scholarly journals Memory like NK cells display stem cell like properties after Zika virus infection

2020 ◽  
Vol 16 (12) ◽  
pp. e1009132
Author(s):  
Weshely Kujur ◽  
Oscar Murillo ◽  
Raju S. R. Adduri ◽  
Ramakrishna Vankayalapati ◽  
Nagarjun V. Konduru ◽  
...  
Keyword(s):  
Stem Cells ◽  
T Cells ◽  
Stem Cell ◽  
Virus Infection ◽  
Nk Cells ◽  
Zika Virus ◽  
Gene Signature ◽  
Hematopoietic Stem ◽  
Zika Virus Infection ◽  
With Memory

NK cells have been shown to display adaptive traits such as memory formation akin to T and B lymphocytes. Here we show that Zika virus infection induces memory like NK cells that express CD27. Strikingly, these cells exhibit stem-like features that include expansion capacity, self-renewal pathway, differentiation into effector cells, longer telomeres and gene signature associated with hematopoietic stem cell (HSC) progenitors. This subset shared transcriptional and epigenetic changes with memory CD8 T cells, stem cells and stem like T cells. These NK cells with memory and stem cell features, which we term “NK memory stem cells”, demonstrated greater antiviral potential than CD27- or naïve CD27+ NK when adoptively transferred to Zika infected mice. Our results also suggest a role for the transcription factor TCF-1 in memory and stemness features of this NK subset. This study defines a unique TCF1hi CD27+ NK subset with memory capacity and stem cell features that play a role in antiviral immunity.

scholarly journals CD34+ Hematopoietic Stem Cells Exert Accessory Function in Lipopolysaccharide-induced  T Cell Stimulation and CD80 Expression on Monocytes

1999 ◽  
Vol 189 (4) ◽  
pp. 693-700 ◽  
Author(s):  
Taila Mattern ◽  
Gundolf Girroleit ◽  
Hans-Dieter Flad ◽  
Ernst T. Rietschel ◽  
Artur J. Ulmer
Keyword(s):  
Stem Cells ◽  
T Cells ◽  
Stem Cell ◽  
T Cell ◽  
Hematopoietic Stem Cells ◽  
Cell Stimulation ◽  
Hematopoietic Stem ◽  
Accessory Function ◽  
T Cell Stimulation ◽  
Blood Stem Cells

CD34+ hematopoietic stem cells, which circulate in peripheral blood with very low frequency, exert essential accessory function during lipopolysaccharide (LPS)-induced human T lymphocyte activation, resulting in interferon γ production and proliferation. In contrast, stimulation of T cells by “conventional” recall antigens is not controlled by blood stem cells. These conclusions are based on the observation that depletion of CD34+ blood stem cells results in a loss of LPS-induced T cell stimulation as well as reduced expression of CD80 antigen on monocytes. The addition of CD34-enriched blood stem cells resulted in a recovery of reactivity of T cells and monocytes to LPS. Blood stem cells could be replaced by the hematopoietic stem cell line KG-1a. These findings may be of relevance for high risk patients treated with stem cells or stem cell recruiting compounds and for patients suffering from endotoxin-mediated diseases.

scholarly journals The Role of T Cells in Hematopoietic Stem Cell Engraftment

2006 ◽  
Vol 6 ◽  
pp. 246-253 ◽  
Author(s):  
Elizabeth Hexner
Keyword(s):  
Stem Cells ◽  
T Cells ◽  
Stem Cell ◽  
Hematopoietic Stem Cell ◽  
Immune Cells ◽  
Immune Reconstitution ◽  
Host Immune System ◽  
Hematopoietic Stem ◽  
Cell Engraftment ◽  
Donor T Cells

Much attention has focused on the immune recovery of donor T cells following hematopoietic stem cell transplantation (HSCT). Termed immune reconstitution, a better understanding of the dynamics of the functional recovery of immune cells following HSCT has important implications both for fighting infections and, in the allogeneic setting, for providing antitumor activity while controlling graft-vs.-host disease (GVHD). The immune cells involved in immune reconstitution include antigen-presenting cells, B lymphocytes, natural killer cells, and, in particular, T lymphocytes, the immune cell that will be the subject of this review. In addition, T cells can play an important role in the process of engraftment of hematopoietic stem cells. The evidence for a T cell tropic effect on hematopoietic engraftment is both direct and indirect, and comes from the clinic as well as the research lab. Animal models have provided useful clues, but the molecular mechanisms that govern the interaction between donor stem cells, donor T cells, the host immune system, and the stem cell niche remain obscure. This review will describe the current published clinical and basic evidence related to T cells and stem cell engraftment, and will identify future directions for translational research in this area.

