Immunotyping of Lymphoma by Fine-needle Aspiration:A Comparative Study of Cytospin Preparations and Flow Cytometry

Abstract

Immunotyping is an essential adjunct to cytomorphology for the diagnosis of lymphoma by fine-needle aspiration (FNA). Two independent techniques, cytospin preparations and flow cytometry, were used for immunotyping studies on 71 patients with histologically confirmed non-Hodgkin's lymphoma (63 B-cell lymphomas and 8 T-cell lymphomas). Diagnostic concordance between the two methods was obtained in 69 patients (97%). kappa, lambda, and CD3 (Leu-4) markers were routinely measured on all cytospins, and additional markers were requested when indicated. The standard panel measured by flow cytometry included 14 markers. In general, mean values of light-chain (kappa and lambda) immunoglobulins were consistently slightly higher by cytospin analysis than by flow cytometry. Light-chain immunoglobulin ratios (kappa/lambda or lambda/kappa) obtained by both methods proved to be reliable independent predictors of polyclonality or monoclonality. Correlation studies using the Spearman rank coefficient revealed good concordance among values of kappa, alpha, CD3, and CD5 obtained by the two techniques, suggesting that subjective quantitation by cytospins yields similar results to objective quantitation by flow cytometry. Cytospin analysis and flow cytometry appear equally capable of immunotyping aspirated lymphoid samples reliably. The advantages of each method are discussed.

Adult T-cell leukemia/lymphoma

“…This has been evidenced by studies showing that nonHodgkin lymphomas can be accurately diagnosed in cytologic specimens utilizing ancillary techniques such as immunocytochemistry and flow cytometry. 4,[15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31][32] In the REAL classification, ATLL belongs to the category of peripheral T-cell and natural killer cell neoplasms. Although several reports of peripheral Tcell lymphoma have been published in the cytology literature, [33][34][35][36][37] we are aware of only a single cytologic study of T-cell lymphoma in HTLV-1 positive patients.…”

Romidepsin and Azacitidine Synergize in their Epigenetic Modulatory Effects to Induce Apoptosis in CTCL

“…After incubation of CTCL cell lines with romidepsin and azacitidine simultaneously for 48 hours, the samples were placed on the slides using the cytospin procedure, as previously published (19). After fixation in 10% acetone, the slides were incubated with the RhoB primary antibody (C-5, mouse monoclonal, Santa Cruz Biotechnology), and then secondary antibody as suggested by manufacturer's protocol.…”

Fine-needle cytology and flow cytometry immunophenotyping and subclassification of non-hodgkin lymphoma

“…1,2,[13][14][15] The possibility of applying a complete panel of antibodies and diagnostic algorithms is the most appreciated advantage of the technique; 4,6,8 consequently, many reports stress the high sensitivity in detecting non-Hodgkin lymphoma (NHL) through the combined application of FNC and FC. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] The most enthusiastic reports hypothesize that, in the near future, histology will not be necessary strictly for the diagnosis of NHL. In fact, in many institutions, FNC coupled with ancillary techniques like FC routinely has replaced histology in the diagnosis of recurrent NHL (rNHL).…”

“…In fact, in many institutions, FNC coupled with ancillary techniques like FC routinely has replaced histology in the diagnosis of recurrent NHL (rNHL). 1,2,4,8,10 However, the possibility of a definite cytologic diagnosis even for primary NHL is related to the possibility of a correct subclassification of the single entities, according to the most recent Revised European-American Classification of Lymphoid Neoplasms (the REAL classification system). 16,17 The possibility of identifying a follicular NHL through CD10/CD19 coexpression without microscopic visualization of follicular structures is a stimulating idea; and, in recent years, there have been some reports using FC or immunocytochemistry to exploit the possibility of subclassifying NHL.…”