Interferon Alpha Treated Patients with Chronic Myeloid Leukemia (CML) In Prolonged Complete Remission Have Increased Numbers of NK-Cells and Clonal Gamma-Delta T-Cells, and a Distinct Plasma Cytokine Profile.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1201-1201 ◽  
Author(s):  
Anna Kreutzman ◽  
Peter Rohon ◽  
Edgar Faber ◽  
Karel Indrak ◽  
Vesa Juvonen ◽  
...  
Keyword(s):  
T Cells ◽  
Healthy Volunteers ◽  
Nk Cells ◽  
Interferon Alpha ◽  
Cytokine Profile ◽  
Healthy Controls ◽  
Disease Relapse ◽  
Immunomodulatory Effects ◽  
Plasma Cytokine ◽  
Clonal Rearrangement

Abstract Abstract 1201 Background. Before the era of tyrosine kinase inhibitors (TKIs), interferon alpha (IFN-α) was the treatment of choice in CML and prolonged survival of responding patients. Recent studies suggest that combination of IFN-α with TKI improves therapy outcome. Significantly, a proportion of IFN-α treated patients in prolonged complete cytogenetic remission (CCyR) have been able to discontinue treatment without disease relapse (Mahon et al. JCO 2002). The mechanism of action of IFN-α therapy is incompletely understood; the drug exerts both direct cytotoxic and immunomodulatory effects on leukemic cells. The aim of this project was to study the immunomodulatory effects of IFN-α in CML patients in prolonged remission and isolate biological markers predicting response. Patients and methods. The study population consisted of CML patients treated with IFN-α monotherapy (n = 10, median therapy time 146 months, range 63–231 months) and CML patients who had discontinued IFN-α therapy but remained in remission for >2 years (n = 9, median therapy time 120 months, range 80–184; median time without therapy 53 months, range 24–96). None of the patients were previously treated with TKI therapy. In addition, non-CML patients (3 patients with essential trombocythemia and one patient with polycythemia vera) treated with IFN-α and healthy volunteers (n = 43) were included as controls. Lymphocyte populations in all four groups were characterized with comprehensive immunophenotyping panels. The clonality of T-cells was analyzed by a TCR γ/δ rearrangement assay by PCR. Lymphocytes were further sorted into CD3+ TCR αβ+ and CD3+ TCR γδ+ populations (n = 8). Additionally, plasma levels of 25 cytokines were measured with a multiplex bead-based cytokine assay (Luminex®). Results. The proportion of NK-cells from lymphocytes was significantly increased in IFN-α discontinued patients (median 26%, range 18–51%) compared to healthy volunteers (11%, 5–21%) or patients on IFN-α therapy (12%, 6–31%)(P=0.0005). Similarly the proportion of CD8+ cells from T-cells was significantly increased in both CML IFN-α groups (55% in IFN-α discontinued patients, 44% in IFN-α treated patients vs. 31% in healthy volunteers; P<0.05 for both groups). Also a larger proportion of T-cells expressed the long-term memory antigen CD45RO in IFN-α patients (74%, 58% vs. 44% in healthy controls, P<0.01). The proportion of regulatory T-cells (CD4+CD25+FoxP3+) was increased in IFN-α groups (6.1%, 5.2% vs. 3.8% in healthy volunteers, P=0.01). Similar changes in immunoprofile were not observed in IFN-treated non-CML patients. Clonal TCR γ/δ rearrangements were observed in 18 of 19 (95%) IFN-treated CML patients as compared to 3 of 22 (14%) in healthy volunteers (P<0.01). In both IFN-α CML patient groups a unique rearrangement pattern was observed: 14/19 (79%) of patients had the Vγ9 gene clonally rearranged. This clonal rearrangement resided in CD3+ γδ+ T-cell population, as assessed by cell sorting. Two of four non-CML patients treated with IFN-α had the same clonal rearrangement, as well as one healthy control (1/22; 5%). Similar clonality patterns have not been observed in dasatinib or imatinib treated CML patients (Kreutzman et al. Blood 2010). IFN-α treatment was associated with a distinct plasma cytokine profile in CML patients. IP-10, IL-6, IL-12, eotaxin, MCP-1, and IFN-γ levels were significantly increased in IFN-α treated CML patients. In particular, eotaxin and MCP-1 levels differed significantly between healthy controls and IFN-α patients who had successfully discontinued IFN-α therapy (428 vs. 1173 pg/ml, P<0.0001 and 107 vs. 459 pg/ml, P=0.0003, respectively). In IFN-α treated non-CML patients, eotaxin or MCP-1 levels were not increased. Conclusions. Our results show that IFN-α treatment induces distinct changes in the immunoprofile of CML patients. Patients who had successfully discontinued IFN-α therapy differed markedly from healthy controls. IFN-α therapy was associated with increased numbers of NK-cells and clonal γδ+ T-cells. These cells possess potent anti-leukemic activity and may contribute to the prolonged therapy responses in this group of patients. Furthermore, plasma cytokine profile could be a helpful biomarker when considering which patients can discontinue the IFN-α treatment without imminent disease relapse. Disclosures: Faber: BMS, Novartis: Consultancy, Honoraria. Porkka:BMS, Novartis: Consultancy, Honoraria, Research Funding. Mustjoki:BMS, Novartis: Honoraria.