scholarly journals CD4+ T Cells Cross-Reactive with Dengue and Zika Viruses Protect against Zika Virus Infection

Cell Reports ◽  
2020 ◽  
Vol 31 (4) ◽  
pp. 107566 ◽  
Author(s):  
Jinsheng Wen ◽  
Ying-Ting Wang ◽  
Kristen M. Valentine ◽  
Rúbens Prince dos Santos Alves ◽  
Zhigang Xu ◽  
...  
Keyword(s):  
T Cells ◽  
Virus Infection ◽  
Zika Virus ◽  
Cd4 T Cells ◽  
Zika Virus Infection

scholarly journals CD4+ T cells promote humoral immunity and viral control during Zika virus infection

2019 ◽  
Vol 15 (1) ◽  
pp. e1007474 ◽  
Author(s):  
Annie Elong Ngono ◽  
Matthew P. Young ◽  
Maximilian Bunz ◽  
Zhigang Xu ◽  
Sararat Hattakam ◽  
...  
Keyword(s):  
T Cells ◽  
Virus Infection ◽  
Humoral Immunity ◽  
Zika Virus ◽  
Cd4 T Cells ◽  
Viral Control ◽  
Zika Virus Infection

Poor Graft Function in Recipients of T Cell Depleted (TCD) Allogeneic Hematopoietic Stem Cell Transplants (HSCT) Is Mostly Related to Viral Infections and Anti-Viral Therapy.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 3147-3147 ◽  
Author(s):  
Roni Tamari ◽  
Sheetal Ramnath ◽  
Deborah Kuk ◽  
Craig S. Sauter ◽  
Doris M Ponce ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
T Cells ◽  
Stem Cell ◽  
Peripheral Blood ◽  
Viral Infections ◽  
Graft Function ◽  
Hematopoietic Stem ◽  
Allogeneic Hsct ◽  
Poor Graft Function

Abstract Abstract 3147 Introduction: Poor graft function (PGF) without immune rejection, defined as persistent cytopenias with hypocellular marrow and full donor myeloid chimerism, can be a life-threatening complication after allogeneic HSCT. It is commonly caused by viral infectious, myelosuppressive drugs like antivirals, and graft-vs-host disease (GvHD). Treatment options include supportive therapy with transfusions and growth factors and in severe cases administration of additional hematopoietic stem cells (HSCs) from the same donor without conditioning (stem cell boost). The incidence, natural history, and the indications for stem cell boost therapy are not well defined. Aims: To assess the incidence, etiologies, and indications for stem cell boost for PGF in a homogeneous group of patients with advanced MDS and AML who underwent TCD HSCT from matched or mismatched related or unrelated donors after conditioning with the same myeloablative regimen. Patients and methods: Poor graft function was defined as persistent neutropenia (ANC <1,000 μL and G-CSF administration x3 in 30 days), thrombocytopenia (platelets <50,000 μL or platelets transfusion × 4 in 30 days), and/or hemoglobin <8 g//dL after engraftment with hypocellular BM and full donor myeloid chimerism. Severe PGF was defined as ANC <500 μL, red cell transfusion-dependent anemia with reticulocytopenia of < 20,000 μL, and platelets <20,000 μL. The patient population in which this study was done included 42 patients enrolled between 09/2009 and 05/2012 in a phase 2 trial of palifermin peri-transplant to reduce transplant-related mortality. The median age was 57.5 years (1–65). All patients received the same myeloablative conditioning regimen with busulfan, melphalan, fludarabine, rabbit ATG and palifermin peri-transplant. G-CSF mobilized donor peripheral blood stem cells underwent CD34+ selection and depletion of T cells using CliniMACS immunomagnetic selection columns (Milteny Biotec). Donors were HLA matched (31; 13 related and 18 unrelated) or mismatched unrelated (11). Chimerism was determined in bone marrow as well as neutrophils, B cells, and T cells by short tandem repeat analysis on DNA extracted from bone marrow and peripheral blood cell subsets. Results: Forty-one patients were evaluable for this analysis; 1 patient was not included as he rejected the allograft shortly after engraftment. There were 8 cases of PGF with a cumulative incidence (CI) at 1 year of 18% (13% HLA matched, 33% HLA mismatch). The etiology was infection in 7 cases, and unknown in the 8th case. This patient presented with presumed autoimmune anemia and thrombocytopenia associated with a hypercellular marrow and did not respond to multiple lines of therapies. Her marrow became later hypocellular and met the criteria for PGF. None of the PGF cases in this series was associated with GvHD at the time of diagnosis of PGF. The infectious etiologies included: 6 viral infections and 1bacterial sepsis + myelosuppressive drugs. The most common viral etiology associated with PGF was CMV (50%). The 1-year CI of PGF in CMV seropositive patients was 25% and in CMV seronegative patients was 14%. Of note, HHV6 viremia was detected in patients with PGF. HHV6 is not routinely monitored, however, making it difficult to establish a causative role. All patients had moderate PGF at diagnosis and 3 cases had worsening of cytopenias and met the criteria for severe PGF. To date, 3 PGF patients have died from EBV-PTLD, adenovirus infection or GVHD (developed after CMV treatment with liposomal cidofovir), 3 continue to suffer from PGF and 2 patients are alive with recovered good blood counts after eradication of CMV. Of the 3 patients with persistent PGF, one received a TCD boost with no response, and 2 continued to be treated for CMV viremia. A stem cell boost was indicated if pancytopenia persisted despite eradication of cause of the PGF. In this small series, there were not enough events to evaluate association between PGF and CD34 cell dose, CD3 cell dose or day 100 T-cell chimerism. Conclusions: In this homogenous population of patients with MDS who underwent TCD allogeneic HSCT, the incidence of PGF is about 20%. The most common cause was viral infection with predominance of CMV. Therefore, strategies to prevent CMV reactivation in patients undergoing allogeneic HSCT has the potential to reduce the risk of PGF and avoid the need for infusion of additional stem cells. Disclosures: No relevant conflicts of interest to declare.

