Asian Journal of Transfusion Science
Home About Journal Editorial Board Search Current Issue Ahead of print Back Issues Instructions Subscribe Login  Users: 146 Print this page  Email this page Small font sizeDefault font sizeIncrease font size 


 
REVIEW ARTICLE Table of Contents   
Year : 2022  |  Volume : 16  |  Issue : 2  |  Page : 159-166
Application of flow cytometry in transfusion medicine: The Sanjay Gandhi Post Graduate Institute of Medical Sciences, India experience


1 Department of Transfusion Medicine, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, Uttar Pradesh, India
2 Department of Transfusion Medicine, Apollo Multispeciality Hospitals, Kolkata, West Bengal, India

Click here for correspondence address and email

Date of Submission20-May-2022
Date of Decision30-May-2022
Date of Acceptance05-Jun-2022
Date of Web Publication28-Sep-2022
 

   Abstract 

The application of flow cytometry (FC) is diverse and this powerful tool in used in multiple disciplines such as molecular biology, immunology, cancer biology, virology, and infectious disease screening. FC analyzes a single cell or a particle very rapidly as they flow past single or multiple lasers while suspended in buffered solution. FC has a great impact in the field of transfusion medicine (TM) due to its ability to analyze individual cell population and cell epitopes by sensitive, reproducible, and objective methodologies. The main uses of FC in TM are detection of fetomaternal hemorrhage, diagnosis of paroxysmal nocturnal hemoglobinuria, quantification of D antigen, detection of platelet antibody, quality control of blood components, for example, residual leukocyte counts and evaluation of CD34-positive hematopoietic progenitor cells in stem cell grafts. In recent years, FC has been implemented as an alternative method for the detection and characterization of red cell autoantibodies in autoimmune hemolytic anemia. Many workers considered FC as a very good complement when aberrant expression of various erythrocyte antigens needs to be elucidated. It has been extensively used in the resolution of ABO discrepancies and chimerism study. FC has also been used successfully in various platelet immunological studies. In the recent past, FC has been used in several studies to assess the platelet storage lesions and elucidate granulocyte/monocyte integrity and immunology. FC analysis of CD34+ stem cells is now the method of choice to determine the dosage of the collected progenitor cells. The technique is vastly used to evaluate residual leukocytes in leukodepleted blood components. We conclude that flow cytometers are becoming smaller, cheaper, and more user-friendly and are available in many routine laboratories. FC represents a highly innovative technique for many common diagnostic and scientific fields in TM. Finally, it is the tool of choice to develop and optimize new cellular and immunotherapeutic trials.

Keywords: Coomb's negative autoimmune hemolytic anemia, fetomaternal hemorrhage, flow cytometry, platelet antibody, transfusion medicine

How to cite this article:
Chaudhary R, Das SS. Application of flow cytometry in transfusion medicine: The Sanjay Gandhi Post Graduate Institute of Medical Sciences, India experience. Asian J Transfus Sci 2022;16:159-66

How to cite this URL:
Chaudhary R, Das SS. Application of flow cytometry in transfusion medicine: The Sanjay Gandhi Post Graduate Institute of Medical Sciences, India experience. Asian J Transfus Sci [serial online] 2022 [cited 2022 Dec 5];16:159-66. Available from: https://www.ajts.org/text.asp?2022/16/2/159/356892



   Introduction Top


The application of flow cytometry (FC) is diverse and this powerful tool in used in multiple disciplines today. Important disciplines include molecular biology, immunology, cancer biology, virology, and infectious disease screening. FC which primarily consists of the three systems, namely fluidics, optics, and electronics is an advanced technology that analyzes a single cell or a particle very rapidly as they flow past single or multiple lasers while suspended in buffered solution [Figure 1].[1],[2],[3] Each particle is analyzed for visible light scatter and one or multiple fluorescence parameters. Visible light scatter is measured in two different directions, the forward direction or the Forward Scatter which measures the relative size of the cell and at 90° or the side scatter which indicates the internal complexity or granularity of the cell. Importantly, the samples under investigation are prepared for fluorescence measurement through transfection and expression of fluorescent proteins, staining with fluorescent dyes, or staining with fluorescently conjugated antibodies.[2],[3],[4],[5] Importantly, the FC allows simultaneous characterization of mixed populations of cells from blood and bone marrow as well as solid tissues that can be dissociated into single cells such as lymph nodes, spleen, mucosal tissues, and solid tumors. In addition to the analysis of populations of cells, a major application of FC is sorting cells for further analysis. The data in FC are analyzed conventionally using the two-parameter histogram or dot plot gating; however with the advancement in instrumentation advanced data analysis tools are applied and explored today.[4],[5],[6],[7]
Figure 1: A schematic diagram of a flow cytometer

Click here to view



   Flow Cytometry in Transfusion Medicine Top


Today transfusion medicine (TM) is a multidisciplinary specialty with various recent developments that continue to challenge immunohematologists. Definitely, FC has a great impact in the field of TM due to its ability to analyze individual cell population and cell epitopes by sensitive, reproducible, and objective methodologies.[8]

