid differentiation by up-regulation of lineage specific genes and by cell proliferation arrest. Epigenetic regulation of CEBPα expression through DNA methylation has been demonstrated in acute myeloid leukemia (AML) (Figueroa et al, Cancer Cell, 2010). However, only limited data are available regarding CEBPA promoter methylation and its expression in B cell precursor acute lymphoblastic leukemia (BCP-ALL).Methylation status of CEBPA promoter (-295 to -593bp upstream of the transcription start site (TSS), 24 CpG dinucleotides) was analyzed by bisulfite sequencing. Five subgroups of BCP-ALLs were analyzed: MLL gene rearranged (n=5), hyperdiploid (n=6), mBCR-ABLpos(n=5), ETV6-RUNX1pos(n=6) and other BCP-ALLs (no hyperdiploidy, MLL gene rearrangement, BCR-ABL or ETV6-RUNX1 fusion gene (“BCP-others”, n=29)). CEBPA promoter was hypermethylated in MLL-rearranged, hyperdiploid and ETV6-RUNX1pos BCP-ALL (5/5, 6/6 and 4/6 respectively). Surprisingly CEBPA promoter was hypomethylated in all mBCR-ABLpos cases (5/5). In subgroup of other BCP-ALLs both hypermethylation (10/29) and hypomethylation of CEBPA promoter (19/29) were detected (Figure 1A). [Display omitted] In previous study we found association of CD2 (LFA-2) aberrant expression and switch to the monocytic lineage during the early phase of treatment in BCP-ALLs (Slamova et al, ASH 2012). We were interested if a possible link between hypomethylation of CEBPA promoter correlates with aberrant expression of CD2. There was a significant association between aberrant expression of CD2 antigen and hypomethylation in CEBPA promoter in BCP-others (Fisher exact test, p<0.0001). Interestingly, in the only hypomethylated ETV6-RUNX1pos case we found aberrant CD2 expression on blasts, which is exceptional in ETV6-RUNX1pos ALL.We next asked whether methylation of CEBPA promoter correlates with CEBPα expression. It is generally accepted that promoter hypomethylation is often associated with increased expression of the relevant gene. Our data prove that in general, this holds true also for BCP-ALL. However, in two genetically defined subsets we observed either high expression despite hypermethylation (MLL-rearranged ALL) or low expression despite hypomethylation (mBCR-ABLpos ALL) (Figure 1B). In BCP-others hypomethylation of CEBPA promoter was significantly associated with upregulation of myeloid antigens (CD14 and/or CD33) and downregulation of B cell marker CD19 on blasts during the first weeks of the treatment (Fisher test, p=0.0009). In summaryMethylation status of CEBPA promoter correlates with genetic subtypes of BCP-ALL. The notion that hypomethylation leads to overexpression was confirmed in majority of BCP-ALLs, while in mBCR-ABLpos and MLL gene rearranged BCP-ALL it did not follow this pattern. Hypomethylation of CEBPA promoter in BCP- others correlates with CD2 expression on blasts and increased CEBPα gene expression. During the early phase of the treatment in other BCP-ALLs with hypomethylated CEBPA promoter increase of myeloid and decrease of B lymphoid markers on blasts was observed.Supported by: GACR P301/10/1877, GACR P304/12/2214, GAUK 914613, UNCE 204012, NT13462, NT12397- 4, project for conceptual development of research organization (Ministry of Health, CZ) 00064203, the FACS Aria instrument was supported by EU-Prague project CZ.2.16/3.1.00/24022 Disclosures:No relevant conflicts of interest to declare.

Immunophenotypic shift of CD4 and CD8 antigen expression in primary cutaneous T‐cell lymphomas: a clinicopathologic study of three cases