Transplantation Of TcRαβ/CD19 Depleted Stem Cells From Haploidentical Donors In Children: Current Results

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 692-692 ◽  
Author(s):  
Peter Lang ◽  
Tobias Feuchtinger ◽  
Heiko-Manuel Teltschik ◽  
Michael Schumm ◽  
Patrick Schlegel ◽  
...  
Keyword(s):  
Stem Cells ◽  
T Cells ◽  
Stem Cell ◽  
T Cell ◽  
T Lymphocytes ◽  
Nk Cells ◽  
Chronic Gvhd ◽  
Cell Depletion ◽  
T Cell Depletion ◽  
Αβ T Cells

Abstract T-cell depletion of the graft is an effective method to prevent or completely avoid Graft-versus-Host Disease (GvHD) in haploidentical stem cell transplantation. In order to increase the T-cell depletion efficacy while maintaining the anti-tumor and anti-infectious properties of the graft, we have investigated a new T-cell depletion method which removes αβ+ T-lymphocytes via a biotinylated anti-TcRαβ antibody followed by an anti-biotin antibody conjugated to magnetic microbeads while retaining γδ+ T-lymphocytes, Natural killer (NK) cells and other cells in the graft. In addition, CD19+ B-lymphocytes were concomitantly depleted for the prevention of posttransplant EBV-associated lymphoproliferative disease. The CliniMACS system was used for manipulation of peripheral stem cell grafts from full haplotype mismatched family donors in 35 patients. Results The overall depletion of αβ+ T-cells was highly effective with 4.6 log (range 3.8–5.0). Patients received a median number of only 14 x 103/kg residual αβ+ T-cells. Recovery of CD34+ stem cells was 72%, and the median number of infused CD34+ stem cells was 12 x 106/kg (range 5-38 x 106/kg). Additionally, the patients received 2 types of potential antileukemic effector cells: 107 x 106/kg (range 35 -192 x 106/kg) CD56+ NK-cells and 11 x 106/kg (range 5–30 x 106/kg) γδ+ T-lymphocytes. Diagnoses were ALL (n=20), AML/MDS/JMML (n=9), nonmalignant diseases (n=4), solid tumors (n=2); disease status: CR2-CR6 (n=17), active disease (n=18). 23 patients received a second or third SCT (65%). A toxicity reduced conditioning regimen (fludarabin 40mg/m² or clofarabin 50mg/m² (day -8 to d -5), thiotepa 10mg/kg (d -4), melphalan 70mg/m² (d -3 and d -2) was used. The anti CD3 specific OKT3 antibody was used as rejection prophylaxis from day -8 to day -1 without affecting cotransfused effector cells because of its short half-life period in the first 7 patients. However, due to its restricted availability, the substance was substituted since 2011 by a reduced ATG-F dose (15mg/kg) given at start of the conditioning regimen in order not to impair NK and γδ+ T-cells of the grafts (1 mg/kg d -12, 4 mg/kg d -11, 5 mg/kg d -10 and -9; n=28 patients). Short course MMF (until day +30) was given in 25 patients. Graft rejection occurred in 14% of the patients. However, after reconditioning and second stem cell donation, final engraftment was achieved in all patients. The median time to reach neutrophil and platelet recovery in patients with primary engraftment was 10 and 11 days respectively. All patients showed a rapid immune reconstitution with 250 (OKT3 conditioning) and 273 (ATG conditioning) CD3+ T-cells/µl, 30 (OKT3) and 47 (ATG) CD3+4+/µl and 300 (OKT3) and 382 (ATG) CD56+ NK-cells/µl at day +30 posttransplant. γδ+ T-cells started to expand faster than αβ+ T-cells in the early post-transplant period (156 vs. 82 cells/µl at day +30) whereas at day +90, αβ+ T-cells were predominant (170 vs. 134 cells/µl). Acute GvHD grade 0-I occurred in 25 patients (71%); 6 patients had GvHD II (17%), 3 patients had GvHD III (9%) and one patients experienced GvHD grade IV (3%). 