Through the years FC has been used to examine red blood cells (RBC) in different immunohematology settings; although it has never completely found its way into the routine immunohematology laboratories. Today, the main use of FC analysis in TM is for the diagnosis of the direct antiglobulin test (DAT) negative autoimmune hemolytic anemia (AIHA), detection of fetomaternal hemorrhage (FMH), diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), quantification of D antigen, detection of platelet antibody, quality control of blood components, for example, residual leukocyte counts, elucidating aberrant antigen expression, and red cell phenotyping.[9],[10]

As cellular therapies are now a part of TM in many places, therefore, FC is widely used in the hematopoietic stem cell transplantation setting for analyzing the content of CD34 positive hematopoietic progenitor cells in stem cell grafts and to measure T-cell content before allogeneic transplantations.[9],[10],[11]

The Department of TM at the Sanjay Gandhi Post Graduate Institute of Medical Sciences (SGPGIMS), Lucknow, India, is an academic facility with a fully-fledged immunohematology laboratory. The laboratory handles numerous requests and samples for FC for the diagnosis of DAT-negative AIHA and PNH; the detection of FMH and platelet antibody and quantification of D antigen. Few of these FC applications performed in the immunohematology laboratory are discussed below.

Flow cytometry in the diagnosis of direct antiglobulin test negative autoimmune hemolytic anemia

In recent years, FC has been implemented as an alternative method for the detection and characterization of red cell autoantibodies in AIHA. Garraty and Arndt cited FC to be a highly sensitive antibody detection technique with the ability to detect as low as 30–40 molecules per RBC.[12] Chaudhary et al. from SGPGIMS, India investigated that AIHA patients with negative DAT results by conventional tube technique or column agglutination techniques (CAT) can be easily diagnosed using FC. Authors commented that FC is a very useful tool in assessing “Coomb's negative AIHA” and should be employed when other DAT methods give discordant results and there is a strong clinical suspicion of AIHA. However to rule out false-positive results in FC due to its extreme sensitivity proper positive and negative controls have to be executed during running test samples in FC for diagnosis of AIHA [Figure 2].[13] In another study, FC was used to confirm the result of CAT in the determination of immunoglobulin G (IgG) subclass and titer. The authors concluded that the advantage of FC allows it to diagnose the so-called “Coomb's negative” AIHA which at times are missed by the CAT technique as it requires approximately 200 molecules per red cells for a positive DAT.[14]
Figure 2: Flow cytometry in the diagnosis of Coomb's negative autoimmune hemolytic anemia

Click here to view


Flow cytometry in fetomaternal hemorrhage

An adequate dose calculation of anti-D Ig in any sensitized mother is possible through the estimation of correct FMH volume. In this regard, the Kleihauer Betke test or acid elution test is the most commonly used one, but it is time-consuming, difficult and not easily amenable to standardization.[15],[16] These factors led many immunohematology laboratories to adopt the new methods of FMH quantifications of which FC has been designated as the most sensitive, reproducible, and rapid technique.[17],[18],[19] Two FC techniques such as the indirect immunofluorescence technique and direct immunofluorscence technique are advocated to determine FMH. While the former technique is based on the labeling of RhD antigen with an anti-D reagent followed by the addition of preconjugated IgG antibody as described previously; the later uses fluorescein isothiocyanate labeled anti-CD45 monoclonal antibody and phycoerythrin (PE) labeled anti-fetal haemoglobin (HbF) antibody to detect HbF.[15] Agarwal et al. from SGPGIMS, India, found both techniques sensitive overall sample dilutions tested. They observed FC as the most sensitive technique that could detect RhD-positive/HbF-positive events accurately even at a cell concentration of 0.06% which at times other techniques fail [Figure 3].[20] It was Nance et al. in 1989 who first reported FC as a simple technique to quantitate FMH.[21] Almost all authors who worked on FC and FMH found the technique readily adaptable to laboratories with a high degree of accuracy and reproducibility.[17],[22] Agarwal et al. also concluded that detection of FMH volume is an important step in the management of maternal RhD iso-immunization, therefore, each laboratory should adopt technique, which is simple, rapid, sensitive, reproducible, and importantly economically affordable.[20]
Figure 3: Flow cytometric analysis of fetomaternal hemorrhage