scholarly journals Characterization of the DNAM-1, TIGIT and TACTILE Axis on Circulating NK, NKT-Like and T Cell Subsets in Patients with Acute Myeloid Leukemia

Cancers ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 2171
Author(s):  
Isabel Valhondo ◽  
Fakhri Hassouneh ◽  
Nelson Lopez-Sejas ◽  
Alejandra Pera ◽  
Beatriz Sanchez-Correa ◽  
...  
Keyword(s):  
T Cells ◽  
Acute Myeloid Leukemia ◽  
T Cell ◽  
Healthy Volunteers ◽  
Nk Cells ◽  
Myeloid Leukemia ◽  
Cell Activation ◽  
Nk Cell ◽  
T Cell Subsets ◽  
Acute Myeloid

Background: Acute myeloid leukemia (AML) remains a major clinical challenge due to poor overall survival, which is even more dramatic in elderly patients. TIGIT, an inhibitory receptor that interacts with CD155 and CD112 molecules, is considered as a checkpoint in T and NK cell activation. This receptor shares ligands with the co-stimulatory receptor DNAM-1 and with TACTILE. The aim of this work was to analyze the expression of DNAM-1, TIGIT and TACTILE in NK cells and T cell subsets in AML patients. Methods: We have studied 36 patients at the time of diagnosis of AML and 20 healthy volunteers. The expression of DNAM-1, TIGIT and TACTILE in NK cells and T cells, according to the expression of CD3 and CD56, was performed by flow cytometry. Results: NK cells, CD56− T cells and CD56+ T (NKT-like) cells from AML patients presented a reduced expression of DNAM-1 compared with healthy volunteers. An increased expression of TIGIT was observed in mainstream CD56− T cells. No differences were observed in the expression of TACTILE. Simplified presentation of incredibly complex evaluations (SPICE) analysis of the co-expression of DNAM-1, TIGIT and TACTILE showed an increase in NK and T cells lacking DNAM-1 and co-expressing TIGIT and TACTILE. Low percentages of DNAM-1−TIGIT+TACTILE+ NK cells and DNAM-1− TIGIT+TACTILE+ CD56− T cells were associated with a better survival of AML patients. Conclusions: The expression of DNAM-1 is reduced in NK cells and in CD4+ and CD8+ T cells from AML patients compared with those from healthy volunteers. An increased percentage of NK and T cells lacking DNAM-1 and co-expressing TIGIT and TACTILE is associated with patient survival, supporting the role of TIGIT as a novel candidate for checkpoint blockade.

Mono/Oligoclonal T and NK Cells Are Common in Philadelphia Chromosome Positive (Ph+) Leukemia Patients at Diagnosis and Expand During Successful Tyrosine Kinase Inhibitor Therapy.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 856-856
Author(s):  
Anna Kreutzman ◽  
Vesa Juvonen ◽  
Veli Kairisto ◽  
Marja Ekblom ◽  
Leif Stenke ◽  
...  
Keyword(s):  
T Cell ◽  
Tyrosine Kinase ◽  
Nk Cells ◽  
Research Funding ◽  
Nk Cell ◽  
Gene Rearrangements ◽  
Imatinib Treatment ◽  
Healthy Controls ◽  
Clonal Rearrangement

