Diagnostic Immunohistochemistry with Manual Tissue Microarray Technique: A Pilot Study on Non-Hodgkin Lymphoma
Keywords:
Non Hodgkin Lymphoma, Immunohistochemistry, Donor Block, Recipient Block, Tissue Microarray
Abstract
BACKGROUND: Non Hodgkin lymphomas are clonal lymphoproliferative disorders that needs to be classified immunologically by variety of immunological markers targeted against specific antigens. Tissue microarray allows for high-throughput molecular profiling of tissue specimens using immunohistochemistry resulting in reduced consumption of time and reagents as compared to conventional immunohistochemistry. METHOD: This three-year single institutional observational study was conducted at Tirunelveli Medical College, Tirunelveli, Tamilnadu. 21 cases of histopathologically diagnosed NHL were subjected to Immunohistochemistry using manual Tissue microarray. Paraffin embedded tissue blocks of all the NHL cases formed donor blocks. Lay out for Tissue microarray was constructed followed by manual transfer of cored tissue from representative areas of donor blocks into recipient block using bone marrow needles. Immunohistochemistry was done using antibody against CD3, CD5, CD10 and CD20. Inadequate lymph node samples, poorly processed samples and extranodal NHL were excluded. RESULT: Of total 21 cases subjected to immunohistochemistry using Tissue microarray technique, only 19 cases were taken for analysis due to tissue loss and histopathological misdiagnosis. Among 19 cases there were 11[57.89%] males and 8[42.11%] females with male to female ratio of 1.37:1. Mean age of study group was 54.7 years. There were 18[94.73%] cases of B-cell NHL with 1[5.27%] case of T-cell NHL. DLBCL constituted for 9[47.36%] cases. Immunohistochemistry using Tissue microarray consumed 1/6th of the reagent volume as that of conventional immunohistochemistry.References
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16. Campo E, Miquel R, Krenacs L, Sorbara L, Raffeld M, Jaffe ES. Primary nodal marginal zone lymphomas of splenic and MALT type. Am J Surg Pathol. 1999 Jan;23(1):59-68.
17. Mucci NR, Akdas G, Manely S, Rubin MA. Neuroendocrine expression in metastatic prostatic cancer: evaluation of high throughput tissue microarrays to detect heterogenous protein expression. Hum Pathol. 2000;31(4):406-414.
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19. Richter J, Wagner U, Kononen J, et al. High-throughput tissue microarray analysis of cyclin E gene amplification and overexpression in urinary bladder cancer. Am J Pathol. 2000;157(3):787-794.
2. Afaf Abdel-Aziz Abdel-Ghafar, Manal Ahmed Shams El Din El Telbany, Hanan Mohamed
Mahmoud, Yasmin Nabil El-Sakhawy. Immunophenotyping of chronic B-cell neoplasms: flow cytometry versus immunohistochemistry. Hematology reports. 2012;4(1):e3.
3. Tumwine LK, Agostinelli C, Campidelli C, et al. Immunohistochemical and other prognostic factors in B cell non Hodgkin lymphoma patients, Kampala, Uganda. BMC Clinical Pathology. 2009;9(11).
4. Naresh KN, Srinivas V, Soman CS. Distribution of various subtypes of non-Hodgkin’s lymphoma in India: a study of 2773 lymphomas using R.E.A.L. and WHO Classifications. Ann Oncol. 2000;11(Suppl 1):63-67.
5. Kalyan K, Basu D, Soundararaghavan J. Immunohistochemical typing of non Hodgkin’slymphoma-comparing working formulation and WHO classification. Indian J Pathol Microbiol. 2006Apr ; 49(2):203-7.
6. Mushtaq S, Akhtar N, Jamal S, et al. Malignant lymphomas in Pakistan according to the WHO classification of lymphoid neoplasms. Asian Pac J Cancer Prev. 2008Apr-Jun;9(2):229-232.
7. Padhi S, Paul TR, Challa S, et al. Primary Extra Nodal Non Hodgkin Lymphoma: A 5 Year Retrospective Analysis. Asian Pac J Cancer Prev. 2012;13(10):4889-4895.
8. Hunt KE, Reichard, KK. Diffuse large B-cell lymphoma. Arch Pathol Lab Med. 2008; 132(1):118-124.
9. Sengar M, Akhade A, Nair R, et al. A retrospective audit of clinicopathological attributes and treatment outcomes of adolescent and young adult non-Hodgkin lymphomas from a tertiary care center. Indian Journal of Medical and Paediatric Oncology. 2011;32:197-203.
10. Jaffe ES. Hematopathology: integration of morphologic features and biologic markers for diagnosis. Mod Pathol. 1999;12(2):109-115.
11. Roy A, Kar R, Basu D, Badhe BA. Spectrum of histopathologic diagnosis of lymph node biopsies: A descriptive study from a tertiary care center in South India over 5½ years. Indian J Pathol Microbiol. 2013;56;103-108.
12. Picker LJ, Weiss LM, Medeiros LJ, Wood GS, Warnke RA. Immunophenotypic criteria for the diagnosis of non-Hodgkin's lymphoma. Am J Pathol. 1987;128(1):181–201.
13. Fang JM, Finn WG, Hussong JW, Goolsby CL, Cubbon AR, Variakojis D. CD10 antigen expression correlates with the t(14;18)(q32;q21) major breakpoint region in diffuse large B-cell lymphoma. Mod Pathol. 1999;12:295–300.
14. Watson P, Wood KM, Lodge A, et al. Monoclonal antibodies recognizing CD5, CD10 and CD23 in formalin-fixed, paraffin embedded tissue: production and assessment of their value in the diagnosis of small B-cell lymphoma. Histopathology. 2000;36(2):145-150.
15. Kurtin PJ, Hobday KS, Ziesmer S, Caron BL. Demonstration of distinct antigenic profiles of small B-cell lymphomas by paraffin section immunohistochemistry. Am J Clin Pathol. 1999;112(3):319-329.
16. Campo E, Miquel R, Krenacs L, Sorbara L, Raffeld M, Jaffe ES. Primary nodal marginal zone lymphomas of splenic and MALT type. Am J Surg Pathol. 1999 Jan;23(1):59-68.
17. Mucci NR, Akdas G, Manely S, Rubin MA. Neuroendocrine expression in metastatic prostatic cancer: evaluation of high throughput tissue microarrays to detect heterogenous protein expression. Hum Pathol. 2000;31(4):406-414.
18. Schraml P, Kononen J, Bubendorf L, et al. Tissue microarrays for gene amplification surveys in many different tumors types. Clin cancer Res. 1999;5(8):1966-75.
19. Richter J, Wagner U, Kononen J, et al. High-throughput tissue microarray analysis of cyclin E gene amplification and overexpression in urinary bladder cancer. Am J Pathol. 2000;157(3):787-794.
Published
2018-10-27
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