CC BY-NC-ND 4.0 · J Lab Physicians 2022; 14(04): 435-442
DOI: 10.1055/s-0042-1748828
Original Article

Flow Cytometric Expression of CD49d in Newly Diagnosed Chronic Lymphocytic Leukemia and Its Correlation with Established Prognostic Markers

Arathi Kunnumbrath
1   Department of Pathology and Laboratory Medicine, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India
,
Neha Singh
1   Department of Pathology and Laboratory Medicine, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India
,
Arvind Kumar Gupta
1   Department of Pathology and Laboratory Medicine, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India
,
Nilotpal Chowdhury
1   Department of Pathology and Laboratory Medicine, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India
,
Uttam Kumar Nath
2   Department of Medical Oncology and Hematology, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India
,
Harish Chandra
1   Department of Pathology and Laboratory Medicine, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India
› Author Affiliations
Funding None.

Abstract

Introduction Chronic lymphocytic leukemia (CLL) is the commonest hematological malignancy in the West but is relatively uncommon in India. The prognosis of CLL is determined by well-established prognostic markers. CD49d has been emerging as a promising prognostic marker in CLL. CD49d expression in CLL has been found to have an aggressive clinical course, shorter time to first treatment, and poorer prognosis. The aim of this study was to analyze the flow cytometric expression of CD49d in newly diagnosed CLL and to correlate its expression with clinico-hematological parameters.

Materials and Methods Twenty-five consecutive patients of CLL, diagnosed on flow cytometry, were included in the study. Patients on treatment or those with relapse were excluded. The panel for flow cytometry included the routine markers used for CLL diagnosis along with CD49d. The expression of CD49d was correlated with clinico-hematological parameters in all patients. “R” software was used for the statistical analysis. Fisher's exact test and Wilcox test were used to assess the correlation of CD49d to categorical and continuous data, respectively.

Results The mean age of the patients was 62.6 ± 12.5 years, and 80% were symptomatic at diagnosis. CD49d expression was found in 44% cases, with a higher proportion being male patients. CD49d and prolymphocyte percentage showed a statistically significant correlation (p = 0.0007). We found a statistically significant correlation between CD49d expression and lymphadenopathy and splenomegaly with p-values of 0.033 and 0.0472, respectively. CD49d positivity correlated significantly with a higher Rai stage (p = 0.0196) and intermediate and high-risk cases according to Binet staging (p = 0.033).

Conclusion CD49d expression in the present study correlated with a higher prolymphocyte percentage, lymphadenopathy, splenomegaly, and higher Rai and Binet stages. CD49d expression on flow cytometry was reproducible and easy to interpret.

Note

This study has been presented as an oral paper at Haematocon 2021. The study was approved by the Institutional Ethics Committee (letter no.: AIIMS/IEC/19/1166).


Authors' Contributions

A.K. contributed to data acquisition, analysis, and literature search; N.S. contributed to the concept, definition of intellectual content, literature search, manuscript preparation, manuscript editing, and manuscript review; A.K.G. contributed to manuscript editing and manuscript review; N.C. contributed to statistical analysis and manuscript review; U.K.N. contributed to clinical details and follow-up of patient and manuscript editing; and H.C. contributed to the definition of intellectual content and manuscript review.