Primary cutaneous T-cell lymphomas (CTCL) comprise a heterogeneous group of neoplasms with diverse clinical behavior. Mycosis fungoides (MF) is the most common type of CTCL. Immunophenotypical shift during progression of the disease is a rare event and its significance is unknown. We present three primary CTCL cases that showed an immunophenotypical shift and poor prognosis. Conventional hematoxylin/eosin and immunohistochemical-stained sections were examined in all the cases. Molecular analysis for rearrangement of the T-cell receptor (TCR) gene was performed in two cases. One case was classified as MF, while the other two lacked epidermotropism, and were considered primary cutaneous peripheral T-cell lymphoma (PTCL), NOS. Two cases were CD3+/CD4+ and one case was CD3+/CD8+ at diagnosis. The first two patients suffered many relapses and eventually, new CTCL lesions with a CD3+/CD8+ phenotype were observed. Both cases revealed identical clonal TCR rearrangements on the initial and late lesions, supporting the interpretation of a single clonal proliferation with different phenotypes. The third case progressed with skin recurrences and pulmonary lesions with a predominant CD3+/CD4+/CD8- phenotype. All cases manifested poor prognosis and two patients died of lymphoma. Immunophenotypical shift between CD4 and CD8 in CTCL seems to be a rare phenomenon that may be associated with disease progression.

The Surface Molecular Signature of Leukemic Cells Is Associated with NPM1 Mutations and FLT3-ITD in Patients with de novo Acute Myeloid Leukemia

Certain molecular mutations are associated with signs of cell morphology and differentiation in acute myeloid leukemia (AML). However, only limited data are available for the detailed analysis of such correlations. In this study, AML patients were classified into 4 subsets according to CD34, HLA-DR and CD11c expression levels. Significantly low CD34 antigen expression was observed in nucleophosmin (NPM1)-mutated patients and in those with FMS-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD). No correlations were observed among NPM1 mutations, FLT3-ITD and monocytic morphology in patients without CD34 expression. Both NPM1 mutations and FLT3-ITD were absent in cluster IIb patients (CD34⁺CD11c^-). The associations among NPM1 mutations, FLT3-ITD and the surface molecular signature of leukemic cells may offer beneficial information about the pathogenesis of AML.

Pathogenesis of human hemangiosarcomas and hemangiomas

Hemangiosarcomas are uncommon aggressive vascular tumors that have recently become the focus of attention because several chemicals and pharmaceuticals increase their incidence in mice. The relevance of these mouse vascular tumors to humans is unclear. In the present study, we semiquantitatively evaluated the expression profiles of hematopoietic stem cell markers (CD117 [c-kit], CD133, CD34, and CD45), endothelial cell markers (vascular endothelial growth factor receptor 2, CD31, and factor VIII-related antigen), and a myeloid lineage cell marker (CD14) in human hemangiosarcoma (n = 12) and hemangioma (n = 10) specimens using immunohistochemistry. CD133 was completely negative in almost all cases of hemangiosarcomas and hemangiomas. Most hemangiosarcomas, but not hemangiomas, stained for CD117 and CD45. Both groups diffusely expressed CD34, vascular endothelial growth factor receptor 2, and factor VIII-related antigen; however, hemangiomas had more intense and diffuse CD34 and factor VIII-related antigen expression compared with hemangiosarcomas, whereas CD31 was positive in all hemangiosarcomas but only half of the hemangiomas. CD14 staining was negative in most hemangiosarcoma and hemangioma cases. Our results indicate that multipotential bone marrow-derived hematopoietic stem cells or early endothelial progenitor cells (EPCs) expressing CD117, CD34, and CD45 are involved in hemangiosarcoma formation, whereas hemangiomas originate from late EPCs or differentiated endothelial cells, which have lost the expression of most hematopoietic stem cell markers. This contrasts with our previous results that demonstrated that both hemangiosarcomas and hemangiomas in mice may be derived from early EPCs that are not completely differentiated.

Peripheral T-cell lymphoma emerging in a patient with aggressive polymyositis: molecular evidence for neoplastic transformation of an oligoclonal T-cell infiltrate

We report a rare case of peripheral T-cell lymphoma arising in a 52-year-old man with biopsy-proven aggressive polymyositis, who had cardiac involvement, progressive bulbar symptoms, and died 11months post diagnosis due to multiorgan failure. Using a multimodality approach including immunohistochemistry, genome-wide single nucleotide polymorphism (SNP)-array analysis, and high-throughput sequencing of the complementary determining region 3 (CDR3) of T-cell receptor beta (TCRβ) genes, our study demonstrates a molecular link between polymyositis and T-cell lymphoma, and provides evidence of the rapid and possibly late occurrence of genomic instability during neoplastic transformation of an oligoclonal T-cell population. Immunohistochemical analysis revealed loss of CD5, CD7, and CD8 antigen expression in autopsy tissue samples, as well as the occurrence of aberrant CD56 expression, not seen in pre-mortem biopsies, supporting the emergence of a neoplastic T-cell population. Multiplex polymerase chain reaction and next-generation sequencing of the TCRβ CDR3 region displayed two unique T-cell clones in both the diagnostic biopsy confirming polymyositis and the autopsy muscle tissue exhibiting T-cell lymphoma, linking the two pathological processes. SNP-array analysis revealed complex genomic abnormalities at autopsy but not in the pre-mortem muscle biopsies displaying polymyositis, confirming malignant transformation of the oligoclonal T-cell infiltrate. Our findings raise the possibility that clinically aggressive polymyositis might represent a preneoplastic condition in some instances, similar to certain other autoimmune and inflammatory disorders.

