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Introduction: New tools have been developed to distinguish the COVID‐19 diagnosis from other viral infections presenting similar symptomatology and mitigate the lack of sensitivity of molecular testing. We previously identified a specific "sandglass" aspect on the white blood cells (WBC) scattergram of COVID‐19 patients, as a highly reliable COVID‐19 screening test (sensitivity: 85.9%, specificity: 83.5% and positive predictive value: 94.3%). We then decided to validate our previous data in a multicentric study. Methods: This retrospective study involved 817 patients with flu‐like illness, among 20 centers, using the same CBC instrument (XN analyzer, SYSMEX, Japan). After training, one specialist per center independently evaluated, under the same conditions, the presence of the "sandglass" aspect of the WDF scattergram, likely representing plasmacytoid lymphocytes. Results: Overall, this approach showed sensitivity: 59.0%, specificity: 72.9% and positive predictive value: 77.7%. Sensitivity improved with subgroup analysis, including in patients with lymphopenia (65.2%), patients presenting symptoms for more than 5 days (72.3%) and in patients with ARDS (70.1%). COVID‐19 patients with larger plasmacytoid lymphocyte cluster (>15 cells) more often have severe outcomes (70% vs. 15% in the control group). Conclusion: Our findings confirm that the WBC scattergram analysis could be added to a diagnostic algorithm for screening and quickly categorizing symptomatic patients as either COVID‐19 probable or improbable, especially during COVID‐19 resurgence and overlapping with future influenza epidemics. The observed large size of the plasmacytoid lymphocytes cluster appears to be a hallmark of COVID‐19 patients and was indicative of a severe outcome. Furthers studies are ongoing to evaluate the value of the new hematological parameters in combination with WDF analysis. [ABSTRACT FROM AUTHOR]

Simple Summary: We train a deep neural network model to identify CD3 expressing cells from Hoechst stained slides only, without the need for costly immunofluorescence. Using interpretability techniques to understand what the model has learned, we find that morphological features in the nuclear chromatin are predictive of CD3 expression. Multiplex immunofluorescence and immunohistochemistry benefit patients by allowing cancer pathologists to identify proteins expressed on the surface of cells. This enables cell classification, better understanding of the tumour microenvironment, and more accurate diagnoses, prognoses, and tailored immunotherapy based on the immune status of individual patients. However, these techniques are expensive. They are time consuming processes which require complex staining and imaging techniques by expert technicians. Hoechst staining is far cheaper and easier to perform, but is not typically used as it binds to DNA rather than to the proteins targeted by immunofluorescence techniques. In this work we show that through the use of deep learning it is possible to identify an immune cell subtype without immunofluorescence. We train a deep convolutional neural network to identify cells expressing the T lymphocyte marker CD3 from Hoechst 33342 stained tissue only. CD3 expressing cells are often used in key prognostic metrics such as assessment of immune cell infiltration, and by identifying them without the need for costly immunofluorescence, we present a promising new approach to cheaper prediction and improvement of patient outcomes. We also show that by using deep learning interpretability techniques, we can gain insight into the previously unknown morphological features which make this possible. [ABSTRACT FROM AUTHOR]

We studied a child with severe viral, bacterial, fungal, and parasitic diseases, who was homozygous for a loss-of-function mutation of REL, encoding c-Rel, which is selectively expressed in lymphoid and myeloid cells. The patient had low frequencies of NK, effector memory cells reexpressing CD45RA (Temra) CD8+ T cells, memory CD4+ T cells, including Th1 and Th1*, Tregs, and memory B cells, whereas the counts and proportions of other leukocyte subsets were normal. Functional deficits of myeloid cells included the abolition of IL-12 and IL-23 production by conventional DC1s (cDC1s) and monocytes, but not cDC2s. c-Rel was also required for induction of CD86 expression on, and thus antigen-presenting cell function of, cDCs. Functional deficits of lymphoid cells included reduced IL-2 production by naive T cells, correlating with low proliferation and survival rates and poor production of Th1, Th2, and Th17 cytokines by memory CD4+ T cells. In naive CD4+ T cells, c-Rel is dispensable for early IL2 induction but contributes to later phases of IL2 expression. The patient's naive B cells displayed impaired MYC and BCL2L1 induction, compromising B cell survival and proliferation and preventing their differentiation into Ig-secreting plasmablasts. Inherited c-Rel deficiency disrupts the development and function of multiple myeloid and lymphoid cells, compromising innate and adaptive immunity to multiple infectious agents. [ABSTRACT FROM AUTHOR]

Background: Extensive burns is one of the most common severe injuries, with a high annual death rate. Previous studies showed that the neutrophil to lymphocyte ratio (NLR) is a prognostic factor for some inflammatory diseases. However, until now, no study has evaluated the clinical prognostic value of NLR in extensively burned patients. The aim of this study was to investigate the prognostic value of NLR in this medical condition to provide clinical guidance.Methods: 271 patients diagnosed with extensive burns were analysed retrospectively between 2005 and 2018 in the Department of Burn Surgery of Changhai Hospital. NLR cut-off values at the first 3 days of hospitalization were calculated by the ROC analysis.Results: Of the 271 patients in this study, the majority (82.3%) were injured by flame. The median total body surface area (TBSA) was 55% (IQR, 40% to 85%) and the median full thickness burn (FTB) was 20% (IQR, 3%-44%). The patients' NLR declined within the first 3 days after admission, and we found that NLR was negatively correlated with the ventilator-free days at day 28 (r = -0.127, P = 0.048). In a multivariate logistic regression analysis, higher admission NLR was independently predictive of higher mortality. According to the ROC curve, the best cut-off values for day 1 (or admission day), day 2 and day 3 NLR were 14, 13 and 7.5, respectively. We then performed a survival analysis, finding that those NLR above the cut-off point had decreased overall survival compared to those with NLR below the cut-off point (p = 0.023, 0.045 and 0.019 for day 1, 2, and 3, respectively).Conclusions: NLR continuously decreased in the first 3 days of hospitalization. Admission NLR above 14 is associated with a decreased survival in patients with extensive burns. These findings demonstrate that NLR has prognostic value in these patients. [ABSTRACT FROM AUTHOR]

Mixed-phenotype acute leukemia (MPAL) is the infrequent type of acute leukemia characterized by immunophenotypic and/or cytochemical features of both lineages, but the diagnosis of this disease still is a challenge. In this study, we analyzed immunophenotyping, cytochemistry and frequency of MPAL patients to better diagnosis of MPAL characteristics according to WHO 2016 criteria for the first time in Iran. In this retrospective study, 27 patients were diagnosed as MPAL based on WHO 2016 criteria during 2014-2017. Flow cytometric immunophenotyping was performed on PB and BM samples evaluation of different CD marker expressions in MPAL subsets. RT-PCR was performed for the analyses of BCR/ABL1 fusion in MPAL subsets. Among 27 cases, (70.4%) 19 cases were B + My, (22.22%) 6 cases were T + My, and 2 cases (7.40%) were B + T + My. CD34, CD19, HLA-DR, TdT, CD22, iMPO were positive in majority of B + My cases. CD45, iMPO, iCD3, CD7, CD2 and CD5 were positive in majority of T + My cases. HLA-DR, TdT, CD10, CD22, iCD79a, iMPO, CD45, iCD3, CD7, CD3, CD2, CD5 were positive in majority of B + T + My cases. BCR/ABL1 fusion was positive for 3 cases (11.1%) of p190 fusion and 2 cases (7.4%) of p210 fusion in B + My cases. WHO 2016 criteria are the current standard for diagnosing MPAL. Also, evaluation of TdT, CD2, CD5, CD7 expressions by flow cytometry in EGIL criteria is useful for the better diagnosis of MPAL subsets. In addition, evaluation of BCR/ABL1 and MLL rearrangements in patients should be part of standard work-up in MPAL. [ABSTRACT FROM AUTHOR]

