Intraepithelial CD15 infiltration identifies high-grade anal dysplasia in people with HIV
Joaquín BurgosAleix Benítez-MartínezCristina ManceboNúria MassanaAntonio Astorga-GamazaJosep CastellvíStefania LandolfiAdrián CurranJorge GarcíaVicenç FalcóMaría J. BuzónMeritxell Genescà
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Abstract:
Men who have sex with men (MSM) with HIV are at high risk for squamous intraepithelial lesion (SIL) and anal cancer. Identifying local immunological mechanisms involved in the development of anal dysplasia could aid treatment and diagnostics. Here we studied 111 anal biopsies obtained from 101 MSM with HIV, who participated in an anal screening program. We first assessed multiple immune subsets by flow cytometry, in addition to histological examination, in a discovery cohort (n = 54). Selected molecules were further evaluated by immunohistochemistry in a validation cohort (n = 47). Pathological samples were characterized by the presence of Resident Memory T cells with low expression of CD103 and by changes in Natural Killer cell subsets, affecting residency and activation. Furthermore, potentially immune suppressive subsets, including CD15+CD16+ mature neutrophils, gradually increased as the anal lesion progressed. Immunohistochemistry confirmed the association between the presence of CD15 in the epithelium and SIL diagnosis, with a sensitivity of 80% and specificity of 71% (AUC 0.762) for the correlation with high-grade SIL. A complex immunological environment with imbalanced proportions of resident effectors and immune suppressive subsets characterizes pathological samples. Neutrophil infiltration, determined by CD15 staining, may represent a valuable pathological marker associated with the grade of dysplasia.Keywords:
CD15
Squamous intraepithelial lesion
Immunophenotyping
The aim of this study was to explore the immunophenotypic characteristics of Philadelphia chromosome positive (Ph(+)) acute lymphoblastic leukaemia (ALL) in adults and to evaluate their significance in predicting prognosis and guiding clinical treatment of diseases. The cell immunophenotypes of leukemic marrow or blood samples from 35 cases of Ph(+) ALL and 59 cases of Philadelphia chromosome negative (Ph(-)) ALL were detected by multiparameter flow cytometry, and their abnormal expressions were analysed. The results showed that the expression of all the Ph(+)ALL cases was found in B-cell lineage. As compared with Ph(-)B-ALL cases, the Ph(+)B-ALL cases displayed the higher expression of CD34 and CD13 (p < 0.05), but lower expression of CD38 (p < 0.05). The coexpressed rates of CD13, CD33 and CD15 in cases of Ph(+)B-ALL and Ph(-)B-ALL were 85.7% and 61.0% respectively. The former was higher than the later (p < 0.05). It is concluded that the Ph(+)ALL cases have the unique immunophenotype. The immunophenotypic analysis of CD34, CD13 and CD38 in adult B-ALL cases contributes to judging the existence of Ph chromosome. Thereby for adult ALL patients having above-mentioned unique immunophenotypes, the detection of bcr/abl fusion gene must be performed. Such phenotypic profile is helpful for predicting the poor outcome of the disease, and for defining patients who require different treatment strategies.
Immunophenotyping
CD15
Philadelphia chromosome
Minimal Residual Disease
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The data of detailed studies of immunophenotype of blast cells in 426 patients with acute leukemias are presented. Diagnostic and prognostic significance of different marker expression has been evaluated in groups of patients with ALL and AML. Frequency distribution of T1, T2, T3, pre-B, B, common, Ia and null subvariants identified according to immunoclassification of Baryshinkov et al. was studied in 250 children with ALL. These subvariants differed both in duration of disease (p = 0.0015) and in duration of first complete remission (p = 0.0031). The use of monoclonal antibodies of VI-series in 90 patients with ALL allowed to describe an immunophenotype of the subvariants in detail. The mosaic expression of myeloid antigens CD11, CD14, CD15, CDw65 identified by MoAbs VIM-12, VIM-13, VIM-D5 and VIM-2, respectively, on blast cells of patients with AML was shown. The expression of CD11 (ICO-GM1) or CD15 (ICO-G2) was prognostically unfavorable in children with AML (p = 0.0028). The expression of T-cell markers (E-receptor, CD7, reactivity with anti-T-cell serum) on blasts was prognostically favorable in children with AML (p = 0.003). So the data of immunophenotyping are of great value for accurate diagnosis and prognosis of acute leukemias.
