Expression of CD56 is a risk factor for acute lymphocytic leukemia with central nervous system involvement in adults

Objective: To gain further insights into the predisposing risk factors for central nervous system (CNS) involvement in patients with acute lymphocytic leukemia (ALL), the impact of CD56 expression in these patients was investigated.

Methods: We reviewed the clinical features of CD56 expression in 588 consecutive ALL patients treated with systemic chemotherapy regimens between 2000 and 2014. The categorical data from CD56⁺ ALL patients were compared with those from CD56^? ALL patients.

Results: Among the 588 patients studied, 18.9% showed CD56 expression. The expression was significantly associated with CD33⁺, CD10^?, CD15⁺, TdT^?, and CD5⁺ immunophenotypes. After systemic chemotherapy, 8.8% patients showed CNS involvement, of which 3.2% exhibited combined recurrences and 5.6% exhibited isolated CNS involvement. The 5-year event-free survival was significantly lower for patients with CD56⁺ immunophenotype compared with patients with CD56^? immunophenotype (22.5% vs. 32.7%, P?=?0.04). Cumulative incidences of CNS involvement were significantly greater in the CD56⁺ cohort compared with the CD56^? cohort (14.4% vs. 7.5%, P?=?0.02). Multivariate analysis revealed CD56 expression to be statistically significant risk factors for CNS involvement.

Conclusion: CD56 expression should be regarded as an independent risk factor for ALL with CNS involvement in adults.     

Decrease in CD4+CD25+ and CD8+CD28+ T cells in interstitial pneumonitis associated with rheumatic disease

Abstract

The expression of CD25 or CD28 on T cells was examined in patients with rheumatic diseases associated with interstitial pneumonitis (IP), in order to investigate the conditions of CD4⁺CD25⁺ regulatory T cells and CD8⁺CD28^? suppressor T cells. Fifty-five patients with various rheumatic diseases and 23 normal controls were enrolled. CD4⁺CD25⁺ T cells of patients with IP were significantly decreased in comparison with non-IP patients, and the ratio of CD8⁺CD28^? T cells in patients with IP was significantly higher than that in non-IP patients or normal controls. These results for CD8⁺CD28^? T cells were in accord with the decrease in CD8⁺CD28⁺ T cells, and may be related to activation-induced CD8⁺CD28⁺ T-cell death. Thus, the abnormality of CD4⁺CD25⁺ regulatory T cells may be related to the pathogenesis of IP, and the survival and activation of CD8⁺ T cells.     

Regulatory T cells and CD20+ B cells in pediatric very severe aplastic anemia: possible clinical markers for evaluating the therapeutic efficacy and prognosis

ABSTRACT

Objectives: To investigate the immune status of children with very severe aplastic anemia (VSAA), and evaluate the frequencies of CD20⁺ B cells and Regulatory T cells (Tregs) as potential markers for evaluating the therapeutic efficacy and prognosis.

Methods: We systematically analyzed CD20⁺ B cells and Tregs using Flow Cytometry in 36 children with VSAA (14 newly diagnosed cases and 22 cases in remission after therapy with HDIVIG?+?r-ATG?+?CSA).

Results: In newly diagnosed VSAA patients, the percentage of CD20⁺ B cells was higher than that in healthy children (P?<?.01), whereas the percentage of Tregs was lower than that in healthy children (P?<?.001). After treatment with HDIVIG?+?r-ATG?+?CSA, the percentage of CD20⁺ B cells in peripheral blood was decreased obviously, and the percentage of Tregs was significantly increased.

Conclusion: There is a moderate negative correlation between the percentage of Tregs and CD20⁺ B cells in our study. Our results shed light on the roles of Tregs and CD20⁺ B cells as therapeutic efficacy and prognostic markers of pediatric VSAA. Moreover, the mechanism underlying the decrease of blood Tregs and increase of CD20⁺ B cells in pediatric VSAA patients have been discussed, indicating that Tregs may suppress B cell responses.     

