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1.
Acta sci. vet. (Impr.) ; 49: Pub. 1831, 2021. graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1363715

ABSTRACT

Lymphoma is a neoplasm of hematopoietic origin that affects canines. The proper establishment of prognosis and rapid institution of treatment are essential for a better quality of life, and immunophenotyping is one of the tools used for this purpose. The objective of this study was to perform a clonality test for immunophenotypic characterization of canine lymphomas using the polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) technique in real-time from samples fixed in formalin and embedded in paraffin. The 23 analyzed samples were fixed in formalin and embedded in paraffin canine lymphoma from the collection Laboratory of Histopathology of the Animal Pathology Area of the Departament of Veterinary Medicine - Federal Rural University of Pernambuco (UFRPE). Samples were processed, their DNA was extracted, quantified, diluted, and standardized at a concentration of 50 ng/µL. After extraction, all samples were subjected to conventional PCR for endogenous control (detection of the IgM target region), in which the extracted DNA was amplified in a final volume of 25 µL. The 128 bp amplified product was detected by 1.5% agarose gel electrophoresis. Of the 23 samples analyzed for the detection of the conserved region referring to the endogenous gene, 91.30% (21/23) amplified the conserved region Cµ by conventional PCR, and two samples 8.70% (2/23) were negative. Endogenous control positive samples were subjected to real-time PCR-PARR for detection of IgH Major and IgH Minor for B lymphocytes (LB), and TCRy for lymphocytes T (LT) target regions. All reactions were performed in duplicate to reduce the risk of false-positive or false-negative results due to technical errors. Samples previously confirmed by immunohistochemistry were used as positive controls for T cell and B cell lymphoma, and MilliQ water was used as a negative reaction control. After amplification, the melting curve gradually increased the temperature by 1o C/5 s to 95o C during continuous fluorescence monitoring. Of the 21 samples analyzed, 100.00% (21/21) demonstrated clonal amplification. Of these, 57.15% (12/21) were positive for phenotype B, and 42.85% (9/21) were positive for phenotype T. Due to the importance of researching and confirming samples from files fixed and embedded in paraffin samples in laboratories, PCR-PARR is a good tool for this purpose. In the present study, real-time PCR analysis demonstrated greater sensitivity in the characterization of the immunophenotype of lymphomas from old samples fixed in formalin and embedded in paraffin. The temperature of melting curve analysis may vary depending on the amount of DNA and its quality. In the present study, it was found that the average melting temperature in the samples varied between ± 3o C when compared to that in the control sample for LB and LT, 83.5o C and 80o C, respectively: in the literature, there is a relative difference in this temperature, which may vary up to 4o C. Real-time PCR-PARR was satisfactory in the characterization of the immunophenotype of canine lymphomas from formalin-fixed and paraffin-embedded samples; therefore, its use is recommended for both retrospective studies. The use of PCR-PARR associated with histopathological and/or cytopathological examination in cases of canine lymphomas strongly helps pathologists, provide a safe establishment of the immunophenotype, minimize errors, and optimize the diagnosis, thus directly contributing to the establishment of the prognosis.(AU)


Subject(s)
Animals , Immunophenotyping/veterinary , Dog Diseases/genetics , Real-Time Polymerase Chain Reaction/veterinary , Lymphoid Tissue , Lymphoma/veterinary , Dogs
2.
Article in Chinese | WPRIM | ID: wpr-912526

ABSTRACT

Flow cytometric immunophenotyping of leukemia has been used as the main method of acute leukemia diagnosis and classification. The revised Classification of Hematopoietic and Lymphoid Tissue Tumors in 2016 improved the diagnostic method for acute leukemia assessment. In China, at present there are still difficulties in the large-scale application and the standardization of this new flow cytometric immunophenotyping of acute leukemia in daily work. A thorough analysis and solution is demanded.

3.
J. Bras. Patol. Med. Lab. (Online) ; 57: e2992021, 2021. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1250137

ABSTRACT

ABSTRACT Introduction: The paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal disease of the hematopoietic stem cells, and it is clinically characterized by chronic intravascular hemolysis, bone marrow failure and hypercoagulability leading to thrombosis. It is a rare disorder of the hematopoietic stem cells that occurs due to a somatic mutation in the gene phosphatidylinositol glycan class A (PIG-A). Objective: Here we reviewed the importance of screening and monitoring of individuals with high risk of developing PNH, since the early diagnosis of the disease is essential for better prognostic and treatment choice for the patient. Method: A review was carried out with great focus on the pathophysiology and diagnosis of PNH, mainly with the use of flow cytometry technique to detect the disease. Results: This gene codifies an enzyme essential to the formation of glycosylphosphatidylinositol (GPI), which acts as a molecular anchor for many membrane proteins. The alteration of GPI synthesis promotes a partial or complete loss of proteins that needs this molecular anchor to bind to the cell surface. Among these proteins are the CD55 and the CD59, which control the activation of the complement cascade. Conclusion: The immunophenotyping exam with flow cytometry is considered the reference test for PNH diagnosis, since the technique is highly sensitive and specific, presenting advantages as the quantitative identification of small populations of cells with PNH phenotype and the capacity to distinguish cells with partial or total deficiency of GPI-anchored proteins.