Abstract Abstract 856 Introduction. Central to current treatment of Ph+ leukemia patients are tyrosine kinase inhibitors (TKIs), which predominantly target the BCR-ABL1 kinase in malignant cells. However, broader-spectrum 2nd generation TKIs, such as bosutinib, dasatinib and nilotinib, also inhibit off-target kinases with important physiological functions. Several in vitro studies have implied that TKIs may have immunosuppressive effects by suppressing activation and proliferation of effector lymphocytes. In contrast, we recently observed immunostimulation during dasatinib therapy in the form of marked expansion of clonal cytotoxic lymphocytes (T- and NK cells) resulting in chronic LGL-type lymphocytosis in peripheral blood (PB). The prevalence, detailed molecular background and clinical implications of clonal lymphocytes during TKI therapy are currently unknown. The aim of this study was to comprehensively analyze clonality and evolution of lymphocyte clones during TKI therapy. Patients and methods. The study population included patients with Ph+ leukemia, both CML (n=28) and ALL patients (n=4) on dasatinib (n=23) and imatinib (n=9) therapies. In addition, samples from 12 healthy controls and diagnostic samples from the nine imatinib treated patients were analyzed. Lymphocyte clonality was determined by analysis of PB mononuclear cells (MNC) for clonal T cell receptor (TCR) γ and δ gene rearrangements by 18 primer pairs covering most known clonal TCR γ and δ rearrangements. Upon positive reaction in heteroduplex analysis, the purified PCR products were sequenced. If clonal rearrangement was observed, allele-spesific PCR primers were designed to allow for quantitative follow-up of lymphocyte clones in each patient. Results. Sequencing-confirmed clonal TCR γ rearrangement was observed only in 1 of 12 healthy controls and no TCR δ gene rearrangements were found in this group. Surprisingly, 7 of 9 (78%) CML patients showed clonal TCR rearrangements at diagnosis. In 3 patients the clonal rearrangement was detected in the TCR δ genes, in 7 patients in the TCR γ genes and 3 patients had rearrangemens both in TCR δ and γ genes. After one year of imatinib treatment the same clones could be detected in 5 of the 7 patients (71%). Although clonal cells were observed, none of the imatinib patients had signs of a concomitant lymphoproliferative disorder and the distribution of lymphocyte subclasses was normal. Next, 23 patients treated with dasatinib were studied, 10 without (LGLneg) and 13 with PB LGL lymphocytosis (LGLpos) including T- or NK-cell expansions. In all LGLpos dasatinib patients (including patients with a CD3neg NK-cell expansion) clonal TCR γ or δ rearrangements were found. In LGLneg dasatinib patients the prevalence of TCR rearrangements was 80%. LGLpos patients had more often clonal rearrangements in TCR δ genes (62%) than LGLneg patients (10%). No differences in clonal rearranged TCR γ genes (77% vs. 80%) were detected. Most patients displayed more than one clonal TCR rearrangement. Quantitative follow-up of LGLpos patients revealed that the expansion of a single predominant lymphocyte clone accounted for LGL lymphocytosis. Intriguingly, quantitative follow-up of lymphocyte clones by PCR showed that the observed clones existed at low levels already before start of dasatinib therapy during imatinib treatment, but no lymphocyte expansions were then seen. Sorting of lymphocytes showed that clonal cells resided in the CD8+ and CD4+ T-cell populations and, strikingly, also among CD16/56+CD3neg NK cells. All dasatinib patients with NK cell expansions (n=3) showed TCR δ rearrangements in their NK cells. Conclusions. Clonal lymphocytes could rarely be found in healthy controls. In contrast, they were frequently present in CML patients at diagnosis and persisted during TKI therapy. In a distinct subgroup of dasatinib treated patients, clonal cells massively expanded during successful therapy. Clonal TCR rearrangements were detected in CD4+, CD8+ and, unexpectedly, also in NK cells. The epitopes and function of clonal, CML-associated lymphocytes are under investigation. Previous studies showed that clonal expansions during dasatinib were associated with excellent, long-lasting therapy responses in advanced leukemia. We therefore hypothesize, that the clonal lymphocytes present at CML diagnosis may be anergic anti-leukemic cells and part of the immune escape mechanisms inherent to leukemogenesis and that dasatinib therapy can reverse this anergy. Disclosures: Ekblom: BMS: Honoraria. Seggewiss:BMS: Honoraria. Porkka:BMS: Honoraria, Research Funding; Novartis: Honoraria, Research Funding. Mustjoki:BMS: Honoraria.