Background: The absolute neutrophil count (ANC), absolute lymphocyte count (ALC), absolute monocyte count (AMC) and neutrophil to lymphocyte ratio (NLR) are known markers of inflammation. We evaluated whether ANC, ALC, AMC and NLR, both before and after treatment with nivolumab, are indicative markers of overall survival (OS) and evaluated change in NLR as a predictive marker of response in non -small cell lung cancer (NSCLC) patients treated with nivolumab. Methods: A total of 109 patients with advanced NSCLC treated with nivolumab were included. ANC, ALC, AMC and NLR were examined at initiation of nivolumab therapy and after two cycles. The prognostic role of ANC, ALC, AMC and NLR with OS and changes in NLR ratio were examined with Kaplan-Meier curves and proportional hazard model. Result: Post-treatment NLR ≥5 after two cycles of nivolumab was associated with poor OS (median OS in NLR = <5 vs NLR = ≥5 was 29.1 (16.2–40.9) vs 24.2(16.1–36.2) months respectively, p<0.001). In addition NLR increased in non-responders after two cycles of nivolumab by 6.6±21.8 as compared to responders (p = 0.027). Conclusions: Post-treatment ANC, ALC and NLR are independent prognostic factors in NSCLC patients treated with nivolumab. Changes in NLR can be an early biomarker for response in NSCLC patients treated with nivolumab. [ABSTRACT FROM AUTHOR]

Background: Splenic hamartoma is a rare benign vascular proliferative lesion composed of unorganized sinusoidlike channels lined with plump or flat endothelial cells and characterized by a CD8-positive immunophenotype of the lining cells. Scattered bizarre stromal cells can be found in some splenic hamartomas. The presence of splenic hamartoma with bizarre stromal cells is extremely rare and these bizarre cells make it possible to be regarded as a malignancy. Recognition of this rare histologic variant will help to avoid diagnostic confusion and overtreatment of this benign entity. Case presentation: We report a case of a 40-year-old man with occasional left-sided waist back pain. A splenic spaceoccupying lesion was detected by ultrasound and magnetic resonance imaging. Microscopically bizarre large cells were scattered throughout the splenic hamartoma. The cells exhibited atypical nuclei, scarcely visible cytoplasm, and vesicular chromatin, and they did not form expansile clusters and lacked mitotic activity. An immunohistochemical panel was performed. The bizarre cells strongly expressed vimentin, and the Ki-67 index was very low. The lesion was diagnosed as a splenic hamartoma with bizarre stromal cells. Conclusions: To the best of our knowledge, this is the first systematic review on a splenic hamartoma with bizarre stromal cells; only six cases have been described in the literature. Proper identification is important to secure adequate treatment. [ABSTRACT FROM AUTHOR]

Background: Early reperfusion of the occluded coronary artery during acute myocardial infarction is considered crucial for reduction of infarcted mass and recovery of ventricular function. Effective microcirculation and the balance between protective and harmful lymphocytes may have roles in reperfusion injury and may affect final ventricular remodeling.Methods/design: BATTLE-AMI is an open-label, randomized trial comparing the effects of four therapeutic strategies (rosuvastatin/ticagrelor, rosuvastatin/clopidogrel, simvastatin plus ezetimibe/ticagrelor, or simvastatin plus ezetimibe/clopidogrel) on infarcted mass and left ventricular ejection fraction (LVEF) (blinded endpoints) in patients with ST-segment elevation myocardial infarction submitted to fibrinolytic therapy before coronary angiogram (pharmacoinvasive strategy). All patients (n = 300, 75 per arm) will be followed up for six months. The effects of treatment on subsets of B and T lymphocytes will be determined by flow-cytometry/ELISPOT and will be correlated with the infarcted mass, LVEF, and microcirculation perfusion obtained by cardiac magnetic resonance imaging. The primary hypothesis is that the combined rosuvastatin/ticagrelor therapy will be superior to other therapies (particularly for the comparison with simvastatin plus ezetimibe/clopidogrel) for the achievement of better LVEF at 30 days (primary endpoint) and smaller infarcted mass (secondary endpoint) at 30 days and six months. The trial will also evaluate the improvement in the immune/inflammatory responses mediated by B and T lymphocytes. Omics field (metabolomics and proteomics) will help to understand these responses by molecular events.Discussion: BATTLE-AMI is aimed to (1) evaluate the role of subsets of lymphocytes on microcirculation improvement and (2) show how the choice of statin/antiplatelet therapy may affect cardiac remodeling after acute myocardial infarction with ST elevation.Trial Registration: ClinicalTrials.gov, NCT02428374 . Registered on 28 September 2014. [ABSTRACT FROM AUTHOR]

Lymphocyte-specific interferon regulatory factor (LSIRF) (now called IRF4) is a transcription factor expressed only in lymphocytes. Mice deficient in IRF4 showed normal distribution of B and T lymphocyes at 4 to 5 weeks of age but developed progressive generalized lymphadenopathy. IRF4-deficient mice exhibited a profound reduction in serum immunoglobulin concentrations and did not mount detectable antibody responses. T lymphocyte function was also impaired in vivo; these mice could not generate cytotoxic or antitumor responses. Thus, IRF4 is essential for the function and homeostasis of both mature B and mature T lymphocytes. [ABSTRACT FROM AUTHOR]

The article presents a study which reveals that distinct bilinear leukemia immunophenotypes are not genetically determined. Topics discussed include mixed phenotype acute leukemia (MPALs) and standard routine molecular and immunologic diagnostics, used high-throughput genomic methods to analyze the genomic landscape of immunophenotypically distinct subpopulations in 2 BLL patients.

We showed previously that ellagitannin-rich cloudberries and anthocyanin-rich bilberries reduce the number of intestinal adenomas in multiple intestinal neoplasia/+ ( Apc Min ) mice. We also found that cloudberries decreased the size of adenomas, whereas bilberries increased it. Here we hypothesized that the difference in adenoma growth could be explained by dissimilar effects of the berries on intestinal immune responses and gut microbiota, potentially driven by the distinct polyphenol compositions of the 2 berries. Our objectives were to investigate lymphocyte subtypes and the predominant cecal bacterial diversity in mice fed with bilberries and cloudberries, and to analyze global gene expression profiles in the intestinal mucosa. Immunostainings of CD3 + T lymphocytes, FoxP3 + regulatory T lymphocytes, and CD45R + B lymphocytes revealed a smaller ratio of intraepithelial to all mucosal CD3 + T lymphocytes in the cloudberry-fed mice compared with controls, suggesting an attenuation of inflammation. Bilberry feeding induced no changes in the density of any of the lymphocyte subtypes. The predominant bacterial diversity in cecal contents, analyzed using polymerase chain reaction–denaturating gradient gel electrophoresis, was higher in the bilberry group than in the control or cloudberry groups. The microbial profiles of cloudberry-fed mice clustered together and were associated with small adenoma size. Pathway analyses of gene expression data showed that cloudberry down-regulated and bilberry up-regulated the expression of energy metabolism–related genes in the intestinal mucosa. In conclusion, attenuation of intestinal inflammation, changes in microbial profiles, and down-regulation of mucosal energy metabolism may account for the smaller adenoma size in cloudberry-fed mice in comparison to bilberry-fed mice. [ABSTRACT FROM AUTHOR]