RESUMEN Introducción: La hemoglobinuria paroxística nocturna (HPN) es una enfermedad clonal adquirida de células madre hematopoyéticas; se caracteriza clínicamente por hemólisis intravascular crónica, insuficiencia medular e hipercoagulabilidad, que conduce a trombosis. Es un trastorno raro de las células madre hematopoyéticas que ocurre debido a una mutación somática en el gen fosfatidilinositol-glicano de clase A (PIG-A). Objetivo: Este estudio tuvo como objetivo revisar la importancia del cribado y seguimiento de individuos con alto riesgo de desarrollar HPN, pues el diagnóstico precoz de la enfermedad es vital para un mejor pronóstico y la elección del tratamiento del paciente. Métodos: Se realizó una revisión con mayor enfoque en la fisiopatología y diagnóstico de la HPN. El foco principal de la investigación fue el uso de la técnica de citometría de flujo para detectar la enfermedad. Resultados: Ese gen codifica una enzima esencial en la formación de glicosilfosfatidil inositol (GPI), que actúa como molécula de anclaje para varias proteínas de membrana en las células hematopoyéticas. Cambiar la síntesis de GPI genera una pérdida parcial o total de proteínas que necesitan esta molécula de anclaje para unirse a la superficie celular. Entre esas proteínas se encuentran CD55 y CD59 presentes en los eritrocitos, que controlan la activación de la cascada del complemento. Conclusión: La técnica de inmunofenotipificación por citometría de flujo se considera la prueba de referencia para el diagnóstico de HPN, ya que es altamente sensible y específica, presenta ventajas como la identificación cuantitativa de pequeñas poblaciones de células con el fenotipo de HPN y la capacidad de distinguir células con deficiencia parcial o total de proteínas ancladas por GPI.


RESUMO Introdução: A hemoglobinúria paroxística noturna (HPN) é uma enfermidade clonal adquirida de células-tronco hematopoiéticas; caracteriza-se clinicamente por hemólise intravascular crônica, falência medular e hipercoagulabilidade, levando a tromboses. É uma rara desordem das células-tronco hematopoiéticas que ocorre devido a uma mutação somática no gene fosfatidilinositol glicano classe A (PIG-A). Objetivo: Este trabalho teve como objetivo revisar a importância do rastreamento e monitoramento de indivíduos com alto risco de desenvolvimento da HPN, pois o diagnóstico precoce da doença é essencial para um melhor prognóstico e a escolha do tratamento para o paciente. Metodologia: Foi realizada uma revisão com mais enfoque na fisiopatologia e no diagnóstico da HPN. O foco principal da pesquisa foi o uso da técnica da citometria de fluxo para a detecção da doença. Resultados: Esse gene codifica uma enzima essencial na formação de glicosilfosfatidil inositol (GPI), a qual atua como molécula âncora de diversas proteínas de membrana nas células hematopoiéticas. A alteração da síntese de GPI gera uma perda parcial ou completa de proteínas que necessitam dessa molécula-âncora para se ligarem à superfície celular. Entre estas proteínas estão o CD55 e o CD59 presente em eritrócitos, que controlam a ativação da cascata do complemento. Conclusão: O exame de imunofenotipagem por citometria de fluxo é considerado o teste de referência para diagnóstico de HPN, pois a técnica é altamente sensível e específica, apresentando vantagens como a identificação quantitativa de pequenas populações de células com fenótipo HPN e a capacidade de distinguir células com deficiência parcial ou total de proteínas ancoradas pela GPI.

4.
Autops. Case Rep ; 11: e2020196, 2021. tab, graf
Article in English | LILACS | ID: biblio-1142407

ABSTRACT

B-cell prolymphocytic leukemia (B-PLL) is an extremely rare disease, accounting for approximately 1% of the lymphocytic leukemias. B-PLL generally occurs in older people. It is characterized by the presence of more than 55% prolymphocytes in the peripheral blood (PB), no or minimal lymphadenopathy, massive splenomegaly, and very high white blood cell counts. The prognosis of B-PLL patients is generally poor, with a median survival of 3 years, although a subset of patients may show a prolonged survival. Herein, we report a case of a 70-year-old male with weakness, generalized lymphadenopathy, and moderate splenomegaly at the initial presentation. Hematologic examination revealed lymphocytic leukocytosis, favoring a chronic lymphoproliferative disorder (CLPD). The key to decoding the precise CLPD was a combination of the clinical profile, morphologic findings on the peripheral blood and the bone marrow, immunophenotypic analysis, and cytogenetic study. The best diagnosis proffered was a de novo chronic lymphocytic leukemia/prolymphocytic leukemia. There was no prior history of lymphoproliferative disorder or lymphocytic leukocytosis. Discriminating this entity from other lymphoproliferative disorders is crucial as the treatment and prognosis are varied compared to the other lymphoproliferative disorders. The diagnostic conundrum encountered and the incredible utility of ancillary studies in such a scenario are highlighted in this study.


Subject(s)
Humans , Male , Aged , Leukemia, Lymphocytic, Chronic, B-Cell , Leukemia, Prolymphocytic , Leukemia, Lymphoid , Leukemia, Prolymphocytic, B-Cell , Rare Diseases , Lymphadenopathy
5.
Clin. biomed. res ; 41(3): 192-198, 20210000. graf
Article in English | LILACS | ID: biblio-1344765

ABSTRACT

Introduction: Treatment of childhood acute lymphoblastic leukemia (ALL) is based on risk stratification. This study aimed to assess the agreement between risk group classifications in the different childhood ALL treatment protocols used in a referral hospital in southern Brazil. Methods: We retrospectively reviewed the medical records of patients aged 1 to 18 years with B-cell ALL treated at a hospital from January 2013 to April 2017. Agreement between risk classifications was assessed by the kappa coefficient. Results: Seventy-five patients were analyzed. There was poor agreement between risk stratification by GBTLI 2009 and BFM 95 protocols (kappa = 0.22; p = 0.003) and by GBTLI 2009 and IC-BFM 2002 protocols (kappa = 0.24; p = 0.002). Risk group distribution was 13.3% for low risk, 32.0% for intermediate risk, and 54.7% for high risk based on stratification by the GBTLI 2009 protocol, and 28.0% for low risk, 42.7% for intermediate risk, and 29.3% for high risk based on stratification by the IC-BFM 2002 protocol. Overall survival was 68.6%. Conclusion: This study provides numerous points to ponder about the treatment of leukemia in Brazil. The percentage of patients classified as high risk in our sample was higher than that reported in the international literature. This difference, however, had no impact on overall survival, which was shorter than that reported in the international literature. (AU)


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Precursor B-Cell Lymphoblastic Leukemia-Lymphoma/mortality , Antineoplastic Combined Chemotherapy Protocols , Risk Factors , Cancer Survivors
6.
Autops. Case Rep ; 10(2): e2020170, Apr.-June 2020. tab, graf
Article in English | LILACS | ID: biblio-1131812

