Immunophenotypic Patterns Research Articles

Immunohistochemical profiles of different subsets of Merkel cell carcinoma

The literature records many examples of Merkel cell carcinoma (MCC) exhibiting aberrant immunohistochemical profiles. These can lead to diagnostic difficulty. The objectives of the current study were (1) to examine the immunohistochemical profile of different subsets of MCC to determine whether predictable subset-specific patterns exist and (2) to establish whether shared immunophenotypic patterns might reveal links between individual subsets, as demonstrated previously at a genetic level. In 52 cases of MCC, stratified by viral status and morphology, we studied 5 markers commonly used in the diagnostic evaluation of these tumors (CK20, CK7, chromogranin, neurofilament and TTF-1). Expression of these proteins was recorded as quantitative (H-scores) and absolute (positive vs negative) variables. In general, our data indicate that the "classical" or expected panel (CK20+, NF+, Chromo+, TTF-1, CK7-) is observed significantly more often in pure Merkel cell polyomavirus (MCPyV)-positive than in MCPyV-negative cases (78% vs 25%; P = .002). Neurofilament was less frequently encountered in MCPyV-negative than in MCPyV-positive tumors (66.7% vs 100%; P = .001) and expression of TTF-1 (37.5% vs 3.6%; P = .003) and CK7 (45.8 vs 14.3; P = .02) was more frequent. No significant immonophenotypic differences were observed between pure and combined MCPyV-negative tumors. Recognition of the more aberrant immunohistochemical profile of MCPyV-negative MCC should inform the diagnostic approach to this tumor. Moreover, the shared aberrant immunophenotype in pure and combined MCPyV-negative tumors supports a link between these entities and serves to separate them from MCPyV-positive tumors.

A study on clinical, immunophenotypic pattern in pediatric acute leukemias in a teaching hospital

Background: Acute leukemia contributes to nearly one third of the pediatric malignancies. For effective management of a curable malignancy, the epidemiology of childhood leukemias and audit of previously treated patients would be required. The objective was to study the clinical and immunophenotypic pattern of acute leukemia in children <12 years of age admitted in a tertiary centre. Methods: Prospective study done on 31 children, diagnosed as leukemia and treated over a period of 3 years in GRH, Madurai. Detailed history analysis, clinical examination, lab investigations including flowcytometry were done.Results: ALL was the most common acute leukemia in children (90.3%). 85.7% were identified as B-cell and 14.3% as T-cell. Of the B-ALL, 87.5% was pre-B, 8.3% pro-B and 4.2% mature B ALL. Myeloid antigen co-expression was seen in 35.7%. Fever was the most common symptom (87%) and hepatosplenomegaly was the most common sign (>90%). Poor outcome was noted in 42.3%. T-cell appeared to have worse prognosis than B-cell but did not retain independent prognostic significance in univariate analysis.Conclusions: B-ALL was more common than T-ALL. Extra medullary organ involvement indicates increased tumor burden and poor outcome. None of the clinical or laboratory parameters appeared to predict poor outcomes in our study and this may be due to small sample size. This study provides an insight into the data regarding the epidemiology, clinicopathologic feature and outcome of acute leukemia in a center with low resource settings.

Plasma Cell Immunophenotyping Improve Prognostic Stratification of Multiple Myeloma Patients

Background: The aims of this study were to establish the clinical value of multi-parametric flow cytomentry (MFC) in multiple myeloma (MM) and monoclonal gammopathy of undetermined significance (MGUS). Methods: We analyzed bone marrow aspirates from 112 MM and 17 MGUS patients by MFC, using 3 combinations of 9 color labeling: a, CD38 / CD138 / CD45 / CD56 / CD19 / CD27 / CD117 / CD20 / CD33; b, CD38 / CD138 / CD45 / cytoplasmic Kappa / cytoplasmic Lambda; and c, CD38 / CD138. MFC data were classified based on clinical features and prognosis factors. Results: Myeloma’s patients compared to MGUS group had plasma cells (PCs) with abnormal immunophenotypic patterns, including high CD56 and CD20 expression and weak or negative CD45, CD19, and CD27 expression without significant median differences in expression of CD33 and CD117. Multiple myeloma patient with low expression of CD19, CD27 or CD45, overexpression of CD56 or with a high proportion of PCs at diagnosis demonstrated shorter overall survival times. Moreover, MM patients with combined abnormal expressions of 4 or 5 antigens demonstrated shorter survival times (P = 0.001). These high-risk MFC patients were associated with poor clinical outcomes, including ISS stage III and DS stage III, low hemoglobin levels, and elevated serum beta2-microglobilin (P = 0.01, P = 0.006, P = 0.01 and P = 0.008, respectively). Conclusions: The present study highlights the benefits of assessing abnormal antigen expression for clinical uses. These measures could facilitate proper diagnosis of disorders and improve risk stratification for a targeted early treatment regimen.