Early Disease Relapse after Tyrosine Kinase Inhibitor Treatment Discontinuation in CML Is Related Both to Low Number and Impaired Function of NK-Cells

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 812-812 ◽  
Author(s):  
Mette Matilda Ilander ◽  
Ulla Olsson-Strömberg ◽  
Hanna Lähteenmäki ◽  
Kasanen Tiina ◽  
Perttu Koskenvesa ◽  
...  
Keyword(s):  
T Cells ◽  
Nk Cells ◽  
Research Funding ◽  
Nk Cell ◽  
Treatment Discontinuation ◽  
Disease Relapse ◽  
Cell Counts ◽  
Tnf Α ◽  
Ifn Γ

Abstract Background: Recent reports suggest that approximately 40% of CML patients who have achieved sustained complete molecular remission are able to stop TKI treatment without disease relapse. However, there are no predictive markers for successful therapy discontinuation. Therefore, we set up an immunological sub-study in the ongoing pan-European EURO-SKI stopping study. Our aim was to identify predictive biomarkers for relapse/non-relapse and to understand more on the mechanisms of immune surveillance in CML. Methods: The EURO-SKI study started in 2012, and patients included were at least three years on TKI and at least one year in MR4 or deeper before the study entry. Basic lymphocyte immunophenotyping (the number of NK-, T- and B-cells) was performed at the time of therapy discontinuation and 1, 6, and 12 months after the TKI stop and in case of relapse (defined as loss of MMR, BCR-ABL1>0.1% IS). In addition, from a proportion of patients more detailed immunophenotypic and functional analyses (cytotoxicity of NK-cells and secretion of Th1 type of cytokines IFN-γ/TNF-α) were done at the same times. Results: Thus far 119 Nordic patients (imatinib n=105, dasatinib n=12, nilotinib n=2) who have discontinued TKI treatment within the EURO-SKI study have been included in the lymphocyte subclass analysis (results are presented from patients who have reached 6 months follow-up). Immunophenotyping analysis demonstrates that imatinib treated patients who were able to maintain remission for 6 months (n=36) had increased NK-cell counts (0.26 vs. 0.15x109cells/L, p=0.01, NK-cell proportion 18.9% vs. 11%, p=0.005) at the time of drug discontinuation compared to patients who relapsed early (before 5 months n=22). Furthermore, the phenotype of NK-cells was more cytotoxic (more CD57+ and CD16+cells and less CD62L+cells), and also their IFN-γ/TNF-α secretion was enhanced (19.2% vs. 13%, p=0.02). Surprisingly, patients who relapsed more slowly (after 5 months, n=16) had similar baseline NK-cell counts (0.37x109cells/L), NK-cell proportion (21.2%), and phenotype and function as patients, who were able to stay in remission. No differences in the NK-cell counts were observed between patients who had detectable or undetectable BCR-ABL1 transcripts at the baseline (0.22 x109cells/L vs. 0.31 x109cells/L, p=0.61). Interestingly, NK-cell count was higher in patients with low Sokal risk score than in patients with intermediate risk (0.33 x109cells/L vs. 0.20 x109cells/L, p=0.04). Furthermore, there was a trend that male patients had a higher proportion of NK-cells than females (21.6% vs. 15.7%, p=0.06). Pretreatment with IFN-α or the duration of imatinib treatment did not have an effect on NK-cell count or proportion. In comparison to the imatinib group, dasatinib treated patients had higher NK-cell counts at the baseline (median 0.52x109cells/L vs. 0.26x109cells/L, p=0.02), and also the proportion of CD27 (median 50% vs. 16%, p=0.01) and CD57 expressing (median 79% vs. 74%, p=0.05) NK-cells was higher. The follow-up time of dasatinib treated patients is not yet long enough to correlate the NK-cell counts with the success of the treatment discontinuation. The absolute number of T-cells or their function did not differ significantly between relapsing and non-relapsing patients at the time of treatment discontinuation. However, both CD4+ and CD8+ T-cells tended to be more mature in patients who stayed in remission compared to patients who relapsed early (CD4+CD57+CD62L- median 5.7% vs. 2.4%, p=0.06, CD8+CD62L+CD45RA+ 13% vs. 26.7%, p=0.05). The analysis of follow-up samples showed that in patients who stayed in remission the Th1 type cytokine (IFN-γ/TNF-α) secretion of CD8+T-cells increased at 6 months compared to baseline (23.6 vs. 18.5%, p=0.07). Same phenomenon was observed in the late relapsing group at relapse compared to baseline (37.9 vs. 13.5%, p=0.03). No similar increase was observed in the early relapsing group. Conclusions: Low NK-cell numbers and poor cytokine secretion may predict early disease relapse after TKI discontinuation. However, patients who relapse later have high numbers of normally functioning NK-cells. Further research (detailed phenotypic analysis of NK- and T-cells including activating and inhibitory receptors and immune checkpoint molecules) and correlation of biomarker data with clinical parameters are ongoing to understand the ultimate determining factors of relapse. Disclosures Själander: Novartis: Honoraria. Hjorth-Hansen:Novartis: Honoraria; Bristol-myers Squibb: Honoraria; Ariad: Honoraria; Pfizer: Honoraria. Porkka:BMS: Honoraria; BMS: Research Funding; Novartis: Honoraria; Novartis: Research Funding; Pfizer: Research Funding. Mustjoki:Bristol-Myers Squibb: Honoraria, Research Funding; Novartis: Honoraria, Research Funding.