ABSTRACT

Histiocytic sarcoma (HS) is a rare hematolymphoid malignant neoplasm with an aggressive clinical course. It can arise de novo or from low-grade B-cell lymphoma. We describe the case of a 16-year-old boy referred to our hospital with generalized lymphadenopathy, weight loss, and decreased appetite for one month. The patient died undiagnosed on the 7th day of hospitalization. Lymph node and bone marrow biopsies were performed one day before the patient died. The lymph node biopsy revealed an architectural effacement with a diffuse proliferation of large pleomorphic neoplastic cells containing large, multilobulated nuclei, coarse vesicular chromatin, prominent nucleoli, and a moderate amount of eosinophilic cytoplasm. The bone marrow aspiration smears and biopsy also showed evidence of infiltration by these above-mentioned cells. Based on the morphology, along with the exclusion of many differential diagnoses by an extensive panel of immunohistochemical markers, a diagnosis of HS was made. This case report aims at evaluating all the clinical and immunophenotypic features of a case of HS with multifocal presentation and an aggressive clinical course in order to give a correct and definite diagnosis at the proper time.


Subject(s)
Humans , Male , Adolescent , Histiocytic Sarcoma/pathology , Autopsy , Immunophenotyping , Lymphoma, B-Cell , Fatal Outcome , Diagnosis, Differential , Lymphadenopathy
7.
J Cancer Res Ther ; 2020 Apr; 16(1): 105-109
Article | IMSEAR | ID: sea-213753

ABSTRACT

Background: Aberrant phenotypes in acute leukemia have been reported with varying frequencies in independent studies and their association with prognostic factors is still a matter of debate. Aim: This study aims to identify the frequency of aberrant immunophenotypes in de novo acute myeloid leukemia (AML) and to evaluate their association with initial clinical and hematological features. Materials and Methods: A total of 181 patients of de novo AML were included during the time (July 2010–June 2012). The immunophenotype of all cases of AML was studied by using flow cytometry. Results: Aberrant lymphoid antigen expression was seen in 43.1% cases. Most frequent aberrant lymphoid antigen was CD7, seen in 26.5% cases. All French-American-British (FAB) subtypes except AML-M3 expressed aberrant lymphoid antigens. The expression was most common in AML-M4 in the current study. CD34 expression in AMLs was significantly associated with the expression of aberrant lymphoid antigens. Lymphoid antigen expression in adult AML was significantly associated with higher white blood cell (WBC) count (>50,000/mm3) and higher number of peripheral blasts (>70%). Conclusion: In summary, CD7 is the most common aberrant lymphoid antigen expressed in AML. FAB subtype AML-M3 is usually not associated with aberrant lymphoid antigen expression. AML cases with CD34 positivity are more likely to express aberrant lymphoid markers. The current study also supports that aberrant lymphoid antigen expression in adult AML is associated with adverse presenting hematological features (WBC count >50,000/mm3, peripheral blasts >70%). Pediatric Ly + AML cases are not associated with adverse presenting clinical and biological features.

8.
Acta cir. bras ; 35(10): e202001003, 2020. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1130614

ABSTRACT

Abstract Purpose: Comparing survival rates of rats subjected to spleen procedures after fecal peritonitis induction. Assessing changes in TCD4 and CD8 lymphocyte rates before and after the procedures. Correlating animal survival with CD4 and CD8 lymphocytes. Methods: Thirty male Wistar rats were distributed into 3 groups of ten: spleen manipulation (SM); total splenectomy (TS); subtotal splenectomy with preservation of the inferior pole (IP). Rats were subjected to surgical procedure depending on the group. Seven days after surgery they underwent induction of peritonitis and survival time was recorded. All animals were subjected to two blood collections: before surgery and 70 days after it for TCD4/TCD8 lymphocyte counting. Results: Mean survival time was longer in the IP and SM groups and shorter in the TS group; there was significant difference between them. The comparison of the median number of CD4 did not present changes, whereas the comparison of the median number of CD8 decreased in the SM and IP groups. The correlation between the median number of TCD4 and TCD8 lymphocytes and the animals' survival was not significant. Conclusion: The maintenance of splenic tissue contributed to increase the survival of rats and there was a change in the number of TCD8 lymphocytes.

9.
Rev. cuba. hematol. inmunol. hemoter ; 35(4): e1123, oct.-dic. 2019. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1093297

ABSTRACT

Introducción: El cáncer epitelial de ovario (CEO) ocupa el sexto lugar en incidencia y mortalidad a nivel mundial yen Cuba, el quinto en incidencia. Este cáncer es inmunogénicoy sus células malignas crecen en interacción conlas células inmunitarias. Su curso clínico depende del infiltrado inflamatorio acompañante del tumor. La citología e histopatología son los métodos diagnóstico de elección. Sin embargo, la citometría de flujo emerge como una tecnología de mayor sensibilidad, objetividad y rapidez. Objetivo: Diseñar un panel multicolor de citometría de flujo para inmunofenotipar el infiltrado linfocitario de tres tipos de muestras de pacientes con CEO. Métodos: Se realizó un diseño experimental, para la creación y evaluación de un panel multicolor de citometríade flujo, en el laboratorio de Inmunología del Instituto Nacional de Oncología y Radiobiología. El panel se diseñó en sangre de 3 sujetos sanos y se optimizó para sangreperiférica en 33 sujetos sanos y, en sangreperiférica, ascitis y tejido tumoral ovárico de tres pacientes con CEO. En cada muestra se inmunofenotiparon varias poblaciones linfocitarias. Resultados: Se seleccionaron 11 marcadores antigénicos para el inmunofenotipo, el panel quedó conformado por 4 tubos de citometría. La metodología se pudo aplicar a las muestras de ascitis y tejido tumoral sin interferencias, se obtuvieron porcentajes de las subpoblaciones linfocitarias dentro de los valores esperados. Conclusiones: El panel diseñado permitió inmunofenotipar linfocitos en distintos tipos de muestras de pacientes con CEO, con resultados confiables y reproducibles. Esta metodología puede extenderse a la realización de inmunofenotipaje en otras enfermedades(AU)