Immune cell metabolism is dynamically regulated in parallel with the substantial changes in cellular function that accompany immune cell activation. While these changes in metabolism are important for facilitating the increased energetic and biosynthetic demands of activated cells, immune cell metabolism also has direct roles in controlling the functions of immune cells and shaping the immune response. A theme is emerging wherein nutrients, metabolic enzymes, and metabolites can act as an extension of the established immune signal transduction pathways, thereby adding an extra layer of complexity to the regulation of immunity. This Review will outline the metabolic configurations adopted by different immune cell subsets, describe the emerging roles for metabolic enzymes and metabolites in the control of immune cell function, and discuss the therapeutic implications of this emerging immune regulatory axis. [ABSTRACT FROM AUTHOR]

Background: Single cell gene expression assays have become a powerful tool with which to dissect heterogeneous populations. While methods and software exist to interrogate such data, what has been lacking is a unified solution combining analysis and visualisation which is also accessible and intuitive for use by non-bioinformaticians, as well as bioinformaticians. Results: We present the Single cell expression visualiser (SCExV), a webtool developed to expedite the analysis of single cell qRT-PCR data. SCExV is able to take any data matrix of Ct values as an input, but can handle files exported by the Fluidigm Biomark platform directly. In addition, SCExV also accepts and automatically integrates cell surface marker intensity values which are measured during index sorting. This allows the user to directly visualise relationships between a single cell gene expression profile and the immunophenotype of the interrogated cell. Conclusions: SCExV is a freely available webtool created to import, filter, analyse, and visualise single cell gene expression data whilst being able to simultaneously consider cellular immunophenotype. SCExV is designed to be intuitive to use whilst maintaining advanced functionality and flexibility in how analyses are performed. [ABSTRACT FROM AUTHOR]

Context: The distribution of different types of lymphoma varies across geographic regions.Aims: The present study was done to understand the occurrence of nodal lymphomas in a referral hospital of Mangalore city.Settings and Design: Descriptive study.Setting: The present study was conducted on 95 lymph node biopsy specimens, received in a referral hospital of Mangalore city.Materials and Methods: All cases of nodal lymphoma diagnosed between January 2007 and June 2010 in a referral hospital of Mangalore were selected for the study. The patients age and clinical details were obtained. Immunophenotyping was done for all cases of NHL and selected cases of HL. IPI score for NHL was noted and correlated with the outcome.Statistical Analysis Used: Chi square test was used.Results: Out of the total 95 cases of nodal lymphoma 37 (39%) were HL while 58 (61%) were NHL. Classical HL was diagnosed in 35 cases (37%) while 2 cases (2%) were diagnosed NLPHL. B cell lymphoma formed 72% of NHL. Follicular lymphoma accounted for 28% of all NHL. T cell lymphoma formed 28% of NHL.Conclusions: In conclusion the following points are significant about the distribution of lymphomas in a referral hospital of Mangalore city. The incidence of NSHL is higher while national data suggests MCHL as the most common subtype. The incidence of FL as well as T cell NHL is much higher when compared to national incidence. The epidemiological factors for this high frequency (either genetic or environmental) needs to be studied further. [ABSTRACT FROM AUTHOR]

Leucocyte profiles, especially the heterophil/lymphocyte ratio (H/L), are widely used to assess immune function and stress in birds. In this study, we investigated whether leucocyte profiles in a small colonial seabird, the little auk ( Alle alle), are related to age (chicks, adults), sex or biometric traits, including body mass. We blood-sampled chicks at 4-8 and 14-18 days of life and chick-feeding adults with 14-20- and 24-30-day-old chicks. Relative numbers of heterophils and H/L generally differed among chicks and adults, with the highest values in adults with 14-20-day-old chicks and the lowest ones in 4-8-day-old chicks. Females had a higher proportion of lymphocytes than males. The relative numbers of eosinophils were similar in all the groups. The percentage of monocytes tended to differ among the age categories. Analyses of chicks revealed that the percentage of heterophils and the H/L ratios were significantly higher in older than in younger chicks. The opposite pattern was recorded for lymphocytes. In 14-18-day-old chicks, H/L (stress level) was negatively related to body mass, head-bill length and bill width. This time period is energetically stressful as nestlings start to exercise their wings outside the nest chamber. This suggests that the origin of stress in nestlings was mainly nutritional. We found a significant positive correlation between body mass at fledging and the relative number of eosinophils at 14-18 days of life, indicating a higher level of stress in lighter nestlings. The significant relationship between fledging age and the percentage of monocytes at 14-18 days of life suggested a poorer state of health in chicks that fledged at an older age. [ABSTRACT FROM AUTHOR]

Following in vivo recombinant adeno-associated virus (rAAV)-based gene transfer, adaptive immune responses specific to the vector or the transgene product have emerged as a potential roadblock to successful clinical translation. The occurrence of such responses depends on several parameters, including the route of vector administration as well as the viral serotype and the genome configuration, either self-complementary (sc) or single-stranded (ss). These parameters influence rAAV vector-associated immunity by modulating the crosstalk between the vector and the host immune system, including vector ability to interact or even transduce lymphoid tissues in general and antigen-presenting cells (APCs) in particular. Little is known about immune cell populations that are targeted in vivo by rAAV vectors. Moreover, the transduction of dendritic cells is still controversial and not directly demonstrated. Here, we show that intramuscular administration of an sc rAAV8 vector in the mouse leads to a rapid distribution of viral genomes in the lymphoid tissues that is associated with transgene expression. Transduced cells were detected in follicular areas of the spleen and the draining lymph nodes. In addition to B and T lymphocytes, transduced professional APCs were detected although at very low frequency. In addition, viral genomes and transgene transcripts were also detected in these cell populations after ss rAAV8 vector administration. Although the functional significance of those observations needs further explorations, our results highlight an early and intricate interaction between the rAAV vector upon its in vivo delivery and the host immune system. [ABSTRACT FROM AUTHOR]

Introduction: Acute leukemia is the most common malignancy in children and acute lymphoblastic leukemia (ALL) accounts for 75% of acute leukemia cases. New treatment protocols have resulted incomplete remission rates up to nearly 100% in children with acute lymphoblastic leukemia. Today, one of the most important prognostic factors in acute lymphoblastic leukemia is intensity of the treatment. Risk stratification is accomplished based on clinical, morphological, immune-phenotypic and cytogenetic findings. The aim of this study was to determine some prognostic factors in children with acute lymphoblastic leukemia. Methods: In this retrospective study information about age at onset of acute leukemia, sex, initial white blood cell count, FAB-subtype, immunophenotype, and clinical course of newly diagnosed acute lymphoblastic leukemia were extracted from medical records of children admitted to pediatric oncology department of ShafaHospital between- 2011and2012. Results: There were 21 male patients (51.2%) and 20 were female patients (48.8%). The mean age was 4.2 ± 6.34 years, and 24 patients (58.5%) had Arab origins, while 17 patients (41.5%) were of non-Arab ethnicity. Age distribution showed higher incidence of ALL in younger children: 1-4 years 47.5%, 5-9 years 27.5% and 25% in patients >10 years.L2 subtype was more common in our patients 51.2% while L1 subtype was reported 46.3%.Only one patient was reported to be L3 subtype (2.4%), yet we did not detect any significant relation between different age groups and trend for incidence for specific subtype. The number of white blood cell (WBC) at the time of admission was reported as: less than 10,000 cells/cm in 30%, between11-50,000 in 37.5% and >50,000 in 32.5% patients. Organ involvement was present in 47.5%, and central nervous involvement, (proved by positive malignant cells in CSF fluid) was detected in 4.9% of our patients. In our study, HLA-DR was 62.5% in ALL patients and CD 20 and CD19 was the most common marker in these patients. In our work the most common markers in L1, which was found in 19 patients were reported CD 19, CD33, CD22,CD35, CD20 and CD9. Also the percentage of markers in L2 subtype had a similarity to L1 group. Conclusion: Conclusion: In this study, FAB-subtype L1 was less than previous studies, while FAB-subtype L2 and pre-B cell immunophenotype was more common than previous studies. Other results were the same as reported in older studies. [ABSTRACT FROM AUTHOR]

