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1.
Methods Mol Biol ; 2559: 231-242, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36180636

RESUMO

Regulatory T cells are an important component of the immune system that plays a key role in maintaining homeostasis. Identification of distinct regulatory T cell subsets is essential to understand their function. Mass cytometry or CyTOF is a technology that enables the simultaneous measurement of up to 50 markers in single cells by using antibodies tagged with heavy metals, which are then detected with time-of-flight mass spectrometry. This chapter describes a mass cytometry approach for phenotypic characterization of regulatory T cells and determination of their master transcription factor Foxp3.


Assuntos
Metais Pesados , Linfócitos T Reguladores , Citometria de Fluxo/métodos , Fatores de Transcrição Forkhead/genética , Imunofenotipagem , Subpopulações de Linfócitos T
2.
Exp Oncol ; 44(3): 198-207, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36325710

RESUMO

BACKGROUND: Acute myeloid leukemia (AML) is a highly heterogeneous disease accompanied by the arrest of myeloid cell lineage differenti-ation due to accumulation of genetic abnormalities and clonal proliferation of myeloid blasts. Finding the differentially expressed molecules and studying their function within AML subgroups may help to improve diagnosis and prognosis with the aim of developing selected therapies for AML subsets. The aim of this study was to reveal the profile of CD150 cell surface expression on bone marrow (BM) cells of AML patients. MATERIALS AND METHODS: The study was performed on samples of BM aspirates from 55 patients with primarily diagnosed AML. Flow cytometry analysis was applied for the evaluation of immunophenotype profile and CD150 cell surface expression on BM cells from AML patients. RESULTS: Four AML subtypes (M1, M2, M3 and M5) were identified. The CD150 expression was found in 14 (25.5%) cases predominantly of AML M3 subtype. CD150 expression was detected on 43.2-83.8% of leukemia cells in AML M3. The frequency of CD150 positive cases of non-M3 AML subtypes was low: all AML M1 cases were CD150-negative, while only 1 (10.0%) of 10 patients with AML M2 and 6 (19.4%) of 31 patients with AML M5 were CD150 positive. The median percentage of CD150 positive leukemia cells and the index of mean fluorescence intensity in AML M3 cases were significantly higher than in non-M3 AML cases (p < 0.05). The CD150 expression was significantly associated with CD11c, CD11b, CD14, CD34, CD36, CD56 and HLA-DR negative expression and CD33, CD38, CD117 positive expression among the examined cohort of patients with AML M3. CONCLUSIONS: High level of CD150 expression is a unique feature of AML M3 subtype and may serve as additional phenotype marker for the identification of blast cells with impaired maturation at the promyelocyte stage and the development of AML M3. At the same time, the revealed negative association of CD150 expression with poor prognostic factor CD56 in AML M3 subtype also allows us to suggest potential prognostic value of CD150 examination in AML patients.


Assuntos
Leucemia Mieloide Aguda , Leucemia Promielocítica Aguda , Humanos , Leucemia Mieloide Aguda/genética , Imunofenotipagem , Citometria de Fluxo , Células da Medula Óssea
3.
Front Immunol ; 13: 976739, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36341325

RESUMO

Background: Previous studies have suggested that patients with pulmonary sarcomatoid carcinoma (PSC)may benefit from immune checkpoint inhibitors (ICIs); however, relevant data are lacking. This study aimed to establish the immunophenotype of PSC by assessing PD-L1 and CD8+ T-cell infiltration. Methods: A retrospective analysis of pathologically confirmed PSC cases from two centers was performed from January 2009 to May 2021. According to the infiltration of CD8+ T cells in different spatial regions, patients were classified into three types: immune-inflamed, immune-excluded, and immune desert. PD-L1 staining was also performed on the intratumoral region and the tumor proportion score (TPS) was used for scoring. Combined with CD8+ T-cell infiltration and PD-L1 expression in the intratumoral region, immunophenotyping can be divided into four types: type I (PD-L1+/CD8+, adaptive immune resistance), type II (PD-L1-/CD8-, immunologic ignorance), type III (PD-L1+/CD8-, intrinsic induction), and type IV (PD-L1-/CD8+, tolerance). Finally, correlation analysis was performed on the immunophenotype, clinicopathological characteristics, and outcomes of the patients. Results: A total of 32 patients with PSC were included in the final analysis. Of these patients, 65.6% (21/32), 15.6% (5/32), and 18.8% (6/32) were classified as immune-inflamed, immune-excluded, and immune-desert, respectively. Notably, the immune-inflamed type is predominantly observed in pleomorphic carcinomas (PC, 66.7%). Moreover, among these participants, 19 (59.4%) were classified as PD-L1 positive according to the TPS score. In particular, 11 (34.4%) patients had PD-L1 TPS scores >50%. Next, we immunophenotyped patients with PSC based on CD8+ T cell infiltration and tumor cell PD-L1 expression (types I-IV). Type I (PD-L1+/CD8+, adaptive immune resistance) was the most prevalent subtype, accounting for 46.9% (15/32), followed by type II (PD-L1-/CD8-, immunological ignorance) (21.9%), type IV (PD-L1-/CD8+, tolerance) (18.7%), and type III (PD-L1+/CD8-, intrinsic induction) (12.5%). Finally, we performed a survival analysis and found that neither immunophenotype was a predictor of prognosis in patients with PSC. Multivariate analysis showed that pneumonectomy increased the risk of death by four times compared with lobectomy (RR: 4.1; 95% CI:1.3-12.4, P=0.014). Conclusion: Patients with PSC are characterized by immune-inflamed type and type I (PD-L1+/CD8+, adaptive immune resistance), explaining the intrinsic reasons for their high response rate to immunotherapy.