Introduction: Epithelial ovarian cancer occupies the 6th place in incidence and mortality in women worldwide. In Cuba, it occupies the 5th place in incidence in females. This cancer is immunogenic and its malignant cells grow in interaction with multiple cells from immune system. Its clinical course depends largely on the type of inflammatory infiltrate accompanying the tumor. Cytology and histopathology are gold standard as diagnostic methods. However, flow cytometry emerges as a technology with greater sensitivity, objectivity and speed. Objective: To design a multicolored flow cytometry panel to immunophenotype the lymphocytic infiltrate of three types of samples for patients with ovarian cancer. Methods: An experimental design was carried out in vitro for the creation and evaluation of a multicolored flow cytometry panel in the Immunology laboratory of the National Institute of Oncology and Radiobiology of Cuba. The panel was designed in the blood of three healthy subjects; then it was optimized for blood in 33 healthy volunteers and blood, ascites and ovarian tumor tissue, from three patients with epithelial ovarian cancer. Several lymphocytes lineages were immunophenotypedin each sample. Results: Eleven markers were selected for the immunophenotype and the panel was made up of four multiparameter cytometry tubes. The methodology created could be applied to the samples of ascites and tumor tissue without interferences and percentages of different lymphocyte subpopulations were obtained within the expected values. Conclusions: The designed panel allowed immunophenotyping of lymphocytes in different types of ovarian cancer patient samples and reliable and reproducible results were obtained. This methodology could be employed for others diseases(AU)


Subject(s)
Humans , Female , Flow Cytometry/methods , Immunophenotyping/methods , Equipment Design/methods , Carcinoma, Ovarian Epithelial/diagnosis
10.
Pesqui. vet. bras ; 39(7): 492-498, July 2019. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1040704

ABSTRACT

Lymphoma is a neoplasm that originates from solid hematopoietic tissues and is one of the most common tumors in dogs. The goal of the present study was to perform a retrospective study of canine lymphomas diagnosed at the "Laboratório Regional de Diagnóstico", at the "Faculdade de Veterinária" of the "Universidade Federal de Pelotas" (LRD-UFPel) from 2000 to 2017, to determine the epidemiology and anatomical distribution, and to evaluate the histopathological and immunohistochemical aspects of each case according to the adapted Kiel classification. The protocols for necropsies and biopsies in the laboratory were reviewed. Lymphoma was diagnosed in 77 dogs. Approximately 37.7% (29/77) of affected dogs had no defined breed, while dogs with defined breeds accounted for 58.4% (45/77) of the diagnoses. The occurrence in males (40/77) was slightly higher than that in females (36/77), and the mean age was 8.1 years (1.4-17 years). The most affected age group was between six and 10 years of age with 31 cases (40.2%). Regarding the anatomical classification, the multicentric form was the most prevalent, accounting for 71.4% (55/77) of the diagnoses. In 40 cases that immunophenotyping was performed, B-cell lymphomas represented 62.5% of the diagnoses (25/40), while T-cell lymphomas corresponded to 37.5% of the diagnoses (15/40). The degree of malignancy according to the modified Kiel classification was low in 35% of lymphomas (14/40) and high in 65% of cases (26/40). The multicentric form was more frequent in the region of influence of the LRD-UFPel. Identification of the immunophenotype can improve the quality of life and survival in affected dogs since it allows the most appropriate treatment for each patient.(AU)


O linfoma é uma neoplasia com origem nos tecidos hematopoiéticos sólidos e é um dos tumores mais frequentes em cães. O objetivo do presente trabalho foi efetuar um estudo retrospectivo dos linfomas caninos recebidos no Laboratório Regional de Diagnóstico, da Faculdade de Veterinária da Universidade Federal de Pelotas (LRD-UFPel) de 2000 a 2017, determinando a epidemiologia e a distribuição anatômica, bem como os aspectos histopatológicos e imuno-histoquímicos de cada caso de acordo com a classificação de Kiel adaptada. Foram revisados os protocolos de necropsias e biópsias recebidos no laboratório identificando-se 77 casos de cães com diagnóstico de linfoma. A doença afetou cães sem raça definida em 37,7% (29/77) dos casos, enquanto os cães com raças definidas tiveram 58,4% (45/77) dos diagnósticos. A ocorrência em machos (40/77) foi discretamente maior do que em fêmeas (36/77) e a idade média foi de 8,1 anos (1,4-17 anos). A faixa etária mais acometida foi entre seis e 10 anos de idade com 31 casos (40,2%). Quanto à classificação anatômica a forma multicêntrica foi a mais prevalente atingindo 71,4% (55/77) dos diagnósticos. Em 40 casos em que a imunofenotipagem foi realizada, os linfomas de células B representaram 62,5% dos casos (25/40), enquanto os linfomas de células T equivaleram a 37,5% dos diagnósticos (15/40). O grau de malignidade de acordo com a classificação de Kiel modificada foi baixo em 35% dos linfomas (14/40) e alto em 65% dos casos (26/40). Conclui-se que a forma multicêntrica é mais frequente na região de influência do LRD-UFPel e que a identificação do imunofenótipo pode melhorar a qualidade de vida e dar maior sobrevida aos cães afetados uma vez que permite o tratamento mais adequado para cada caso.(AU)


Subject(s)
Animals , Dogs , Immunophenotyping/veterinary , Lymphoma/veterinary , Brazil , Lymphoma/epidemiology
11.
Rev. bras. ginecol. obstet ; 41(4): 213-219, Apr. 2019. tab, graf
Article in English | LILACS | ID: biblio-1013607