Acute myeloblastic leukemia accounts for approximately 20% of acute leukemias in children. The days the microscope represented the main tool In the diagnosis and classification of Acute Myeloblastic Leukemia seem to be very far. This review summarizes the current diagnosis of this malignancy, where the morphological, cytochemical, Immunophenotyping, cytogenetic and molecular characterization represents the basement of a risk group related therapy. [ABSTRACT FROM AUTHOR]

The aim was to determine the clinical and cytogenetic characteristics of non-Hodgkin's lymphoma (NHL) in Shanghai. A retrospective analysis was conducted in 516 patients with NHL. Patient clinical data, including age, sex, diagnosis, immunophenotypes, and karyotypes, were collected. The median age was 58 years. There was a male predominance in all NHL, except extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue. Patients with B cell NHL (1.5%) expressed CD3. T cell NHL patients (11.5%) expressed CD20. Epstein-Barr virus latent integral membrane protein 1, BCL6, CD10, Bcl-2, CD68, myeloperoxidase, CD99, CD30, CD15, and CD43 were present in various types of NHL. Complex karyotypes accounted for 92.3% of the 73.7% patients with abnormal karyotypes. Immunoglobin heavy chain gene translocation was present in 60.3% of B cell and 23.7% of T/NK cell neoplasms. Understanding the complex clinicopathological and molecular features of NHL may help with prognosis and serve as targets for treatments. [ABSTRACT FROM AUTHOR]

Background This study assesses the relationships between lymphocyte and monocyte subsets and intelligence quotient (IQ) scores in antiretroviral therapy (ART)-naive, HIV-infected Thai children without advanced HIV disease. Findings Sixty-seven ART-naive Thai children with CD4 between 15-24% underwent cognitive testing by Weschler intelligence scale and had 13 cell subsets performed by flow cytometry including naive, memory and activated subsets of CD4+ and CD8+ T cells, activated and perivascular monocytes and B cells. Regression modelling with log10 cell count and cell percentage transformation was performed. Median age (IQR) was 9 (7-10) years, 33% were male, CDC stages N:A:B were 1:67:31%, median CD4% and count (IQR) were 21 (18-24)%, 597(424-801) cells/mm3 and HIV RNA (IQR) was 4.6 (4.1-4.9) log10 copies/ml. Most (82%) lived at home, 45% had a biological parent as their primary caregiver, and 26 (49%) had low family income. The mean (SD) scores were 75 (13) for full scale IQ (FIQ), 73 (12) for verbal IQ (VIQ) and 80 (14) for performance IQ (PIQ). Adjusted multivariate regression analysis showed significant negative associations between B cell counts and FIQ, VIQ and PIQ (p < 0.01 for all); similar associations were found for B cell percentages (p < 0.05 for all). Conclusions High B cell counts and percentages were strongly associated with poorer FIQ, VIQ and PIQ scores. Prospective, long-term assessment of cell subsets and determination of relevant B cell subpopulations could help further elucidate associations between lymphocyte subsets and neurocognitive development. [ABSTRACT FROM AUTHOR]

Human multipotent mesenchymal stromal cells (MMSCs) were cocultured with allogenic blood-born mononuclear cells (MNCs). The MNCs consisted of cells that differed in their maturity or functional state, such as lymphocytes from adult peripheral blood vs. umbilical cord blood (cb) or nonstimulated vs. phytohemagglutinin (PHA)-activated lymphocytes from peripheral blood, respectively. The share of T, B, and natural killer (NK) cells or T cell subsets within the initial MNCs or cbMNCs were within physiological reference range for adult peripheral blood. After coculturing with the MMSCs, the populations of B cells decreased in both MNCs and cbMNCs, whereas the populations of the T and NK cells decreased among cbMNC only ( p < 0.05). A decrease in the subset of T-NK cells was observed in the T cells of both MNCs and cbMNCs. In the coculture of MMSCs and PHA-MNCs, we found decrease in the number of CD8 and HLA-DR cells and an increase in the number of CD25 lymphocytes compared to monocultured PHA-MNCs. Our data show that the interaction with MMSCs did not substantially modify the composition of allogenic lymphocytes independent of their maturation (MNCs vs. cbMNCs) or activation (MNCs vs. PHA-MNCs), and the means were within the physiological limits. Moreover, exposure to the MMSCs did not reduce the viability of lymphocytes and even promoted the survival of cells in case of cbMNCs. [ABSTRACT FROM AUTHOR]

Kornegoor R, Verschuur-Maes A H J, Buerger H, Hogenes M C, de Bruin P C, Oudejans J J, Hinrichs B & van Diest P J (2012) Histopathology Immunophenotyping of male breast cancer Aims: Male breast cancer is a rare disease, and knowledge of carcinogenesis is limited. Conflicting results, based on small series, have been reported for clinically relevant biomarkers. Methods and results: One hundred and thirty-four cases of male breast cancer were immunohistochemically stained on tissue microarrays for oestrogen receptor (ER), progesterone receptor (PR), androgen receptor, human epidermal growth factor receptor 2 (HER2), BRST2, cyclin D1, bcl-2, p53, p16, p21, Ki67, cytokeratin (CK) 5/6, CK14, and epidermal growth factor receptor. Data were correlated with clinicopathological features and patient outcome. High mitotic count and high grade were correlated with high Ki67, HER2 amplification/overexpression, p53 accumulation, high p21 expression, low PR expression, and low bcl-2 expression. PR negativity ( P = 0.009) and p53 accumulation ( P = 0.042) were correlated with decreased 5-year survival and were independent markers for patient outcome in Cox regression. In unsupervised hierarchical clustering, four groups were identified that were correlated with distinctive clinicopathological features. The hormone negative/ER-positive/high-grade cluster was significantly associated with decreased survival ( P = 0.011) and was an independent prognostic factor in Cox regression. Conclusions: Several tissue biomarkers are associated with an aggressive phenotype in male breast cancer. PR and p53 are the most promising individual prognostic markers. On the basis of immunophenotype, four distinctive and prognostically relevant male breast cancer groups were identified, indicating that protein expression profiling may be clinically useful in male breast cancer. [ABSTRACT FROM AUTHOR]

Epidemiological studies indicate that in chronic HD patients, bacterial and viral infection rank second place in mortality and morbidity, behind cardiovascular disease. Chronic hepatitis C virus (HCV) is very prevalent in some Hemodialysis (HD) centers. We have investigated lymphocyte subtype count and immunoglobulin levels in hemodialyzed patient with HCV. We studied 55 patients with end stage renal disease (ESRD) on chronic HD and 21 healty subjects. Patients group included 34 female and 21 male with mean age 46.5±16.1(18-77) years. Serum concentrations of IgG, IgM, IgA, CD4, CD8, CD19, CD16-56 lymphocytes were measured. Kt/V values were calculated according to DOQI guideline. Lymphocyte and lymphocyte subgroups values of HD patients were significant lower than healty persons. IgG, IgM and ALT levels of HCV positive HD patients compared with significant higher than HCV negative HD patients. CD4/CD8 ratios within groups were no differences. Lymphocyte subtype count of between HCV positive and HCV negative HD patients was no differences. IgG and IgM levels of HCV positive HD patients were higher than HCV negative HD patients. High Ig levels were also associated with ALT levels. [ABSTRACT FROM AUTHOR]