Click here to view


Flow cytometry in paroxysmal nocturnal hemoglobinuria

PNH is an acquired clonal stem cell disorder, resulting in intravascular hemolysis, cytopenia of variable degree, and thrombotic events. The hallmark of PNH blood cells is a deficiency or absence of proteins that utilize the glycosylphosphatidylinositol (GPI) anchor for their attachment to the plasma membrane. The absence of certain GPI-anchored complement regulatory proteins, such as CD55 and CD59, accounts for the complement-mediated hemolysis that characterizes PNH. FC can detect cells with absent or reduced expression of GPI-anchored proteins on cell surfaces. FC immunophenotyping can discriminate between cell populations with differential expression or the absence of one or more GPI-anchored proteins on erythrocytes, leukocytes, and platelets. In most cases, the technique uses PE-conjugated monoclonal antibodies (MoAb), namely CD55 and CD59 [Figure 4].[23],[24],[25]
Figure 4: Flow cytometry in paroxysmal nocturnal hemoglobinuria

Click here to view


Gupta et al. from SGPGIMS compared four techniques for the diagnosis of PNH and investigated that small PNH clones (≤1%), differential expression of GPI-anchored proteins, and defective leukocytes could be easily identified by FC using MoAb to GPI-anchored proteins [Figure 4].[23] Other workers on PNH also commented that the use of sensitive techniques like FC enables the detection of a small PNH clone, which does not produce symptoms or signs of hemolysis, in a variety of clonal hematological disorders, such as aplastic anemia, myelodysplasia, myeloproliferative, and lymphoproliferative disorders as well as in normals.[24],[25],[26] Kashyap et al. also concluded that PNH is not a rare disorder in the Indian subcontinents; however, a high index of clinical suspicions and evaluation of erythrocytes, granulocytes, and monocyte by FC is necessary for its detection to understand the PNH pathophysiology more closely.[27]

Flow cytometry in the quantification of D antigen

FC is considered as a very good complement when aberrant expression of various erythrocyte antigens needs to be elucidated. The use of FC for the analysis of protein/glycoprotein in blood group antigens/systems such as RH and KEL has been discussed.[28],[29],[30],[31]

Genes of the Rhesus (Rh) blood group system, namely RHD and RHCE encode for the D, C, c, and E, e antigens, of which D antigen is the most immunogenic, followed by c and E. It has been observed that as low as 0.1–1 ml of D-positive red cells can induce anti-D formation in D-negative recipients. This may cause severe transfusion reactions and hemolytic disease of the fetus and newborn.[32] The strength of antigenic determinants present on the surface of red cells is one of the factors which determine the immunogenicity of an antigen.[33] The surface of normal RhD-positive red cells expresses about 10,000–30,000 D antigens per cell while weak D red cells have antigen densities between 70 and 4000 D antigens per cell.[34] Red cells with <30 D antigen sites have now been shown to induce alloimmunization in RhD-negative individuals.[35],[36],[37] Several studies on the quantification of D antigens have been reported in normal D positive, weak D and partial D cases, and various Rh phenotypes.[38],[39],[40],[41].

Various quantitative methods have been used for the estimation of D antigenic sites on red cells including the sensitive FC. FC is a valuable tool to study D antigens on red cell surfaces, as it is a rapid, reliable, and efficient method and can detect very small amounts of D antigens.[41]

Using FC, van Bockstaele et al. demonstrated the differences in the D antigen density of various Rh phenotypes but showed that Weak D (Du) was not differentiated from D negative (cde/cde).[42] In an unpublished thesis work by Verma et al. from SGPGIMS it was possible to distinguish between Weak D and D negative antigen densities. The mean channel fluorescence (MCF) of D negative was 5.08 in contrast to the MCF of Weak D which was 15.27 (p< 0.001) [Figure 5]. The D antigenic sites (MCF) in weak D in their study were much higher compared to other studies.[43] However, the result of Verma et al. was found consistent with those obtained by Cunningham et al. who analyzed the binding of polyclonal radiolabelled anti-D antibodies and showed that the number of antigenic sites differed by a factor of 10–15 between the D positive and the weak D (Du) red cells.[44] In the study by Verma et al. the MCF of D positive red cells was 108.06 in contrast to 15.27 in weak D. They could not actually calculate the density of D antigens as it required standard red cells with a known number of D antigens. They have calculated the MCF as an indirect indicator of D antigen density. This approach has also been used in previous studies.[43],[45] Most authors concluded that evaluation by FC is less time-consuming, does not need radiolabeling, and could also be applied for routine analyses.[42],[43],[44],[45]
Figure 5: Flow cytometry in D antigen detection

Click here to view


Flow cytometry in platelet antibody detection and platelet immunology

More than 80% of platelet refractoriness (PR) are due to nonimmune causes such as splenomegaly, fever, sepsis, antibiotics, and disseminated intravascular coagulation.[46] Immune causes, occurring in <20% of the cases, involve alloimmunization against human leukocyte antigens (HLA) and, to a lesser extent, human platelet antigens (HPA) following exposure through transfusion, pregnancy, or transplantation. Among the immune causes, HLA antibodies are responsible for approximately 80%–90% of PR and HPA antibodies for approximately 10%–20% of cases.[47]