ABSTRACT

Abstract Objective To describe the immunological and hematological reference intervals of low-risk pregnant women. Methods A cross-sectional retrospective database analysis of a basic and translational study analyzing the hematological evaluation blood counts and immunophenotyping of TCD3 + , TCD4 + , TCD8 + , B, and natural killer (NK) cells of the peripheral blood in 79 low-risk pregnant women and of 30 control women from the state of Pernambuco, Brazil, was performed. Results No significant differences were detected between the hematological profiles of the 2nd and 3rd trimesters. Nevertheless, the median level of B cells decreased significantly in the 2nd (174 x 103 μL; p < 0.002) and 3rd trimesters (160 x 103 μL; p < 0.001), compared with the control group (296 x 103 μL). Similarly, the median level of NK cells was lower in the 2nd (134 x 103 μL; p < 0.0004) and 3rd trimesters (100 x 103 μL, p < 0.0004), compared with the control group (183 x 103 μL). In contrast, relative TCD4+ and TCD8+ levels increased in the 2nd and 3rd trimesters compared with the controls (TCD4 + : 2nd trimester = 59%; p < 0.001; 3rd trimester = 57%; p < 0.01; control = 50%; and TCD8 + : 2nd trimester = 31%; p < 0.001; 3rd trimester = 36%; p < 0.01; control = 24%). Conclusion Low-risk pregnant women have ~ 40% less B and NK cells in the peripheral blood, compared with non-pregnant women. These parameters may improve health assistance for mothers and contribute to define reference values for normal pregnancies.


Resumo Objetivo Descrever o intervalo de referência imunológico e hematológico de gestantes de baixo risco. Métodos Realizou-se uma análise retrospectiva, de um estudo básico e translacional, analisando o perfil hematológico e a imunofenotipagem das células TCD3 + , TCD4 + , TCD8 + , B e natural killer (NK) do sangue periférico de 79 gestantes de baixo risco e de 30 mulheres (controles) do estado de Pernambuco, Brasil. Resultados Não observamos diferenças significativas entre os perfis hematológicos do 2° e 3° trimestres. No entanto, houve redução das células B no 2° (média = 174 x 103 μL; p < 0,002) e no 3° trimestres (160 x 103 μL; p < 0,001), comparado como grupo controle (296 x 103 μL). A mediana das células NK foi menor no 2° (134 x 103 μL; p < 0,0004) e no 3° trimestres (100 x 103 μL; p < 0,0004), comparado com o grupo controle (183 x 103 μL). Porém, o percentual de TCD4+ e de TCD8+ aumentou no 2° e 3° trimestres em relação aos controles (TCD4 + : 2° trimestre = 59%; p < 0,001; 3° trimestre = 57%; p < 0,01; controle = 50%; e TCD8 + : 2° trimestre = 31%; p < 0,001; 3° trimestre = 36%; p < 0,01; controle = 24%). Conclusão Mulheres grávidas de baixo risco têm ~ 40% menos células B e NK no sangue periférico em comparação com mulheres não grávidas. Estes parâmetros podem melhorar a assistência à saúde das mães e contribuir para a definição de valores de referência para gestações normais.


Subject(s)
Humans , Female , Adolescent , Adult , Young Adult , Pregnancy/immunology , Killer Cells, Natural/physiology , T-Lymphocytes/physiology , Pregnancy Trimesters , Reference Values , Pregnancy/blood , Cross-Sectional Studies , Retrospective Studies , Databases, Factual
12.
Article | IMSEAR | ID: sea-194235

ABSTRACT

Background: Acute Myeloid Leukemia (AML) is a malignancy of the cells of myeloid series characterized by the rapid growth of Myeloblasts. The diagnosis of AML is established by demonstration of more than 20% of the blood and/or bone marrow by leukemic myeloblasts. Immunophenotyping is one of the most useful tool for the confirmation, lineage assignment and subtyping of leukemias. This study was aimed to phenotype and classify acute leukemias by flow cytometry using commonly used markers for leukemia diagnosis and to establish whether CD 117 can be considered as a lineage specific marker in diagnosis and subclassification of AML.Methods: Flow Cytometric Immunophenotyping was employed for the study. The myeloid antibodies employed in AML in our study included - CD117, CD11c, CD13, CD15, CD33, CD34, CD36, CD41, CD65 and MPO.Results: In our study AMLs constituted 46% of all acute leukemias. CD117 positivity was seen in 86.56% of the French American British (FAB) category of AML. The blasts gated using CD45 v/s SSC revealed variable expression of CD34, CD13 and CD33. The expression of CD117 was consistent particularly in AML-M0, AML-M1 and AML M2.Conclusions: CD117 is virtually a myeloid blast marker with a high sensitivity, specificity and positive predictive value. Among the various myeloid markers like cMPO, CD13, CD33 and CD117, it is just CD117 that has got a tremendous reproducibility in AMLs. Besides CD117 is a surface marker unlike MPO thus easier to process, time saving and less prone to nonspecific binding.

13.
Article | IMSEAR | ID: sea-188800

ABSTRACT

Immunophenotyping, although has emerged indispensable in the diagnosis and classification of lymphoid neoplasms, has to be used cautiously with knowledge of the antibodies used. No antigen is totally lineage or lymphoma specific, and for this reason, immunostaining must be performed in the context of a panel. In addition, familiarity with the diagnostic criteria and differential diagnosis of each lymphoid tumor and ultimately correlation with morphology, and clinical history are essential to enhance the diagnostic accuracy and reproducibility. AIM: The present retrospective study aimed to analyse the differentiation and grading of Non-Hodgkin’s lymphoma by immunohistochemical expression of CD20, CD3 and Ki-67 in lymph node. Methods: A total of 50 samples of NHL were included in the study. Written informed consent of the patient was taken where ever required in the vernacular. Relevant history of the patient was also taken as per the proforma attached along with. The tissues were stained with H and E staining, CD20, CD 3 and Ki-67 immunostaining. The positive immunostained slides were then evaluated and scored both qualitatively and quantitatively. Results: In the present study a total of 50 samples of NHL were included with an age range from 12 to 72 years and a mean age of 46.54 years and male predominance.45 cases showed immunopositivity for CD 20, showing that they belong to B cell phenotype and only 5 cases showed immunopositivity for CD 3, thus showing T cell phenotype. The mean Ki-67 for B cell lymphoma patients was 47.86 ±28.04,with a minimum score of 2 and a max score of 92 and for T cell lymphoma patients was 61.4±18.02, with a minimum score of 40 and a max score of 81, but there was no significant correlation between them (P=0.382). Conclusion: Ki-67 expression in NHL can help in monitoring of patients at risk and can to some extent also aid in detecting the degree of aggressiveness of the disease to give suitable treatment but Ki-67 alone cannot be a risk factor in NHL patients and other factors such as age, sex and type of NHL can be affective, too. The outcome of further analyzing the association between Ki-67 expression and the prognosis of various subtypes of lymphoma should be supported.