Assuntos
Carcinoma , Neoplasias Pulmonares , Humanos , Antígeno B7-H1/metabolismo , Linfócitos do Interstício Tumoral , Imunofenotipagem , Estudos Retrospectivos , Neoplasias Pulmonares/patologia , Carcinoma/metabolismo
4.
Blood Cancer J ; 12(11): 147, 2022 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-36323674

RESUMO

Pure erythroid leukemia (PEL), also known as acute erythroid leukemia (AEL), is recognized as a distinct morphologic entity by both the 2016 and 2022 World Health Organization (WHO) classification system. By contrast, the 2022 International Consensus Classification (ICC) includes PEL under a broader category of "acute myeloid leukemia with mutated TP53". We identified 41 Mayo Clinic cases of PEL (mean age 66 years, range 27-86; 71% males) and provide a comprehensive account of bone marrow morphology, immunophenotype, cytogenetic and mutation profiles. PEL was primary in 14 cases, therapy-related in 14, secondary in 12, and undetermined in one. All cases expressed biallelic TP53 alterations, including TP53 deletion/single TP53 mutation (68%), two TP53 mutations (29%) or two TP53 deletions (3%); additional mutations were infrequent. Karyotype was complex in all cases and monosomal in 90%. Treatment details were available in 29 patients: hypomethylating agent (HMA) alone (n = 5), HMA + venetoclax (n = 12), intensive chemotherapy (n = 4), supportive care/other (n = 8); no responses or allogeneic stem cell transplants were documented, and all patients died at a median 1.8 months (range 0.2-9.3). The current study highlights a consistent and reproducible set of morphologic and genetic characteristics that identify PEL as a distinct AML variant whose dismal prognosis requires urgent attention.


Assuntos
Leucemia Eritroblástica Aguda , Leucemia Mieloide Aguda , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Análise Citogenética , Imunofenotipagem , Leucemia Eritroblástica Aguda/genética , Leucemia Eritroblástica Aguda/terapia , Leucemia Mieloide Aguda/genética , Mutação , Proteína Supressora de Tumor p53/genética
5.
Sci Rep ; 12(1): 19644, 2022 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-36385161

RESUMO

The host immune response to a viral immune stimulus has not been examined in children during a life-threatening asthma attack. We determined whether we could identify clusters of children with critical asthma by functional immunophenotyping using an intracellular viral analog stimulus. We performed a single-center, prospective, observational cohort study of 43 children ages 6-17 years admitted to a pediatric intensive care unit for an asthma attack between July 2019 to February 2021. Neutrophils were isolated from children, stimulated overnight with LyoVec poly(I:C), and mRNA was analyzed using a targeted Nanostring immunology array. Network analysis of the differentially expressed transcripts for the paired LyoVec poly(I:C) samples was performed. We identified two clusters by functional immunophenotyping that differed by the Asthma Control Test score. Cluster 1 (n = 23) had a higher proportion of children with uncontrolled asthma in the four weeks prior to PICU admission compared with cluster 2 (n = 20). Pathways up-regulated in cluster 1 versus cluster 2 included chemokine receptor/chemokines, interleukin-10 (IL-10), IL-4, and IL-13 signaling. Larger validation studies and clinical phenotyping of children with critical asthma are needed to determine the predictive utility of these clusters in a larger clinical setting.