BRAFV600E mutation in hairy cell leukemia: A single-center experience.

BRAFV600E mutation has been reported as a unique genetic lesion of hairy cell leukemia (HCL), a subset of which lacks this lesion and shows adverse outcomes. To determine the prevalence of BRAFV600E in HCL from our center and derive clinicopathological correlation, if any. A 9-year retrospective analysis of 46 consecutive cases of HCL diagnosed on morphology and immunophenotyping was done. Stained smears were used as samples for amplification refractory mutation system polymerase-chain reaction using fluorescent primers for mutation detection. BRAFV600E mutation was detected in 41/46 patients (89.1%) while absent in control samples of chronic lymphocytic leukemia. Cases mimicking HCL-variant clinically or immunophenotypically too showed the presence of this mutation. HCL with mutated BRAF presented at a younger age. No statistical difference in blood counts, tumor load, and immunophenotype patterns existed among BRAF mutated and unmutated group. Nine patients (45%) with mutated BRAF had residual disease following treatment with cladribine. BRAFV600E mutation analysis has a definitive role in the diagnosis of HCL.

Prognostic value of multicenter flow cytometry harmonized assessment of minimal residual disease in acute myeloblastic leukemia

The assessment of minimal residual disease (MRD) in acute myeloblastic leukemia is of growing interest as a prognostic marker of patients' outcome. Multiparameter flow cytometry (MFC), tracking leukemia-associated immunophenotypic patterns, has been shown in several studies to be a useful tool to investigate MRD. Here, we report a multicenter prospective study which allowed to define a harmonized analysis strategy, as well as the efficacy of MFC MRD to predict outcome. This study included 276 patients, in 10 different MFC centers, of whom 268 had at least 1 MRD check point. The combination of a CD45, CD34, and CD33 backbone, with the addition of CD117, CD13, CD7, and CD15 in 2 five-color tubes allowed to define each patient's multiparameter immunophenotypic characteristics at diagnosis, according to a Boolean combination of gates. The same individual diagnosis gating strategy was then applied at each MRD time point for each patient. MRD levels were stratified according to log by log thresholds, from 5×10-2 (the classical morphological threshold to define remission) down to <5×10-5 . MRD was found to be constantly negative (<5×10-5 ) for 148 patients. Survival analyses significantly associated MRD negativity with a good prognosis and any positive value with poorer outcome. All P values were <0.0001 both for disease-free and overall survival at the earliest time point (post-induction, MRD1) as well as when considering all time points together. Finally, MRD levels were independent of cytogenetics and allowed in fact to further stratify all cytogenetics risk groups. In summary, this multicenter study demonstrates that a simple combination of immunophenotypic markers successfully allows for the detection of MRD in acute myeloblastic leukemia patients, with a strong correlation to outcome.

Intermediate and nonclassical monocytes show heterogeneity in patients with different types of acute coronary syndrome.

This study was performed to gain further insight in the heterogeneity of monocytes in the different categories of acute coronary syndrome (ACS), especially between patients with unstable angina pectoris, ST-elevation myocardial infarction (STEMI), and non-ST-elevation myocardial infarction (NSTEMI). For this purpose, blood samples were collected in the acute phase from patients presenting with an ACS. These samples were examined with multiparameter flow cytometry to identify the different monocyte subsets and to analyze the expression of monocyte-associated molecules. Leukocytes, as well as an absolute number of monocytes, showed a clear and significant increase in patients with STEMI. This increase was seen in all subtypes of monocytes. The classical monocytes (CD14++CD16-) of patients with an NSTEMI had a significantly increased CD11b expression when compared to the control group, while these cells showed a decreased expression pattern in STEMI patients. This increased CD11b-expression was also seen in the intermediate monocytes of NSTEMI, while it was almost completely downregulated on the intermediate monocytes of STEMI. Finally, CX3CR1, which is almost exclusively expressed on intermediate and nonclassical monocytes, showed a significant decrease in expression in patients with STEMI. In conclusion, intermediate and nonclassical monocytes have a different immunophenotypic pattern in patients with STEMI versus NSTEMI. These differences reflect the pro-inflammatory state of the monocytes in NSTEMI and can be used as target molecules for novel therapeutic strategies to diminish the migration of proinflammatory monocytes into the myocardial tissue. © 2017 International Society for Advancement of Cytometry.