The immune causes of PR can be diagnosed by laboratory tests and testing for HPA antibodies is technically demanding. The platelet antibody detection tests available include microcytotoxicity and the platelet immunofluorescence test (PIFT) either by microscopy or FC. PIFT is the gold standard technique that permits the identification and quantification of platelet-specific antibodies however this technique is very laborious and time-consuming.[48],[49],[50] This typical assay involves incubating patient's serum with intact blood Group O platelets, followed by washing away unbound antibody and detection of platelet-bound antibodies with fluorescent-labeled anti-human IgG or immunoglobulin M (IgM) reagents by FC [Figure 6].[51]
Figure 6: Flow cytometry in the detection of platelet antibodies

Click here to view


Bub et al. found the FC-based PIFT to be efficient, fast, and feasible as an initial screening to detect platelet antibodies and a useful tool to crossmatch platelets for the transfusional support of patients with refractoriness. They investigated that the sensitivity of FC-PIFT was 86.11% and the specificity was 75.00% with a positive predictive value of 75.61% and a negative predictive value of 85.71%. The accuracy of the method was 80.26%.[52] FC has also been used successfully to detect autoantibodies in a direct assay of the patient's platelets.[53] Platelet autoantibodies testing in immune thrombocytopenia (ITP) can discriminate acute from chronic forms of the disease and is helpful in follow-up of patients. Determination of platelet-associated Ig (PAIg) like PAIgM in the combination of PAIgG has been found to be of interest in the investigation of ITP. Aref et al. concluded that FC is a sensitive method of the detection of platelet autoantibodies that could be used in the screening of suspected ITP [Figure 7].[54]
Figure 7: Flow cytometry in acute ITP = Immune thrombocytopenia

Click here to view


Regarding HLA antibody screening, Buakaewand Promwong observed that the FC technique detected antibodies more frequently than the lymphocytotoxicity test (LCT) and solid-phase red cell adherence assay (SPRCA). In addition, the FC technique was found to be quick, required only a small amount of sample and was easy to perform and simple to interpret. They found the FC technique to be suitable for routine investigations of alloimmune causes of platelet transfusion refractoriness including neonatal alloimmune thrombocytopenia, post-transfusion purpura, and HLA antibody screening in a potential organ transplant recipient.[55]

Freedman and Hornstein compared LCT and SPRCA with the FC technique in platelet crossmatching and found that the FC technique had the best sensitivity and specificity.[56] Köhler et al. reported that sensitivity/specificity of FC technique was 94.7% and 96.3%, respectively, when monoclonal antibody-specific immobilization of platelet antigens (MAIPA) was taken as a reference indicating that sensitivity/specificity of FC is approaching that by the MAIPA assay.[57] This is a reason why most studies used FC as the reference method in the investigation of platelet antibodies.[57]

In addition, heparin-induced thrombocytopenia (HIT) can be diagnosed by flow cytometric detection of antiheparin/PF4 antibodies. Several publications have evaluated this method for the diagnosis of HIT.[58],[59],[60]


   Conclusion Top


We conclude that today FC is used in both routine and research laboratories. In addition to the important application of FC discussed above many diverse roles of FC has been discussed in the literature [Table 1]. FC plays an important role in various investigations of patient samples and is a vital addition in research-focused studies. Flow cytometers are becoming smaller, cheaper, and more user-friendly and are available in many routine laboratories. Hence, the method should be considered a valuable tool to use in addition to serology and genomic typing. In addition, FC represents a highly innovative technique for many common diagnostic and scientific fields in TM. Finally, it is the tool of choice to develop and optimize new cellular and immunotherapeutic trials.
Table 1: Application of flow cytometry in transfusion medicine

Click here to view


Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
   References Top

1.
Barteneva NS, Fasler-Kan E, Vorobjev IA. Imaging flow cytometry: Coping with heterogeneity in biological systems. J Histochem Cytochem 2012;60:723-33.  Back to cited text no. 1
    
2.
Han Y, Wang S, Zhang Z, Ma X, Li W, Zhang X, et al. In vivo imaging of protein-protein and RNA-protein interactions using novel far-red fluorescence complementation systems. Nucleic Acids Res 2014;42:e103.  Back to cited text no. 2
    
3.
Leipold MD, Newell EW, Maecker HT. Multiparameter phenotyping of human PBMCs using mass cytometry. Methods Mol Biol 2015;1343:81-95.  Back to cited text no. 3
    
4.
Mei HE, Leipold MD, Maecker HT. Platinum-conjugated antibodies for application in mass cytometry. Cytometry A 2016;89:292-300.  Back to cited text no. 4
    
5.
Matz Mikhail V, Fradkov AF, Labas Yulii A, Savitsky Aleksandr P, Zaraisky Andrey G, Markelov Mikhail L, et al. Fluorescent proteins from nonbioluminescent Anthozoa species. Nat Biotechnol 1999;17:969-73.  Back to cited text no. 5
    