Assuntos
Asma , Estado Asmático , Criança , Humanos , Adolescente , Neutrófilos , Imunofenotipagem , Estudos Prospectivos , Asma/genética , Expressão Gênica
6.
PLoS One ; 17(11): e0278040, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36409726

RESUMO

OBJECTIVES: Myelodysplastic syndrome (MDS) is a heterogeneous hematopoietic stem cell disorder with thrombocytopenia. Flow cytometric immunophenotyping of blood cells has been instrumental in diagnosis as co-criteria, but the data regarding platelets remains lacking. This study aims to determine if there is a difference in surface antigen levels on platelets by comparing surface antigen levels in MDS patients and healthy control subjects. Concurrently, as flow cytometric gating can reveal the diameter of cells, this study will investigate differences in giant platelet percentage by comparing these percentages in high- and low-risk MDS patients. STUDY DESIGN: Twenty newly diagnosed MDS patients were enrolled in this study. Platelet surface antigen levels were determined by measuring the binding capacity of antibodies with flow cytometry. RESULTS: Platelets of MDS patients were shown to have a lower level of CD61 and higher levels of CD31 and CD36 than healthy controls. Judged by forward scatter (FSC), MDS patients' platelets appeared to be larger than those of healthy control subjects, whereas the MFI adjusted by diameter (MFI/FSC ratio) of CD31, CD41a, CD42a, CD42b and CD61 on platelets were lower in MDS patients than in healthy control subjects. There was a significant quantity of giant platelets found in MDS patients, and the high-risk MDS patients tended to have a higher percentage of giant platelets than low-risk patients. Conclusions: All the results indicate that MDS patients exhibit a lower antigen presentation (MFI) adjusted by diameter on platelets than healthy controls and the giant platelets detected by flow cytometry might correlate with the condition of MDS.


Assuntos
Antígenos de Plaquetas Humanas , Síndromes Mielodisplásicas , Humanos , Imunofenotipagem , Plaquetas/metabolismo , Projetos Piloto , Síndromes Mielodisplásicas/diagnóstico , Antígenos de Superfície/metabolismo
7.
Front Immunol ; 13: 991928, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36300109

RESUMO

Human T-cell leukemia virus type 1 (HTLV-1), a retrovirus which mainly infects CD4+ T cells and causes adult T-cell leukemia/lymphoma (ATL), is primarily transmitted via direct cell-to-cell transmission. This feature generates a wide variety of infected clones in hosts, which are maintained via clonal proliferation, resulting in the persistence and survival of the virus. The maintenance of the pool of infected cells is achieved by sculpting the immunophenotype of infected cells and modulating host immune responses to avoid immune surveillance. Here, we review the processes undertaken by HTLV-1 to modulate and subvert host immune responses which contributes to viral persistence and development of ATL.


Assuntos
Vírus Linfotrópico T Tipo 1 Humano , Leucemia-Linfoma de Células T do Adulto , Adulto , Humanos , Carcinogênese , Imunofenotipagem , Linfócitos T
8.
Anal Chem ; 94(43): 14906-14916, 2022 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-36256869

RESUMO

Multiplex immunophenotyping of cell surface proteomes is useful for cell characterization as well as providing valuable information on a patient's physiological or pathological state. Current methods for multiplex immunophenotyping of cell surface proteomes still have associated technical pitfalls in terms of limited multiplexing capability, challenging result interpretation, and large equipment footprint limited to use in a laboratory setting. Herein, we presented a portable surface-enhanced Raman spectroscopy (SERS) assay for multiplex cell surface immunophenotyping. We synthesized and functionalized customizable SERS nanotags for cell labeling and subsequent signal measurement using a portable Raman spectrometer. We extensively evaluated and validated the analytical assay performance of the portable SERS immunophenotyping assay in two different cellular models (red blood cells and breast cancer cells). In terms of analytical specificity, the cell surface immunophenotyping of both red blood cells and breast cancer cells correlated well with flow cytometry. The portable SERS immunophenotyping assay also has comparable analytical repeatability to flow cytometry, with coefficient of variation values of 21.89-23.33% and 6.88-17.32% for detecting red blood cells and breast cancer cells, respectively. The analytical detection limits were 77 cells/mL for red blood cells and 1-17 cells/mL for breast cancer cells. As an alternative to flow cytometry, the portable SERS immunophenotyping assay demonstrated excellent analytical assay performance and possessed advantages such as quick sample-to-result turnaround time, multiplexing capability, and small equipment footprint.