Preventing relapse after haematopoietic stem cell transplantation for acute leukaemia: the role of post-transplantation minimal residual disease (MRD) monitoring and MRD-directed intervention.

Relapse is the main cause of treatment failure after allogeneic haematopoietic stem cell transplantation (allo-HSCT) for acute leukaemia (AL). Post-transplantation minimal residual disease (MRD) monitoring enables risk stratification and identifies AL patients at higher risk of relapse. MRD assessment primarily involves the determination of leukaemia-associated immunophenotypic patterns using multiparameter flow cytometry, and the polymerase chain reaction (PCR)-based evaluation of expression levels of leukaemia-related genes (specific reciprocal gene rearrangements and other mutation types). In addition, next generation sequencing and digital PCR may further enrich current MRD detection. Several MRD-directed interventions have demonstrated the ability to reduce the risk of relapse with acceptable treatment-related toxicities. Donor lymphocyte infusion (DLI) is the most important intervention for MRD-positive patients, while several modified strategies, such as granulocyte colony-stimulating factor-mobilized peripheral blood cells followed by short term immune suppression and escalating dose regimen, further improve the safety and efficacy of DLI. Interferon therapy, targeted drugs, and hypomethylating agents have also been introduced for MRD-directed interventions. Referring to the issues of whether and who would benefit from pre-emptive intervention according to MRD, in this review, we summarized this rapidly evolving area of MRD monitoring and MRD-directed interventions in AL patients after allo-HSCT.

Acquired Cystic Disease-Associated Renal Cell Carcinoma: Review of Pathogenesis, Morphology, Ancillary Tests, and Clinical Features.

Acquired cystic disease-associated renal cell carcinoma (ACD-RCC) is a recently described subtype of RCC found in individuals with ACD of the kidney. Because of underrecognition, information regarding this lesion is sparse but continues to accumulate with each new report. Herein, a thorough literature review amassing the current understanding of this unique neoplasm is presented. Discussion focuses on clinical features, pathogenesis, disease outcome, and relation to the duration of dialysis. The macroscopic and characteristic microscopic features are described with illustrations. Compared with previous opinion, compiled immunohistochemical data may now allow for recognition of a unique immunophenotypic pattern of ACD-RCC. Distinction of ACD-RCC from clear cell and papillary RCCs based on molecular genetic information is deliberated, including a summary of the most frequently detected cytogenetic abnormalities. The key morphologic and immunophenotypic patterns used to distinguish this entity from a comprehensive differential diagnosis are provided.

Comparison of clinicopathological characteristics between T-cell prolymphocytic leukemia and peripheral T-cell lymphoma, not otherwise specified.

T-cell prolymphocytic leukemia (T-PLL) is a very rare, aggressive T-cell neoplasm. Peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) is also a highly aggressive lymphoma. These two diseases can often be confused with each other; therefore, we aimed to determine the clinical and pathological differences between T-PLL and PTCL-NOS. We analyzed 15 T-PLL and 91 PTCL-NOS patients and also compared clinical features between T-PLL and PTCL-NOS with leukemic presentation. Peripheral blood images and biopsy specimens were analyzed, and treatment responses were determined via imaging modalities. The clinicopathological characteristics were statistically compared. T-PLL cells were smaller in size than those of PTCL-NOS with leukemic presentation (P=.0068); moreover, PTCL-NOS cells with leukemic presentation were smaller than those of PTCL-NOS without leukemic presentation (P=.0017). Immunophenotypic patterns in T-PLL and PTCL-NOS were similar. Five-year overall survival rates of T-PLL and all PTCL-NOS patients were 57.5% and 36.8%, respectively. No significant differences were found in clinical manifestations or prognoses; T-PLL and PTCL-NOS with leukemic presentation had essentially equivalent characteristics. T-PLL and PTCL-NOS may share common biological and clinical characteristics in Japanese patients.