scholarly journals NKG2C+CD57+ Natural Killer Cells May Play an Important Role in Maintaining Virus-Specific Immune Reconstitution Post-Allogeneic HSCT

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 3327-3327
Author(s):  
Debora Queiros ◽  
Susanne Luther-Wolf ◽  
Eva M Weissinger ◽  
Arnold Ganser
Keyword(s):  
T Cells ◽  
Healthy Volunteers ◽  
Nk Cells ◽  
Natural Killer ◽  
Peripheral Blood ◽  
Immune Reconstitution ◽  
Blood Samples ◽  
Allogeneic Hsct ◽  
Cell Counts ◽  
Cmv Reactivation

Abstract Background: Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative treatment for malignant hematological diseases in adults. Due to the delayed immune reconstitution after HSCT, human cytomegalovirus (CMV) can reactivate, leading to prolonged hospitalization and increased morbidity and even mortality. Natural Killer (NK) cells have recently been described to undergo persistent reconfiguration in response to CMV-reactivation. Here we analyzed the presence and expansion of CMV-specific NK cells in patients after allogeneic HSCT. Methods: A multicolor flow cytometry panel for monitoring the CMV-specific NK cell (NKG2C+CD57+) reconstitution and expression of activating receptors was established. Reconstitution of CMV-specific NK cells was assessed in peripheral blood samples from 67 CMV-seropositive patients. The samples were collected and analyzed between day 0 and 100 post-HSCT at intervals of 7-10 days. Monitoring of CMV-reactivation by CMV-pp65 expression and reconstitution of CMV-specific T cells (CMV-CTLs) was done routinely in our laboratory, using 7 commercially available, certified CMV-tetramers, allowing for comparison of CMV-CTL and NKG2C+CD57+ NK cells. For further immunological tests, PBMCs from CMV-seropositive healthy volunteers were isolated by density gradient centrifugation. NK cells were negatively selected by magnetic bead separation. Additional purification of NKG2C+CD57+ NK cellswas achieved by cell sorting. Selected NK cells were expanded by co-culture with irradiated allogeneic PBMCs as feeder cells and the medium was supplemented with PHA and IL-2. Expanded CD57+NKG2C+ NK cells were KIR-typed. Results: Our patient cohort consisted of 67 patients after allogeneic HSCT with a median age of 59 years (range: 20-75). Forty-two patients (62.7%) were transplanted for acute leukemia, 54 (80.6%) received reduced intensity conditioning (RIC) and 62 (92.5%) received anti-thymocyte-antibodies globulin (ATG). GvHD-prophylaxis was cyclosporine A (CsA) in combination with mycophenolate motefil (MMF) for 82.1% of the patients and 77.6% were transplanted from matched donors. Thirty-three (49.2%) patients reactivated CMV (median age: 59.5 years, range 28-75; median day of reactivation: 38 days post-HSCT, range: 19-54). A significant increase in the absolute cell counts of NKG2C+CD57+ NK cells was observed after CMV reactivation, when compared to patients who did not reactivate CMV (p<0.0001). Interestingly, we observed a decreased expression of the CD8-molecule on NK cells during CMV-reactivation. CD8-expression on NK cells was previously described to be associated with a more cytotoxic phenotype of NK cells, this decrease may be a consequence of apoptosis following lytic activity. Monitoring for an additional activation marker, NKG2D, showed a significant increased expression after CMV reactivation (p=0.006), demonstrating not only the activating regulation of NK cells, but also, the co-stimulatory effects on T cell proliferation and cytokine production. Remarkably, when comparing NKG2C+CD57+ NK cells with CMV-specific T cells (Figure 1), both cell populations show similar kinetics of expansion, with an increase in the absolute cell counts during and after CMV-reactivation. NKG2C+CD57+ preliminary expansion-studies were performed using peripheral blood samples from CMV-seropositive healthy volunteers. After two weeks in culture, an expansion of up to 3100-fold was achieved. Further studies to assess the proliferative capacity of NKG2C+CD57+ subpopulation and its functional properties post-HSCT, are ongoing. In addition, an extensive panel of cytokines and chemokines excreted by the NKG2C+CD57+cells will be studied in order to evaluate their recruitment ability of other cell-types. Conclusion: Taken together, our results indicate that NK cells undergo a dynamic modulation and expansion of this population occurs in response to CMV-reactivation. Additionally, NKG2C+CD57+ NK cells may substitute for missing CMV-specific T cells shortly after HSCT and may play an important role in sustaining the immune reconstitution after CMV-reactivation. This study shows that NKG2C+CD57+ NK cells can be selected and expanded in vitro, holding promise for adoptive transfer in patients with recurrent CMV-reactivations. Disclosures Ganser: Novartis: Membership on an entity's Board of Directors or advisory committees.