CD56 expression in T-cell acute lymphoblastic leukemia is associated with non-thymic phenotype and resistance to induction therapy but no inferior survival after risk-adapted therapy

Background

Expression of CD56 has been associated with poor prognosis in acute myeloid leukemia and aggressive lymphoma.

Design and Methods

We analyzed the impact of CD56 expression in a cohort of 452 newly diagnosed adult T-cell acute lymphoblastic leukemia (T-ALL) patients; clinical data were available for 306 patients. Treatment was according to the GMALL study protocols 06/99 and 07/03 stipulating stratification into standard (thymic T-ALL) and high risk (pre- and mature T-ALL) groups.

Results

CD56 expression was detected in 63/452 (13.9%) patients. CD56⁺ T-ALL were predominantly of non-thymic (pre-T 35%, mature 41%) immunophenotypic subtypes, whereas 53% of the CD56⁻ cases were thymic T-ALL (p=0.00002). CD13, CD33, CD34 and HLA-DR were significantly more frequently expressed. A clonal T-cell receptor rearrangement was detected in 22/23 CD56⁺ ALL. No major clinical differences were observed at presentation. Treatment of CD56⁺ ALL resulted in a lower rate of complete remissions (70% vs. 88%) (p=0.001) and a higher rate of resistant disease (21% vs. 8%) (p=0.004). CD56 expression had no significant influence on overall (48% vs. 59%) and disease free survival (67% vs. 57%) at three years.

Conclusions

CD56 is expressed on a subset of adult T-ALL with distinct immunophenotypical features and higher resistance to therapy. Most CD56⁺ ALL were treated in the high-risk arm of the GMALL study protocols owing to their non-thymic phenotype. Thus after risk adapted treatment a prognostic impact of CD56 expression was not detectable.     

Peripheral and bone marrow CD34+ cell levels on chronic myeloproliferative disease

Purpose: The aim of the present study was to examine the relationship between peripheral CD34⁺ and bone marrow CD34⁺ levels and the clinicopathologic characteristics and laboratory parameters of myeloproliferative disease (MPD) patients.

Patients and methods: A total of 103 MPD patients were enrolled in this study. We examined the relationship between bone marrow CD34⁺ and peripheral CD34⁺ levels and the patients’ clinicopathologic and laboratory parameters.

Results: There were no significant correlations between the peripheral CD34⁺ levels and the JAK-2 V617F mutation, thrombosis, white blood cells (WBC), lactate dehydrogenase (LDH), transferrin saturation (TS), ferritin, or bone marrow cellularity. In addition, there were no significant correlations between bone marrow CD34⁺ levels and the JAK-2 V617F mutation, thrombosis, WBC, LDH, TS, ferritin, or bone marrow cellularity (P?>?0.05). We did not identify any significant relationship between peripheral CD34⁺ and bone marrow CD34⁺ levels (P?>?0.05). However, there were significant correlations between peripheral CD34⁺ levels and bone marrow fibrosis (P?+ levels and constitutional symptoms (P?+ levels and bone marrow fibrosis (P?Conclusion: We did not find any significant relationship between the clinicopathologic and laboratory characteristics and peripheral and bone marrow CD34⁺ cells from bone marrow fibrosis patients. There was also no significant relationship between bone marrow CD34⁺ cells and peripheral CD34⁺ cells. Some peripheral CD34⁺ cells may originate from the spleen rather than the bone marrow, which may given us different result of some parameters.     

The level of peripheral blood circulating CD34+ cells is higher in acute promyelocytic leukemia patients with adverse prognostic factors

Introduction: The increased flow cytometry enumeration of peripheral blood circulating CD34+ cells in patients with acute leukemia has been found in our previous work. In this study, we also demonstrated that acute promyelocytic leukemia (APL) patients not only had elevated CD34+ cell count, but also had some clinical features.