Natural killer (NK) cells and natural killer T (NKT) cells are subsets of lymphocytes that share some phenotypical and functional similarities. Both cell types can rapidly respond to the presence of tumour cells and participate in antitumour immune responses. This has prompted interest in the development of innovative cancer therapies that are based on the manipulation of NK and NKT cells. Recent studies have highlighted how the immune reactivity of NK and NKT cells is shaped by the environment in which they develop. The rational use of these cells in cancer immunotherapies awaits a better understanding of their effector functions, migratory patterns and survival properties in humans. [ABSTRACT FROM AUTHOR]

The heterogeneity in the healthy human immune system, and the immunological changes that portend various diseases, have been only partially described. Their comprehensive elucidation has been termed the 'Human Immunology Project'. The accurate measurement of variations in the human immune system requires precise and standardized assays to distinguish true biological changes from technical artefacts. Thus, to be successful, the Human Immunology Project will require standardized assays for immunophenotyping humans in health and disease. A major tool in this effort is flow cytometry, which remains highly variable with regard to sample handling, reagents, instrument setup and data analysis. In this Review, we outline the current state of standardization of flow cytometry assays and summarize the steps that are required to enable the Human Immunology Project. [ABSTRACT FROM AUTHOR]

Objective: The balance between the bacillus and host defense mechanisms seems to be important for manifestations of tuberculosis infection. Not only subtypes of T-lymphocytes but their activation status also is important in the immunity of tuberculosis infection. The aim of this study is to address the relationship between T-lymphocyte subtypes, activation and natural killer cells with different clinical forms of mycobacterium tuberculosis infection. Material and Method: Twenty-one active pulmonary tuberculosis, 10 inactive pulmonary tuberculosis, 10 tuberculous pleurisy patients and 18 healthy subjects were involved in the study. Immunophenotyping was carried out in peripheral blood for T cell subtypes, natural killer cells and B cells by flow-cytometry, and HLA-DR and CD25 positivity were studied to determine T-lymphocyte activation. Results: The CD4+ T lymphocyte rates was found significantly lower in bilateral active pulmonary tuberculosis compared to healthy controls. T-lymphocyte activation was found to be increased in tuberculous pleurisy cases. The CD3/CD25+ T lymphocyte rate was significantly higher in tuberculous pleurisy cases compared both to healthy controls and inactive pulmonary tuberculosis cases. Conclusion: CD4+ T-lymphocyte rate may be decreased in pulmonary tuberculosis patients with bilateral pulmonary infiltration. Prominent T-lymphocyte activation in tuberculous pleurisy might be a result of a hypersensitivity reaction against the bacillus. [ABSTRACT FROM AUTHOR]

Background and Objectives: Classic Hodgkin lymphoma (CHL), anaplastic large cell lymphoma (ALCL) and some cases of diffuse large B cell lymphoma (DLBCL) have overlapping morphologic features. Since they all represent distinct clinico-pathologic entities, we explored the differential diagnostic impact of immunophenotyping to discriminate between them. Materials and Methods: We included 61 cases diagnosed as CHL, ALCL, and anaplastic variant of DLBCL. We reviewed morphologic microscopic findings by conventional staining and immunohistochemistry (IHC) with antibodies against PAX-5, CD30, CD15, CD45, EMA, ALK-1, and LMP-1. Results: Fifty cases corresponded to CHL (81.97%), 4 cases to ALCL (6.56%), and 4 cases to DLBCL (6.56%) excluding 3 cases, which remained unclassifiable (4.92%). PAX-5 was expressed in 94% of CHL and 100% of DLBCL cases. LMP-1 was expressed in 52% of CHL and 25% of DLBCL cases. EMA was invariably expressed in all 4 cases of ALK+ ALCL. It was expressed in 4/50 cases (8%) of CHL and in 2/4 cases (50%) of DLBCL. CD45 was expressed in all cases of ALCL and DLBCL but also in 3/50 cases (6%) of CHL. Conclusion: The differentiation between ALCL and CHL based on EMA and CD45 is not reliable. Utilization of PAX-5 in combination with other markers such as CD15 and LMP-1 is recommended. CD20 and PAX-5 are not too helpful in the differentiation of CHL and DLBCL, while CD15 and CD79a were found to be quite useful discriminative markers for this purpose. [ABSTRACT FROM AUTHOR]

We determined the utility of leukocyte cell population data (CPD) for the screening of sepsis and fungemia. Blood culture-positive CBC samples, 117 bacteremia and 27 fungemia, and 134 CBC samples from healthy controls were analyzed using the DxH800 and CPD of neutrophils, lymphocytes, and monocytes were analyzed. Immature granulocytes (IG) were counted using Sysmex XE-2100. The neutrophils and monocytes volume were increased significantly, and the neutrophils light scattering values were reduced significantly in the sepsis samples. ROC curves evidenced excellent sensitivity in the lymphocyte SD parameters (sensitivity 78-89%, specificity 78-87%), monocytes volume (at 177.5, sensitivity 88.2% specificity 87.3%), and monocytes volume SD (at 22.16, sensitivity 93.1% specificity 91.0%) for sepsis. The IG value was significantly higher in sepsis and the ROC curve evidenced a sensitivity of 82.8% and a specificity of 90.8% for sepsis. Only lower angle light scatter of lymphocytes SD value evidenced good sensitivity and specificity in the discrimination of fungemia from bacteremia (sensitivity 74.1%, specificity 72.4% at 12.6). Many of the leukocyte CPD have been identified as useful parameters of sepsis. Hopefully, these parameters can ultimately be incorporated into a decision rule for the screening of sepsis samples and to discriminate fungemia from bacteremia. [ABSTRACT FROM AUTHOR]

The article focuses on lymphocytes. It discusses the three main classification and their roles in the immune response of the body, including B lymphocytes (B cells), T lymphocytes (T cells) and natural killer (NK) cells. It presents a case study of a patient diagnosed with chronic lymphocytic leukemia (CLL) to highlight the vulnerability of patients with decreased production or increased destruction of lymphocytes. INSETS: Five infection fighters;How immunoglobulins come into play.

The t(14;19)(q32;q13) involving the IGH@ and BCL3 loci is an infrequent cytogenetic abnormality detected in B-cell malignancies. We describe the clinicopathologic, cytogenetic, and molecular genetic characteristics of 14 cases of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) with t(14;19)(q32;q13). All patients (10 men and 4 women) had lymphocytosis; 10 had lymphadenopathy. Blood and bone marrow lymphocytes were predominantly small, but cytologically and immunophenotypically atypical. In all cases, t(14;19) was found in the neoplastic stem line; it was the sole abnormality in 4. Ten cases showed additional cytogenetic abnormalities, including trisomy 12 in 9 and complex karyotypes in 7. Fluorescence in situ hybridization demonstrated IGH@/BCL3 fusion gene in all cases. In all cases, the IGHV genes were unmutated, but only 7 expressed ZAP70. Seven cases preferentially used IGHV4-39. Our results indicate that t(14;19)(q32;q13) identifies a subset of CLL/SLL with distinctive clinicopathologic and genetic features. Furthermore, t(14;19) may represent an early, possibly primary, genetic event. [ABSTRACT FROM AUTHOR]