scholarly journals Circulating cytokines and lymphocyte subsets in patients who have recovered from COVID-19

2020 ◽  
Author(s):  
Hasi Chaolu ◽  
Xinri Zhang ◽  
Xin Li ◽  
Xin Li ◽  
Dongyan Li
Keyword(s):  
T Cells ◽  
B Cells ◽  
Nk Cells ◽  
Cd8 T Cells ◽  
Cd4 T Cells ◽  
Lymphocyte Subsets ◽  
Healthy Controls ◽  
Tnf Α ◽  
Ifn Γ ◽  
Circulating Cytokines

To investigate the immune status of people who previously had COVID-19 infections, we recruited patients 2 weeks post-recovery and analyzed circulating cytokines and lymphocyte subsets. We measured levels of total lymphocytes, CD4+ T cells, CD8+ T cells, CD19+ B cells, CD56+ NK cells, and the serum concentrations of interleukin (IL)-1, IL-4, IL-6, IL-8, IL-10, transforming growth factor beta (TGF-β), tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ) by flow cytometry. We found that in most post-recovery patients, levels of total lymphocytes (66.67%), CD3+ T cells (54.55%), CD4+ T cells (54.55%), CD8 + T cells (81.82%), CD19+ B cells (69.70%), and CD56+ NK cells(51.52%) remained lower than normal, whereas most patients showed normal levels of IL-2 (100%), IL-4 (80.88%), IL-6 (79.41%), IL-10 (98.53%), TNF-α (89.71%), IFN-γ (100%) and IL-17 (97.06%). Compared to healthy controls, 2-week post-recovery patients had significantly lower absolute numbers of total lymphocytes, CD3+ T cells, CD4+ T cells, CD8+ T cells, CD19+ B cells, and CD56+ NK cells, along with significantly higher levels of IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ and IL-17. Among post-recovery patients, T cells, particularly CD4+ T cells, were positively correlated with CD19+ B cell counts. Additionally, CD8+ T cells positively correlated with CD4+ T cells and IL-2 levels, and IL-6 positively correlated with TNF-α and IFN-γ. These correlations were not observed in healthy controls. By ROC curve analysis, post-recovery decreases in lymphocyte subsets and increases in cytokines were identified as independent predictors of rehabilitation efficacy. These findings indicate that the immune system has gradually recovered following COVID-19 infection; however, the sustained hyper-inflammatory response for more than 14 days suggests a need to continue medical observation following discharge from the hospital. Longitudinal studies of a larger cohort of recovered patients are needed to fully understand the consequences of the infection.