Assuntos
Neoplasias da Mama , Nanopartículas Metálicas , Humanos , Feminino , Análise Espectral Raman/métodos , Proteoma , Imunofenotipagem , Citometria de Fluxo , Neoplasias da Mama/diagnóstico
9.
Clin Lab ; 68(10)2022 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-36250828

RESUMO

BACKGROUND: Aberrant phenotypes in acute myeloid leukemia have variable frequencies and their prognostic value with adverse hematological and other biological prognostic factors is still controversial, despite several reports of clinical significance. To date, no study has been reported evaluating the incidence of these phenotypic aberrations in the Moroccan population. The aim is to evaluate the incidence of aberrant phenotype expressions in acute myeloid leukemia and correlate their presence with the different AML subtypes, and clinical and biological characteristics in Moroccan patients. METHODS: Fifty-four AML patients were diagnosed according to WHO classification 2016 criteria. Immunopheno-typing by flow cytometric analysis was performed to evaluate aberrant phenotypes on myeloblasts. RESULTS: The occurrence of lymphoid antigens in AML was higher (51.8%), which was closer to that reported in the literature. CD7 has been revealed to be the most commonly expressed lymphoid antigen. Besides, CD19 was expressed in all 3.7% of M2 AML subtype. However, we could find no statistically significant differences between these aberrant phenotypes regarding FAB subtypes or clinical and biological outcomes. The great majority of AML cases showed asynchronous expression (57%) with significant differences regarding FAB subtypes. CONCLUSIONS: Aberrant phenotypes might be associated with different leukemia subtypes that should be studied for a better understanding of their biological significance and adding important information for prognosis and, at the same time, could be of help when looking for minimal residual disease during morphologic remission.


Assuntos
Hematologia , Leucemia Mieloide Aguda , Antígenos CD/análise , Antígenos CD/genética , Hospitais , Humanos , Imunofenotipagem , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/genética , Marrocos/epidemiologia , Fenótipo , Prognóstico
10.
Clin Lab ; 68(10)2022 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-36250836

RESUMO

BACKGROUND: Lymphocyte immune-phenotyping is considered as a useful tool in diagnosis and monitoring different medical conditions. This study aimed to establish a reference value of different lymphocyte subsets in healthy individuals from southern Iran. METHODS: Flow cytometric method was used to determine the frequency of T cells, helper T cells, cytotoxic T cells, activated T cells, NK cells, NKT cells, and B cells in peripheral blood of 86 healthy subjects from southern Iran. RESULTS: Regardless of gender, the mean percentage of the lymphocyte subsets have been observed as T cells (69.2 ± 8.84%), B cells (11.88 ± 4.14%), T helper cells (39.85 ± 7.75%), T cytotoxic cells (33.97 ± 6.64%), activated T cells (6.67 ± 3.60%), NK cells (12.56 ± 7.32%), and NKT cells (5.07 ± 2.83%). The ratio of CD4+/CD8+ was found to be 1.22 ± 0.4. An insignificant difference was found between lymphocytes of male and female. The percentage of helper T-cells has demonstrated a positive correlation and the percentage of cytotoxic T-cells and NK cells have il-lustrated a negative correlation with age. CONCLUSIONS: The results provide normal reference values of peripheral blood lymphocyte subsets in healthy individuals from southern Iran with the age range of 15 - 55 years old. Moreover, the results support this approach that reference values of lymphocyte subsets should be provided for each population. The reference range value of circulating lymphocytes is a powerful tool for monitoring and/or clinical management of immune related disorders.


Assuntos
Subpopulações de Linfócitos , Subpopulações de Linfócitos T , Adolescente , Adulto , Feminino , Citometria de Fluxo , Humanos , Imunofenotipagem , Irã (Geográfico) , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Valores de Referência , Adulto Jovem
11.
Ann Clin Lab Sci ; 52(5): 846-849, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36261189

RESUMO

Granular acute lymphoblastic leukemia (ALL) is defined by the presence of intracytoplasmic granules in lymphoblastic blasts, mimicking acute myeloblastic leukemia. The disease is extremely rare in adults, and hence, the characteristics thereof are poorly understood. We report a case of a 70-year-old man diagnosed with granular ALL. Bone marrow examination showed blasts with azurophilic granules in the cytoplasm, but immunophenotyping showed B-ALL with aberrant expression of myeloid antigens CD13 and CD33. Karyotyping revealed monosomy 7, and targeted NGS showed DNMT3A mutation, which suggested poor prognosis. Despite conventional chemotherapy treatment, the patient did not achieve complete remission. He declined further chemotherapy treatment and was maintained on only supportive care. This is the first report of adult granular ALL with DNMT3A mutation and monosomy 7.


