Indian Journal of Pathology and Microbiology
Home About us Instructions Submission Subscribe Advertise Contact e-Alerts Login 
Users Online: 73
Print this page  Email this page Bookmark this page Small font sizeDefault font sizeIncrease font size


 
REVIEW ARTICLE Table of Contents   
Year : 2008  |  Volume : 51  |  Issue : 2  |  Page : 161-166
Report of proceedings of the national meeting on "Guidelines for Immunophenotyping of Hematolymphoid Neoplasms by Flow Cytometry"


1 Department of Pathology, Tata Memorial Hospital, Mumbai, India
2 Department of Pathology, Christian Medical College, Vellore, India
3 Department of Hematology, Bombay Hospital, Mumbai, India
4 Department of Hematology, PDJ Hinduja Hospital, Mumbai, India
5 Indian Institute of Immunohematology, KEM Hospital, Mumbai, India
6 Jaslok Hospital, Mumbai, India
7 Ranbaxy India Limited, Mumbai, India

Click here for correspondence address and email
 

   Abstract 

Background: Immunophenotyping of hematolymphoid neoplasms is being done in many laboratories in India. The first national meeting on "Guidelines for Immunophenotyping of Hematolymphoid Neoplasms by Flow Cytometry" was held on 14 March 2008 in Mumbai, India. Aim: To achieve uniformity in the laboratory practice regarding antibody panel selection in diagnosing hematolymphoid neoplasms. Settings and Design: Members of the Inter-Laboratory Comparison Program (ILCP) group in Mumbai prepared a draft regarding immunophenotypic panel selection for acute leukemias (ALs) and chronic lymphoproliferative disorders (CLPDs), which was further circulated among national and international cytometrists, hematopathologists, and oncologists for their written inputs, suggestions, proposed modifications; as well as their indications, if any, of the recommendations not being acceptable. Practice-based questionnaire was circulated among all the participants. Results: Consensus was attained, and the panel recommended the use of a minimal screening panel, followed by a secondary directed panel. The aim of the minimal screening panel would be to provide a diagnosis of all commonly occurring hematolymphoid neoplasms without the need of additional antibodies in most cases. Conclusion: Thus we could attain a consensus for our guidelines in selecting panels for ALs and CLPDs. The guideline is an attempt to formulate a minimal panel for immunophenotyping of hematolymphoid neoplasms. Laboratories are encouraged to add additional antibodies to the above panel to increase the sensitivity; however, they should refrain from immunophenotyping with fewer antibodies. This national guideline hopefully brings about uniformity and comparability in reporting of leukemia and lymphoma and bridges the divide between low-cost reporting and an accurate diagnosis.

Keywords: Flow cytometry, guidelines, immunophenotyping

How to cite this article:
Gujral S, Subramanian P G, Patkar N, Badrinath Y, Kumar A, Tembhare P, Vazifdar A, Khodaiji S, Madkaikar M, Ghosh K, Yargop M, Dasgupta A. Report of proceedings of the national meeting on "Guidelines for Immunophenotyping of Hematolymphoid Neoplasms by Flow Cytometry". Indian J Pathol Microbiol 2008;51:161-6

How to cite this URL:
Gujral S, Subramanian P G, Patkar N, Badrinath Y, Kumar A, Tembhare P, Vazifdar A, Khodaiji S, Madkaikar M, Ghosh K, Yargop M, Dasgupta A. Report of proceedings of the national meeting on "Guidelines for Immunophenotyping of Hematolymphoid Neoplasms by Flow Cytometry". Indian J Pathol Microbiol [serial online] 2008 [cited 2014 Nov 26];51:161-6. Available from: http://www.ijpmonline.org/text.asp?2008/51/2/161/41602



   Introduction Top


WHO classification has divided non-Hodgkin lymphoma into B-cell and T/NK cell subtypes, which are further subclassified into precursor and peripheral lymphomas. [1] Immunophenotyping by flow cytometry (FCM) is an essential aid for accurately diagnosing and prognosticating leukemia and lymphoma. It serves various purposes, mainly identifying and differentiating neoplastic population from normal population; defining number of neoplastic cells with their phenotypes; addressing and giving pertinent answers to, specific questions like remission status, etc. For all the above-mentioned purposes, it requires a strict quality control system in place with standardized methodology and practice, panel of antibodies and their suitable combinations, data acquisition, analysis and reporting and best possible turnaround time. Most of these issues have been addressed in published literature. Antibody panel selection is a critical step in specimen processing and plays a vital role in obtaining an accurate diagnosis. However, there is a great degree of variability on various issues, including panel selection, antibody combinations, fluorochrome and clone selection, gating strategies. There are guidelines that have addressed antibody panels for immunophenotyping of hematolymphoid neoplasms. Majority of these have been formulated by countries in North America and Europe. Diversity of panels is mirrored in these guidelines.