Methods: Fifty APL patients and 19 healthy volunteers were included in the study. The enumeration of circulating CD34+ cells, cytogenetic subgroup, immunophenotype analysis, and leukemic-related gene mutation detection were performed.

Results: Some APL patients with higher count of CD34+ cells (≤10 × 10⁶/l) usually possessed one or more poor prognostic factors (higher WBCs count, PML/RARa gene complex fusion, chemotherapy-related APL, normal karyotype/complex karyotype abnormalities, CD56/CD34 antigen positive expression, FLT3-ITD positive mutation, myelofibrosis, and marrow necrosis). A cut-off value of 10 × 10⁶/l CD34+ cells may have the power to distinguish APL patients with above adverse clinical prognostic factor from other APL subjects.

Conclusion: The circulating CD34+ cell count appears to increase in some APL patients and a higher CD34+ cell count may be indicative of inferior survival and serve as an adverse biomarker for APL.     

CD56+ NK lymphomas:clinicopathological features and prognosis

The surface molecule CD56 marks a category of malignant lymphoma of putative natural killer (NK) cell origin. We conducted a retrospective analysis of 24 cases of CD56⁺ NK lymphoma/leukaemia to define the clinicopathologic and prognostic features of this specific group of lymphomas. 56 cases of nasal lymphomas and 204 cases with an initial diagnosis of peripheral T-cell lymphoma were retrospectively analysed. To specifically examine lymphomas of putative NK origin, only those that were negative for surface expression of CD3 but positive for CD56 were analysed. 24 cases were identified. The initial predominant sites of involvement were nasal (n =18), palate (n = 1), nodal (n = 1) and multi-organ (n =4). Clinically, in patients with disease localized to one anatomical site (n = 20), most had symptoms confined to the nose, with a high percentage in early stage (I: 91%; IV: 9%). The marrow was not involved in any of these cases. However, patients with multi-organ involvement at presentation (n = 4) behaved differently. All presented acutely with pancytopenia, hepatosplenomegaly, and marrow infiltration with haemophagocytosis. A leukaemic phase was observed in one case. Anthracycline containing combination chemotherapy resulted in complete remission in 75% of patients with localized disease, but only in 25% with multi-organ involvement. The median survival of patients with localized disease was 12 months, compared with 2 months in the multi-organ group (P =0.06); the disease-free survival was significantly better in the former (P <0.01). The overall median survival of all patients was still poor at 11 months. We conclude that CD56⁺ NK lymphomas could be divided into two main patterns of disease presentations: localized (predominantly nasal), and multi-organ involvement. Each has different clinicopathologic and prognostic features. Conventional chemotherapy appeared ineffective for the majority of patients, and innovative treatment modalities are needed to improve outcome.     

CD5 Positive B Lymphoblastic Leukemia: Report of a Case with Review of Literature

We report a rare CD5 positive B cell acute lymphoblastic leukemia (B-ALL) with a review of the clinopathological features and prognosis of previously reported cases in the literature. The aberrant expression of CD 5 antigen is uncommon in B-ALL; the morphological differential diagnosis includes blastic mantle cell lymphoma, denovo CD5⁺ diffuse large B cell lymphoma and secondary diffuse large cell lymphoma/Richter’s transformation. CD5⁺ B cell ALL is commonly reported in younger patients (<18 years). Though the expression of T cell antigens is reported to have poor prognosis, the experience with CD5⁺ B-ALL is limited to draw any firm conclusion regarding its prognosis.     