14.
Blood Research ; : 63-73, 2019.
Article in English | WPRIM | ID: wpr-739432

ABSTRACT

BACKGROUND: Acute leukemia (AL), not clearly assigned to myeloid, B-lymphoid, or T-lymphoid lineage, is classified as either biphenotypic acute leukemia (BAL) based on the European Group for Immunological Classification of Leukemias (EGIL) or acute leukemia of ambiguous lineage (ALAL) encompassing acute undifferentiated leukemia (AUL) and mixed-phenotype acute leukemia (MPAL) based on the World Health Organization (WHO) criteria. METHODS: Medical records of children newly diagnosed with BAL or ALAL, based on the EGIL or the 2008/2016 WHO criteria, respectively, admitted at Chonnam National University Hospital in 2001–2017 were retrospectively reviewed. RESULTS: Twelve (3.2%) of 377 AL patients satisfied the BAL or ALAL definitions based on the EGIL or the WHO criteria, respectively. Among 12 patients including 11 with BAL and another with undefined case based on the EGIL criteria, 7 (1.9%) had ALAL based on more stringent 2016 WHO criteria (AUL, 2; MPAL, 5). One patient had MPAL with t(9;22)(q34;q11.2), BCR-ABL+, and two had MLL gene abnormality. ALL-directed regimen was associated with better complete remission rate compared with AML-directed regimen (100.0% vs. 16.7%; P=0.015). The 5-year overall survival (OS) and event-free survival (EFS) were 51.1±15.8% and 51.9±15.7%, respectively. AUL was associated with poor OS and EFS compared with MPAL (0.0% vs. 75.0±21.7%; P=0.008). CONCLUSION: Due to the rarity of the cases, future multicenter, prospective studies incorporating large number of cases are urgently warranted to identify the clinical, biologic, and molecular markers for the prediction of prognosis and determine the best tailored therapy for each patient.


Subject(s)
Child , Classification , Disease-Free Survival , Humans , Immunophenotyping , Leukemia , Leukemia, Biphenotypic, Acute , Medical Records , Prognosis , Prospective Studies , Retrospective Studies , World Health Organization
15.
Article in Chinese | WPRIM | ID: wpr-732684

ABSTRACT

Objective To analyze the immunophenotypic characteristics of the patients with minimal residual disease (MRD) positive acute B lymphocytic leukemia (B-ALL). Methods The leukemia-associated immunophenotype (LAIP) of 106 cases with MRD positive B-ALL from Department of Hematology, Tianjin KingMed Diagnois Center between June 2014 and January 2016 were retrospectively analyzed. CD10, CD13/CD33, CD19, CD38, CD58, CD45 and other antibodies were used to analyze the MRD of B-ALL. Results All the patients were positive for CD19. CD34 was negatively or weakly positive expressed in 27 cases (25.4%). CD10 was negatively or weakly positive expressed in 23 cases (21.7%). CD10 was strongly positive in 24 cases (22.6%). Totally, CD10 was weakly or strongly expressed in 47 cases (44.3%). CD58 was strongly positive in 98 cases (92.5%). CD13/CD33 was positively or weakly positive expressed in 64 cases (60.4%). CD38 was negative or weakly expressed in 33 cases (31.1%). CD45 was negative in 21 cases (19.8%). 15 cases (14.1%) were positive for 6 types of LAIP; 30 (28.3%) cases were positive for 5 types of LAIP; 42 (39.6%) cases were positive for 4 types of LAIP; 13 (12.3%) cases were positive for 3 types of LAIP;5 cases (4.7%) were positive for 2 types of LAIP; only one case (0.9%) was positive for 1 type of LAIP. Conclusion The combination of CD58, CD13/CD33, CD10, CD38 and CD34 antibodies can distinguish the neoplastic blast/immature B lymphocytes from progenitor B cells. This strategy has a high accuracy for the judgement of MRD in B-ALL.

16.
Chinese Journal of Pathology ; (12): 951-954, 2019.
Article in Chinese | WPRIM | ID: wpr-800346

ABSTRACT

Objectives@#To investigate the clinicopathological features, therapy and prognosis of primary cardiac CD5-positive diffuse large B-cell lymphoma with C-MYC and bcl-2 double expression.@*Methods@#Two cases diagnosed at Guangdong Provincial People′s Hospital were included, the clinical data were collected; the tumor morphology, immunophenotypic profiles, therapy and prognosis were analyzed.@*Results@#Case 1 was a 55-year-old man and case 2 was a 61-year-old women. Intraoperatively, both cases showed large masses in the right atrium or ventricle, involving adjacent tissue. Pathologically, the tumors were composed of diffusely infiltrating large lymphoid cells with high mitotic activity and apoptosis. The tumor cells were positive for CD20, CD5, bcl-6, MUM1, C-MYC and bcl-2, and the Ki-67 index was equal or greater than 90%. Case 1 had bcl-6, but not bcl-2 or MYC gene rearrangements. No MYC, bcl-2 or bcl-6 gene rearrangements were detected in case 2. Case 1 defaulted chemotherapy after operation and died 1 month after diagnosis. Case 2 was treated with 4 cycles of rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP) therapy after surgery and attained partial remission, and was then treated with apatinib and ibrutinib, and remained stable 18 months after initial diagnosis.@*Conclusion@#Primary cardiac CD5-positive diffuse large B-cell lymphoma with C-MYC and bcl-2 double expression usually shows large infiltrative mass in the right atrium or ventricle, non-germinal center like immunophenotype and high proliferation index, and this may contribute to the aggressiveness of primary cardiac lymphoma.