Aim

To propose guidelines for a minimal panel of antibodies to diagnose acute leukemias (ALs) and peripheral lymphomas/chronic lymphoproliferative disorders (CLPDs) by FCM without compromising on accuracy. This guideline should enable uniformity in reporting of hematolymphoid neoplasms.


   Materials And Methods Top


Immunophenotyping of hematolymphoid neoplasms is being done in many laboratories in India. However, no national-level external quality assurance program existed. Two and half years back, 6 clinical cytometry laboratories from Mumbai joined hands and formed a group to start an Inter-Laboratory Comparison Program (ILCP) for immunophenotyping of hematolymphoid neoplasms by FCM. Though started as a small group, the aim was to establish a national-level program (external quality assurance program). ILCP group members selected Hematopathology Laboratory at the Tata Memorial Hospital, Mumbai, as a nodal center for sending samples to participating laboratories. The approval of Institute Review Board (Hospital Scientific Review Committee and Hospital Ethics Committee) was obtained. Peripheral blood samples were sent every 3 months for 2 years. Inputs were followed by formal meetings. It was realized that there was a wide variation among laboratories in various aspects, starting from sample collection to processing, analysis and dispatch of final report. Thus we planned our first national meeting on "Guidelines for Immunophenotyping of Hematolymphoid Neoplasms by Flow Cytometry", with an aim to achieve uniformity in the laboratory practice regarding antibody panel selection in diagnosing hematolymphoid neoplasms.

These guidelines were formulated step by step at various levels. An initial consensus on panel selection was reached by ILCP group in Mumbai. This document was based on the experience of ILCP group members and on the 4-year data (2003-2006) of ALs (2511 cases) and CLPDs (416 cases) from Tata Memorial Hospital, Mumbai. This document was circulated on January 8, 2008, via electronic mail to national and international experts from referral, hospital-based, government-funded and private laboratories. It was circulated for their written inputs, suggestions, proposed modifications; as well as their indications, if any, of the recommendations not being acceptable. Inputs were obtained from cytometrists, hematopathologists, hematologists, medical oncologists and pediatric oncologists from within India and from international experts. Acceptable suggestions were incorporated, while the remaining ones were attached as appendages for further discussion. Revised document (number 2) was re-circulated on February 20, 2008. A practice-based questionnaire was also circulated among all the participants. It included a checklist of antibodies/reagents used in current practice. After incorporating inputs/suggestions, revised document (number 3) was circulated among all the delegates and faculty on March 5, 2008, for discussion in the meeting.

This revised document (number 3) with suggested panel for ALs and CLPDs was presented during the national meeting on "Guidelines for Immunophenotyping of Hematolymphoid Neoplasms by Flow Cytometry" held on March 14, 2008. This meeting was jointly co-organized by Tata Memorial Hospital (a government hospital with Department of Atomic Energy), Indian Institute of Immunohematology - Indian Council of Medical Research, The Cytometry Society (of India), Clinical Cytometry Society (of USA), International Society of Analytical Cytology and Specialty Ranbaxy Limited, India. Corporate sponsorship was obtained from Beckman Coulter, BD Biosciences, Rea Metrix, Abbott Diagnostics and Unique Diagnostics. Distinguished national and international faculties from USA, UK, Portugal, Turkey and Australia were invited. In addition various national hematopathologists, medical oncologists, pediatric oncologists and hematologists were invited. There were altogether 180 participants, including 11 international delegates, from various countries, including Pakistan, Nepal, Sri Lanka, Bangladesh, Kuwait, Doha, Dubai, Portugal, UK and USA (list attached).

Cytometrists from India, rest of Asia, Europe, Australia and America presented their perspective on panel selection for ALs and CLPDs. Consensus was attained during these 2 days and a final draft (revised document number 4) was presented on March, 15, 2008.

The basic idea of this guideline was to reach an agreement on selection of antibody panels for ALs and CLPDs. It was felt that the literature sufficiently addressed other issues like sample collection, transport, viability, adequacy of cell yield and storing of samples. It was agrred in principle to select panels of antibodies as recommended in the literature. [2],[3]

Morphology-based evaluation of a hematolymphoid neoplasm preferably should precede antibody panel selection. Morphological evaluation helps in panel selection, gating strategy and interpretation, as well as validation of FCM data. In rare instances where the morphology is like that of a blast but a large cell lymphoma is suspected, a combination of AL panel and CLPD panel should be used (including Tdt). Morphology, along with clinical information, may help in deciding a panel of antibodies.

