Issue Highlights—May 2013

Abstract

Cytometry Part B: Clinical CytometryVolume 84B, Issue 3 p. 133-134 Issue HighlightsFree Access Issue Highlights—May 2013 First published: 19 April 2013 https://doi.org/10.1002/cyto.b.21093 [email protected] AboutSectionsPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Taking the sting out of functional basophil testing In this issue, we see manuscripts covering a wide assortment of topics of interest to clinical cytometrists. Among these excellent articles is one by Nullens and colleague (1) describing the measurement of intracellular histamine in basophils during the course of venom immunotherapy therapy (VIT). For individuals who suffer systemic reactions to wasp stings, VIT offers a specific treatment to reduce the odds of recurrent severe reactions (2). In this paper, a method to detect histamine release as well as cell surface markers of basophils is used to monitor the effects of VIT therapy. Although this study is limited in the number of patients studied, it offers examination of both function and phenotyping in rare cells as a method of monitoring the efficacy of a specific immunotherapy. Historically basophils were an often overlooked subset of leukocytes in the peripheral circulation, but in the past ten years, our ability to detect basophils and study their function has markedly increased (3-5). The readers are referred to several recent articles published in this journal for the latest developments in basophil assays in the clinical cytometry arena. Doing the FOXP3 trot Regulatory T (Tregs) cells are key components in down-regulating immune responses, and have been shown to play a role in diseases ranging from cancer to autoimmune diseases and infections (6). Tregs are CD4 T cells most often characterized by the expression of forkhead box P3 (FoxP3). These cells also regarded as having have bright expression of CD25 and low expression of CD127, although a recent publication has shown a great heterogeneity in the phenotypes of Tregs found in patients with chronic lymphocytic leukemia (7). In this issue, Demaret et al demonstrate a one step technique for staining FoxP3 in order to identify these cells (8). FoxP3 is an intracellular antigen, and as such, fixation/permeabilization techniques are required for staining. In the approach presented here, a commercial no wash kit is used for staining these cells with FoxP3 as well as surface markers with good results. This expedited protocol might obviate the need to use surrogate staining, such as CD25high and CD127 low on CD4 cells to identify Tregs (9). One might hope that this expedient method would facilitate samples in clinical trials being run on fresh rather than cryopreserved samples, as cryopreservation has been shown to negatively impact the detection and quantification of Tregs (10). Seeing double trouble One of the most common uses of flow cytometry in the clinical laboratory is for the detection of leukemic cells in the bone marrow or peripheral circulation. While clinicians most often suspect a single lineage of leukemic cells, it is not altogether uncommon for the cytometry results to reveal a bilineal, or mixed phenotypic acute leukemia (MPAL). In the current issue, Rahman and colleagues report an unusual case of a patient presenting with both T and B lineages among the leukemic blasts (11). Even among MPAL this phenotype is rare, with most cases displaying a myeloid component in addition to the lymphocytic lineage (12, 13). Those who find the current case report of interest might also want to review a previous case report in which three simultaneous lymphoproliferative disorders (HCL, B-CLL, and T-LGL) were detected in both peripheral blood and bone marrow (14). More double trouble Double hit lymphomas (DHL), those with concurrent translocations of MYC and BCL2are rare, but well documented clinical entities (15). The issue of whether or not flow cytometry can be of use in identifying these lymphomas is the subject of investigation by Platt et al (15). Phenotypic abnormalities have been associated with DHL, including dim expression of CD20 and/or CD19. In the current report, the authors find these abnormalities, but not in such frequencies to permit these to be used as unequivocal biomarkers of DHL. Rather, they suggest that the immunophenotypic findings be used only in conjunction with clinical and other laboratory findings to recommend cytogenetic testing. A case of double hit lymphoma was presented as a ‘Case Study Interpretation (CSI)’ during the annual meeting of the International Clinical Cytometry Society in 2011 for attendees to test their diagnostic skills. For those who were unable to attend, this case is available in this journal (16). J. Philip McCoy Jr., PhD Bethesda, MD LITERATURE CITED 1 Nullens S, Sabato V, Faber M, Leysen J, Bridts CH, De Clerck LS, Falcone FH, Maurer M, Ebo DG. Basophilic histamine content and release during venom immunotherapy: Insights by flow cytometry. Cytometry Part B Clin Cytom 2013 84B:( 173– 178. 