Assuntos
Leucemia Mieloide Aguda , Leucemia-Linfoma Linfoblástico de Células Precursoras , Masculino , Adulto , Humanos , Idoso , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Deleção Cromossômica , Leucemia Mieloide Aguda/genética , Indução de Remissão , Imunofenotipagem
13.
Cytometry B Clin Cytom ; 102(6): 427-439, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36314855

RESUMO

BACKGROUND: High-quality data on bone marrow involvement (BMI) assessed by flow cytometry (FC) in follicular lymphoma (FL) is lacking. AIMS: We set up a prospective protocol with a 10-color tube and acquisition of 500.000 leukocytes on a Nav flow cytometer for evaluation of BMI in FL by FC. MATERIALS AND METHODS: FC was compared with a combination of histopathology and IGH gene rearrangement, which were considered the gold standard. We also compared BMI by FC with PET. RESULTS: Fifty-two patients were included (median 67 years, 54% female). BMI by FC was seen in 35 (67%), with a median involvement of 1.2% (interquartile range: 0.3%-7%) of leukocytes. Comparison with the gold standard revealed two false negatives and two false positives (potentially true involvement undetected by the gold standard). BMI by PET was seen in 14/46 (30%). Immunophenotype of FL in the bone marrow was highly heterogeneous. The most common phenotypic abnormality was dim expression of CD19 (>0.5 log loss in 30% of patients). CD10 was negative in 13 (37%) and incompletely positive (overlap with the negative population) in a further 8 (28%) while entirely positive only in 14 (48%). Other abnormalities (loss of CD20, gain or loss of CD79b, expression of CD43, and substantial loss of CD45) were rare. Computational analysis by means of FlowSOM confirmed the heterogeneous phenotype, with FL from different patients clustering in unrelated metaclusters. CONCLUSION: BMI by FL was frequent and immunophenotype was heterogeneous. However, this protocol enabled detection of FL in bone marrow in the vast majority of patients with bone marrow involvement by the gold standard.


Assuntos
Linfoma Folicular , Feminino , Humanos , Masculino , Linfoma Folicular/genética , Citometria de Fluxo/métodos , Medula Óssea/patologia , Estudos Prospectivos , Imunofenotipagem
14.
Sci Rep ; 12(1): 16880, 2022 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-36207336

RESUMO

Graves' disease (GD) is an organ-specific autoimmune disease, but there are a few studies that have evaluated how immunophenotypes are related to clinical symptoms and intractable pathology, or the effects of treatment on immunophenotypes. We performed peripheral blood immunophenotyping in GD. We assessed the proportion of functional subsets of T helper cells (such as Th1, Th17, Treg and Tfh cells), B cells (Naïve, IgM memory, Class-switched, IgD-CD27- double negative and Plasmablasts cells), Monocytes, Dendritic cells and NK cells, and evaluated the relationship of immunophenotypes with clinical indices, disease activity, risk of relapse, and changes in immunophenotypes after treatment with antithyroid drugs. The activated Th17 cells, activated T follicular helper (Tfh) cells, and IgD-CD27- double-negative B cells were higher in newly onset GD compared with healthy participants. Th17 cells were associated with thyroid autoantibodies, thyroid function, thyroid enlargement, and Graves' Recurrent Events After Therapy (GREAT) score; while double-negative B cells were associated with thyroid autoantibodies. Treatment with antithyroid drugs decreased the activated Tfh cells in parallel with the improvement in thyroid function. However, activated Th17 cells were not associated with clinical improvement and remained unchanged. Peripheral blood immunophenotyping identified the differential involvement of T and B cell subsets in the pathogenesis of GD. Abnormalities in the differentiation of Th17, Tfh, and double-negative B cells reflected the clinical pathology associated with autoantibody production and excess thyroid hormones. And Th17 cells are significantly associated with the marker for resistance to treatment. These results suggest the involvement of Th17 cell activation in the intractable pathology associated with potential immune abnormalities in GD. Clinical trial registration: #UMIN000017726 (Date: June 1st, 2015).