Experimental model for the study of the effects of platelet-rich plasma on the early phases of muscle healing.

There is abundant evidence suggesting that growth factors may play a key role in the healing process, especially in the early stages of inflammation. Despite the reported clinical successes with the use of growth factors there is still a lack of knowledge on the biological mechanism underlying the activity of platelet-rich plasma during the process of muscle healing. The aim of this study was to analyse the early effects of platelet- rich plasma in an easily reproducible animal model. Wistar male adult rats (n=102) were used in this study. The muscle lesion was created with a scalpel in the flexor sublimis muscles. Platelet-rich plasma was applied immediately after surgery. Treated, untreated and contralateral muscles were analysed by morphological evaluation and western blot assay. Leucocyte infiltration was significantly greater in muscles treated with platelet-rich plasma than in both untreated and contralateral muscles. The latter showed greater leucocyte infiltration when compared to the untreated muscles. Platelet-rich plasma treatment also modified the cellular composition of the leucocyte infiltration leading to increased expression of CD3, CD8, CD19 and CD68 and to decreased CD4 antigen expression in both platelet-rich plasma treated and contralateral muscles. Blood vessel density and blood vessel diameters were not statistically significantly different between the three groups analysed. The results of this study showed that treatment with platelet-rich plasma magnified the physiological early inflammatory response following a muscle injury, modifying the pattern of cellular recruitment. Local platelet-rich plasma treatment may exert a direct or, more plausibly, indirect systemic effect on healing processes, at least in the earliest inflammatory phase.

Use of CD30 expression in human malignant mesothelioma as a novel therapeutic target.

e18526 Background: CD30 antigen is important in the diagnosis of Hodgkin’s disease (HD) and anaplastic large cell lymphoma (ALCL). CD30 is a cytokine receptor that belongs to the tumor necrosis factor superfamily (TNFRSF8) and acts as a positive regulator of apoptosis. The expression of CD30 in malignant mesothelioma is unknown. Given the remarkable success of brentuximab vedotin, an antibody-toxin conjugate directed against CD30 antigen, in treating patients with relapsed HD and ALCL, we undertook a study to validate the incidence of CD30 expression in mesothelioma and to examine its clinical correlates. Furthermore, we performed studies using cultured mesothelioma and non-small cell lung cancer (NSCLC) cell lines to determine the expression and potential targeting of CD30 antigen in vitro. Methods: Patient mesothelioma tissue specimens (n=52) were examined for CD30 expression by immuno-histochemistry (IHC) with an anti-BerH2 antibody (Cell Marque) used for the diagnosis of HD. Staining was assessed by a thoracic pathologist. Mesothelioma (H28, H2052, H2452, 211H) and NSCLC (A549, H1299) cell lines were also grown in culture and examined for CD30 expression by FACS analysis and confocal microscopy using anti-CD30 antibody (Novus Biologicals). Cells were permeabilized with Triton X-100. Results: Positive CD30 expression was noted in six out of 52 total mesothelioma specimens. Five of the six positive tumors demonstrated epithelial histology and were scored as low or intermediate grade. Membrane-associated as well as diffuse cytoplasmic staining was observed. The percentage of tumor cells stained positive varied greatly. Interestingly, no high grade epithelial tumors were scored as CD30 positive. The remaining CD30 positive tumor was a high grade, biphasic metastatic tumor. Confocal analysis of both mesothelioma cultured cells localized CD30 antigen to the cell membrane. Flow cytometry of intact and permeabilized mesothelioma cells supported the idea that CD30 antigen was located on the cell-surface. Conclusions: Our studies validate the presence of CD30 antigen on the cell surface of epithelial-type mesothelioma and indicate that selected mesothelioma patients may derive benefit from brentuximab vedotin (Adcetris).