INTRODUCTION: Primary central nervous system lymphoma (PCNSL) is rare and accounts for 1-2% of all primary intracranial tumors (ICT). There are conflicting reports regarding the increased incidence of PCNSL over the last two decades in both immunocompromised and immunocompetent patients. AIM: This study was designed to study the clinicopathological characteristics of PCNSL and to access the trend of PCNSL at our institute. MATERIALS AND METHODS: All the histopathologically proven cases of PCNSL were reviewed from January 1997 to December 2009 (13 years). Immunophenotyping was performed on available paraffin-embedded tissue blocks. Immune status was evaluated and human immunodeficiency virus (HIV) serology was performed in all cases. Cerebrospinal fluid (CSF) findings were recorded whenever available. Possibility of secondary involvement by a systemic lymphoma was excluded in every case. Statistical analysis was done using ?2-test. RESULTS: During the study period (13 years), a total of 4715 cases of ICT were diagnosed, out of which 66 cases were PCNSL, which accounted for 1.4%. The age ranged from 10 to 75 years with a median age of 46 years. All the patients were immunocompetent. Frontal lobe was the most common site of involvement. Diffuse large B-cell lymphoma was the histological pattern in all the cases. CSF involvement was seen in only one case. CONCLUSIONS: In this study, no significant increase in the incidence of PCNSL was found at our institute. Association of PCNSL cases with HIV or acquired immunodeficiency syndrome was not found in our study. [ABSTRACT FROM AUTHOR]

Composite lymphoma (CL) is a rare occurrence of 2 or more morphologically and immunophenotypically distinct lymphoma clones in a single anatomic site. A retrospective analysis of 1,722 solid tissue samples clinically suggestive of lymphoma was carried out in our institute during a 12-year period to evaluate the efficacy of flow cytometry (FC) in identifying CL. We report 17 CL cases. A strong correlation between morphologic findings and FC was observed in 13 cases (76%). In the 4 cases diagnosed as non-Hodgkin lymphoma plus Hodgkin lymphoma, although FC did not detect Reed-Sternberg cells, it accurately identified the neoplastic B- or T-cell component. In 3 cases, FC indicated the need to evaluate an additional neoplastic component that was not morphologically evident. Our data demonstrate that FC immunophenotyping of tissues may enhance the performance of the diagnostic morphologic evaluation of CL. To the best of our knowledge, this is the first report in the literature of a wide series of CL studied also by FC. [ABSTRACT FROM AUTHOR]

Lymphocyte subpopulations present circadian variation of some of their subsets and this variation may influence the magnitude and expression of immune responses. The aim of this study was to analyze the dynamics of variation of specific lymphocyte subsets. Lymphocyte subpopulation analyses were performed on blood samples collected every four hours for 24 hours from 15 healthy subjects aged 37-54 years. A clear circadian rhythm was validated for CD8 with acrophase at noon and for CD3 and CD4 with acrophase at night. Specific lymphocyte subsets present different profiles of nyctohemeral changes and this might explain time-related variations of immune responses. [ABSTRACT FROM AUTHOR]

Introduction: Discordant lymphomas are rare entities characterized by the simultaneous presence of two distinct types of lymphomas in different anatomic sites. We describe a very rare case of simultaneous occurrence of splenic mantle cell lymphoma and marginal zone lymphoma involving the bone marrow and peripheral blood.Case Presentation: We report the case of a 60-year-old asymptomatic Caucasian woman in whom discordant lymphomas were discovered when a slight lymphocytosis and a conspicuous splenomegaly were observed. The different morphological, immunophenotypical and immunohistochemical features found in the different pathologic samples obtained from peripheral blood, bone marrow and spleen sections made it possible to differentiate two types of non-Hodgkin B-cell lymphomas: a mantle cell lymphoma infiltrating the spleen and a marginal zone lymphoma involving both the bone marrow and peripheral blood. Since a similar IgH gene rearrangement was found both in the bone marrow and in the spleen, the hypothesis of a common origin, followed by a different clonal selection of the neoplastic lymphocytes may be taken into consideration.Conclusion: Our case emphasizes the usefulness of investigating simultaneous specimens from different anatomic sites from the same patient and the relevant diagnostic role of splenectomy. [ABSTRACT FROM AUTHOR]

The newly proposed glioma stem cell (GSC) hypothesis may re-model the way we diagnose and treat the tumor, which highlights the need for a complete knowledge on the genetic and epigenetic “blueprints” of GSCs. To identify the true “stemness” signatures, pure GSC populations are primarily needed. Reliable in vitro methods enriching for GSCs and thereby identifying the key stem-like characteristics constitute the preliminary step forward. We discuss in this review the current widely used methods for enriching and isolating GSCs, namely neurosphere assay, CD133 Immunophenotyping and side population assay, and detail their limitations and potential pitfalls that could complicate interpretation of corresponding results. [ABSTRACT FROM AUTHOR]

A number of biomarkers, particularly proteins that contribute to prognosis in diffuse large B cell lymphoma (DLBCL), have been identified. However, translation into accepted standards to predict survival has not yet been accomplished, primarily due to contradictory reports in the literature resulting from, among other factors, arbitrary methodologies used to set cut-off values for determining positivity. Some of these problems might be resolved by application of rational statistical methods for determination of cut-off scores. Herein, we critically address issues on in situ phenotypic prognostic tumor-related biomarkers in DLBCL with a particular and practical emphasis on tools for cut-off level determination, especially receiver operating characteristic curve analysis. Moreover, we candidly illustrate the application of these tools for efficient disease-specific survival prognostication on a tissue microarray collective of 240 primary DLBCL using the common prognostic biomarkers Bcl-2, Bcl-6, CD10, FOXP1, MUM1, and Cyclin E. Comparison of the results relative to disease-specific survival unequivocally showed the superior discriminatory power of the cut-off levels calculated by receiver operating curves and the Youden's index, compared to arbitrary cut-off values from the literature, advocating fundamental application of rational methods for determination of clinically relevant prognostic biomarkers' cut-off scores. [ABSTRACT FROM AUTHOR]

CD4+ and CD8+ T lymphocytes are powerful components of adaptive immunity, which essentially contribute to the elimination of tumors. Due to their cytotoxic capacity, T cells emerged as attractive candidates for specific immunotherapy of cancer. A promising approach is the genetic modification of T cells with chimeric antigen receptors (CARs). First generation CARs consist of a binding moiety specifically recognizing a tumor cell surface antigen and a lymphocyte activating signaling chain. The CAR-mediated recognition induces cytokine production and tumor-directed cytotoxicity of T cells. Second and third generation CARs include signal sequences from various costimulatory molecules resulting in enhanced T-cell persistence and sustained antitumor reaction. Clinical trials revealed that the adoptive transfer of T cells engineered with first generation CARs represents a feasible concept for the induction of clinical responses in some tumor patients. However, further improvement is required, which may be achieved by second or third generation CAR-engrafted T cells. [ABSTRACT FROM AUTHOR]

Background: Dry type localized cutaneous leishmaniasis, one of the most prevalent cutaneous parasitic infections in Kerman Province, is presented as a polarized disease in which cytokine profiles secreted by immune cells play a major role in its presentation. In order to clarify the idea, immunohistochemical study of skin biopsies were performed to elucidate the cytokine release capabilities of immune cells. Methods: Skin biopsies of acute, chronic nonlupoid, and chronic lupoid recidivans lesions of dry type localized cutaneous leishmaniasis were studied by immunohistochemical staining methods for immunophenotypic patterns (CD4, CD8, CD14, CD19, CD56, CD11a, CD18, CD1a, HLA-DR, CD54) and cytokines (INF-gamma, IL-12, IL-4, TNF-alpha) released by immune inflammatory cells. Results: The descriptive analysis of data showed that the mean percentage of positive immunostained cells of CD4, CD8, and CD14; antigen-presenting cells (CD1a, HLA-DR); and markers of the extravasated positive memory T cells (CD11a, CD18, CD54) are more frequent in lupoid recidivans than in acute active and chronic nonlupoid lesions, in order of frequency. Conclusion: Based on the results, it seems that Th1-like response is predominant in acute active form and lupoid recidivans while Th2-like response is predominant in chronic nonlupoid lesions. It seems that lupoid recidivans is a type IV hypersensitivity reaction to the reactivation of hidden antigens. [ABSTRACT FROM AUTHOR]