Flow cytometric analysis of intracellular CD6 8 molecule expression in normal and malignant haemopoiesis

Summary. CD68 molecules are heavily glycosylated lysosomal membrane constituents of unknown function with strong expression in monocytes and macrophages. Using flow cytometry, we quantified expression levels of CD68 molecules in normal and malignant haemopoietic cells. CD68 molecules are intensely expressed in the cytoplasm and weakly on the surface of mature CD14⁺ monocytes. CD68 expression seems to start very early during granulo-monopoietic differentiation. Virtually all myeloperoxidase (MPO)⁺ bone marrow cells coexpress CD68 and similar proportions of CD34⁺ progenitor cells weakly express CD68 or MPO molecules. During further differentiation, CD68 expression is strongly up-regulated in early MP0⁺ precursor cells which lack lactoferrin (LF) and CD14 molecules. Compared to these, more mature MPO⁺LF⁺ bone marrow and peripheral blood granulocytes express considerable lower levels of CD68. In-line with this broad expression, all investigated acute myeloid leukaemia (AML) cases, classified as FAB M1-M5, were CD68 positive, and compared to normal CD34⁺ bone marrow cells, CD34⁺ AML blast cells expressed increased levels. CD68 expression is, however, not restricted to cells of myeloid origin, because a subset (40 – 15%, n = 6) of CD19⁺ peripheral blood B-lympho-cytes and 50% of B-ALL are also weakly positive. In contrast, normal CD3+ lymphocytes lack (<3%. n = 6) CD68 and only low proportions (6 – 3%, n= 6) of CD56⁺ NK cells are CD68⁺. Also, all investigated T-ALL cases (n = 6) lacked CD68.     

The prognostic impact of multiparameter flow cytometry immunophenotyping and cytogenetic aberrancies in patients with multiple myeloma

Objectives: The aim of this study was to evaluate the prognostic impact of immunophenotyping in patients with multiple myeloma (MM), as well as other markers of disease, such as serum hyaluronan and cytogenetic aberrancies.

Methods: We have prospectively analyzed the prognostic impact of antigenic markers, assessed by multiparametric flow cytometry (MFC), in a series of newly diagnosed MM patients (n?=?79).

Results and discussion: Our results show that the expression of CD44, CD45, and CD28 and the absence of CD117 were associated with a significantly shorter progression free-survival (PFS). Clinical characteristics were collected; Cytogenetic aberrancies were assessed in 40 patients. Multivariate survival analyses identified that the CD117^?, CD28⁺, CD45⁺, and the percentage of bone marrow plasma cells by MFC are survival predictor, along with the International Staging System stage. Interestingly, the CD117^? patients were associated with chromosomal aberrancies, including del (17p), +1q21, and IgH translocations.

Conclusion: The incorporation of multiparameter flow cytometry immunophenotyping into the routine diagnostic evaluation of MM patients can help to identify patients at a high risk of progression.     

CD66c expression in B‐cell acute lymphoblastic leukemia: strength and weakness

Introduction: In B‐cell acute lymphoblastic leukemia (B‐ALL), testing at diagnosis for BCR/ABL1 gene rearrangements is mandatory for prognostic stratification and treatment decisions. Several diagnostic methods have been proposed using flow cytometry to identify BCR/ABL1⁺ B‐ALL. Methods: We evaluated expression of the myeloid antigen CD66c by flow cytometry in B‐ALL. We studied 94 patients with B‐ALL. The t(9;22)(q34;q11) or BCR/ABL1 rearrangement was detected by cytogenetic analysis or RT/PCR. Myeloid antigens CD66c, CD13, CD33, CD117, Myeloperoxidase, CD15 and CD65 were determined by flow cytometry. Results: Of these 94 cases, 17 (18%) cases displayed BCR/ABL1 gene rearrangements and 38 (40%) cases were CD66c positive. CD66c was the most common myeloid antigen expressed on malignant lymphoblasts. Its expression was correlated with BCR/ABL1 rearrangements (P = 0.0001): sensitivity 82%, specificity 69%, positive predictive value 37% and negative predictive value 95%. Co‐expression of CD66c⁺ CD13⁺ was more frequent in BCR/ABL1⁺ B‐ALL (29%) than BCR/ABL1^? cases (4%) (P = 0.0044). Some BCR/ABL1^? B‐ALL cases (including hyperdiploid or cases with normal karyotype) were CD66c positive (31%). Conclusion: CD66c expression is correlated, but not specifically, with BCR/ABL1 rearrangement. It would seem better to interpret the absence of CD66c expression with a lack of BCR/ABL1 rearrangement. This myeloid antigen could be interesting in the detection of minimal residual disease.     