There are various factors influencing panel selection for a particular test. It may depend upon type of laboratory, laboratory throughput, technical expertise available, number of antibodies used and other factors like indication for which immunophenotyping is being done, e.g., initial diagnosis, subtyping, follow-up studies and detection of minimal residual disease; or the type of flow instrument, for example, three or more colors, etc.

Our panel has been defined based on the fact that most laboratories use 3 colors for immunophenotyping of hematolymphoid neoplasms.

Various approaches to antibody panel selection are as follows:

  1. General comprehensive panel: This uses an extensive panel of monoclonal antibodies. This approach has the best turnaround time (TAT), saves technical labor and requires minimal judgment from the flow cytometry operator. Laboratories that do not have financial constraints use roughly 40 to 55 monoclonal antibodies per panel. The obvious impediment is the cost involved. [3]
  2. Use of a primary screening panel followed by a secondary directed panel: This approach is more economical; however it uses more time and labor. Also, the pathologist may have to make "real-time" decisions. This panel is not necessarily always cost effective, as an error in deciding the directed antibody panel will require additional antibodies and will also increase the TAT. [3]
  3. Directed approach: Here, a combination of clinical information and morphology is used to suggest a panel of antibodies. This approach, although cost effective, can be impractical at times due to lack of clinical information. At times, morphology can be misleading and subjective. [3]
  4. Indication-based approach: This is quite different from the aforementioned approaches and uses a combination of a lineage-sensitive panel of monoclonal antibodies (B cells, T cells, plasma cells, myeloid and myelomonocytic cells) based upon the clinical presentation and the medical indication rather than the disease in question. Similar approach was used in the 2006 Bethesda International Consensus Conference. [4]


Recommendations

We propose an approach of minimal screening panel followed by directed additional markers. The aim of minimal screening panel is to provide a diagnosis of all commonly occurring hematolymphoid neoplasms without the need of additional antibodies. Laboratory must ensure that adequate cells/sample are/is available for additional markers to avoid re-sampling.

Acute leukemia

Cases which are unequivocally myeloid, for example,  Auer rods More Details seen and/or positive cytochemical stains like myeloperoxidase (MPO) or Sudan black B, may not be processed for FCM. All other subtypes of AL are diagnosed by FCM with the suggested minimal screening panel and cytogenetics (as and when required). In those cases where a diagnosis is not clearly established, additional markers should follow. In cases of aberrant expression of two or more antigens, a biphenotypic acute leukemia should be ruled out using lineage-specific cytoplasmic markers. Panels are decided after doing a meticulous morphological examination of the peripheral blood/bone marrow/fluid smears. Recommended minimal screening panels for immunophenotyping of acute leukemia include CD10, CD19, CD7, CD5, CD13, CD33, CD117, CD34, HLA-DR, CD45. The selection of antibodies in the additional markers may be based on the results of the minimal panel. Panel recommended following antibodies in the additional panel: cCD3, cCD22/cCD79a, anti-MPO (lineage-specific cytoplasmic markers for T, B and myeloid series cells), SIg (K/'λ), Tdt, CD41, CD61, CD56, CD38, CD138, CD64, CD2, CD4, CD8.

If a case is morphologically suggestive of Burkitt's lymphoma, surface immunoglobulin (K/'λ), IgM and Tdt may be added to the primary panel. In cases of pleural effusion or mediastinal mass, cCD3, Tdt, CD4 and CD8 may be added to the primary panel.

Acute biphenotypic leukemia has been defined by the Immunological Characterization of Leukemias (EGIL) group and has been recommended by the WHO. [1],[5] Another criteria for work-up of these neoplasms has been defined by Campana et al. [6] The latter define true mixed-lineage leukemia giving more credence to cytoplasmic expression of highly lineage-specific markers (e.g., cCD22/cCD79a, cCD3 and anti-MPO). Both these criteria have their own limitations. The laboratories can use any of the above two criteria for the diagnosis of biphenotypic acute leukemia.