6.
Tsien RY. The green fluorescent protein. Annu Rev Biochem 1998;67:509-44.  Back to cited text no. 6
    
7.
McKinnon KM. Flow cytometry: An overview. Curr Protoc Immunol 2018;120:5.1.1-5.1.11.  Back to cited text no. 7
    
8.
Freedman J, Lazarus AH. Application of flow cytometry in transfusion medicine. Transfus Med Rev 1995;9:87-109.  Back to cited text no. 8
    
9.
Nance SJ. Flow cytometry related to red cells. Transfus Sci 1995;16:343-52.  Back to cited text no. 9
    
10.
Arndt P, Garraty G. Flow cytofluorometric analysis in red blood cell immunology. Transfus Med Hemother 2004;31:163-74.  Back to cited text no. 10
    
11.
Whitby A, Whitby L, Fletcher M, Reilly JT, Sutherland DR, Keeney M, et al. ISHAGE protocol: Are we doing it correctly? Cytometry B Clin Cytom 2012;82:9-17.  Back to cited text no. 11
    
12.
Garratty G, Arndt PA. Applications of flow cytofluorometry to red blood cell immunology. Cytometry 1999;38:259-67.  Back to cited text no. 12
    
13.
Chaudhary R, Das SS, Gupta R, Khetan D. Application of flow cytometry in detection of red-cell-bound IgG in Coombs-negative AIHA. Hematology 2006;11:295-300.  Back to cited text no. 13
    
14.
Lynen R, Krone O, Legler TJ, Kohler M, Mayer WR. A newly developed gel centrifugation test for quantification of RBC bound IgG antibodies and their subclasses IgG1 and IgG3: Comparison with flow cytometry. Transfusion 2002;42:612-8.  Back to cited text no. 14
    
15.
Gomez-Arbones X, Pinacho A, Ortiz P, Macia J, Gallart M, Araguas C, et al. Quantification of foetomaternal haemorrhage: An analysis of two cytometric techniques and a semiquantitative gel agglutination test. Clin Lab Haem 2002;24:47-53.  Back to cited text no. 15
    
16.
Bowman JM, Pollock JM. Failures of intravenous Rh immune globulin prophylaxis: An analysis of the reasons for such failures. Transfus Med Rev 1987;1:101-2.  Back to cited text no. 16
    
17.
Bayliss KM, Kueck BD, Johnson ST, Fueger JT, McFadden PW, Mikulski D, et al. Detecting fetomaternal hemorrhage: A comparison of five methods. Transfusion 1991;31:303-7.  Back to cited text no. 17
    
18.
Duguid JK, Bromilow IM. Laboratory measurement of fetomaternal hemorrhage and its clinical relevance. Transfus Med Rev 1999;13:43-8.  Back to cited text no. 18
    
19.
Duguid JK, Bromilow IM, Eggington J. Kleihauer testing and flow cytometry. A comparative study for assessment of feto-maternal haemorrhage. Haematology 1996;1:79-83.  Back to cited text no. 19
    
20.
Agarwal P, Das SS, Gupta R, Khetan D, Chaudhary R. Quantification of fetomaternal hemorrhage – An analysis of three techniques. Gynaecol Obstet Investig 2011;71:47-52.  Back to cited text no. 20
    
21.
Nance SJ, Nelson JM, Arndt PA, Labm HT, Garratty G. Quantitation of fetal-maternal hemorrhage by flow cytometry. A simple and accurate method. Am J Clin Pathol 1989;91:288-92.  Back to cited text no. 21
    
22.
Bayliss KM, McFadden PW, Kueck BD. Flow cytometric quantitation of fetal-maternal hemorrhage: A technique designed for the clinical laboratory. Am J Clin Pathol 1990;93:450.  Back to cited text no. 22
    
23.
Gupta R, Pandey P, Choudhry R, Kashyap R, Mehrotra M, Naseem S, et al. A prospective comparison of four techniques for diagnosis of paroxysmal nocturnal hemoglobinuria. Int J Lab Hematol 2007;29:119-26.  Back to cited text no. 23
    
24.
Araten DJ, Nafa K, Pakdeesuwan K, Luzzatto L. Clonal populations of hematopoietic cells with paroxysmal nocturnal hemoglobinuria genotype and phenotype are present in normal individuals. Proc Natl Acad Sci U S A 1999;96:5209-14.  Back to cited text no. 24
    
25.
Meletis J, Terpos E. Recent insights into the pathophysiology of paroxysmal nocturnal hemoglobinuria. Med Sci Monit 2003;9:RA161-72.  Back to cited text no. 25
    
26.
Ware RE. Is there a little PNH in all of us? Blood 2005;105:3760-1.  Back to cited text no. 26
    
27.
Kashyap R, Awasthi NP, Gupta R. Clinical and flow cytometric analysis of paroxysmal nocturnal hemoglobinuria in Indian patients. J Appl Hematol 2018;9:85-90.  Back to cited text no. 27
  [Full text]  
28.
Svensson L, Hult AK, Stamps R, Ångström J, Teneberg S, Storry JR, et al. Forssman expression on human erythrocytes: Biochemical and genetic evidence of a new histo-blood group system. Blood 2013;121:1459-68.  Back to cited text no. 28
    