Publication History

Article published online:
10 June 2022

© 2022. The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

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  • References

  • 1 Campo E, Ghia P, Montserrat E. et al. Chronic lymphocytic leukaemia/small lumphocytic lymphoma. In: Swerdlow SH, Campo E, Harris NL, et al., eds. WHO Classification of Tumors of Haematopoietic and Lymphoid Tissues. Revised 4th ed. Lyon, France:: International Agency for Research on Cancer (IARC);; 2017: 216-221
  • 2 Saxena R, Kumar R, Sazawal S, Mahapatra M. CLL in india may have a different biology from that in the west. Blood 2016; 128 (22) 5574
  • 3 Rai KR, Sawitsky A, Cronkite EP, Chanana AD, Levy RN, Pasternack BS. Clinical staging of chronic lymphocytic leukemia. Blood 1975; 46 (02) 219-234
  • 4 Binet JL, Auquier A, Dighiero G. et al. A new prognostic classification of chronic lymphocytic leukemia derived from a multivariate survival analysis. Cancer 1981; 48 (01) 198-206
  • 5 Oscier DG, Gardiner AC, Mould SJ. et al. Multivariate analysis of prognostic factors in CLL: clinical stage, IGVH gene mutational status, and loss or mutation of the p53 gene are independent prognostic factors. Blood 2002; 100 (04) 1177-1184
  • 6 Boonstra JG, van Lom K, Langerak AW. et al. CD38 as a prognostic factor in B cell chronic lymphocytic leukaemia (B-CLL): comparison of three approaches to analyze its expression. Cytometry B Clin Cytom 2006; 70 (03) 136-141
  • 7 Gattei V, Bulian P, Del Principe MI. et al. Relevance of CD49d protein expression as overall survival and progressive disease prognosticator in chronic lymphocytic leukemia. Blood 2008; 111 (02) 865-873
  • 8 Crespo M, Villamor N, Giné E. et al. ZAP-70 expression in normal pro/pre B cells, mature B cells, and in B-cell acute lymphoblastic leukemia. Clin Cancer Res 2006; 12 (3 Pt 1): 726-734
  • 9 Wiestner A, Rosenwald A, Barry TS. et al. ZAP-70 expression identifies a chronic lymphocytic leukemia subtype with unmutated immunoglobulin genes, inferior clinical outcome, and distinct gene expression profile. Blood 2003; 101 (12) 4944-4951
  • 10 Hemler ME. VLA proteins in the integrin family: structures, functions, and their role on leukocytes. Annu Rev Immunol 1990; 8 (01) 365-400
  • 11 Bosch F, Muntañola A, Giné E. et al. Clinical implications of ZAP-70 expression in chronic lymphocytic leukemia. Cytometry B Clin Cytom 2006; 70 (04) 214-217
  • 12 Rassenti LZ, Huynh L, Toy TL. et al. ZAP-70 compared with immunoglobulin heavy-chain gene mutation status as a predictor of disease progression in chronic lymphocytic leukemia. N Engl J Med 2004; 351 (09) 893-901
  • 13 Ibrahem L, Elderiny WE, Elhelw L, Ismail M. CD49d and CD26 are independent prognostic markers for disease progression in patients with chronic lymphocytic leukemia. Blood Cells Mol Dis 2015; 55 (02) 154-160
  • 14 Shanafelt TD, Geyer SM, Bone ND. et al. CD49d expression is an independent predictor of overall survival in patients with chronic lymphocytic leukaemia: a prognostic parameter with therapeutic potential. Br J Haematol 2008; 140 (05) 537-546
  • 15 Bulian P, Shanafelt TD, Fegan C. et al. CD49d is the strongest flow cytometry-based predictor of overall survival in chronic lymphocytic leukemia. J Clin Oncol 2014; 32 (09) 897-904
  • 16 Molica S. Sex differences in incidence and outcome of chronic lymphocytic leukemia patients. Leuk Lymphoma 2006; 47 (08) 1477-1480
  • 17 Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin 2020; 70 (01) 7-30
  • 18 Agrawal N, Naithani R, Mahapatra M. et al. Chronic lymphocytic leukemia in India—a clinico-hematological profile. Hematology 2007; 12 (03) 229-233
  • 19 Gogia A, Gupta R, Sharma A, Kumar L, Raina V, Rani L. Chronic lymphocytic leukemia: an Indian experience. J Clin Oncol 2019; 37 (15, suppl): e19007-e19007