Assuntos
Doença de Graves , Células Th17 , Antitireóideos , Autoanticorpos , Humanos , Imunoglobulina D , Imunoglobulina M , Imunofenotipagem , Hormônios Tireóideos
15.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 30(5): 1305-1310, 2022 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-36208227

RESUMO

OBJECTIVE: To retrospectively analyze the laborotary test results and clinical data of 31 patients with mixed phenotype acute leukemia (MPAL) in order to summarize and discuss the biological characteristics, curative effect, and prognosis of each subtype of MPAL based on immunophenotype results. METHODS: MPAL patients diagnosed and treated in our hospital from July 2013 to January 2019 were selected to analyze the data of cell morphology, immunophenotyping, cytogenetics, molecular biology (MICM), and routine blood at initial diagnosis. Follow-up was carried out until the last discharge time. RESULTS: Among 31 patients, there were 19 males and 12 females, with a median age of 41(12-76) years old. According to the results of immunophenotyping and EGIL score, there were 16 cases of myeloid-T lymphoid mixed phenotype (myeloid-T group), 9 cases of myeloid-B lymphoid mixed phenotype (myeloid-B group), 5 cases of T-B lymphoid mixed phenotype (T-B group), and 1 case of myeloid-T-B lymphoid mixed phenotype. Compared between different subtypes, the antigen expression characteristics were the highest positive rate and expression rate of HLA-DR in myeloid-B group, and the positive rate of CD2 in T-B group was significantly higher than that in the myeloid-T group. Meanwhile, the expression rates of CD7 and cCD3 (cytoplasmic CD3) in T-B group were higher than those in myeloid-T group, and cCD79a was positive in all cases of myeloid-B group and T-B group. The median WBC of T-B group was 81.92×109/L, which was significantly higher than that of the other two groups (P<0.05). The quantitative results of WT1 were higher than 10-4 in 92.6% of the patients, and the WT1 expression level in myeloid-B group was significantly lower than the other two groups (P<0.01). Among the 9 patients with myeloid-B mixed phenotype, 5 cases showed BCR-ABL positive. Among 28 patients followed up, 21 cases achieved complete remission (CR), the median time to first obtain CR was 32.5(9-75) days, and the median follow-up time was 16 months (range from 21 days to 6 years). The CR rate and median overall survival (OS) time in myeloid-B group were 88.9% and 40 months, which were higher than the other two groups. The CR rate and 3-year OS rate in T-B group were relatively lower (50.0%, 0). CONCLUSION: WT1 gene is highly expressed in patients with MPAL, and each subgroup of MPAL based on immuophenotype has its unique antigen expression characteristics. Compared with myeloid-T group and T-B group, myeloid-B group can acquire higher remission rate and have better prognosis.


Assuntos
Leucemia , Doença Aguda , Feminino , Antígenos HLA-DR , Humanos , Imunofenotipagem , Masculino , Fenótipo , Prognóstico , Estudos Retrospectivos
16.
Front Immunol ; 13: 935879, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36189252

RESUMO

Innate myeloid cell (IMC) populations form an essential part of innate immunity. Flow cytometric (FCM) monitoring of IMCs in peripheral blood (PB) has great clinical potential for disease monitoring due to their role in maintenance of tissue homeostasis and ability to sense micro-environmental changes, such as inflammatory processes and tissue damage. However, the lack of standardized and validated approaches has hampered broad clinical implementation. For accurate identification and separation of IMC populations, 62 antibodies against 44 different proteins were evaluated. In multiple rounds of EuroFlow-based design-testing-evaluation-redesign, finally 16 antibodies were selected for their non-redundancy and separation power. Accordingly, two antibody combinations were designed for fast, sensitive, and reproducible FCM monitoring of IMC populations in PB in clinical settings (11-color; 13 antibodies) and translational research (14-color; 16 antibodies). Performance of pre-analytical and analytical variables among different instruments, together with optimized post-analytical data analysis and reference values were assessed. Overall, 265 blood samples were used for design and validation of the antibody combinations and in vitro functional assays, as well as for assessing the impact of sample preparation procedures and conditions. The two (11- and 14-color) antibody combinations allowed for robust and sensitive detection of 19 and 23 IMC populations, respectively. Highly reproducible identification and enumeration of IMC populations was achieved, independently of anticoagulant, type of FCM instrument and center, particularly when database/software-guided automated (vs. manual "expert-based") gating was used. Whereas no significant changes were observed in identification of IMC populations for up to 24h delayed sample processing, a significant impact was observed in their absolute counts after >12h delay. Therefore, accurate identification and quantitation of IMC populations requires sample processing on the same day. Significantly different counts were observed in PB for multiple IMC populations according to age and sex. Consequently, PB samples from 116 healthy donors (8-69 years) were used for collecting age and sex related reference values for all IMC populations. In summary, the two antibody combinations and FCM approach allow for rapid, standardized, automated and reproducible identification of 19 and 23 IMC populations in PB, suited for monitoring of innate immune responses in clinical and translational research settings.