3 patients experienced chronic GvHD (8%). Incidence of acute GvHD was not influenced by the number of residual T cells or by the type of serotherapy. 1 year EFS for patients with acute leukemias was 66% (any CR) and 14% (active disease).TRM at 1 year was 20%. Conclusions These data indicate that transplantation of TcR αβ+/CD19 depleted cells from a haploidentical donor results in sustained engraftment, remarkably fast immune reconstitution and low incidence of both acute and chronic GvHD. OKT3 could be substituted by ATG without negative effects. The anti-leukemic efficacy of this approach in comparison to other methods of T-cell depletion needs to be evaluated with a longer patient follow-up. Disclosures: No relevant conflicts of interest to declare.

Hematopoietic stem cell dose correlates with the speed of immune reconstitution after stem cell transplantation

Blood ◽  
2004 ◽  
Vol 103 (11) ◽  
pp. 4344-4352 ◽  
Author(s):  
Benny J. Chen ◽  
Xiuyu Cui ◽  
Gregory D. Sempowski ◽  
Jos Domen ◽  
Nelson J. Chao
Keyword(s):  
Stem Cells ◽  
T Cells ◽  
Stem Cell ◽  
T Cell ◽  
B Cells ◽  
Hematopoietic Stem Cells ◽  
Immune Reconstitution ◽  
Hematopoietic Stem ◽  
Time Points ◽  
Cell Dose

Abstract In the current study, we tested whether higher numbers of hematopoietic stem cells correlate with the speed of immune reconstitution in a congenic transplantation model (C57BL/Ka, CD45.1, Thy1.1→C57BL/6, CD45.2, Thy1.2) using purified hematopoietic stem cells (c-Kit+Thy1.1lowLin-/lowSca-1+). There were 3 different doses of stem cells used (400, 1000, and 5000). Phenotypic analyses in peripheral blood and spleen demonstrated that higher numbers of infused stem cells are associated with more rapid regeneration of T cells (CD4+, CD8+, naive CD4+, naive CD8+) and B cells at early time points. The numbers of T and B cells eventually became equivalent between different dose groups at late time points. Production of interleukin-2 and inter-feron-γ per T cell was similar regardless of stem cell dose even when tested at the time when there were significant differences in peripheral T-cell counts. The improved immune recovery was attributed to a more rapid regeneration of donor-type immune cells. Higher numbers of total thymocytes and signal joint T-cell receptor excision circles were observed in the higher dose stem cell recipients, suggesting that accelerated regeneration of T cells was due to enhanced thymopoiesis. (Blood. 2004;103:4344-4352)

scholarly journals Costimulatory Molecule DNAM-1 Is Essential for Optimal NK Education Post Allogeneic Hematopoietic Stem Cell Transplantation

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 3291-3291
Author(s):  
Qiannan Shang ◽  
Xingxing Yu ◽  
Xunhong Cao ◽  
Xuefei Liu ◽  
Zhengli Xu ◽  
...  
Keyword(s):  
T Cells ◽  
Stem Cell ◽  
Hematopoietic Stem Cell Transplantation ◽  
Nk Cells ◽  
Cd8 T Cells ◽  
Nk Cell ◽  
Costimulatory Molecule ◽  
Point Of View ◽  
Post Transplantation ◽  
Hematopoietic Stem