Activated CD8+ T cells exhibit blunted effector functions in early HIV-1 disease (P3035)

Abstract Elevated, persistent CD8+ T cell activation is a hallmark of HIV-1 infection. CD8+ T cell activation is measured by expression of CD38 and HLA-DR antigens on CD8+ T cells from the blood. How CD8+ T cell activation state may define impairments to CD8+ T cell function remains unclear. ERK1/2 signaling is critical for multiple T cell functions, and interruption of this pathway due to immune activation and inflammation may blunt CD8+ T cell function. We previously demonstrated that a major proportion of activated (CD38+HLA-DR+) CD8+ T cells from recently HIV-1-infected antiretroviral-treatment-naïve adults are refractory to phosphorylation of the ERK1/2 kinases (p-ERK1/2-refractory). Here we hypothesized that p-ERK1/2-refractory CD8+ T cells would exhibit reduced effector function compared to CD8+ p-ERK1/2-responsive cells. We combined single-cell phospho-kinase flow cytometry with intracellular cytokine staining into a single assay (PFICS), to examine IFN-γ and perforin responses in CD8+ T cells by ERK1/2 signaling ability. On a per cell basis, p-ERK1/2-refractory cells produced less IFN-γ in response to polyclonal or HIV-1 Gag peptide stimulation (% IFN-γ+ cells was 5.5-fold lower, p=0.006 for polyclonal and 1.5-fold lower, p=0.048 for Gag) and expressed reduced levels of perforin (2.1 and 1.6-fold lower perforin MFI). Blunted CD8+ T effector functions, secondary to ERK1/2 signaling deficits concentrated within activated CD8+ T cells, may contribute to HIV immunodeficiency.

Persistent CD30 Expression by Embryonal Carcinoma in the Treatment Time Course: Prognostic Significance of a Worthwhile Target for Personalized Treatment

CD30 is expressed by untreated embryonal carcinoma, supporting the rationale for a targeted approach. However, the reported chemotherapy induced switching off of CD30 noted on immunohistochemistry may affect its therapeutic potential for disease relapse. We evaluated persistent CD30 expression and its prognostic meaning in cases of post-chemotherapy residual disease. Paraffin blocks of surgical samples that yielded nonteratomatous viable cells after 1 or more cisplatin based chemotherapy treatments were retrieved and reassessed by 2 pathologists blinded to the study purpose. Multivariable analysis was done for prespecified factors. A total of 49 cases of pure embryonal carcinoma or mixed germ cell tumor from August 1991 to August 2012 had full clinical data and suitable tissue available for analysis. Of the 35 cases (71.4%, 95% CI 56.7-83.4) with preserved CD30 positivity 14 (40.0%) showed residual disease after a median of 1 regimen (IQR 1-2). Five-year overall survival in CD30 positive and negative cases was 37.0% (95% CI 22.1-61.8) and 50.1% (95% CI 27.9-90.0, p=0.078), while after first line treatment it was 23.2% (95% CI 8.6-62.5) and 47.6% (95% CI 18.8-100, p=0.025), respectively. On multivariable analysis CD30 positivity was a significant prognostic factor for progression-free survival (HR 2.32, 95% CI 1.04-5.19) and overall survival (HR 2.77, 95% CI 1.05-7.29). CD30 was retained even after an intensive pretreatment load, confirming that it is a reliable treatment target. Its expression was associated with a significantly poorer prognosis in multiple relapse/chemoresistant cases and it was an independent prognostic factor for survival.