Chronic lymphoproliferative disorders/Peripheral lymphomas

All suspected cases of peripheral lymphomas/CLPDs involving bone marrow/peripheral blood (e.g., absolute lymphocyte count > 5 × 10 9 /L) should be immunophenotyped. Other cases in appropriate clinical setting may need immunophenotyping; for example, a case of pancytopenia with relative lymphocytosis may require a panel including markers for hairy cell leukemia. A primary minimal panel of 9 antibodies is suggested (CD3, CD5, CD19, CD23, CD10, CD20, FMC7, Kappa, Lambda), which enables us to reach a diagnosis of chronic lymphocytic leukemia (CLL), follicular lymphoma and mantle cell lymphoma. However, aberrant expression of markers is well known in these common hematolymphoid neoplasms. [1],[2],[5] If hairy cell leukemia is suspected clinically and/or morphologically, additional markers (CD25, CD103) must be added in the primary panel. In case a diagnosis is not established, additional markers may be done to rule out other common lymphomas like hairy cell leukemia (CD25, CD103), plasma cell dyscrasia (CD38, CD138, CD56). Other markers may include CD11c, IgM and IgD (for splenic marginal zone lymphoma). If the tumor cells express only CD5 and are negative for CD19, then a T/NK cell neoplasm should be ruled out. Additional markers for CLPDs may include CD45, CD2, CD4, CD7, CD8, TCRα/b, TCR γ/δ, CD25, CD103, cyclin D1, CD43, CD38, CD138, CD16, CD56, CD57, CD11c, IgM and IgD. In cases of primary effusion lymphomas, CD38 and CD138 may be included in the minimal screening panel.

FCM immunophenotyping has become a gold standard in the diagnosis of AL and CLPD. Today this technique is gaining immense acceptance in various clinical laboratories in India.

This document deals with optimal selection of reagents; however, it does not address other issues like selection of clone, combination of markers, methods of analysis, quality control procedures. Most of these are well standardized and laboratory should comply with the published standards. [2] Study of patterns of maturation of the neoplastic cells is not possible in the limited reagent panel recommended. Analyses of myelodysplastic syndrome, minimal residual disease and fine needle aspirate examination of lymph nodes have also not been addressed. It is suggested that clinical indications may be considered for panel selection on a case-to-case basis, in addition to morphology. Laboratories having proficiency in doing cytoplasmic markers may replace one or more of primary panel markers by cytoplasmic markers (cCD22/cCD79a, cCD3, AMPO).

In an ideal setting, CD45 gating (CD45/side scatter histogram) is recommended in every case of AL; however, this considerably adds to the cost. Though it may be left to the laboratory to decide regarding the gating strategy, we recommend that CD45 gating be used in cases of AL/myelodysplastic syndrome where the population of blasts is not predominant (using an arbitrary figure of <50% blasts). For majority of rest of the cases, forward scatter (FSC)/side scatter (SSC) gating shall suffice. In case of CLPDs, a FSC/SSC plot usually suffices; however, when neoplastic cells are scanty, specific gating strategies like CD19, CD3 and CD138 gate may be used for B, T and plasma cell neoplasm respectively.

In addition to the above, there were panel discussions on indications for immunophenotyping, training of cytometrists, stem cell counts and quality control issues. These were oriented to comprise a component of Indian, European, Australian and American perspective. It was agreed that there is a growing need of formal training and education of cytometrists in India as the number of FCM laboratories is increasing at a fast pace. Six-month certified training course and 1-month observer-ship for technologists and pathologists/hematologists were suggested.

Selection of fluorochrome conjugates is important. It is based on the principle that fluorochrome with high-quantum yield must be used if antigen expression is low. [2],[4] For CD13, CD33 and CD19, phycoerythrin (PE) / per chloroperdinin (Per CP) (bright) conjugated antibodies are preferred. Strongly expressed antigens like CD45 and HLADR may be analyzed with more dim conjugates as FITC. Selection of a clone of an antibody is important. Different clones of antibodies which recognize the same antigen may differ in their performance. Combination of antibodies should be selected so as to give maximum information regarding target population.


   Discussion Top


Acute leukemia

Most of the guidelines published have found difficulty in reaching a consensus for an ideal panel required for the diagnosis of AL. These guidelines have recommended panels for the diagnosis of AL and CLPD. An algorithm can be devised in CLPD to reach a diagnosis. However, this practice does not hold true for AL. This is due to antigenic heterogeneity and inconsistent antigen expression shown by the leukemic blasts, e.g., CD2 expression in AMLM3 and loss of CD7 expression in cases of T-cell lymphomas. So an increase in antibodies, at least till a certain level, enables an accurate diagnosis. This statement, however, holds true only till a certain point. Surveys of clinical laboratories in the United States revealed that laboratories, on an average, usually utilized 16 to 19 antibodies, a figure ranging from 5 to 45 antibodies. [7],[8],[9] The US-Canadian consensus (1997) recommended that a diagnostic panel should be divided in two parts: an initial core panel containing a small number of reagents designed to identify the type of disease (leukemia, lymphoma, etc) and the predominant cell lineage in a population, followed by a supplemental panel. [3] It recommended that CD2, CD5, CD7, CD10, CD13, CD14, CD19, CD33, CD34, HLA-DR and kappa/lambda be added to the core panel. At the ISAC Consensus Meet in 2000, a questionnaire-based consensus was reached amongst US and European experts. [9] It recommended that CD10, CD19, CD33, CD34, CD45, CD7, CD14, CD3 and HLA-DR be used for the initial characterization of leukemia, along with at least one antibody of B-cell (CD20, CD79a, CD22, IgM), T-cell (CD1a, CD2, CD4, CD5, CD8), myeloid (CD11b, CD15, CD64, CD117, MPO), erythroid (CD36, CD71, glycophorin-A) and megakaryocytic (CD41, CD61) lineage. The General Hematology Task Force of British Committee for Standards in Hematology (2002) recommended that cCD22, cCD79a, CD19, CD10, cCD3, CD2, CD117, CD13, anti-MPO and Tdt be added to the first-line panel. [10] The second Latin American consensus guideline (2005) recommended that a large panel of antibodies compromising of cCD3, CD2, CD7 (T lineage), CD19, CD79a, CD10, surface Ig, cytoplasmic K/'λ (B lineage), anti-MPO, CD13, CD33, CD117 and CD15 (myeloid lineage), along with HLA-DR, CD45, Tdt and CD34, be added to their panel. [11]