Upregulation of CD72 expression on CD19+CD27+ memory B cells by CD40L in primary immune thrombocytopenia

CD72 is a co‐receptor of B cells and regulates B cell activation. Although aberrant expression of CD72 has been reported in primary immune thrombocytopenia (ITP), it is uncertain whether this aberrant expression is restricted to specific B cell subsets. Furthermore, the mechanisms that regulate CD72 expression are unknown. In this study, we found higher frequency of CD19⁺ B cells, CD19⁺CD27⁺ memory B cells and lower frequency of CD19⁺CD27⁻ naive B cells in active ITP patients compared with controls and patients in remission. CD72 expression on CD19⁺CD27⁺ cells was upregulated in active ITP patients and correlated with platelet count and anti‐platelet autoantibodies. In vitro, CD40L could specifically induce CD72 upregulation on CD19⁺CD27⁺ B cells. In combination with CD40L, interleukin (IL) 10 and BAFF (also termed TNFSF13B) further enhanced CD72 expression on CD19⁺CD27⁺ B cells, whereas IL21 reduced CD72 upregulation. CD72mRNA expression after CD40L stimulation was increased in ITP patients and controls. Significant increase of CD40L on CD4⁺ T cells was correlated with CD72 expression on CD19⁺CD27⁺ B cells in ITP patients. In conclusion, upregulation of CD72 expression on CD27⁺memory B cells might take part in the pathogenesis of ITP. Elevated CD40L on CD4⁺ cells combined with cytokines might contribute to the upregulation of CD72 expression on CD27⁺memory B cells.     

Extracellular vesicle-associated soluble CD163 and CD206 in patients with acute and chronic inflammatory liver disease

Abstract

Background: Extracellular vesicles (EVs) are implicated in intercellular communication in liver diseases. An EV-associated fraction of the macrophage biomarker soluble CD163, denoted EV-CD163, was recently identified. EV-CD163 may be released during later phases of the inflammatory response as opposed to the acute shedding of CD163 ectodomain (Ecto-CD163). Total sCD163 is a well-described biomarker in liver inflammation, and we investigated the distribution of CD163 fractions along with EV-associated soluble CD206 (EV-CD206) in patients with acute and chronic alcoholic liver inflammation.

Methods: Patients with acute alcoholic hepatitis (AH) (n?=?48) and alcoholic cirrhosis (AC) (n?=?26) were enrolled. Patients with AH were followed for 30?days after diagnosis. Healthy blood donors (n?=?30) served as a reference group. Fractions of sCD163 and sCD206 were separated using ExoQuick? and measured by ELISA.

Results: We demonstrated a possible EV-associated fraction of CD206 in plasma, correlating with levels of EV-CD163 (r_s = 0.46, p?<?.001). The distribution of biomarker fractions was skewed toward EVs in chronic cirrhosis for both biomarkers (median: 35.8% EV-CD163, 58.8% EV-CD206) as compared to AH patients (median: 26.2% EV-CD163 p?<?.0001, 48.8% EV-CD206, p?<?.01). In AH patients, total sCD163 and Ecto-CD163 at inclusion were related to survival, whereas EV-CD163 was not.

Conclusion: Extracellular vesicles of macrophage origin associated with membrane receptors CD163 and CD206 are present in liver disease. We observed a shift in the distribution towards an increased EV fraction in chronic liver cirrhosis. These data support that Ecto and EV fractions may be markers of different inflammatory processes, possibly resulting from a switch in macrophage phenotype.     