29.
Seltsam A, Grüger D, Just B, Figueiredo C, Gupta CD, Deluca DS, et al. Aberrant intracellular trafficking of a variant B glycosyltransferase. Transfusion 2008;48:1898-905.  Back to cited text no. 29
    
30.
Yazdanbakhsh K, Rios M, Storry JR, Kosower N, Parasol N, Chaudhuri A, et al. Molecular mechanisms that lead to reduced expression of duffy antigens. Transfusion 2000;40:310-20.  Back to cited text no. 30
    
31.
Hustinx H, Poole J, Bugert P, Gowland P, Still F, Fontana S, et al. Molecular basis of the Rh antigen RH48 (JAL). Vox Sang 2009;96:234-9.  Back to cited text no. 31
    
32.
Basu S, Kaur R, Kaur G. Hemolytic disease of the fetus and newborn: Current trends and perspectives. Asian J Transfus Sci 2011;5:3-7.  Back to cited text no. 32
[PUBMED]  [Full text]  
33.
Jones JW, Lloyd-Evans P, Kumpel BM. Quantitation of Rh D antigen sites on weak D and D variant red cells by flowcytometry. Vox Sang 1996;71:176-83.  Back to cited text no. 33
    
34.
Beckers EA, Faas BH, Ligthart P, Overbeeke MA, von dem Borne AE, et al. Lower antigen site density and weak D immunogenicity cannot be explained by structural genomic abnormalities or regulatory defects of the RHD gene. Transfusion 1997;37:616-23.  Back to cited text no. 34
    
35.
Flegel WA, Curin-Serbec V, Delamaire M, Donvito B, Ikeda H, Jørgensen J, et al. Section 1B: Rh flow cytometry. Coordinator's report. Rhesus index and antigen density: An analysis of the reproducibility of flow cytometric determination. Transfus Clin Biol 2002;9:33-42.  Back to cited text no. 35
    
36.
Flegel WA, Khull SR, Wagner FF. Primary anti-D immunization by weak D type 2 RBCs. Transfusion 2000;40:428-34.  Back to cited text no. 36
    
37.
Ansart-Pirenne H, Asso-Bonnet M, Le Pennec PY, Roussel M, Patereau C, Noizat-Pirenne F. RhD variants in Caucasians: Consequences for checking clinically relevant alleles. Transfusion 2004;44:1282-6.  Back to cited text no. 37
    
38.
Gorick B, McDougall DC, Ouwehand WH, Overbeeke MA, Tippett P, Jones H, et al. Quantitation of D sites on selected 'weak D' and partial D red cells. Vox Sang 1993;65:136-40.  Back to cited text no. 38
    
39.
Fletcher A, Bryant JA. The analysis of Rh phenotype by flowcytometry. Biotest Bull 1997;5:495-502.  Back to cited text no. 39
    
40.
Wagner FF, Frohmajer A, Ladewig B, Eicher N, Lonicer CB, Muller TH, et al. Weak D alleles express distinct phenotypes. Blood 2000;95:2699-708.  Back to cited text no. 40
    
41.
Kulkarni S, Mohanti D, Gupte S, Vasantha K, Joshi S. Flow cytometric quantitation of antigen D sites on red blood cells of partial D and weak D variant in India. Transfus Med 2006;16:285-9.  Back to cited text no. 41
    
42.
van Bockstaele DR, Berneman ZN, Muylle L, Cole-Dergent J, Peetermans ME. Flow cytometric analysis of erythrocytic D antigen density profile. Vox Sang 1986;51:40-6.  Back to cited text no. 42
    
43.
Hasekura H, Ota M, Ito S, Hasegawa Y, Ichinose A, Fukushima H, et al. Flow cytometric studies of the D antigen of various Rh phenotypes with particular reference to Du and Del. Transfusion 1990;30:236-8.  Back to cited text no. 43
    
44.
Cunningham NA, Zola AP, Hui HL, Taylor LM, Green FA. Binding characteristics of anti-Rh0(D) antibodies to Rh0(D)-positive and Du red cells. Blood 1985;66:765-8.  Back to cited text no. 44
    
45.
Wagner FF. Influence of Rh phenotype on the antigendensity of C, c and D: Flow cytometric study using a frozen standard red cell. Transfusion 1994;34:671-5.  Back to cited text no. 45
    
46.
Pavenski K, Freedman J, Semple JW. HLA alloimmunization against platelet transfusions: Pathophysiology, significance, prevention and management. Tissue Antigens 2012;79:237-45.  Back to cited text no. 46
    
47.
Doughty HA, Murphy MF, Metcalfe P, Rohatiner AZ, Lister TA, Waters AH. Relative importance of immune and non-immune causes of platelet refractoriness. Vox Sang 1994;66:200-5.  Back to cited text no. 47
    