Background: DNAM-1 (DNAX accessory molecule-1, CD226) is a costimulatory molecule that is constitutively expressed by NK cells and CD8+ T cells. Interaction between DNAM-1 on NK cells and CD8+ T cells with its ligands on target cells triggers cell-mediated cytotoxicity and cytokine production. Previous mouse model had showed that the expression of DNAM-1 is associated with NK education. Moreover, Monika's group found that DNAM-1 serves as an intrinsic, readout-independent marker for NK cell education in health donor. Our previous study had demonstrated that when both donors and hosts present all the KIR ligands for donor KIRs, reconstituted NK cells would achieve better functional education and contribute to least relapse for the patients post allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, the roles of DNAM-1 in NK education post allo-HSCT were unknown. Aims: In this study, we have investigated the contribution of DNAM-1 expression to NK education post transplantation. Methods :We prospectively enrolled 114 patients undergoing haplo-SCT between May 2016 and April 2017 to explore the NK cell dynamic education at days 30, 90 and 180 post-transplant. Peripheral blood mononuclear cells of each sample were analyzed by 15-colors flow cytometry to evaluate of the phenotype as well as functional recovery of NK cells with different maturation expression levels of NKG2A, KIR, and CD57, as well as of activating receptors (NKp30, NKp46, NKG2D, DNAM-1) and CD25, CD122 as well as CD107a and IFN-gamma on NK cells. To study the correlation between the expression of DNAM-1 and effect of donor and host HLA on NK cell education, the expression of DNAM-1 on single-KIR+ NK cells (exhibiting NKG2A, KIR2DL1, KIR2DL2/KIR2DL3, or KIR3DL1 as their sole receptor) was compared based on the combination between donor/host HLA and donor inhibitory KIR. Results: The DNAM-1 expression on single-KIR+ NK cell ligated by sole donor HLA, or sole host HLA, or both donor and host HLA is higher compared to those single-KIR+ NK cells lacking ligands from donor or host or both. From the donor point of view, when donor only presented C1C1 ligand for KIR2DL2/L3, the MFI of DNAM-1 on single KIR2DL2/L3+ NK cells was significantly higher than KIR2DL1+ NK cells and KIR3DL1+ NK cells at 90day (P<0.0001, P=0.046) and 180day (P<0.0001, P=0.034) post transplantation. However, when donor presented C1C1 ligand for KIR2DL2/L3 and Bw4 ligand for KIR3DL1, the MFI of DNAM-1 on single KIR2DL1+ NK cells was significantly lower than KIR2DL2/l3+ NK cells and KIR3DL1+ NK cells at 90day (P<0.0001, P=0.0002) and 180day (P<0.0001, P<0.0001) post transplantation. There was no difference between single KIR2DL1+ NK cells and single KIR3DL1+ NK cells. When donor expressed all HLA(Bw4C1C2) for KIR2DL2/L3, KIR2DL1, KIR3DL1, there was no difference in the expression of DNAM-1 among three single KIR+ NK cells. No matter from the host point or from the combination of donor and host point of view, the DNAM-1 expression differences were same to from donor point of view. There was a clear hierarchy of DNAM-1 expression among NK populations when both of donors and hosts presenting all HLA (Bw4C1C2) for donor KIRs. NK cells with sole KIR or sole NKG2A exhibited higher DNAM-1 expression compared with NKG2A-KIR- NK cells. NK cells with two or three inhibitory KIRs exhibited higher DNAM-1 expression compared with NKG2A+KIR- NK cells. NK cells with 3 inhibitory KIRs and NKG2A expression exhibited maximum DNAM-1 expression at day 30, 90, 180 post transplantation. Meanwhile, the expression of DNAM-1 on NK cells correlated with different ways of education through NKG2A and KIR. There were two fundamental forms of HLA haplotype -21 HLA-B dimorphism: one preferentially supplying CD94:NKG2A ligands (-21M HLA-B), the other preferentially supplying KIR ligands (-21T HLA-B). The expression of DNAM-1 on NKG2A-KIR2DL1+ NK cells was significantly lower when donor or patient was Bw6C1C1 (M/M) compared with when donor or patient was Bw4C2Cx (T/T). However, the expression of DNAM-1 on NKG2A+KIR- NK cells showed no significant difference when donor or patient was Bw6C1C1 (M/M) compared with when donor or patient was Bw4C2C2 (T/T). Summary: This study demonstrated that no matter from donor or/and host point of view, DNAM-1 expression contributes to optimal NK cells education post allo-HSCT. Disclosures No relevant conflicts of interest to declare.