Effect of thrombopoietin on proliferation of blasts from CD7-positive acute myelogenous leukaemia

We investigated the effect of thrombopoietin (TPO) on the growth of leukaemic blasts from 30 acute myelogenous leukaemia (AML) patients according to the surface expression of CD7 and CD34: 10 patients were CD7 positive (CD7⁺), nine were CD7 negative/CD34⁺(CD7^?/CD34⁺) and the remaining 11 were CD7^?/CD34^?. Significant growth response of leukaemic blasts to TPO was observed in 10/10 CD7⁺, 5/9 CD7^?/CD34⁺ and 2/11 CD7^?/CD34^? AML cases using ³H-thymidine incorporation. Synergistic stimulatory effects of TPO with stem cell factor (SCF), interleukin-3 (IL-3), granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor were observed in both TPO-responding cases (9/17) and TPO-non-responding cases (8/13). In a leukaemic blast colony assay, significant growth response to TPO was observed in 5/6 CD7⁺ and 4/17 CD7^? AML cases examined. However, the effect of TPO on the growth of CD7⁺ leukaemic blasts was not so potent as that of IL-3 and SCF, both of which support the proliferation of primitive haemopoietic progenitors. Expression of c-mpl (TPO receptor) was significantly higher in CD7⁺ AML cases than in CD7^? cases, suggesting a relationship between expression of c-mpl and proliferative response to TPO. These data indicate that CD7⁺ leukaemic blasts express functional TPO receptors and proliferate in response to TPO. These observations also imply that CD7 expression on AML blasts may indicate involvement of leukaemic progenitors at an early stage of multipotent haemopoietic stem cells.     

Increased CD4+CD16+ and CD4+CD56+ T cell subsets in Behçet's disease

Behçet's disease is a systemic vasculitis of unknown etiology. Various immune abnormalities have previously been shown in Behçet's disease. We investigated T lymphocyte subsets associated with cytotoxic activity and natural killer (NK) cells by flow cytometry in 37 patients with Behçet's disease, 38 healthy controls, and 17 diseased control patients. Compared to the healthy controls, CD4⁺CD16⁺ and CD4⁺CD56⁺ subsets were found to be higher in the Behçet's disease group as well as in the disease control group (CD4⁺CD16⁺: BD=5?±?3, DC=14?±?14, HC= 3?±?2, P=0.001; CD4⁺CD56⁺: BD=11?±?5, DC= 18?±?17, HC=8?±?6, P=0.01). CD8⁺CD16⁺ and CD8⁺CD56⁺ T cell subsets were at normal levels in Behçet's disease but found to be elevated in disease controls. Similarly, NK cells (CD16⁺CD56⁺) were high only in the disease control group. Significant increases in CD4⁺CD16⁺ and CD4⁺CD56⁺ cell subsets in Behçet's patients and disease controls suggest that T cell activation patterns of these subsets in Behçet's disease are similar to those in other inflammatory disorders.     

CD31+, CD38+, CD44+, and CD103+ lymphocytes in peripheral blood,bronchoalveolar lavage fluid and lung biopsy tissue in sarcoid patients and controls