48.
Ferreira AA, Zulli R, Soares S, Castro VD, Moraes-Souza H. Identification of platelet refractoriness in oncohematologic patients. Clinics (Sao Paulo) 2011;66:35-40.  Back to cited text no. 48
    
49.
He Y, Zhao YX, Zhu MQ, Wu Q, Ruan CG. Detection of autoantibodies against platelet glycoproteins in patients with immune thrombocytopenic purpura by flow cytometric immunobead array. Clin Chim Acta 2013;415:176-80.  Back to cited text no. 49
    
50.
van Velzen JF, Laros-van Gorkom BA, Pop GA, van Heerde WL. Multicolor flow cytometry for evaluation of platelet surface antigens and activation markers. Thromb Res 2012;130:92-8.  Back to cited text no. 50
    
51.
Curtis BR, McFarland JG. Detection and identification of platelet antibodies and antigens in the clinical laboratory. Immunohematology 2009;25:125-35.  Back to cited text no. 51
    
52.
Bub CB, Martinelli BM, Avelino TM, Gonçalez AC, Barjas-Castro Mde L, Castro V. Platelet antibody detection by flow cytometry: An effective method to evaluate and give transfusional support in platelet refractoriness. Rev Bras Hematol Hemoter 2013;35:252-5.  Back to cited text no. 52
    
53.
Hézard N, Simon G, Macé C, Jallu V, Kaplan C, Nguyen P. Is flow cytometry accurate enough to screen platelet autoantibodies? Transfusion 2008;48:513-8.  Back to cited text no. 53
    
54.
Aref S, Selim T, Ibrahim L, Abd-Elghaffar H, Ashery RE. Flow cytometry detection of platelets autoantibodies in children with idiopathic thrombocytopenic purpura. Indian J Hematol Blood Transfus 2009;25:96-103.  Back to cited text no. 54
    
55.
Buakaew J, Promwong C. Platelet antibody screening by flow cytometry is more sensitive than solid phase red cell adherence assay and lymphocytotoxicity technique: A comparative study in Thai patients. Asian Pac J Allergy Immunol 2010;28:177-84.  Back to cited text no. 55
    
56.
Freedman J, Hornstein A. Simple method for differentiating between HLA and platelet-specific antibodies by flow cytometry. Am J Hematol 1991;38:314-20.  Back to cited text no. 56
    
57.
Köhler M, Dittmann J, Legler TJ, Lynen R, Humpe A, Riggert J, et al. Flow cytometric detection of platelet-reactive antibodies and application in platelet crossmatching. Transfusion 1996;36:250-5.  Back to cited text no. 57
    
58.
Poley S, Mempel W. Laboratory diagnosis of heparin-induced thrombocytopenia: Advantages of a functional flow cytometric test in comparison to the heparin-induced platelet-activation test. Eur J Haematol 2001;66:253-62.  Back to cited text no. 58
    
59.
Hughes M, Hayward CP, Warkentin TE, Horse-wood P, Chorneyko KA, Kelton JG. Morphological analysis of microparticle generation in heparin- induced thrombocytopenia. Blood 2000;96:188-94.  Back to cited text no. 59
    
60.
Tomer A, Masalunga C, Abshire TC. Determination of heparin-induced thrombocytopenia. A rapid flow cytometric assay for direct demonstration of antibody-mediated platelet activation. Am J Hematol 1999;61:53-61.  Back to cited text no. 60
    
61.
Velliquette RW, Hue-Roye K, Lomas-Francis C, Gillen B, Schierts J, Gentzkow K, et al. Molecular basis of two novel and related high-prevalence antigens in the Kell blood group system, KUCI and KANT, and their serologic and spatial association with K11 and KETI. Transfusion 2013;53:2872-81.  Back to cited text no. 61
    
62.
Ji Y, Veldhuisen B, Ligthart P, Haer-Wigman L, Jongerius J, Boujnan M, et al. Novel alleles at the Kell blood group locus that lead to Kell variant phenotype in the Dutch population. Transfusion 2015;55:413-21.  Back to cited text no. 62
    
63.
Berneman ZN, Van Bockstaele DR, Uyttenbroeck WM, Van Zaelen C, Cole-Dergent J, Muylle L, et al. Flowcytometric analysis of erythrocytic blood group A antigen density profile. Vox Sang 1991;61:265-74.  Back to cited text no. 63
    
64.
Hult AK, Olsson ML. Many genetically defined ABO subgroups exhibit characteristic flow cytometric patterns. Transfusion 2010;50:308-23.  Back to cited text no. 64
    
65.
Hagberg IA, Lyberg T. Blood platelet activation evaluated by flow cytometry: Optimised methods for clinical studies. Platelets 2000;11:137-50.  Back to cited text no. 65
    