Carcinoma of Ampulla of Vater: Carcinogenesis and Immunophenotypic Evaluation

The ampulla of Vater represents the epithelial junction of the main pancreatic duct and the distal bile duct, surrounded by the pancreatic head and duodenum parenchyma. Ampullary carcinoma can arise either from the intestinal epithelium or the epithelium covering the pancreatobiliary ducts. Many tumors develop from pre-existing adenomas, others evolve from precursor intraepithelial neoplasia supporting that two histological subtypes have different carcinogenesis. Thus, the question arises of classifying ampullary adenocarcinomas as the immunophenotypic pattern allowing a better understanding of the clinical implications of this neoplasm.

Can exome scans be expected to be part of real‐time decision‐making in patients with haematological cancers?

The laboratory aspects of diagnosis of patients with haematological malignancies are forever changing. Microscopic examination of blood and bone marrow smears, cytogenetics, flow cytometry and single-assay molecular diagnostics have been and are still essential tools in cancer diagnostics. Flow cytometry has brought the unprecedented possibility of rapid multiplexing and characterization of complex immunophenotypic patterns. However, the advent of next generation sequencing in the haematology laboratory brings a whole new perspective on multiplexing and potentially lowers the cost per analysis effectively. These informative methods still require skilled technicians and bioinformaticians, evolve at a rapid pace, and call for clinical guidelines and best practice. Here, we discuss the potential and caveats of whole exome sequencing as it moves closer to routine laboratory practice. The question is: Will exome sequencing be performed, real-time, in the standard haemodiagnostic laboratory?

Minimal residual disease monitoring and preemptive immunotherapy in myelodysplastic syndrome after allogeneic hematopoietic stem cell transplantation.

This study investigated the efficacy of minimal residual disease (MRD) monitoring and MRD-directed preemptive immunotherapy in high-risk myelodysplastic syndrome (MDS) patients who received allogeneic hematopoietic stem cell transplantation (HSCT). MRD assessment consisted of Wilms' tumor gene 1 (WT1) detection with PCR and leukemia-associated immunophenotypic pattern examination with multiparameter flow cytometry (FCM). Post-HSCT, 31 patients were positive for WT1, and 8, for FCM; positivity for WT1 (18.6 vs. 6.1%, P = 0.040) or FCM (62.5 vs. 3.6%, P < 0.001) indicated a higher 2-year relapse rate. Twenty-one patients met our combined criteria for MRD, and the presence of MRD was associated with a higher 2-year relapse rate (27.3 vs. 4.5%, P = 0.003). Preferentially expressed antigen of melanoma (PRAME) expression alone was not an appropriate MRD marker; however, it suggested that the MRD-positive patients may fail to respond to preemptive immunotherapy. In patients positive for both PRAME and MRD, the relapse rate was 60% despite preemptive immunotherapy. Multivariate analysis confirmed the association between the increased relapse rate and positivity for both PRAME and MRD (hazard ratio = 42.8, P = 0.001). MRD monitoring predicted relapse in high-risk MDS post-HSCT patients, and PRAME- and MRD-positive patients did not benefit from preemptive immunotherapy.

An analysis of immunophenotyping of peripheral lymphocytes in adult patients with infectious mononucleosis and chronic active Epstein-Barr virus infection