BackgroundThe mechanisms driving the transition from inflammation to fibrosis in sarcoidosis patients are poorly understood; prognostic features are lacking. Immune cell profiling may provide insights into pathogenesis and prognostic factors of the disease. This study aimed to establish associations in simultaneous of lymphocyte subset profiles in the blood, bronchoalveolar lavage fluid (BALF), and lung biopsy tissue in the patients with newly diagnosed sarcoidosis.MethodsA total of 71 sarcoid patients (SPs) and 20 healthy controls (HCs) were enrolled into the study. CD31, CD38, CD44, CD103 positive T lymphocytes in blood and BALF were analysed. Additionally, the densities of CD4, CD8, CD38, CD44, CD103 positive cells in lung tissue biopsies were estimated by digital image analysis.ResultsMain findings: (I) increase of percentage of CD3⁺CD4⁺CD38⁺ in BALF and blood, and increase of percentage of CD3⁺CD4⁺CD44⁺ in BALF in Löfgren syndrome patients comparing with patients without Löfgren syndrome, (II) increase of percentage of CD3⁺CD4⁺103⁺ in BALF and in blood in patients without Löfgren syndrome (comparing with Löfgren syndrome patients) and increase of percentage of CD3⁺CD4⁺103⁺ in BALF and in blood in more advanced sarcoidosis stage. (III) Increasing percentage of BALF CD3⁺CD4⁺CD31⁺ in sarcoidosis patients when comparing with controls independently of presence of Löfgren syndrome, smoking status or stage of sarcoidosis. Several significant correlations were found.ConclusionsLymphocyte subpopulations in blood, BALF, and lung tissue were substantially different in SPs at the time of diagnosis compared to HCs. CD3⁺CD4⁺CD31⁺ in BALF might be a potential supporting marker for the diagnosis of sarcoidosis. CD3⁺CD4⁺CD38⁺ in BALF and blood and CD3⁺CD4⁺CD44⁺ in BALF may be markers of the acute immune response in sarcoidosis patients. CD4⁺CD103⁺ T-cells in BALF and in blood are markers of the persistent immune response in sarcoidosis patients and are potential prognostic features of the chronic course of this disease.     

Prognostic significance of PD-L1 expression and CD8+ TILs density for disease-free survival in surgically resected lung squamous cell carcinoma: a retrospective study

BackgroundThis study sought to depict the genomic landscape of patients with surgically resected lung squamous cell carcinoma (LUSC) and its relationship with clinical outcome indicators.MethodsWe retrospectively collected the clinical data of 180 patients from the electronic medical records and applied targeted sequencing and immunohistochemistry (IHC) to depict the genomic landscape, including the tumor mutation burden (TMB), programmed cell death-ligand 1 (PD-L1), and cluster of differentiation CD8⁺ tumor-infiltrating lymphocytes (CD8⁺ TILs). And comparative analysis and survival analysis of these parameters were conducted to find prognostic factors for clinical outcome.ResultsPD-L1+ tumor cells were observed in 75 (41.7%) of the patients, the median rate of CD8⁺ TILs was 11.5 [4, 24], and the median TMB was 9.4 (7.5–13.7) mutations per megabase (mut/Mb). Patched receptor 1 (PTCH1) gene mutation frequency was significantly associated with CD8⁺ TILs density (12% vs. 1%; P=0.024). High PD-L1 expression and CD8⁺ TILs+ were significantly associated with longer disease-free survival (DFS), and a further subgroup analysis revealed that both were significantly correlated with the DFS of stage I/II patients but not stage III patients.ConclusionsThe results suggest that only PTCH1 gene mutation frequency was correlated with CD8⁺ TILs density. Additionally, intense CD8⁺ TILs density and high PD-L1 expression were found to be associated with longer DFS. Our findings provide insights into the precise treatment strategy for surgically resected LUSC patients.     

Comparative quantitative expression of bcl-2 by normal and leukaemic myeloid cells