66.
Vucetic D, Ilic V, Vojvodic D, Subota V, Todorović M, Balint B. Flow cytometry analysis of platelet populations: Usefulness for monitoring the storage lesion in pooled buffy-coat platelet concentrates. Blood Transfus 2018;16:83-92.  Back to cited text no. 66
    
67.
Dzik WH. Leukocyte counting during process control of leukoreduced blood components. Vox Sang 2000;78 Suppl 2:223-6.  Back to cited text no. 67
    
68.
Cardigan R, Phipps A, Seghatchian J, Bashir S, Aynsley S, Beckman N, et al. The development of a national standardized approach for the enumeration of residual leucocytes in blood components. Vox Sang 2002;83:100-9.  Back to cited text no. 68
    
69.
Kao GS, Wood IG, Dorfman DM, Milford EL, Benjamin RJ. Investigations into the role of anti- HLA class II antibodies in TRALI. Transfusion 2003;43:185-91.  Back to cited text no. 69
    
70.
Stroncek DF, Shankar R, Litz C, Clement L. The expression of the NB1 antigen on myeloid precursors and neutrophils from children and umbilical cords. Transfus Med 1998;8:119-23.  Back to cited text no. 70
    
71.
Taniguchi K, Kobayashi M, Harada H, Hiraoka A, Tanihiro M, Takata N, et al. Human neutrophil antigen-2a expression on neutrophils from healthy adults in western Japan. Transfusion 2002;42:651-7.  Back to cited text no. 71
    
72.
Lim J, Kim Y, Han K, Kim M, Lee KY, Kim WI, et al. Flow cytometric monocyte phagocytic assay for predicting platelet transfusion outcome. Transfusion 2002;42:309-16.  Back to cited text no. 72
    
73.
Chang A, Ma DD. The influence of flow cytometric gating strategy on the standardization of CD34+cell quantitation: An Australian multicenter study. Australasian BMT Scientists Study Group. J Hematother 1996;5:605-16.  Back to cited text no. 73
    
74.
Gratama JW, Kraan J, Levering W, Van Bockstaele DR, Rijkers GT, Van der Schoot CE. Analysis of variation in results of CD34+hematopoietic progenitor cell enumeration in a multicenter study. Cytometry 1997;30:109-17.  Back to cited text no. 74
    
75.
Inaba T, Shimazaki C, Tatsumi T, Yamagata N, Hirata T, Goto H, et al. Expression of differentiation-associated antigens and adhesion molecules on CD34-positive cells harvested from peripheral blood and bone marrow. Prog Clin Biol Res 1994;389:331-7.  Back to cited text no. 75
    
76.
De Bruyn C, Delforge A, Bron D, Bernier M, Massy M, Ley P, et al. Comparison of the coexpression of CD38, CD33 and HLA-DR antigens on CD34+purified cells from human cord blood and bone marrow. Stem Cells 1995;13:281-8.  Back to cited text no. 76
    
77.
D'Arena G, Cascavilla N, Musto P, Greco M, Di Mauro L, Carella AM, et al. Flow cytometric characterization of CD34+hematopoietic progenitor cells in mobilized peripheral blood and bone marrow of cancer patients. Haematologica 1996;81:216-23.  Back to cited text no. 77
    
78.
Zarcone D, Tilden AB, Cloud G, Friedman HM, Landay A, Grossi CE. Flow cytometry evaluation of cell-mediated cytotoxicity. J Immunol Methods 1986;94:247-55.  Back to cited text no. 78
    
79.
Fischer K, Mackensen A. The flow cytometric PKH-26 assay for the determination of T-cell mediated cytotoxic activity. Methods 2003;31:135-42.  Back to cited text no. 79
    
80.
Berger TG, Feuerstein B, Strasser E, Hirsch U, Schreiner D, Schuler G, et al. Large-scale generation of mature monocyte-derived dendritic cells for clinical application in cell factories. J Immunol Methods 2002;268:131-40.  Back to cited text no. 80
    
81.
Gottfried E, Krieg R, Eichelberg C, Andreesen R, Mackensen A, Krause SW. Characterization of cells prepared by dendritic cell tumor cell fusion. Cancer Immunol 2002;2:15.  Back to cited text no. 81
    

Top
Correspondence Address:
Dr. Sudipta Sekhar Das
Department of Transfusion Medicine, Apollo Multispeciality Hospitals, Kolkata - 700 054, West Bengal
India
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ajts.ajts_61_22

Rights and Permissions


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7]
 
 
    Tables

  [Table 1]



 

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
    Flow Cytometry i...
   Conclusion
    References
    Article Figures
    Article Tables

 Article Access Statistics
    Viewed1003    
    Printed54    
    Emailed0    
    PDF Downloaded153    
    Comments [Add]    

Recommend this journal

Association Contact us | Sitemap | Advertise | What's New | Copyright and Disclaimer | Privacy Notice


2006 - Asian Journal of Transfusion Science | Published by Wolters Kluwer - Medknow
Online since 10th November, 2006