A review of all these guidelines demonstrates that only CD10 and CD19 are common to all the 5 primary panels. This highlights the diversity in panel selection for ALs. Furthermore, many antigens initially considered specific for a lineage showed expression in other lineages, e.g., NK cells expressing CD2; myeloid blasts expressing CD7, CD19 and CD56. Most of these guidelines have tried to avoid cytoplasmic markers in the primary panel as it is more cumbersome to do them in routine practice and it also affects the light scatter patterns.

The British Committee for Standards in Haematology (BCSH) guideline by Bain et al. [11] has recommended that immunophenotyping is not essential if the lineage is unequivocally myeloid based on morphology and cytochemistry. Many of our patients cannot afford standard treatment for AML. Hence it was agreed that there was no purpose gained by immunophenotyping all MPO-positive AMLs. If the patient intends to be put on a standard treatment protocol, it is recommended to immunophenotype the disease at diagnosis. However, it is essential to immunophenotype all MPO-negative ALs. In other words, all cases of ALL; and other subtypes of AML, including AMLM0 and AMLM7 and other rare subtypes like biphenotypic acute leukemia, are diagnosed by FCM only.

Consensus was to add CD117, in addition to CD13 and CD33 (myeloid markers), in the minimal panel as it would reduce the number of cases where additional processing is required. Thus there was an addition of 3 more antibodies (CD117, CD7 and CD5), while 1 antibody (CD3) was removed from the minimal panel of our first proposed document. This panel of 10 antibodies covers most of the common ALs, including AML, B-ALL and T-ALL. This final panel of 10 antibodies as a minimal panel for ALs was accepted in the consensus guideline meeting with a caution to add additional markers in those rare instances where a diagnosis or subtyping could not be achieved. Morphology and clinical details are taken into account to decide the primary panel. Addition of CD4, CD8 and Tdt (especially in pleural fluids) helps diagnose T-cell lymphoblastic lymphoma. Few of the international cytometrists emphasized the importance of maturation myeloid markers, including CD11b, CD15 and CD64.

Unlike all the aforementioned guidelines, which use a panel of antibodies to diagnose a specific disease condition, e.g., AL or CLPD, 2006 Bethesda international consensus guidelines use a panel of antibodies that are sensitive to pick up cells of a particular lineage. [4] It defines a consensus reagent panel suitable for both AL and CLPD for the initial evaluation of hematolymphoid neoplasm based on medical indications. [12] A combination of these markers (for example B-cell, T cell and myeloid markers) is used for a particular medical indication or symptom (for example, lymphadenopathy or blasts in the blood). This guideline recommended that an initial evaluation panel comprising of B cell (CD5, CD10, CD19, CD20, CD45, kappa, lambda), T/NK cells (CD2, CD3, CD4, CD5, CD7, CD8, CD45, CD56), myelomonocytic cells (CD7, CD11b, CD13, CD14, CD15, CD16, CD33, CD34, CD45, CD56, CD117 and HLA-DR) and plasma cells (CD19, CD38, CD45 and CD56) be used. This was followed by a more extensive lineage-specific panel of antibodies, depending upon the results of the primary panel. [4]