Phagocytic plasma cells in a patient with smoldering myeloma

A 43-year-old man, originally from Cuba, was referred in June 2012 because of a 6-month history of fatigue and weight loss. Physical examination was unremarkable. Laboratory findings revealed a IgG Lambda monoclonal protein level of 3.6 g/L serum IgG. Quantitative immunoglobulin determinations showed levels of IgG 14.8 g/L, IgA 1.4 g/L, and IgM 0.8 g/L. Lactate dehydrogenase (LDH) was slightly elevated at 284 U/L. Other biochemical data, including hemoglobin, reticulocyte count, platelet count, and leukocyte count, were within normal limits. Urine immunoelectrophoresis was negative for Bence Jones protein. Skeletal X-rays showed no lytic lesions. The bone marrow was normocellular, with normal maturing trilineage hematopoiesis. The aspirate showed 18 % infiltration of plasma cells, some of which contained several Russell bodies. Active phagocytosis, mainly of granulocytes, by plasma cells was a remarkable feature (Fig. 1). Immunophenotypic characterization of plasma cells showed expression of CD 38, CD 138, and CD56 antigens. The patient was diagnosed with smoldering myeloma. As recommended in the guidelines, the patient has been followed without starting chemotherapy. Under normal circumstances, plasma cells have no phagocytic function. Phagocytic plasma cells have been reported in association with lymphoproliferative disorders, and occasionally in patients with multiple myeloma. Phagocytosis by plasma cells in patients with smoldering myeloma is extremely rare and has only been described twice previously. In 1986 and 1987, two patients with asymptomatic multiple myeloma were described in which phagocytic plasma cells were present in the bone marrow. None of these reports provided an explanation for this phenomenon, and its clinical significance remains unknown. There have been speculations that the presence of CD15, a myeloid marker expressed on phagocytic myeloma cells, is associated with phagocytosis. There are other non-B cell antigens, such as CD56, that malignant plasma cells might rarely express. Normal plasma cells are typically CD56-negative. CD56 antigen is expressed in NK/T cell lymphoma, multiple myeloma, and AML. CD56 is a signaling receptor that impacts cellular adhesion, proliferation, differentiation, and apoptosis. Overexpression of CD56 in malignant neoplasms is associated with an aggressive tumor type, progression of multiple myeloma, and reduced overall survival in patients with AML and ALL. Jekarl et al. describe an association between CD56 antigen expression level and the percentage of hemophagocytosis in AML with t(16;21), but the mechanisms underlying this phenomenon remain unknown. A longitudinal retrospective study showed rapid progression to multiple myeloma in 85 % of CD56-positive MGUS patients. We speculate that the presence of CD56 antigen in our patient (Fig. 2) may not only be related to hemophagocytosis, but also lead to the rapid evolution of symptomatic multiple myeloma. The prognosis and time of progression to symptomatic multiple myeloma of patients with smoldering myeloma and phagocytic plasma cells are unknown. Further studies are needed to clarify the mechanisms leading to hemophagocytosis in plasma cells. T. Sayar Bahri (&) F. Weerkamp M. B. L. Leys Department of Internal Medicine, Maasstad Hospital, Rotterdam, The Netherlands e-mail: tanaz.bahri@gmail.com

Increased expression of CD69 antigen on human peripheral blood natural killer cells in patients with allergic rhinitis.

Allergic rhinitis (AR) is an inflammatory disorder of the nasal mucosa with high morbidity and prevalence. Natural killer (NK) cells might have a role in AR. We aimed to evaluate the changes of the markers and receptors on NK cells in AR patients compared to the non-atopic controls.Flow cytometric analysis was used with double staining of the Peripheral Blood Mononuclear Cells (PBMCs) to examine the expression of CD25 and CD69 markers, and NKG2D and NKG2A receptors on NK cells of 20 patients with AR and 20 non-atopic controls. The serum total IgE level was measured by Enzyme-linked Immunosorbent Assay.The expression of CD69 antigen on NK cells in AR patients was significantly higher than that of healthy group (p=0.03). No significant changes were observed between CD25, NKG2D and NKG2A expression on the surface of NK cells from healthy and AR subjects. Our study also showed that there was no significant correlation between the expression of CD69, CD25, NKG2D and NKG2A and level of serum total IgE in AR patients and normal subjects.These results indicated that the expression of CD69 antigen on NK cells of AR patients was increased. The high expression of CD69 on NK cells in AR patients suggested that these cells were activated, probably due to the cytokines secreted from allergen-stimulated T cells and activated monocytes.

Guzkowy Rozrost Limfoidalny—Rzadki Przypadek Choroby Limfoproliferacyjnej w Płucach