  • 20 Gogia A, Sharma A, Raina V. et al. Assessment of 285 cases of chronic lymphocytic leukemia seen at single large tertiary center in Northern India. Leuk Lymphoma 2012; 53 (10) 1961-1965
  • 21 Okaly GVP, Nargund AR, Venkataswamy E, Jayanna PK, Juvva CR, Prabhudesai S. Chronic lymphoproliferative disorders at an Indian tertiary cancer centre—the panel sufficiency in the diagnosis of chronic lymphocytic leukaemia. J Clin Diagn Res 2013; 7 (07) 1366-1371
  • 22 Baumann T, Delgado J, Santacruz R. et al. CD49d (ITGA4) expression is a predictor of time to first treatment in patients with chronic lymphocytic leukaemia and mutated IGHV status. Br J Haematol 2016; 172 (01) 48-55
  • 23 Strati P, Parikh SA, Chaffee KG. et al. CD49d associates with nodal presentation and subsequent development of lymphadenopathy in patients with chronic lymphocytic leukaemia. Br J Haematol 2017; 178 (01) 99-105
  • 24 Criel A, Michaux L, De Wolf-Peeters C. The concept of typical and atypical chronic lymphocytic leukaemia. Leuk Lymphoma 1999; 33 (1-2): 33-45
  • 25 Frater JL, McCarron KF, Hammel JP. et al. Typical and atypical chronic lymphocytic leukemia differ clinically and immunophenotypically. Am J Clin Pathol 2001; 116 (05) 655-664
  • 26 Matutes E, Oscier D, Garcia-Marco J. et al. Trisomy 12 defines a group of CLL with atypical morphology: correlation between cytogenetic, clinical and laboratory features in 544 patients. Br J Haematol 1996; 92 (02) 382-388
  • 27 Buggins AGS, Levi A, Gohil S. et al. Evidence for a macromolecular complex in poor prognosis CLL that contains CD38, CD49d, CD44 and MMP-9. Br J Haematol 2011; 154 (02) 216-222
  • 28 Baldini L, Cro L, Calori R, Nobili L, Silvestris I, Maiolo AT. Differential expression of very late activation antigen-3 (VLA-3)/VLA-4 in B-cell non-Hodgkin lymphoma and B-cell chronic lymphocytic leukemia. Blood 1992; 79 (10) 2688-2693
  • 29 Nematollahi P, Shakery M, Kefayat A, Goli P. Association of CD49d expression with clinicopathological features of chronic lymphocytic leukemia patients in the Iranian population. Int J Physiol Pathophysiol Pharmacol 2020; 12 (01) 32-39
  • 30 Al-Rubaie HA, Thabit ZA, Jawad AM. CD49d as prognostic marker in B-cell chronic lymphocytic leukemia in correlation with the expression of CD38 ZAP-70 and clinical binet stage. Iraqi Postgrad Med J 2016; 15 (04) 486-492
  • 31 Rossi D, Zucchetto A, Rossi FM. et al. CD49d expression is an independent risk factor of progressive disease in early stage chronic lymphocytic leukemia. Haematologica 2008; 93 (10) 1575-1579
  • 32 Oscier D, Else M, Matutes E, Morilla R, Strefford JC, Catovsky D. The morphology of CLL revisited: the clinical significance of prolymphocytes and correlations with prognostic/molecular markers in the LRF CLL4 trial. Br J Haematol 2016; 174 (05) 767-775
  • 33 van der Velden VHJ, Hoogeveen PG, de Ridder D. et al. B-cell prolymphocytic leukemia: a specific subgroup of mantle cell lymphoma. Blood 2014; 124 (03) 412-419
  • 34 Criel A, Verhoef G, Vlietinck R. et al. Further characterization of morphologically defined typical and atypical CLL: a clinical, immunophenotypic, cytogenetic and prognostic study on 390 cases. Br J Haematol 1997; 97 (02) 383-391
  • 35 Abdel-Aziz AF, Yahya RS, Abdel-Messih HM, Ata AM. Diagnostic value of CD49d expression in patients with chronic lymphocytic leukemia. Biochem Physiol 2019; 8 (01) 249
  • 36 Cartron G, Linassier C, Bremond JL. et al. CD5 negative B-cell chronic lymphocytic leukemia: clinical and biological features of 42 cases. Leuk Lymphoma 1998; 31 (1-2): 209-216
  • 37 Zucchetto A, Caldana C, Benedetti D. et al. CD49d is overexpressed by trisomy 12 chronic lymphocytic leukemia cells: evidence for a methylation-dependent regulation mechanism. Blood 2013; 122 (19) 3317-3321
  • 38 Léger OJ, Yednock TA, Tanner L. et al. Humanization of a mouse antibody against human alpha-4 integrin: a potential therapeutic for the treatment of multiple sclerosis. Hum Antibodies 1997; 8 (01) 3-16