Assuntos
Anticorpos , Células Mieloides , Anticoagulantes , Citometria de Fluxo , Humanos , Imunofenotipagem , Valores de Referência
17.
Oxid Med Cell Longev ; 2022: 7985596, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36193083

RESUMO

Aging is a complex process often associated with a chronic inflammatory profile that alters several biological functions, including the immune system and cognitive and physical capacity. The practice of physical activity is increasingly gaining popularity as a method of preventing infections, depression, and other disorders that affect the quality of life of the elderly. Thus, this work analyzes the profile of cytokines and molecular markers expressed in immune cells of elderly people who practice physical activities or not, evaluating their impacts on the immune system and quality of life. For this, 48 individuals were recruited, and peripheral blood samples were collected for hemogram analysis, cytokine determination, and immunophenotyping. Elderly people were separated into two groups: practitioners with low-intensity physical activity and non-practitioners. Quality of life was assessed using the Whoqol-Old instrument, and depression was assessed using the Beck II Depression Inventory. When comparing the scores of the Whoqol-Old and Beck questionnaires, we observed a significant negative correlation between these two factors. The perception of a higher quality of life was present in the elderly who exercised and was related to greater autonomy and sensory abilities, whereas the presence of depression was lower. In the hemogram, we observed higher basophil and segmented counts in the sedentary elderly, whereas lymphocytes and monocytes had lower counts. Elderly practitioners of physical activities had higher levels of IFN-γ, IL-4, and IL-10; increased expression of CD69, PD1, and TIM-3 in CD4+ T lymphocytes and increased CD14+CD80+ and CD14+CD86+ monocytes. Elderly people with an increased perception of quality of life had higher levels of IFN-γ, higher expression of CD14+CD80+CD86+, and decreased levels of TRAIL. An increase in TRAIL was observed in individuals with depression, in addition to an increased expression of CD14+CD86+. These results show a clear correlation between the quality of life, level of depression, physical activity, and immune system function. Although some cytokines with a typical proinflammatory profile (IFN-γ) were observed, the results point to a protective state with benefits reflected in the general well-being of the elderly who exercise.


Assuntos
Interleucina-10 , Qualidade de Vida , Idoso , Linfócitos T CD4-Positivos , Citocinas/metabolismo , Depressão , Exercício Físico , Receptor Celular 2 do Vírus da Hepatite A , Humanos , Imunofenotipagem , Interleucina-4 , Monócitos/metabolismo
19.
Diagn Pathol ; 17(1): 74, 2022 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-36199105

RESUMO

BACKGROUND: The diagnosis of mixed phenotype acute leukemia (MPAL) with T/megakaryocyte or T/myeloid lineages accompanied by t(3;3) is always a challenge. Therefore, multiple experimental methods are usually required to avoid misdiagnosis. In this report, we presented a rare case of MPAL with T/myeloid lineages accompanied by t(3;3) and discussed the experience of differential diagnosis and our appreciation of the MPAL with T/megakaryocyte and T/myeloid lineages accompanied by t(3;3). CASE PRESENTATION: A 31-year-old woman was admitted to our hospital due to recurrent fever for 20 days. Two distinct blast populations were detected by flow cytometry analysis: one population fulfills the immunophenotypic criteria for T-lymphoblastic leukemia, while the other population is highly suggestive of megakaryoblasts. These immunophenotypic features support the diagnosis of MPAL (T/megakaryocyte), which is rarely reported​. Interestingly, a complex karyotype was detected afterward by cytogenetics with t(3;3)(q21;q26.2), indicating a diagnosis of AML with t(3;3), a subset of which is also characterized by megakaryocytic markers such as CD41 and CD61. It seems that the second blast population detected by flow cytometry could not be classified into either diagnosis based on the morphology, immunophenotyping, and even cytogenetic findings, posing a real diagnostic problem because of the lack of clear-cut cytogenetic morphological defined criteria to distinguish between acute megakaryocytic leukemia and AML with t(3;3). Combining all of the examination data, this case was ultimately diagnosed as MPAL (T + My)-NOS with t(3;3) through differential diagnosis. Before the cytogenetic results were available, the patient received an acute lymphoblastic leukemia (ALL) regimen for MPAL treatment, but the effect was unsatisfactory. After the diagnosis was clear, she received an AML-like regimen with azacitidine for 7 days and venetoclax for 14 days, and achieved complete morphological remission. CONCLUSION: MPAL with either T/megakaryocyte or T/myeloid lineages accompanied by t(3;3) is rare, and it is difficult to make a clear diagnosis. Thus, comprehensive examinations, including bone marrow cell morphology, flow cytometry analysis, cytogenetics, and molecular analysis are recommended to avoid misdiagnosis. AML-like regimen including azacitidine and venetoclax may be effective for treating MPAL (T + My)-NOS with t(3;3).