Nodular lymphoid hyperplasia (NLH) belongs to a very rare, mild, lymphoproliferative disease of unestablished aetiology historically included in the group of pseudolymphomas. Its existence was controversial for many years, until modern techniques of pathomorphological diagnosis approved it as a separate entity of lung disease. It manifests in the form of well limited nodules localized in the lungs, which are mostly identified accidentally. Clinical symptoms are rare and nonspecific; the disease usually occupies only one lung. Pathomorphological diagnosis requires immunohistochemical designation of expressions of numerous antigens in order to exclude malignant lymphoma of the lungs. Surgical resection is used in cases of larger nodules; the smaller ones require periodic observation, and the prognosis is good. The authors describe the case of 65-year-old woman with pulmonary nodules which were detected accidentally in the right lung. The patient was qualified for right-sided videothoracoscopy and removal of the lung nodule. In classic HE staining of the histological material, the presence of lymphoid infiltration of the lungs was revealed, which formed lymph follicles with reactive germinal centres. In order to differentiate from the malignant lymphatic expansion, immunohistochemical designations were made, which showed positive expression of CD20 antigen in the B cell zone, positive expression of the CD3 antigen in the T cells zone, positive expression of CD23 antigen in the lymph follicles, negative expression of bcl-2 in the lymph follicles, and positive expression of MIB-1 in the germinal centres of lymph follicles. Such a histopathological and immunohistochemical picture provided the basis for diagnosis of nodular lymphoid hyperplasia of the lung.

Melanoma Cells Inhibit NK Cell Functions—Letter

We have read with interest the recent paper by Pietra and colleagues who show that melanoma cells inhibit NKp30, NKp44, and NKG2D expression by allogeneic natural killer (NK) cells and their cytolytic activity (1). These results provide potential mechanistic insights for a number of observations previously made by others and by ourselves (2–5). Indeed, we found that NK cell infiltration is poor, if at all detectable, in various types of human cancers. By taking advantage of a panel of tumor tissue microarrays (TMA), including hundreds of 0.6 mm punches from clinically annotated biopsies, in agreement with a recently published paper (4), we observed that no CD56+ cell infiltrate was detectable in 203 (71.4%) of 284 melanoma. Furthermore, among the 81 remaining punches, surface expression of CD56 antigen was exclusively detected on melanoma cells in 24 specimens (Fig. 1B). We also found that 310 (92%) of 336 hepatocellular carcinomas (HCC), 376 (97%) of 385 breast cancers (Sconocchia and colleagues, unpublished data), and 108 (92%) of 117 renal cell carcinomas (RCC; ref. 3) showed no evidence of CD56+ cell infiltration.On the other hand, CD56+ cell infiltration with a mean number of 10 cells per punch or more was observed in 460 (38%) of 1,203 punches in colorectal cancers (CRC). Even in this case, however, CD56+ cell infiltration was devoid of prognostic significance (5). Similar results were obtained by using CD57 as a NK/NK-T cell marker. Notably, phenotypic analysis of freshly excised and enzymatically digested CRC specimens clearly indicated that CD56 and NKp46 were expressed in comparably (P = 0.09) low percentages of tumor infiltrating cells (5), suggesting that our findings could not be merely attributed to a downregulated CD56 expression.Importantly, more than 90% of RCC, HCC, CRC, and melanoma cells from the tumors included in the TMAs under investigation expressed high amounts of MHC class I-related chain molecule A/B (MICA/B; refs. 3, 5; Fig. 1A), thereby representing potentially excellent NK cell targets.In this respect, it is of interest that functional impairments of NK cell activities, possibly because of soluble factors released by tumor cells, including TGF-β1, have recently been observed also in patients with advanced breast cancers (2, 7).Further studies using autologous cancer cells are warranted to clarify whether molecular mechanisms underlying impaired NK cell functions in a tumor setting are specifically related to neoplastic transformation (3). Nevertheless, the data by Pietra and colleagues, Mamessier and colleagues, and Erdag and colleagues (1, 2, 4, 7), together with our observations question the relevance of NK cells in the control of solid tumor progression in humans.See the Response, p. 5430No potential conflicts of interest were disclosed.Conception and design: G. Sconocchia, S. Ferrone, G.C. SpagnoliDevelopment of methodology: G. SconocchiaAcquisition of data (provided animals, acquired and managed patients, provided facilities, etc.): R. Arriga, L. TornilloAnalysis and interpretation of data (e.g., statistical analysis, biostatistics, computational analysis): G. Sconocchia, R. Arriga, L. Terracciano, G.C. SpagnoliWriting, review, and/or revision of the manuscript: G. Sconocchia, R. Arriga, S. Ferrone, G.C. SpagnoliAdministrative, technical, or material support (i.e., reporting or organizing data, constructing databases): G. Sconocchia, R. ArrigaStudy supervision: G. Sconocchia, L. Terracciano, G.C. SpagnoliGiuseppe Sconocchia is supported by the AIRC grant: IG10555 grant.