17.
Chinese Journal of Pathology ; (12): 767-771, 2019.
Article in Chinese | WPRIM | ID: wpr-796830

ABSTRACT

Objective@#To investigate the clinicopathological characteristics of gastric mixed adenoneuroendocrine carcinoma (MANEC).@*Methods@#The clinical and pathologic data of 36 cases of gastric MANEC collected from January 2011 to December 2018 in the Department of Pathology, Fujian Provincial Hospital were retrospectively analyzed. Light microscopy and EnVision immunohistochemical (IHC) staining were used in the study. The results were compared with 40 cases of gastric neuroendocrine carcinoma collected within the same period.@*Results@#The 36 cases were collected, there were 29 males and 7 females. The patients′ age ranged from 43 to 87 years (mean 66 years). The tumor diameter ranged from 1.0 to 9.0 cm (mean 5.0 cm). Clinical staging showed that four cases were at T1+T2 stages and 32 cases were at T3+T4 stages. The main clinical manifestations were upper abdominal pain, abdominal distension and dysphagia. Complete follow-up data were obtained in 20 (eleven living and nine dead) patients. Pathologic analysis showed that all tumors were composed of neuroendocrine carcinoma and adenocarcinoma and each of the components was more than 30%. IHC staining showed that CK was expressed in adenocarcinoma; whereas the neuroendocrine carcinoma mainly expressed CD56 and Syn. There was no significant difference in age, sex, T stage and prognosis between gastric MANEC and gastric neuroendocrine carcinoma(P>0.05). Gastric neuroendocrine carcinoma predominantly occurred in the esophagogastric junction and the gastric body but only rarely in the gastric antrum; gastric MANEC mainly occurred in the esophagogastric junction and the gastric antrum but rarely in the gastric body. Gastric MANEC was more prone to lymph node metastasis than gastric neuroendocrine cancer(P<0.05).@*Conclusions@#Most of the gastric MANEC patients are middle-aged and elderly males, and the tumors predominantly occur in the esophagogastric junction and the gastric antrum. Most of the patients are found to have higher stages, and most of them have lymph node metastases and poor prognosis.

18.
Journal of Leukemia & Lymphoma ; (12): 468-472, 2019.
Article in Chinese | WPRIM | ID: wpr-751426

ABSTRACT

Objective To explore the effects of different storage time of bone marrow specimens on the expressions of different antigens in normal plasma cells (nPC) and clone plasma cells (cPC) by flow cytometry. Methods The bone marrow samples of 12 patients with multiple myeloma (MM) who were treated in Beijing Chaoyang Hospital from September 2017 to January 2018 were selected as MM group. The minimum residual disease (MRD) level in MM group was 10-3-10-2. The bone marrow samples of 12 patients without plasma cell diseases were used as control group. Bone marrow samples were anticoagulated with ethylenediaminetetraacetic acid dipotassium (EDTA-K2) and stored at 2-8 °C. The fluorescent antibodies CD56, CD138, CD45, CD38, CD117, CD81 and cκ, cλ, CD45, CD38, CD19, CD27 were labeled at 0, 24, 48 and 72 h, respectively. The average fluorescence intensity (MFI) of the above 10 antigens expressed in nPC and cPC was analyzed by Diva software. The proportion and absolute count of nPC in control group and cPC in MM group were analyzed. Results In control group, when stored for 24 h, compared with 0 h, the difference of MFI of antigens in nPC was not statistically significant (P > 0.05). When stored for 48 h, compared with 0 h, the MFI of CD38, CD138, CD27, cκ and cλ in nPC decreased, and the differences were statistically significant (28 943±6 591 vs. 23 569±7 587, P= 0.018; 1 412±399 vs. 817±223, P= 0.014;12 855±3 734 vs. 9 210±3 660, P= 0.005; 26 712±9 025 vs. 17 247±5 078, P= 0.026; 17 707±8 633 vs. 8 307±3 158, P = 0.049); the MFI of CD45 increased, and the difference was statistically significant (7 694± 2 525 vs. 9 184±1 332, P = 0.037). When stored for 72 h, compared with 0 h, the MFI of cκ and cλ increased, but the differences were not statistically significant (both P > 0.05). In MM group, when stored for 24 h, compared with 0 h, the difference in MFI of antigens in cPC was not statistically significant (P> 0.05). When stored for 48 h, compared with 0 h, the MFI of CD38, CD138, CD81, cκ and cλ decreased, and the differences were statistically significant (16 664±11 744 vs. 10 130±10 026, P= 0.003; 2 041±1 145 vs. 1 371±696, P= 0.047; 2 679±784 vs. 1 524±1 153, P= 0.025; 29 102±18 138 vs. 18 372±10 327, P=0.038; 16 314±12 728 vs. 9 752±6 271, P=0.034). When stored for 72 h, compared with 0 h, the MFI of cκ and cλ increased, but the differences were not statistically significant (both P> 0.05). The absolute count of nPC and cPC gradually decreased with the prolongation of the storage time, and the difference was statistically significant (both P<0.05) when stored for 0 h and 24 h. There was no significant difference in the percentage of nPC and cPC among different storage time (all P > 0.05). Conclusion Different storage time of bone marrow samples has effects on the MFI of antigens and absolute count of nPC and cPC, and the detection should be completed within 48 h.