To determine the immunophenotypic features of peripheral lymphocytes in adult patients with Epstein-Barr virus(EBV)-associated infectious mononucleosis(IM) and chronic active EBV infection (CAEBV). Eighteen IM patients, 12 CAEBV patients and 18 healthy donors were included. Lymphocyte subsets including CD3(-)CD19(+) B cells, CD3(-)CD16/56(+) NK cells, CD4(+) and CD8(+) T cells in peripheral blood were measured by flow cytometry. The expression of activation markers (HLA-DR and CD38) on CD8(+) T cells and CD28 expression on T cells were also determined. Kruskal-Wallis H and Mann-Whitney U tests were used to compare variables among groups. IM patients had dramatically increased CD8(+) T cell counts than healthy donors (5.22×10(9)/L vs 0.54×10(9)/L, P<0.001). B cell counts moderately reduced in patients with IM than in healthy donors. No difference was found in absolute CD4(+) T cell and NK cell counts between IM and healthy donors. The levels of HLA-DR and CD38 on CD8(+) T cells significantly increased in IM patients compared with those in healthy controls. The intensity of CD28 on CD8(+) T cells significantly decreased, which was not seen on CD4(+) T cells. The median cell counts of B, NK, CD4(+) T and CD8(+) T subsets in CAEBV patients were 0.02×10(9)/L, 0.06×10(9)/L, 0.26×10(9)/L and 0.21×10(9)/L respectively, which were significantly lower than those in healthy donors (0.22×10(9)/L, 0.38×10(9)/L, 0.78×10(9)/L, 0.54×10(9)/L)and IM patients (0.12×10(9)/L, 0.40×10(9)/L, 0.91×10(9)/L, 5.22×10(9)/L). The positive rates of HLA-DR and CD38 on CD8(+) T cells in CAEBV patients were higher than those in healthy controls, but lower than those in IM patients. The immunophenotypic pattern in adult patients with IM is characterized by a dramatic increase of extensively activated CD8(+) T cells, a moderate reduction of CD19(+) B cells and no significant change of CD4(+) T cells and CD16/56(+) NK cells. CAEBV is featured by an immunosuppression status as demonstrated by significantly decreased B, NK, CD4(+) T and CD8(+) T subsets.

Molecular Features of Three Children Diagnosed With Early T-Cell Precursor Acute Lymphoblastic Leukemia.

Molecular Features of Three Children Diagnosed With Early T-Cell Precursor Acute Lymphoblastic Leukemia.

Characterization of Common Chromosomal Translocations and Their Frequencies in Acute Myeloid Leukemia Patients of Northwest Iran.

Detection of chromosomal translocations has an important role in diagnosis and treatment of hematological disorders. We aimed to evaluate the 46 new cases of de novo acute myeloid leukemia (AML) patients for common translocations and to assess the effect of geographic and ethnic differences on their frequencies. In this descriptive study, reverse transcriptase-polymerase chain reaction (RT-PCR) was used on 46 fresh bone marrow or peripheral blood samples to detect translocations t (8; 21), t (15; 17), t (9; 11) and inv (16). Patients were classified using the French-American-British (FAB) criteria in to eight sub-groups (M0-M7). Immunophenotyping and biochemical test results of patients were compared with RT-PCR results. Our patients were relatively young with a mean age of 44 years. AML was relatively predominant in female patients (54.3%) and most of patients belonged to AML-M2. Translocation t (8; 21) had the highest frequency (13%) and t (15; 17) with 2.7% incidence was the second most frequent. CD19 as an immunophenotypic marker was at a relatively high frequency (50%) in cases with t (8; 21), and patients with this translocation had a specific immunophenotypic pattern of complete expression of CD45, CD38, CD34, CD33 and HLA-DR. Similarities and differences of results in Iran with different parts of the world can be explained with ethnic and geographic factors in characterizations of AML. Recognition of these factors especially in other comprehensive studies may aid better diagnosis and management of this disease.

Microgranular acute promyelocytic leukemia presenting with leukopenia and an unusual immunophenotype

The microgranular variant (M3v) of acute promyelocytic leukemia (APL) is rare, and the diagnosis can be delayed due to variability in how this condition presents. M3v blasts often have folded nuclei, but unlike traditional APL blasts, they often possess faint granules without Auer rods. In addition, microgranular APL often presents with an elevated or normal white blood cell count in contrast with the leukopenia seen in traditional APL. In APL, delayed diagnosis can lead to early death from disseminated intravascular coagulation (DIC), which is the main cause of mortality in an otherwise treatable, and often curable, leukemia. We describe a 19-year-old male with microgranular APL who presented with leukopenia and many blasts resembling non-APL AML blasts with an unexpected immunophenotypic pattern. He was treated for DIC and initiated on all-trans-retinoic acid and arsenic trioxide; he achieved complete molecular remission after induction therapy. Suspicion for APL should always remain high in the presence of clinical manifestations of the disease in order that appropriate treatment can be initiated rapidly to prevent early death.

Blastic plasmacytoid dendritic cell neoplasm with leukemic presentation: 10-Color flow cytometry diagnosis and HyperCVAD therapy.