Alpha-Interferon Secreting Blastic Plasmacytoid Dendritic Cells Neoplasm

We report a new case of blastic plasmacytoid dendritic cell neoplasm (BPDCN) with extensive immunophenoptyping, genotyping (karyotype, array-comparative genomic hybridization, and fluorescent in situ hybridization), and long-term tumor cells culture. BPDCN is a very rare and aggressive disease clinically characterized by a skin revealing localization more or less rapidly disseminating to the bone marrow and other organs with or without and leukemia. The disease was initially phenotypically characterized by the expression of both CD4 and CD56 antigens, whereas lymphoid and myeloid lineage antigens were negative. A phenotypic link with alpha-interferon (IFN-I)-producing plasmacytoid dendritic cells was demonstrated. The data collected in this case report provide additional biological and genotypical data on tumor cells of BPDCN. This study confirms the capability of tumor cells to secrete IFN-I, demonstrated by biological IFN-I activity of cultured cells and immunohistochemical expression of Mx-1 protein. Although a common genetic profile involving chromosomes 5, 6, 9, 12, 13, and 15 has been identified, no specific genetic marker has been demonstrated that is specific to BPDCN. The demonstration of ETV6 gene deletion in this case deserves further investigations as a putative BPDCN marker.

Clinical and laboratory features of adult biphenotypic acute leukemia

To analyze the clinical and biological characteristics of adult BAL. We identified 22 cases (4.4%) of BAL from 496 newly diagnosed acute leukemia patients and compared them with patients with acute myloid leukemia (AML) and acute lymphoblastic leukemia (ALL). CD34 positivity was higher in BAL (87%) than in ALL (60%, P = 0.04) and AML patients (63%, P = 0.05). The median age in the BAL group was greater than in ALL (43 vs 31 years, P = 0.04). Enlargement of liver, spleen and lymphonodes was more common in BAL (81.8%) than AML (46%, P = 0.003). Cytogenetic analysis revealed abnormal karyotypes in 56% of BAL patients. The overall completed remission rate in the BAL group (65%) was lower than in ALL (86%, P = 0.04). BAL patients showed a much higher incidence of older age, CD34 antigen expression, presence of Philadelphia chromosome, liver, spleen and lymphonodes infiltration and resistance to therapy.

CD20 Antigen Expression by Lymphoproliferative Disorders after Kidney Transplant is Independently Associated with a Poor Outcome: PTLD.Int Survey

Antigen expression by neoplastic cells is important because of its effects on the behavior and survival of patients. We sought to gather data on renal transplant recipients who had developed posttransplant lymphoproliferative disorders in their posttransplant era, and had a documented report on CD20 antigen testing. A comprehensive search of the literature was done for reports that indicate test results for the CD20 antigen in kidney recipients having lymphoproliferative disorders after transplant. Their demographics, disease characteristics, and prognoses were analyzed. CD20-positive posttransplant lymphoproliferative disorder patients had a significantly shorter time from transplant to developing posttransplant lymphoproliferative disorder (P < .001). None of patients who had early onset posttransplant lymphoproliferative disorder was CD20 negative. Bone marrow involvement was significantly more prevalent among CD20-negative patients (P < .05) with no CD20-positive patient developing a bone marrow metastasis. Log-rank test showed a relatively worse survival for renal recipients expressing the CD20 antigen (P = .07). CD20-positive posttransplant lymphoproliferative disorder lesions in kidney transplant patients are significantly more likely to develop early after transplant and represent an inferior outcome. We suggest that all renal transplant recipients who develop posttransplant lymphoproliferative disorder within their early time after surgery should be given anti-CD20 therapy. Future prospective studies are required to confirm our conclusions.

Pretreatment of leukemic cells with low-dose decitabine markedly enhances the cytotoxicity of gemtuzumab ozogamicin

Prognosis in relapsed/refractory acute myeloid leukemia (AML) is poor due to drug resistance. Gemtuzumab ozogamicin (GO) is a molecularly targeted drug, which consists of anti-CD33 antibody and calicheamicin. The high expression of CD33 antigen on most AML cells and exceptionally marked cytotoxicity of calicheamicin have attracted strong interest in GO, but clinical results on GO monotherapy have been disappointing, and methods of overcoming resistance to GO have accordingly been sought.