Immunophenotyping
CD15
Clinical Significance
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Conventional intestinal-type dysplasia (CID) is well described in the setting of ulcerative colitis (UC). Other types of dysplasia are less reported. Our aim was to investigate the prevalence and significance of these less known dysplasias in the setting of UC. Colonic resections from UC patients were divided into two cohorts—group 1 (n = 9), which had invasive adenocarcinoma, and group 2 (n = 15), without CID or carcinoma. Archival H&E slides were reviewed, with emphasis on identifying epithelial changes. A characteristic epithelial change was noted, including distinct areas of enlarged glands with architectural distortion and reduced or absent intraepithelial inflammation, associated with increased density and size of goblet cells in the glands. These changes were identified in all 9 cases from group 1 (100%) and 9 of 15 cases from group 2 (60%). In group 1, these findings were seen in close association to CID and invasive tumor. We designated these changes as mucinous dysplasia (MD) rather than metaplasia due to its proximity to tumor. Additionally, dysplastic changes were also noted away from the tumor (MD in 7 cases, CID in 2 cases, and serrated dysplasia in 1 case). Although MD in UC has been recognized by some authors, the description of this spectrum of mucosal changes has not been previously reported. Our study indicates that this is probably an earlier neoplastic manifestation in the setting of UC, before CID arises. It also appears to be a common finding in the setting of neoplastic transformation in UC. Identification of this entity as a dysplastic change in surveillance biopsies of UC patients can probably help prevent subsequent development of carcinoma, by prophylactic colonic resection. The study is limited due to its sample size. A larger cohort is required to confirm the findings of this study.
Intestinal metaplasia
Neoplastic transformation
Metaplasia
Malignant Transformation
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Hodgkin's lymphoma (HL) has a unique immunophenotype derived from immunohistochemistry (positive for CD15, CD30, and Pax‐5; negative for CD3, CD20 in most cases, and CD45). The knowledge gained over recent years enables better diagnosis, prognosis, and treatment of HL. Flow cytometry as a tool for the diagnosis of classic HL has not been useful in the past due to the difficulty in isolating Reed–Sternberg cells as they are admixed in a rich inflammatory background which consists mainly of T cells, B cells, eosinophils, histiocytes, and plasma cells. However, in the recent past, several studies have tried to identify Reed–Sternberg cells using flow cytometry on fine needle aspiration or tissue biopsy of lymph nodes to confirm or supplement immunohistochemistry staining in diagnosis. Newer and more sensitive tools such as flow cytometry can be used for diagnosis, technology that may have been difficult in the past for diagnosis of this lymphoma subtype. Using flow cytometry, diagnosis is faster and could lead to point‐of‐care technology especially where we have typical immunophenotype signatures. © 2018 International Clinical Cytometry Society
Immunophenotyping
CD15
Cytometry
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Aim of the study – characterization of immunophenotype in infant acute myeloid leukemia (AML). 90 patients (40 boys and 50 girls) with acute leukemia (AL) aged up to 365 days were included in the current study. AML was found more frequently in infants than in older children (26.67 % and 10.83 % respectively; p = 0.0002). Significant immunophenotypic differences were observed in patients with and without MLL gene rearrangements. Number of cases in those tumor cells expressed CD99, CD61, CD133, CD15, NG2 varied between MLL-positive and MLL-negative groups. CD61-negativity, high CD99, CD15, CD133 and NG2 expression were immunophenotypic signatures of MLLrearranged infant AML, although CD99 and NG2 had the highest diagnostic efficacy. Thus infants’ AML immunophenotype differs significantly due to the presence of MLL gene rearrangements. Diagnostic immunophenotyping of infants’ AML allows predicting presence of MLL rearrangements by either CD99 or NG2 expression.
Immunophenotyping
CD99
CD15
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Carcinoma in situ
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To analyse the immunophenotype of acute leukaemia (AL) after myelodysplastic syndromes (MDS) (MDS-AL) and to compare the immunophenotypic profile of acute myeloblastic leukaemia (AML) secondary to MDS (MDS-AML) with that of "de novo"-AML.Twenty patients with MDS-AL and 29 patients with "de novo"-AML were studied. Morphocytochemical and flow cytometric studies were done in each case.All the MDS-AL studied displayed a myeloid phenotype (MDS-AML). The main difference between MDS-AML and "de novo"-AML was a significantly higher frequency of CD34 expression in the first group. Differences concerning the expression of other non-lineage related or myeloid-associated markers were not statistically significant, although the percentage of cases CD15(+) was lower in MDS-AML. The overall frequency of expression of lymphoid-associated markers was similar in both groups, T-cell markers being more frequently detected.Our findings support the usefulness of immunophenotyping studies to characterize MDS-AL and suggest some immunophenotyping differences between MDS-AML and "de novo"-AML which might have biological and prognostic significance.