Few studies describe the comprehensive immunophenotypic pattern of blastic plasmacytoid dendritic cell neoplasm (BPDCN) in the bone marrow and its treatment. This retrospective analysis evaluates the diagnostic flow cytometry (FCM) pattern and outcome of nine patients diagnosed with BPDCN. A four-tube 10-color FCM panel used for diagnosis of acute leukemia (AL), showed cells in the blast gate (CD45dim/low SSC) and were positive for CD4(bright), CD33(dim), CD56(heterogenous), CD123(bright), CD36, CD38, HLA-DR, CD71. Seven patients received front-line induction therapy with HyperCVAD with an overall response rate of 86%. Five of six responders underwent planned allogeneic hematopoietic cell transplantation (allo-HCT). For a median follow up of 13.3 months, the 1-year disease free survival and overall survival were 56 and 67%, respectively. An accurate diagnosis of BPDCN can be made by 10-color FCM using a four-tube AL panel demonstrating a characteristic pattern of antigen expression. Front-line induction chemotherapy with HyperCVAD can yield high remission rates, but allo-HCT is required for long-term durable remissions.

The embryo-placental CD15-positive “vasculogenic zones” as a source of propranolol-sensitive pediatric vascular tumors

ObjectivePropranolol-induced involution is a unique biological feature of some pediatric vascular tumors, for instance infantile hemangioma (IH), cerebral cavernoma or chorioangioma. Currently, the cellular origin of these distinct tumors is unclear. In this study, we tested the hypothesis that propranolol-responsive vascular tumors are derived from common vessel-forming CD15 + progenitor cells which occur in early gestation. The aim of this study was to identify the tumor-relevant CD15 + progenitors at the early stages of embryo-placental development. Materials and methodsHuman embryo-placental units of 4–8 weeks gestation and pediatric vascular tumors were tested for expression of the tumor-relevant markers CD15, CD31 and CD34. ResultsPlacental vessel-forming progenitors were characterized by immunostaining for CD15, CD31, and CD34. In embryonic tissue, a discontinuous CD15+/CD31+/CD34 + progenitors was detected in immature vessels of the skin, neural tube, spinal and cerebral meninges. Similarly, vessels in IH and chorioangioma exhibited a co-expression of CD15, CD31, and CD34. In contrast, the majority of embryonic vessels presented a CD31+/CD34+, but CD15-negative immunophenotypic pattern. DiscussionOur results suggest the existence of a CD15+ “vasculogenic zones” in the embryo-placental unit as well as in IH and chorioangioma. A site-specific correlation between normal embryo-placental and tumoral vessel-forming CD15 + progenitors was demonstrated. ConclusionHence, site- and stage-specific CD15 + progenitors of vascular wall could be considered as propronalol-sensitive targets and source of pre- and postnatal vascular tumors. We propose, that the CD15+ “vasculogenic zones” are a site-specific reserve of multi-lineage progenitors that could be recruited in pre- and postnatal emergency situations.

Flowcytometric Evaluation and Morphological and Cytochemical correlation of 150 cases of Acute Leukemia

Background : Immunophenotyping by flowcytometry is well conceived & fundamental tool to diagnose & subtype hematological malignancy, especially acute leukemia. By detecting various antigens presenting in various parts of cell, it is possible to know cell lineage & immaturity of the cell or group of cells. Apart from diagnostic importance, this specialized tool is also useful in prediction of prognosis & detection of minimal residual disease. Now, immunophenotyping can diagnose and type also those acute leukemias where morphology and cytochemistry fail. Aim : Study of Immunophenotypic patterns and their correlation with morphology and cytochemistry in North Indian Population. Materials and methods : Short clinical details and complete blood count of 150 patients were noted in the department of hematology of tertiary health centre. Sample of each patient was processed as per protocol and run on FACS CALIBUR OF BD BIOSCIENCES, USA . Dot plot data of each patient was analyzed and result was released. Results: AML, B-ALL and T-ALL comprised 38%, 49%, and 13% of all cases. Almost all blasts were expressing dimCD45 with no significant differences between the subtypes. CD34 have different expressions in AML subtypes, usually negative in APML. Aberrant expression of CD7 and CD19 were expressed in 5% and 3.4%of all cases of AML respectively. In 40% cases, morphology and Cytochemical studies clinched the diagnosis. 60% cases essentially needed Flowcytometric evaluation for diagnosis and subtyping of acute leukemias. Conclusion: Flowcytometric analysis of the patterns and intensity of antigen expression in blasts improved the diagnosis of AML and ALL in our centre. All cases do not require Immunophenotyping for diagnosis. Simultaneous use of conventional morphology, cytochemistry and flowcytometry reduce diagnostic cost of acute leukemia. Immunophenotyping results of our acute leukemia patients were comparable to international published studies.