BCL6 is a transcriptional repressor that has important functions in lymphocyte differentiation and lymphomagenesis, but there have been no reports of BCL6 expression in gastric cancers. In the present study, we investigated the BCL6 function in gastric cancers. Treatment with TPA resulted in BCL6 degradation and cyclin D2 upregulation. This phenomenon was inhibited by the suppression of the nuclear translocation of HB-EGF-CTF (C-terminal fragment of pro-HB-EGF). The HB-EGF-CTF nuclear translocation leads to the interaction of BCL6 with HB-EGF-CTF and the nuclear export of BCL6, and after that BCL6 degradation was mediated by ubiquitin/proteasome pathway. Real-time RT-PCR and siRNA targeting BCL6 revealed that BCL6 suppresses cyclin D2 expression. Our data indicate that BCL6 interacts with nuclear-translocated HB-EGF-CTF and that the nuclear export and degradation of BCL6 induces cyclin D2 upregulation. We performed immunohistochemical analyses of BCL6, HB-EGF and cyclin D2 in human gastric cancers. The inverse correlation between BCL6 and cyclin D2 was also found in HB-EGF-positive human gastric cancers. BCL6 degradation caused by the HB-EGF-CTF also might induce cyclin D2 expression in human gastric cancers. Inhibition of HB-EGF-CTF nuclear translocation and maintenance of BCL6 function are important for the regulation of gastric cancer progression. [ABSTRACT FROM AUTHOR]

The neutrophil/lymphocyte ratio (NLR) has recently been described as a predictor of mortality in patients who undergo percutaneous coronary intervention. The aim of this study was to investigate the utility of admission NLRs in predicting outcomes in patients with acute coronary syndromes (ACS). A total of 2,833 patients admitted to the University of Michigan Health System with diagnoses of ACS from December 1998 to October 2004 were followed. Patients were divided into tertiles according to NLR. The primary end point was all-cause in-hospital and 6-month mortality. The ACS cohort comprised 564 patients with ST-segment elevation myocardial infarctions and 2,269 patients with non–ST-segment elevation ACS. Patients in tertile 3 had higher in-hospital (8.5% vs 1.8%) and 6-month (11.5% vs 2.5%) mortality compared with those in tertile 1 (p <0.001). After adjusting for Global Registry of Acute Coronary Events risk profile, patients in the highest tertile were at an exaggerated risk for in-hospital (odds ratio 2.04, p = 0.013) and 6-month (odds ratio 3.88, p <0.001) mortality. Admission NLR is an independent predictor of in-hospital and 6-month mortality in patients with ACS. This relatively inexpensive marker of inflammation can aid in the risk stratification and prognosis of patients diagnosed with ACS. [Copyright &y& Elsevier]

Abstract: The great shortage of human hepatic cells makes it desirable to generate extrahepatic stem or precursor cells. In recent years, it has been reported that human multipotential mesenchymal stem cells (hMSCs) differentiate into hepatocyte-like cells. The fetal lung is one of the largest organs containing many MSCs that can be easily obtained. Whether MSCs from fetal lung can differentiate into hepatocytes or bile duct cells is an important issue in basic medicine and clinical application. We isolated fetal lung cells, and expanded and analyzed them. At passage 4, their morphologic, immunophenotyping and cytokine secretions were similar to adult bone marrow-derived MSCs. We conclude that these cells from fetal lung are MSCs, indicating that human fetal lung is an ideal source of hMSCs. hMSCs from fetal lung induced in special differentiation medium showed homogeneous and small polygonal endothelial-like morphology, expressing weak mRNA, as well as Alb and AFP. This implies that hMSCs from fetal lung can differentiate into hepatocyte-like cells. [Copyright &y& Elsevier]

Background and purpose: The cause of Tourette syndrome (TS) is not precisely known, although several lines of evidence point at an involvement of the immune system in its pathogenesis. Results: Here, we report the results of a pilot study investigating frequently analysed lymphocyte surface markers in 20 adult patients with TS (16 males; 37.3 ± 15.8 years) and 20 matched controls (16 males; 37.5 ± 15.3 years). Statistical analysis revealed significant differences for the investigated lymphocyte surface markers. The difference in CD69+/CD22+-B cells (23.0 ± 10.5% vs. 13.1 ± 6.1%; P = 0.001) and in CD95+/CD4+-T cells (41.5 ± 12.1% vs. 24.6 ± 10.0%; P = 0.0001) was still significant after Bonferroni–Holm correction. Conclusion: Our preliminary data indicate that TS may be associated with an increased peripheral immune activity. [ABSTRACT FROM AUTHOR]

Since the discovery of epithelial hair follicle stem cells (eHFSCs) in the bulge of human hair follicles (HFs) an important quest has started: to define useful markers. In the current study, we contribute to this by critically evaluating corresponding published immunoreactivity (IR) patterns, and by attempting to identify markers for the in situ identification of human eHFSCs and their niche. For this, human scalp skin cryosections of at least five different individuals were examined, employing standard immunohistology as well as increased sensitivity methods. Defined reference areas were compared by quantitative immunohistochemistry for the relative intensity of their specific IR. According to our experience, the most useful positive markers for human bulge cells turned out to be cytokeratin 15, cytokeratin 19 and CD200, but were not exclusive, while β1 integrin and Lhx2 IR were not upregulated by human bulge keratinocytes. Absent IR for CD34, connexin43 and nestin on human bulge cells may be exploited as negative markers. α6 integrin, fibronectin, nidogen, fibrillin-1 and latent transforming growth factor (TGF)-beta-binding protein-1 were expressed throughout the connective tissue sheath of human HFs. On the other hand, tenascin-C was upregulated in the bulge and may thus constitute a component of the bulge stem cell niche of human HFs. These immunophenotyping results shed further light on the in situ expression patterns of claimed follicular ‘stem cell markers’ and suggest that not a single marker alone but only the use of a limited corresponding panel of positive and negative markers may offer a reasonable and pragmatic compromise for identifying human bulge stem cells in situ. [ABSTRACT FROM AUTHOR]

The immunophenotype of lymphoid cytologic samples obtained by laser scanning cytometry (LSC) and flow cytometry (FC) was compared in 72 cases composed of a series of 23 cases with simultaneous LSC and FC immunophenotyping and a second series of 49 cases in which nonsimultaneous immunophenotyping was performed. In both series, no discordance in the population immunophenotype was found that would result in changes in diagnostic classification, although minor discordance in some antigens was found, predominantly affecting FMC7, CD11c, and CD23. The immunophenotype obtained by LSC shows a high degree of concordance with that obtained by FC and generates results that are diagnostically equivalent. Potential explanations for the discordant markers include differences related to the techniques, differences in the fluorochrome-labeled antibodies, technical factors, differences in antigen expression related to anatomic sites, temporal variations, and interpretive variances. [ABSTRACT FROM AUTHOR]