2 Krishna MT, Ewan PW, Diwakar L, Durham SR, Frew AJ, Leech SC, Nasser SM. Diagnosis and management of hymenoptera venom allergy: British Society for Allergy and Clinical Immunology (BSACI) guidelines. Clin Exp Allergy. 2011 41( 9): 1201– 1220. 3 González-de-Olano D, Álvarez-Twose I, Morgado JM, López MIE, Castro AV, Díaz de Durana MDA, Sánchez-Muñoz L, Matito A, de la Hoz Caballer, B, Sanz ML, Orfao A, Escribano L. Evaluation of basophil activation in mastocytosis with Hymenoptera venom anaphylaxis. Cytometry Part B Clin Cytom 2011, 80B: 167– 175. 4 Sturm EM, Kranzelbinder B, Heinemann A, Groselj-Strele A, Aberer W, Sturm GJ. CD203c-based basophil activation test in allergy diagnosis: Characteristics and differences to CD63 upregulation. Cytometry Part B Clin Cytom 2010; 78B: 308– 318. 5 Verweij MM, Sabato V, Nullens S, Bridts CH, De Clerck LS, Stevens WJ, Ebo DG. STAT5 in human basophils: IL-3 is required for its FcεRI-mediated phosphorylation. Cytometry Part B Clin Cytom 2012; 82B: 101– 106. 6 Benoist C, Mathis D. Treg cells, life history, and diversity. Cold Spring Harb Perspect Biol 2012; 1; 4( 9): a007021. 7 Biancotto A, Dagur PK, Fuchs JC, Wiestner A, Bagwell CB, McCoy JP Jr. Phenotypic complexity of T regulatory subsets in patients with B-chronic lymphocytic leukemia. Mod Pathol 2012 25( 2): 246– 259. 8 Demaret J, Saison J, Venet F, Malcus C, Poitevin-Later F, Lepape A, Ferry T, Monneret G. Assessment of a novel flow cytometry technique of one-step intracellular staining: Example of FOXP3 in clinical samples. Cytometry B Clin Cytom 2013; 84B: 187– 193. 9 Saison J, Demaret J, Venet F, Chidiac C, Malcus C, Poitevin-Later F, Tardy JC, Ferry T, Monneret G. CD4+CD25+CD127- assessment as a surrogate phenotype for FOXP3+ regulatory T cells in HIV-1 infected viremic and aviremic subjects. Cytometry B Clin Cytom 2013; 84B( 1): 50– 54. 10 Sattui S, de la Flor C, Sanchez C, Lewis D, Lopez G, Rizo-Patrón E, White AC Jr, Montes M. Cryopreservation modulates the detection of regulatory T cell markers. Cytometry B Clin Cytom 2012; 82B( 1): 54– 58. 11 Rahman K, George S, Tewari A, Mehta A. Mixed phenotypic acute leukemia with two immunophenotypically distinct blast populations: Report of an unusual case. Cytometry B Clin Cytom 2013; 84B: 198– 201. 12 Song S. A case report: Concurrent chronic myelomonocytic leukemia and T-cell large granular lymphocytic leukemia-type clonal proliferation as detected by multiparametric flow cytometry. Cytometry B Clin Cytom 2011; 80B( 2): 126– 129. 13 McCoy JP Jr, Johnson E, Catalano E, Blumstein L, Overton WR, Gryn J, Donaldson MH. Detection and monitoring of a concomitant atypical myeloproliferative disorder and chronic lymphocytic leukemia by flow-cytometric immunophenotyping. Arch Pathol Lab Med 1995; 119( 11): 1038– 43. 14 Garrido P, Jiménez P, Sánchez C, Valero F, Balanzategui A, Almagro M, López P, de Pablos JM, Navarro P, Cabrera A, González M, Jurado M, Ruiz-Cabello F. Molecular and flow cytometry characterization during the follow-up of three simultaneous lymphoproliferative disorders: hairy cell leukemia, monoclonal B-cell lymphocytosis, and CD4(++) /CD8(+/- dim) T-large granular lymphocytosis--a case report. Cytometry B Clin Cytom 2011; 80B( 3): 195– 200. 15 Aukema SM, Siebert R, Schuuring E, van Imhoff GW, Kluin-Nelemans HC, Boerma EJ, Kluin PM. Double-hit B-cell lymphomas. Blood 2011; 117( 8): 2319– 2331. 16 Platt MY, DeLelys ME, Preffer FI, Sohani AR. Flow cytometry identification is of limited utility in the early identification of ‘Double-Hit’ B-cell lymphomas. Cytometry Part B Clin Cytom 2013; 84B: 143– 148. 17 Grier DD, Montoya K, Pang C. Case study interpretation--Portland: Case 3. Double hit lymphoma. Cytometry B Clin Cytom 2012; 82B( 3): 183– 185. Volume84B, Issue3May 2013Pages 133-134 ReferencesRelatedInformation