Assuntos
Leucemia Mieloide Aguda , Leucemia-Linfoma Linfoblástico de Células Precursoras , Doença Aguda , Azacitidina , Compostos Bicíclicos Heterocíclicos com Pontes , Linhagem da Célula , Feminino , Humanos , Imunofenotipagem , Leucemia Mieloide Aguda/diagnóstico , Megacariócitos , Fenótipo , Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico , Sulfonamidas
20.
J Anim Sci ; 100(11)2022 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-36198005

RESUMO

This study aimed to investigate the effects of chronic heat stress on the immunophenotyping of lymphocytes in immune organs of growing pigs. A single-factor randomized block design was used, and 15 healthy growing large white barrows (5 litters, 3 pigs/litter) with similar body weight (40.8 kg) were assigned into 3 groups (5 pigs in each group). Groups were: control group (Con, in 23 °C environmental control chamber, fed ad libitum), heat stress group (HS, in 33 °C environmental control chamber, fed ad libitum), and pair-fed group (PF, in 23 °C environmental control chamber, fed diets according to the feed intake of HS group). After a 7-d adaption, the experiment lasted for 21 d. The results showed as follows: (1) activated T cells in the thymus of HS pigs were higher than those in PF pigs (P < 0.05). Monocytes and dendritic cells in the thymus of HS pigs were significantly higher than that in Con and PF pigs (P < 0.05), while the proportions of these 2 lymphocytes in the thymus of Con pigs did not differ from PF pigs (P > 0.05). Compared with Con pigs, the proportion of CD4+ (P < 0.05) and CD8+ T cells (P < 0.10) in the thymus was increased in HS pigs, while the proportion of CD4+ and CD8+ T cells in PF pigs did not differ from Con pigs (P > 0.05). (2) Compared with Con pigs, significantly decreased T cells, increased B cells and monocytes were found in the spleen of pigs exposed to heat stress (P < 0.05); the proportions of these 3 types of lymphocytes were not significantly different between Con and PF pigs (P > 0.05). The proportions of CD4+ T cells and Treg cells in the spleen of pigs exposed to heat stress tended to be lower than those in the Con pigs (P < 0.10). (3) The proportion of lymphocytes in the tonsils of pigs exposed to heat stress did not differ from Con pigs (P > 0.05); compared with PF pigs, the proportion of Treg cells was significantly decreased in HS pigs (P < 0.05). In conclusion, chronic heat stress stimulates the development and maturation of T cells in the pig thymus toward CD4+ and CD8+ T cells and increases the proportion of monocytes and dendritic cells; under the condition of chronic heat stress, the immune response process in the spleen of pigs is enhanced, but chronic heat stress impairs the survival of CD4+ T cells in the spleen.


Chronic heat stress (HS) has become a common hazard to livestock and poultry as global warming intensifies and breeding densities increase, which undoubtedly causes enormous economic losses to animal husbandry annually. Furthermore, it could also negatively impact the immune function of poultry and vaccines, resulting in various animal diseases. Until now, very few studies have focused on how HS affects the immune system of growing pigs, especially the immunophenotyping of lymphocytes in their immune organs (thymus, spleen and tonsils). In this study, the spleen and thymus are more severely affected by chronic HS than tonsils in growing pigs. Chronic HS stimulates the development and maturation of CD4+ and CD8+ T lymphocytes in the thymus. Under chronic HS, the immune response process in the spleen is enhanced, that is, the proportion of monocytes and B lymphocytes supporting immune responses increased, while the proportion of Treg cells decreased; yet long-term HS damaged the survival of CD4+ T lymphocytes in spleen.


Assuntos
Transtornos de Estresse por Calor , Doenças dos Suínos , Suínos , Animais , Imunofenotipagem/veterinária , Temperatura Alta , Transtornos de Estresse por Calor/veterinária , Resposta ao Choque Térmico , Dieta , Linfócitos
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