Chronic lymphoproliferative disorder

At the ISAC Consensus Meet (2000), 75% of the participants agreed upon CD5, CD10, CD19, CD23, CD45, CD20, kappa and lambda (8 antibodies) to be used for the initial work-up of all CLPDs. [9] They concluded that depending upon the results of this panel, additional markers may be required. The US-Canadian consensus (1997) recommendations recommended that CD3, CD4, CD5, CD7, CD8, CD10, CD19, CD20, CD45, kappa and lambda (11 antibodies) be added to the core panel. It also recommended a separate panel of 8 antibodies for body fluids (CD5, CD10, CD19, CD20, CD23, CD45, kappa and lambda). [3] Depending upon the results of the core panel, it recommended a large number of antibodies in the supplemental panel. The General Hematology Task Force of British Committee for Standards in Hematology (2002) recommended that CD2, CD5, CD19, CD22, CD23, FMC-7, CD79b, kappa and lambda (9 antibodies) be added to the first-line panel. [10] This group also relied on a scoring system for forming an algorithm for diagnosis of CLPD. The second Latin American consensus guideline (2005) recommended an initial screening panel comprising of CD3, CD4, CD8, CD19, CD56 and kappa/lambda light chains (7 antibodies) followed by a more specific panel, depending on the results of the primary panel. [11]

B-cell NHL comprises an overwhelming majority of NHLs. This is the rationale for selecting B-cell-centric minimal panels for CLPD. CLPD panels were initially meant for characterizing chronic lymphocytic leukemia (CLL) differentiating it from non-CLL. We initially proposed a primary panel comprising of 8 antibodies. An additional T-cell marker CD3 was suggested for the minimal panel so as to not miss the rare T-cell lymphomas, thereby making it a panel of a total of 9 antibodies. This will enable a diagnosis of CLL, mantle cell and follicular lymphoma, three of the common CLPDs. If a result is not obtained from the primary panel, then additional antibodies are required. Subtyping of rare T/NK/ plasma cell neoplasms requires additional antibodies. Though the CLPD panel is recommended for lymphocytosis, the same may be used to subtype lymphomas of large cell morphology which can be mistaken as blasts. The common lymphomas involving body cavity fluids are T-cell lymphoblastic lymphoma, diffuse large B cell lymphoma, Burkitt's lymphoma and plasmablastic lymphoma. The panel may be modified accordingly on a case-to-case basis.

Thus a final panel with 9 antibodies in the minimal panel was accepted in the consensus guideline meeting. Additional antibodies may be used in those rare instances where a diagnosis or subtyping could not be achieved. Cases with morphological/clinical suspicion of HCL are dealt with accordingly. Phenotypic heterogeneity of B cells in patients with CLPD is well known. [13] Additional markers may be employed in those cases where diagnosis is not reached. Use of cyclinD1 is not popular amongst American cytometrists; however, CD43 is extremely popular in CLPD panel amongst European cytometrists.


   Conclusion Top


Thus we could attain a consensus in formulating guidelines in selecting panels for ALs and CLPDs. Laboratories are encouraged to add additional antibodies to minimal primary panel to increase the sensitivity; however, they should refrain from immunophenotyping with fewer antibodies. This guideline hopefully brings about uniformity and comparability in reporting of leukemia and lymphoma and bridges the divide between low-cost reporting and an accurate diagnosis. To summarize, this effort is the first of its kind - to lay down a minimum standard for the reporting of hematolymphoid neoplasms in India. Any changes in practice based on this guideline may be monitored by our proposed external quality assurance program.


   Acknowledgments Top


List of participants

Faculty

Brent Wood, Michael Borowitz, Phil McCoy, Dario Campana, Elaine Coustan-Smith and Awtar Krishan (USA); Estella Matutes (UK); Maria Arroz (Portugal); Mary Sartor (Australia); Martin Adelmann (Switzerland); Gulderen Demirel (Turkey); Anita Borges, Kanjaksha Ghosh, H. Krishnamurthy, M. B. Agarwal, Rajive Kumar, Swati Pai, Hari Menon, Sudeep Gupta, Ashish Bakshi, Sripad Banavali, Shenaz Khodaiji, Amar Dasgupta, Badrinath Y., Ashok Kumar, Subramanian P. G. and Sumeet Gujral (India).