19.
Journal of Leukemia & Lymphoma ; (12): 390-395, 2019.
Article in Chinese | WPRIM | ID: wpr-751414

ABSTRACT

Objective To investigate the clinical and biological features of patients with mixed﹣phenotype acute leukemia (MPAL). Methods The clinical data of 24 de novo adult patients with MPAL who were admitted to Fujian Medical University Union Hospital from January 2012 to October 2018 were retrospectively analyzed. These patients were diagnosed according to the World Health Organization (WHO) 2016 criteria. The clinical and biological characteristics of the patients were analyzed by morphological and cytochemical staining, immunophenotyping, cytogenetics and molecular biology. Results Of the 24 patients, 16 were male and 8 were female, and the median age of the patients at diagnosis was 27 years old (5-66 years old). The average blasts of bone marrow were (57.41 ±23.20)% . Thirteen cases (54.2% ) were diagnosed as MPAL morphologically, while 5 cases (20.8% ) were diagnosed as acute myeloid leukemia (AML), 5 cases (20.8%) were diagnosed as acute lymphoblastic leukemia (ALL) and 1 case (4.2%) was inconclusive. Eighteen patients (75.0%) co﹣expressed B﹣lymphoid and myeloid markers, while 5 patients (20.8%) with T﹣lymphoid and myeloid markers and 1 patient (4.2%) with B﹣lymphoid and T﹣lymphoid markers, respectively. The positive rate [median (range)] of CD38, HLA﹣DR and CD34 was 90.5% (0.1%-99.7%), 90.1% (1.1%-98.8% ) and 81.3% (0.1%-97.8%), respectively. Eighteen cases underwent chromosome examination, of which 5 cases carried with t(9;22)(q34;q11), 3 cases with t(v;11q23.3), 2 cases with complex karyotypes, and 2 cases with t(9;22)(q34;q11) and complex karyotypes, respectively. Twenty﹣one cases underwent genetic examination, of which 6 cases were positive for BCR﹣ABL, 3 cases were positive for MLL, 1 case was positive for MLL and BCR﹣ABL, 1 case was positive for BCR﹣ABL and TP53, and 1 case was positive for PHF6 and ASXL1 respectively. Of the 24 patients, 7 refused chemotherapy and 17 received induction chemotherapy. Of the patients receiving chemotherapy, 9 cases achieved complete remission (CR), 1 case was partial remission (PR), and 7 cases were not relieved (NR). In 11 patients treated by ALL﹣type induction regimen and 6 patients treated by ALL and AML﹣type induction regimen, 8 cases and 1 case achieved CR, the difference in CR rate was statistically significant (P<0.05). In 6 patients with Philadelphia chromosome (Ph) positive and 11 patients with Ph negative, 1 case and 8 cases achieved CR, the difference in CR rate was statistically significant (P<0.05). The median follow﹣up time was 5.5 months (0-36 months). The 3﹣year overall survival (OS) rate was 17.5% and the median OS time was 6 months. The 3﹣year OS rates in the allogeneic hematopoietic stem cell transplantation and non﹣transplanted groups were 75.2% and 0, respectively, and the median OS time was not reached and 4 months (P< 0.05). Conclusions MPAL is rare, it mostly co﹣expresses lymphoid and myeloid antigens and shows a much higher incidence of CD34, CD38 and HLA﹣DR. MPAL is often associated with Ph positive and complex karyotypes. MPAL has a low remission rate and poor prognosis, and a reasonable and effective treatment plan should be further explored.

20.
Chinese Journal of Pathology ; (12): 92-97, 2019.
Article in Chinese | WPRIM | ID: wpr-810444

ABSTRACT

Objective@#To investigate the expression of immunomarkers CK7, CK20, CK17, CDX2, MUC1 and MUC2 in primary adenocarcinoma of the ampulla of Vater, to explore the role of these markers in the histopathologic subclassification of ampullary carcinoma; and to provide biologic basis for precision treatment of patients with different types of ampullary carcinoma.@*Methods@#Forty-two cases of primary ampullary carcinoma were collected at Peking University People′s Hospital, from 2012 to 2018 year. There were 22 males and 20 females. Aged range 42 to 88 years old, with mean aged (62±11) years. Among the patients, 6 was high differentiation, 19 median differentiation, and 17 low differentiation. Immunohistochemical studies on the expression of CK7, CK20, CK17, CDX2, MUC1 and MUC2 were performed in 42 cases of primary ampullary carcinoma. The relationship between different ampullary carcinoma subtypes and clinicopathologic survival data was analyzed using SPSS 16.0 statistical software.@*Results@#Three histopathologic subtypes were observed. Among 42 cases, 8(19.0%)were classified as intestinal subtype, which showed a positive expression rate of 8/8 for both CK20 and CDX2, and 5/8 for MUC2. Both CK7 and CK17 were weakly expressed in one case (1/8). No expression was observed for MUC1 in this subtype. Twenty-two (52.4%,22/42) cases were classified as pancreaticobiliary subtype, which showed a positive expression rate of 100.0%(22/22) for both CK7 and MUC1, and 90.9% (20/22) for CK17. No expression was observed for CK20, CDX2 and MUC2.The remaining 12 (28.6%) cases were classified as mixed subtype, which showed variable expression patterns. The expression frequencies of these 6 immunomarkers in different subtypes of ampullary carcinoma did not correlate with various clinicopathologic factors such as patient gender and age, tumor size, histologic differentiation, pancreatic and bile duct invasion, or the depth of duodenal invasion. However, stage Ⅲ+Ⅳ diseases were more commonly seen in pancreaticobiliary type (63.6%,14/22) than intestinal type (2/8) and mixed type (3/9; χ2=6.508, P=0.039). Follow-up data showed a trend of better survival rate for patients with intestinal subtype than those with mixed and pancreaticobiliary subtypes.@*Conclusions@#Ampullary carcinoma can be subclassified into three different subtypes using a panel of six immunomarkers, especially for the identification of subtypes of poorly differentiated carcinoma. CK7, CK17 and MUC1 are major markers of pancreaticobiliary subtype, whereas CK20, CDX2 and MUC2 are useful markers for intestinal subtype. The mixed subtype variably expresses these markers. The prognosis of patients with intestinal subtype appears better than that of pancreaticobiliary and mixed subtypes. Accurate subtyping of ampullary carcinoma is clinically important to patient management and prognosis assessment.

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