Aberrant antigen expression in acute myeloid leukemia (AML) has been extensively studied in the West with limited reports from Taiwan. We carried out this retrospective study to characterize the frequency and significance of aberrant antigen expression of AML in Taiwan. Among 111 cases, 58 (52%) showed aberrant antigen expression, most frequently CD7 (27%) and CD56 (23%). Aberrant CD7 expression was observed in all non-AML-M3 subtypes, most frequently in AML-M7 (4/6, 67%); while CD19 expression was only observed in AML-M2 (5/36, 14%). CD56 expression was most common in AML-M5 (4/8, 50%). The relative frequency of CD19 and CD56 expression in AML with t(8;21) was higher than those with other chromosomal abnormalities or normal karyotype ( P = 0.011 and 0.005, respectively). In non-M3 AML, aberrant antigen expression was identified in 56/96 (58%) cases, in contrast to 2/15 (13%) AML-M3 cases ( P = 0.001). CD7, CD19 and CD56 expression was not correlated with remission rate. We concluded that aberrant immunophenotype was more frequent in non-M3 leukemias in Taiwan. The relative frequency of CD19 and/or CD56 expression in AML with t(8;21) was significantly higher than those without this translocation and co-expression of these two antigens may serve as the surrogate markers for AML with t(8;21). [ABSTRACT FROM AUTHOR]

Context.-Beginning with the immunologic classifications of Lukes and Collins and Kiel and culminating in the Revised European-American Lymphoma and World Health Organization classifications, the diagnosis of lymphoid tumors relies heavily on the determination of cell lineage, maturation, and function, based on antigen expression in addition to morphology and clinical features. Technologic advances in immunology, antibody production, genetic analysis, cloning, and the identification of new genes and proteins by microarray and proteomics have provided pathologists with many antibodies to use in routine diagnosis. Objective.-To provide guidance to the practicing pathologist in the appropriate selection of an antibody panel for the diagnosis of lymphoma based on morphology and relevant clinical data and to avoid pitfalls in the interpretation of immunohistochemical data. Attention is given to some of the newer antibodies, particularly against transcription factors, that are diagnostically and prognostically useful. Data Sources.-The information presented in this article is based on review of the literature using the OVID database (Ovid MEDLINE 1950 to present with daily update) and 20 years of experience in diagnostic hematopathology. Conclusions.-Immunophenotyping is required for the diagnosis and classification of lymphoid malignancies. Many paraffin-reactive antibodies are available to the pathologist but most are not specific. To avoid diagnostic pitfalls, interpretation of marker studies must be based on a panel and knowledge of a particular antigen's expression in normal, reactive, and neoplastic conditions. [ABSTRACT FROM AUTHOR]

Background: The diagnosis of Merkel cell carcinoma (MCC) can be rather challenging; therefore, the immunohistochemical profile is important in confirming the microscopic diagnosis. Characteristic of the neuroendocrine and epithelial differentiation of MCC, antibodies to cytokeratin (CK) 20, CK7, epithelial membrane antigen, and neuron-specific enolase among others, are used in confirming the diagnosis. As reported in the literature, the majority of MCC express CK20 and are CK7 negative. Herein, we present a case series of seven patients with CK20−/CK7+ primary cutaneous neuroendocrine carcinoma. Methods: All cases of MCC with specific CK20−/CK7+ staining on file at a large Veterans’ hospital and tertiary referral dermatopathology service were reviewed. All seven cases were analyzed for clinical, pathologic and immunophenotypic attributes. All specimens were submitted for immunohistochemical staining and interpreted by a single dermatopathologist. Results: All the cases showed diffuse cytoplasmic staining for CK7 and positive staining for synaptophysin. CK20 and thyroid transcription factor-1 staining was negative. Conclusions: Herein we have presented a hitherto unreported group of patients with CK20−/CK7+ primary neuroendocrine carcinoma of the skin. The purpose of this case series is to describe a new immunophenotypic variant of MCC, while further expanding the differential diagnosis of tumors included in the subset of neoplasms showing CK20−/CK7+ staining. [ABSTRACT FROM AUTHOR]

Objective. The aim of this study was to determine the basic clinical characteristics of the lymphomas in a Mexican population, as well as the different morphological subtypes and their immunophenotype. Method. All the hematolymphoid neplasms with immunostains were retrieved from the files of the departments of pathology of the Hospital Central Militar, Hospital Español and CEDEHO, from January 1999 through November 2006. They were classified within two groups. Hodgkin lymphoma (HL) and non Hodgkin lymphoma (NHL); NHL were subclassified in B and T cells, according to the WHO classification. The clinical information submitted in each case was also examined. Results. 1,047 hematolymphoid neoplasm were retrived; from these, 898 (85.76%) correspond to NHL, 146 (13.94%) to HL, and 3 (0.28%) to null type. From NHL, the immunophenotype B was the most frequent, and from this, the type most common was diffuse large B-cell lymphoma. From HL, the subtype most frequent was nodular sclerosis. There was a predominance of males, with 555 cases (53%) in men, and 492 cases (47%) in women, with a relation of 1.2:1. The average of edge was 41.3 years, and the location most frequent was lymph nodes. Conclusions. The behavior of lymphomas in our study is very similar to worldwide literature, as well as the necessity of carrying out immunostains for the diagnosis of lymphomas. [ABSTRACT FROM AUTHOR]

Abstract: Evidence from animal models suggests that the expression of CD8αα homodimer on CD8+ T cells plays a key role in the generation of long-lived memory cells. Here, we studied the quantitative alterations of CD8+ T cell subsets expressing CD8αα, interleukin-7 receptor (IL-7Rα) and activation markers in HIV-1-infected individuals including aviremic, viremic and slow progressor subjects using eight-color flow cytometry. Compared to slow progressor subjects, expression of CD8αα was significantly reduced in aviremic and viremic patients and this reduction occurred mainly within central memory cell subsets and not in naive and effector memory compartments. Persistence of antigenemia leads to IL-7Rα loss mainly on central and pre-terminal memory CD8+ T cell subsets in viremic patients but not in slow progressor subjects. Compared to aviremic and viremic patients, slow progressor subjects had lower levels of IL-7 and reduced activated cells. The expression of CD8αα was not significantly related to IL-7Rα although negative associations were evidenced within all CD8+ T cell subsets. Collectively, these results further advance the characterization of immunophenotypic patterns of CD8+ T cell subsets expressing CD8αα/IL-7Rα and provide new insights into the ability of HIV-1 infection to alter memory cell population. [Copyright &y& Elsevier]

SummaryDifferential immunophenotyping is essentialfor diagnosis and follow-up. Routine analysis requiresa minimum of 250 γl blood, and neither a detailedimmunophenosubtyping nor absolute cell count areavailable. We present two concepts using slide-based cytometryin order to break these limitations. Materials andMethods: 10 γl EDTA blood were stained with DRAQ5and either CD3-PE, CD4-Alexa Fluor 488 and CD8-PE-Cy5or CD45-FITC and CD14-PE. A no-lyse-no-wash' versus alyse-no-wash' method (Quicklysis) was performed. 20 γlof the suspension were applied to a Neubauer chamber.Leukocytes were analyzed by laser scanning cytometry(LSC) twice with a time interval of 15 min. Anotheraliquot of blood was taken for routine analysis. Erythrocytecount was performed after another dilution. Results:Brévais' regression coefficient showed a good internalcorrelation of LSC (r > 0.85) with no significant differencebetween routine analysis and LSC (lyse-no-wash': r = 0.9,p = 0.009, α = 0.05; no-lyse-no-wash': r = 0.98, p < 0,0001,α = 0.05). Immunophenosubtyping by LSC (CD3/CD4/CD8, CD45/CD14) was unequivocal. Conclusion: A detailedimmunophenosubtyping and absolute cell count ispossible by using the Neubauer chamber for slide-basedanalysis. This method requires minimal amounts ofblood, is very cost-efficient, and data show no significantdifference to routine laboratory. This concept mightprove versatile in patients with low blood volume andwhere absolute cell count is essential. [ABSTRACT FROM AUTHOR]