Delegates

Nikhil Patkar, Prashant Tembhare, Sophia George, Prashant Khadse, Archana Vazifdar, Kunal Sehgal, Sunita Pollampalli, Kiran Thorat, Vishal Raj, Purvish Parikh, Suresh Pai, Reena Nair, Prabhash Kumar, Brijesh Arora, Manju Shengar, Mary Ann Muckden, Siddharth Lashkar, Tanuja Shet, Sridar Epari, Nirmala Jambhekar, Shubada Kane, Pratibha Amre, Chandrika Nair, Sandeep Dighe, Sharmila Ghosh, Mita Shah, Hemanti Buch, Lourenco Maria Enna, Dimple Bhatia, Charusmitha Modi, Dhanashree Rahalkar, Jyoti Bajaj, Peter Ubaldo, Sharmila Barve, Shaila Shinde, Anuradha Chougule, Shilpa Kushte, Sitaram Ghogle, Sangeeta Masurkar, Sujatha Kurane, Shashikant Channavar, Saurabh Malpani, Jyoti Kode, Usha Agarwal, Mamta Yargop, Manisha Madkaikar, Sujatha N., Bijal Kulkarni, C. S. Soman, Farah Jijina, Zinat Currimbhoy and Rekha Gaur (Mumbai); Biren Parikh, Bipin Patel, Hemagini Vora and Dinesh Rathod (Ahmedabad); Aruna Jain and Suhas Kothari (Bhopal); Jammie Narasimhan, Sridhar Ramanathan, Karuna Rameshkumar, Mohamed Aslam, Ebinezer and Jayashree Kulkarni (Bangalore); Jasmina Ahluwalia and Anita Tahlan (Chandigarh); Sreedevi Dantala and Gaurav Rastogi (Secunderabad); Manu Goyal and Gayathri K. (Hyderabad); Arpita Jindal (Jaipur); Debasis Banerjee and Arnab Chattopadhaya (Kolkata); Shano Naseem, Vikas Shrivastava, Sanjay Mehrotra and Anil Maurya (Lucknow); Maneesh Bagai (Mohali); Alpana Srivastava, Anita Chopra, Nivedita Pathak, Hara Pati, Dipti Kalita, Manish Singhal, Sobuhi Iqbal, Dinesh Pendarkar, Sumant Gupta, Maheswar Sahoo, Tejinder Singh, Aruna Rangan, Dipanjan Panda, Monica Sharma, Sumita Saluja, Naveen Kumar Agarwal, Gurudeep Buxi, Neelu Chabbara, Neeraj Prakash, Deepak Mishra, Kausambhi Shukla, Paresh Jain, Arvinder Singh, Manorama Bhargava and Anil Handoo (New Delhi); Deepak Sadwani (Noida); Prasad Shenoy (Panaji); Ravindra Karle (Pravara); Neeti Jalnapurkar, Vinanti Golwilkar, Pawan Gupta and Ravi Shankar Sarkar (Pune); Prashant Naik (Surat); Rekha Nair and Nileena Nayak (Trivandrum); Shahid Pervez, Naila Kayani and Sowlat Naqvi (Pakistan); Yunus (Bangladesh); Dina Al-Salafi (Dubai); Kanchan Jeswani and Malini Chawla (Kuwait); Suresh Venugopal (Sultanate of Oman); Baddika Jayaratne (Sri Lanka); Muffaddal Moonim (UK); Suchitra Swaminathan (US).

 
   References Top

1.World Health Organization classification of tumors. In : Jaffe ES, Harris NL, Stein H, Vardiman JW, editors. Pathology and genetics of tumors of haematopoietic and lymphoid tissues. Lyons, France: IARC Press; 2001. p. 75-118.  Back to cited text no. 1    
2.Clinical and laboratory standards institute. Clinical flow cytometric analysis of neoplastic hematolymphoid cells: Approved guideline - 2 nd ed. CLSI document H43-A2 [ISBN 1-56238-000-0]. Wayne, Pennsylvania: Clinical and Laboratory Standards Institute; 2006. p. 19087-1898.  Back to cited text no. 2    
3.Stewart CC, Behm FG, Carey JL, Cornbleet J, Duque RE, Hudnall SD, et al. US-Canadian consensus recommendations on the immunophenotypic analysis of hematologic neoplasia by flow cytometry: Selection of antibody combinations. Cytometry 1997;30:231-5.  Back to cited text no. 3  [PUBMED]  [FULLTEXT]
4.Wood BL, Arroz M, Barnett D, DiGiuseppe J, Greig B, Kussick SJ, et al. 2006 Bethesda international consensus recommendations on the immunophenotypic analysis of hematolymphoid neoplasia by flow cytometry: Optimal reagents and reporting for the flow cytometric diagnosis of hematopoietic neoplasia. Cytometry 2007;72:S14-22.  Back to cited text no. 4  [PUBMED]  [FULLTEXT]
5.Bene MC, Castoldi G, Knapp W, Ludwig WD, Matutes E, Orfao A, et al. Proposals for the immunological classification of acute leukemias: European Group for the Immunological characterization of Leukemias (EGIL). Leukemia 1995;9:1783-6.  Back to cited text no. 5  [PUBMED]  
6.Campana D, Behm FG. Immunophenotyping of leukemia. J Immunol Met 2000;243:59-75.  Back to cited text no. 6    
7.Hasset J, Parker J. Laboratory practices in reporting flow cytometery phenotyping results for leukemia/lemphoma specimens: Results of a survey. Cytometry 1995;22:264-81.  Back to cited text no. 7    
8.McCoy JP Jr, Overton WR. A survey of current practices in clinical flow cytometry. Am J Clin Pathol 1996;106:82-6.  Back to cited text no. 8  [PUBMED]  
9.Braylan RC, Orfao A, Borowitz MJ, Davis BH. Optimal number of reagents required to evaluate hematolymphoid neoplasias: Results of an international consensus meeting. Cytometry 2001;46:23-7.  Back to cited text no. 9  [PUBMED]  [FULLTEXT]
10.Bain BJ, Barnett D, Linch D, Matutes E, Reilly JT. General haematology task force of the british committee for standards in haematology (BCSH), British Society of Haematology. Revised guideline on immunophenotyping in acute leukemia and chronic lymphoproliferative disorders. Clin Lab Hematol 2002;24:1-13.  Back to cited text no. 10    
11.Ruiz-Arguelles A, Rivadeneyra-Espinoza L, Duque RE, Orfao A. Report on the second Latin American consensus conference for flow cytometric immunophenotyping of hematological malignancies. Cytometry 2005;70:39-44.  Back to cited text no. 11    
12.Davis BH, Holden JT, Bene MC, Borowitz MJ, Braylan RC, Cornfield D, et al. 2006 Bethesda International Consensus Recommendations on the flow cytometric immunophenotypic analysis of hematolymphoid neoplasms: Medical Indications. Cytometry 2007;72:S5-13.  Back to cited text no. 12  [PUBMED]  [FULLTEXT]
13.Almeida J, Vidriales B, López-Berges MC, Garcνa-Marcos MA, Moro MJ, Corrales A, et al. Incidence of phenotypic aberrations in a series of 467 patients with B chronic lymphoproliferative disorders: Basis for the design of specific four-color stainings to be used for minimal residual disease investigation. Leukemia 2002;16:1460-9  Back to cited text no. 13    