scholarly journals Screening Donor Samples Using Multicolor Flow Cytometry Prior to Allogeneic Hematopoietic Stem Cell Transplantation Can Improve Disease Monitoring Post-Transplantation

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 39-40
Author(s):  
Hui Wang ◽  
Aixian Wang ◽  
Meiwei Gong ◽  
Junyi Zhen ◽  
Xueying Wu ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Stem Cell ◽  
Hematopoietic Stem Cell Transplantation ◽  
Stem Cell Transplantation ◽  
Nk Cells ◽  
Incidence Rate ◽  
Multicolor Flow Cytometry ◽  
Hematopoietic Stem ◽  
Abnormal Cells

Introduction Multicolor flow cytometry (MFC) has been frequently adopted as a method for minimal residual disease (MRD) detection, and is also a promising technique to detect post-transplant lymphoproliferative disorder. Some abnormal donor origin cells might be found when detecting MRD following an allogeneic hematopoietic stem cell transplantation (allo-HSCT). To minimize the effects from donor cells, using MFC prior to allo-HSCT to screen donor peripheral blood (PB) or bone marrow (BM) might be feasible. Methods We performed 3395 allo-HSCTs between January 2013 and December 2019 at Lu Daopei Hospital in Langfang, China. MRD was detected in recipients' BMs according to a conventional two-tube 8 or 9-color MFC panel. Abnormal cells were observed in BMs from three patients in complete remission (CR) one to four months post allo-HSCT. Abnormal neutrophils lacking CD16 expression were found in a patient with secondary acute myeloid leukemia (AML) that developed from a myelodysplastic/ myeloproliferative neoplasm (MDS/MPN). After ruling out MDS and paroxysmal nocturnal hemoglobinuria (PNH), we hypothesized that an Fcγ receptor IIIB (FcγRIIIB) gene deletion was the most likely reason. Abnormal natural killer (NK) cells were detected in the BM from an allo-HSCT recipient with T-cell acute lymphoblastic leukemia (ALL), and monoclonal B lymphocytosis (MBL) in allo-HSCT recipient with B-cell ALL. These three patinets' PBs were detected using MFC after the new finding to decide the cell origin. Besides, 4.54%(in WBC) CD4+ and CD8+ double positive T- cells which were monoclonal cells of the TCRVβ repertoire were detected in a PB sample from a donor prior to allo-HSCT. To evaluate the incidence rate The immunophenotypings were studied in the BMs from 79 NK lymphoma patients. Results Identical phenotypes were recognized in PBs obtained from the three respective donors. The fourth donor did not donate her cells for allo-HSCT, yet. The incidence rate of abnormal cells in donor samples was 0.1% (4/3395 cases), but this rate might be underestimated because MFC screening was not a routine procedure for donors. Additionally, only abnormal immunophenotyping related to patient diagnosis might have been found using an MRD panel as this panel only included markers related to diagnosis. Among general population, the incidence rate of suspicious FcγRIIIB deletion was 0.2% (11/5256 cases), the incidence rate of NK cells without CD2+ and homogeneously expressed CD159c was 0.05% (1/2000 cases) and none among the 79 NK lymphoma samples. The rate of MBL was 0.75% (15/2000 cases) and 1.36% in older than 40 years old people and the rate of monoclonal CD4/CD8 DP T-cells was 0.05% (1/2000 cases). All of these abnormal cells or polymorphism could be analyzed using a two tube MFC panel-- ckappa/clambda/(CD34)/CD19/ CD5/CD20/CD38 /CD45/CD56 and CD16/(CD117)/CD3/ CD4/CD5/ CD8/CD56/CD45/CD2. Conclusion Donor original abnormal cells or phenotypic polymorphisms could have an effect on MFC-based MRD or PTLD detection of recipients following allo-HSCT. These patients might be mis-diagnosed as being MRD positive or having PTLD if the technician lacks experience. To avoid mis-diagnosis and minimize the risk of allo-HSCT, it might be promising to utilize a suitable MFC panel to screen donor PB or BM samples prior to transplantation. Disclosures No relevant conflicts of interest to declare.

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