Summary. Expression of the bcl-2 oncoprotein by AML blasts has previously been demonstrated to be heterogenous with high levels of bcl-2 expression being associated with a low complete remission rate and poor survival. We have quantified bcl-2 expression in AML blasts in relation to expression of the CD34 antigen and in comparison to CD34-positive cells from normal bone marrow. When expressed as molecules of equivalent soluble fluorochrome (MESF) per cell, AML blast cell bcl-2 expression varied from 11.1 to 99.9 × lO³ (median 39.4 × 10³, n=56) with 28.5% of patients expressing high MESF values (>50 × 10³) and 16% of patients expressing low MESF values (<20 × 10³), the remainder expressing intermediate values. There was no significant difference between intensity of bcl-2 expression and FAB classification in the de novo AML cases, and there was no significant differences between de novo and secondary AML cases. Blasts from CD34⁺ AML patients expressed significantly higher levels of bcl-2 (mean MESF 43.6 × 10³, n =36) than CD34^? AML patients (mean MESF 31.7 × 10³, n=19). In five cases of CD34⁺ AML, bcl-2 expression was determined on purified CD34⁺ and CD34^? blast cell populations. In all cases CD34⁺ blasts were found to express significantly higher bcl-2 MESF values compared to the CD34^? fraction. Purified CD34⁺ cells from normal bone marrow consistently expressed high levels of bcl-2 (MESF >75 × 10³ n = 4), which was comparable to that found on CD34⁺ AML cells. Our results suggest that the poor prognosis previously associated with AML blasts expressing the CD34 antigen may in part be related to high expression of bcl-2. Also the ability to measure bcl-2 in AML blasts quantitatively by flow cytometry and to categorize patients into discrete groups may be of value as a prognostic indicator in AML.     

Increased surface receptor Fas (CD95) levels on CD4+ lymphocytes in patients with primary intestinal lymphangiectasia

Objective. Exudative enteropathy secondary to primary intestinal lymphangiectasia (PIL) is characterized by lymphopenia, hypogammaglobulinemia and hypoalbuminemia resulting from leakage of lymph fluid into the intestinal tract. The objective of this study was to better characterize the lymphopenia of PIL-confirmed patients. Material and methods. T-cell markers and T-cell proliferation/capacities (differentiation, activation and death) were analyzed for phenotype in 9 patients (6 F, 3 M, aged from 18 to 72 years). Results. Mean counts of CD4 and CD8 subsets were significantly decreased, 174±123/µl and 134±77/µl compared with controls, 858±260/µl and 482±164/µl, respectively (p<0.0001). Significant depletion of naïve (CD45RA⁺ CD62L⁺) CD4⁺ T cells was noted, with a mean expression of 7±4% compared with controls, 45±1% (p<0.0001). Both CD4⁺ and CD8⁺ T-cell mean subsets were activated as assessed by their proportion expressing the late activation markers HLA-DR, 18±7% and 19±9% compared with controls, 6±3% and 10±6%, respectively (p<0.0001 and p<0.01). The mean expression of CD95/Fas on CD4⁺ T cells was significantly higher in patients than in controls, 83±16% versus 45±13% (p<0.0001). No major abnormality of T-cell proliferation/capacities was observed. Conclusions. Our results suggest that the T-cell loss in PIL patients is probably due to various mechanisms including enteric lymphocytes loss and activation of residual T cells, leading to death. Moreover, this loss is not compensated by a sufficient increase in T-cell thymic production.     

Neutrophil CD64 is upregulated in RA patients with lymphoma but not in other solid cancers

Objective: To evaluate the utility of quantifying CD64 expression on neutrophils in rheumatoid arthritis patients with malignancy, especially its diagnostic role in lymphoma.

Methods: We used flow cytometry to quantify CD64 expression on neutrophils from patients diagnosed with malignancy during the follow-up period prior to initiating treatment.

Results: Neutrophils from 18 patients with lymphoma expressed significantly higher levels of CD64 (9635.6?±?2123.7 molecules/cell) than those from 32 patients with other solid cancers (carcinoma) (1250.5?±?91.1 molecules/cell) (p?<?0.001). When the cutoff value was set at 2060 molecules/cell, the sensitivity and specificity of CD64 for diagnosing lymphoma was 88.9% and 94.4%, respectively.

Conclusions: The quantitative measurement of neutrophil CD64 by flow cytometry may be useful as a subsidiary diagnostic marker in patients with suspected lymphoma. Although neutrophil CD64 is currently a well-known marker of infection, it is necessary to bear in mind that lymphoma is also a candidate in differential diagnosis when CD64 expression on neutrophils is upregulated.