Top
Correspondence Address:
Sumeet Gujral
Department of Pathology, Tata Memorial Hospital, Mumbai
India
Login to access the Email id


DOI: 10.4103/0377-4929.41602

PMID: 18603672

Get Permissions




This article has been cited by
1 NSE/aNAE positivity in B-lineage acute lymphoblastic leukemia: revisiting a potential cytochemical diagnostic pitfall
P Sharma,S Tyagi
Biotechnic & Histochemistry. 2013; : 1
[Pubmed]
2 Flow Cytometry in Hematological Disorders
Hara Prasad Pati,Sonal Jain
The Indian Journal of Pediatrics. 2013; 80(9): 772
[Pubmed]
3 Immunophenotyping of mature T/NK cell neoplasm presenting as leukemia
Gujral, S., Polampalli, S., Badrinath, Y., Kumar, A., Subramanian, P., Nair, R., Sengar, M., Nair, C.
Indian Journal of Cancer. 2010; 47(2): 189-193
[Pubmed]
4 Diversity amongst various guidelines for immunophenotyping
Sumeet Gujral,P.G. Subramanian,A. Dasgupta
Cytometry Part A. 2009; 75A(7): 560
[Pubmed]
5 Study of diagnostic and prognostic parameters (including flow cytometry) in chronic lymphoproliferative disorders: The Indian perspective
Kumar, N., Varma, N., Varma, S., Malhotra, P.
Analytical and Quantitative Cytology and Histology. 2009; 31(5): 296-303
[Pubmed]
6 Diversity amongst various guidelines for immunophenotyping
Gujral, S., Subramanian, P.G., Dasgupta, A.
Cytometry Part A. 2009; 75(7): 560-561
[Pubmed]
7 Hypergranular precursor B-cell acute lymphoblastic leukemia in a 16-year-old boy
Tembhare, P.R., Subramanian, P.G., Sehgal, K., Yajamanam, B., Kumar, A., Gujral, S.
Indian Journal of Pathology and Microbiology. 2009; 52(3): 421-423
[Pubmed]
8 Immunophenotyping of mature B-cell non Hodgkin lymphoma involving bone marrow and peripheral blood: critical analysis and insights gained at a tertiary care cancer hospital
Sumeet Gujral, Sunita Narayan Polampalli, Y. Badrinath, Ashok Kumar, Subramanian P. G., Reena Nair, Sudeep Gupta, Manju Sengar, Chandralekha Nair
Leukemia and Lymphoma. 2009; 50(8): 1290
[VIEW]



 

Top
 
  Search
 
  
  
    Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
    Email Alert *
    Add to My List *
* Registration required (free)  


    Abstract
    Introduction
    Materials And Me...
    Discussion
    Conclusion
    Acknowledgments
    References

 Article Access Statistics
    Viewed7049    
    Printed167    
    Emailed6    
    PDF Downloaded961    
    Comments [Add]    
    Cited by others 8    

Recommend this journal