Immunophenotyping
CD15
Acute myeloblastic leukemia
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Objective To evaluate the value of immunophenotype in diagnosis of myelodysplastic syndrome (MDS).Methods The immunophenotype of bone marrow cells in 27 patients with MDS were detected by monoclonal antibody by flow cytometry.Results As the progression of the disease,CD34 positive cells gradually increased:refractory anemia/ring sideroblasts refractory anemia (RA/ AS) 7.43 %,refractory anemia with excess of blasts (RAEB) 36.81%,refractory anemia with excess of blasts transformed (RAEB-T)56.45 %,and the differences were statistically significant (P ＜0.05); the expressions of CD33+,CD13 and HLA-DR increased gradually,the expressions of CD14 and CD15 antigens gradually decreased,the difference of three groups was statistically significant (P ＜0.05),the differences between RA/RAS and RAEB-T,RAEB and RAEB-T were statistically significant (P ＜0.05); the expression of CD19 and CD10 decreased and the expression of CD7 increased (RA/RAS 2.63 %,RAEB 10.79 % and RAEB-T 11.00 %) with the progression of the disease,the difference of three groups was statistically significant (F =10.439,P ＜0.05),the differences between RA/RAS and RAEB,RA/RAS and RAEB-T were statistically significant (P ＜0.05).Conclusion The detection of immunophenotype of bone marrow cella in patients with MDS contributes to the diagnosis,classification and prognosis of MDS.
Key words:
Myelodysplatic syndromes; Immunophenotyping; Flow cytometry
Immunophenotyping
CD15
Refractory anemia
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Objective To study the character of immunophenotype in children with acute lymphoblastic leukemia(ALL).Methods To assay the immunophenotype in 45 children with acute leukemia by flow cytometer.Results ① In 45 cases ALL T-ALL was 11.1%,B-ALL was 79.9%;ALL1 20%,ALL2 60.0%,ALL3 11.1%;② In 45 cases ALL,four children was diagnosed HAL by immunophenotype and the positive rate of myeloid antigen was 55.6%.While the positive rate of CD13 was 44.4%,CD33 was 22.2%,CD15 was 22.2%,respectively.The positive rate of myeloid antigen in B-ALL(55.6%) was significantly higher compare with T-ALL(20%)(P0.05);the positive rate of myeloid antigen in L2(70.4%) was significant higher than in L1(22.2%) and L3(40%)(P0.05).Conclusion Immunophenotyping,combined with morphology can provide more reliable and valuable evidence in the diagnosis of children with lymphoblastic leukemia.
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CD15
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Other studies have shown that the immunophenotype of Reed- Sternberg and Hodgkin’s (RS-H) cells in Hodgkin’s disease commonly changes over time, as shown by examination of multiple biopsy specimens obtained from an individual patient. In this study we analyzed 96 sequential biopsy specimens (>1 month apart) obtained from 44 patients (nodular sclerosis, 34 specimens; mixed cellularity, 5; lymphocyte depletion, 1; unclassified, 4) using fixed, paraffin-embedded sections; heat-induced epitope retrieval (HIER); a panel of antibodies specific for the CD3, CD15, CD20, CD30, CD43, CD45/45RB, and CD79a antigens and Epstein-Barr virus latent-membrane protein; and a streptavidin - biotin method. In selected cases in which immunophenotypic changes occurred, studies were repeated using enzyme predigestion instead of HIER. There was no change in the immunophenotype of the RS-H cells in 36 (82%) of 44 patients. In 8 patients (18%), the immunophenotype of the RS-H cells varied in expression of one or two antigens. The antigens that varied were as follows: CD30, 3 patients; CD15, 3 patients; CD20,1 patient; and CD15 and CD30,1 patient. We conclude that the immunophenotype of RS-H cells in Hodgkin’s disease is relatively stable over time and that CD15 and CD30 are the most common antigens that change. The frequency of immunophenotypic changes, 18%, is substantially lower than that reported previously. One likely explanation for this discrepancy is that we used HIER, a relatively recent innovation in diagnostic immunohistochemistry that has been shown to reduce artifacts attributable to inconsistent fixation and processing. The significance of immunophenotypic variation in eight cases (18%) is uncertain. This phenomenon may represent true biologic changes in RS-H cells. Alternatively, these changes may be attributable to artifacts secondary to inconsistent fixation or processing that HIER cannot overcome.
Immunophenotyping
CD15
Reed–Sternberg cell
Antigen retrieval
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