Sézary Syndrome: Clinicopathological and Immunophenotypical Characteristics and Outcomes

BackgroundSézary syndrome (SS) is a rare malignant T-cell lymphoproliferative disorder derived from mature CD4+ T-helper/inducer cells. Patients (pts) with SS usually manifest with generalized erythroderma, peripheral blood and lymph node involvement and severe pruritus. Current therapy usually consists of extracorporeal photopheresis, interferon alpha, alemtuzumab, histone deactylase inhibitors (HDACi) and monochemotherapy. SS is considered incurable with conventional skin directed or conventional systemic therapy. Reports in literature suggest overall survival (OS) between 2-5 years.MethodsBetween 2002 and 2015, 50 pts with SS were evaluated and treated at Moffitt Cancer Center (MCC). Patient demographics, disease/treatment characteristics, responses, and outcomes were collected from our CTCL database. We characterized four groups based on immunophenotype, aberrant loss of CD26 and CD7, or both. Responses to treatment were assessed using standard criteria: complete response (CR), partial response (PR), stable disease (SD), and progressive disease (PD). Descriptive statistics were used to report baseline characteristics. Chi square and t-test were used for comparison of categorical and continuous variables respectively. Kaplan-Meier graphs were used for estimation of OS from time of diagnosis.ResultsWe identified 50 patients with SS with median follow-up of 52 months. Median age was 70 years. Male to female ratio was 1.6:1. Most pts were Caucasians (88%),). A majority of patients (76%) had advanced stage disease (IVA+IVB). Measurable adenopathy was identified in 35 pts (70%). Median number of Sezary cell count in peripheral blood was 1,747/ul. Extracorporeal Photopheresis (ECP) was used in 43 pts (86%). Overall response rate (ORR) after 6 months of therapy was the following: CR 3(6%), PR 9 (18%), SD 7 (14%), PD 24 (48%). The only agents with activity achieving were interferon (ORR 6%, CR 2%), alemtuzumab (ORR 8%, CR6%), MTX (ORR 6%, CR 2%), and bexarotene (ORR10%, CR 2%), Responses to HDAC inhibitors were: romidepsin (ORR 6%, all PR) and vorinostat (ORR 4%, CR 2%). Median OS was 96 months (95% CI 70-121). Based on aberrant immunophenotype patterns, pts were stratified into 4 groups: 1) CD4+CD7+CD26- 26% (13), 2) CD4 + CD 7- CD26+ in 12% (6), 3) CD4+ CD7- CD26- in 34% (17) and 4) CD4+ CD7+ CD26+ in 24% (12). The median OS for CD4+ CD 7- CD26- was 84 months versus not reached (NR) for other groups. This difference was statistically significant when groups 3 and 2 (p <0.05), and groups 3 and 4 (p< 0.02) were compared. However, there was only a trend observed when groups 3 and 2 were compared (p=0.3). The median OS was 86 month if absolute Sezary cell count was >1700/ul compared to 96 mo if < 1700/ul (P< 0.4).ConclusionsPts with SS in this study showed an improved survival compared to historical studies. Advent of ECP, novel targeted agents such as HDACi inhibitors and monoclonal antibodies, earlier diagnosis, and better supportive care have most probably contributed to this phenomenon; except ECP, CR rates were very rare. A majority of the pts require multimodality induction therapy followed by maintenance therapy. Pts with Sezary cell immunophenotype characterized by aberrant loss of both CD26 and CD7 antigens had worse outcome. There was also a tendency for worse OS in pts with higher circulating absolute Sezary cell count. DisclosuresKomrokji:Celgene: Consultancy, Research Funding; Incyte: Consultancy; Novartis: Research Funding, Speakers Bureau; Pharmacylics: Speakers Bureau. Sokol:Seatle Genetics: Research Funding; Celgene: Consultancy; Spectrum: Consultancy.