Dasatinib Has a Dual Mode of Action: Direct BCR-ABL1 Mediated Anti-Leukemic Effects Are Complemented by Promotion of Th1-Type and NK-Cell Mediated Cellular Immune Responses,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3760-3760 ◽  
Author(s):  
Anna Kreutzman ◽  
Jukka Vakkila ◽  
Kimmo Porkka ◽  
Satu Mustjoki
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Nk Cells ◽  
Cd8 T Cells ◽  
K562 Cells ◽  
Leukemic Cells ◽  
Healthy Controls ◽  
Median Percentage ◽  
Blood Smears ◽  
Cellular Immune

Abstract Abstract 3760 Background. Tyrosine kinase inhibitors (TKIs; imatinib, dasatinib, nilotinib) have dramatically improved outcome of CML. Besides inhibiting target kinases in leukemic cells, off-target kinases in immune effector cells are also affected. We have previously described that dasatinib therapy induces an oligoclonal expansion and mobilization of large granular lymphocytes (LGLs; CD8+ T-cells or NK-cells) in Ph+ leukemia patients. Importantly, LGL expansion is associated with improved therapy responses, but the actual mechanisms are unknown. In this study, we explored the function and anti-leukemic properties of LGLs. Methods. Peripheral blood samples from CML patients treated with dasatinib (n=10), imatinib (n=4), or nilotinib (n=7), or healthy controls (n=6) were used to analyze the activation and cytotoxicity of T- and NK-cells. Samples were collected before and after drug intake. The number of LGLs was determined from MGG stained blood smears and compared with granzyme B (GrB) positivity analyzed by flow cytometry. Th1-type cytokine (TNF-a, IFN-g) production was measured by flow cytometry after stimulation of mononuclear cells (MNCs) with a-CD3/CD28-antibodies. Unpurified and purified NK cells were cultured with K562 cells, and degranulation (CD107 analysis) and cytotoxicity were measured. Results. As GrB positivity correlated well (r=0.95, p<0.0001, n=17) with the number of LGLs counted from MGG stained blood smears, a GrB specific antibody was used to identify LGLs in further analyses. At diagnosis CML patients had more GrB+CD8+ T-cells than healthy controls (38 % vs. 11%, p=0.028). Also GrB+CD4+ T-cells were slightly increased, but did not differ significantly from healthy controls (3.6% vs. 0.8%, p=0.08). During dasatinib treatment the proportion of GrB+CD4+ (median at 6 months 28.1%, p=0.03) and GrB+CD8+ (70.9%, p=0.03) T cells increased significantly, whereas similar increase was not observed during imatinib (1.2% GrB+CD4+ and 30.0% GrB+CD8+ T-cells) or nilotinib (4.4% and 41.8%, respectively) therapies. In patients on dasatinib therapy, GrB+CD3+cells were more sensitive to CD3/CD28-antibody stimulation and a larger proportion of cells (13.7%) produced Th1-type cytokines (TNF-a+IFN-g) compared to imatinib (2.4%) or nilotinib patients (5.5%) or healthy controls (5%) under same conditions (p=0.015). As Th-1 cytokine-producing T cells are important in promoting cell-mediated immune responses, we next assessed whether dasatinib also enhances the cytolytic activity of NK cells. When MNC fraction was used as effector population (ratio 20:1), the median percentage of dead K562 cells was 18% in samples taken before dasatinib intake and 32% in samples taken 1h after dasatinib intake (p=0.004). Pre-dasatinib killing did not differ significantly from healthy volunteers (p=0.12). No increase in NK-cytotoxicity was observed after imatinib (11% vs. 8%) or nilotinib (10% vs. 10%) intake. Similar results were also obtained with purified NK-cells: the median percentage of dead K562 cells was 12% pre-dasatinib and 29% in post-dasatinib samples (p=0.06), whereas no differences were noticed with imatinib (30% vs. 28%) or nilotinib (14% vs. 15%) patients. The median percentage of dead K562 cells after incubation with pure NK-cells from healthy volunteers was 20%. Interestingly, the cytolytic ability of NK-cells differed significantly among dasatinib treated patients. When the patients were divided into two groups based on therapy response, patients who had achieved CMR within 12 months (n=4) had significantly higher cytotoxic capability compared to patients who had not (n=6): 46% vs. 28% of dead K562 cells in post-dasatinib samples (p=0.02). Conclusions. Dasatinib therapy resulted in increased numbers of GrB+ T-cells and generation of a Th1-type cellular immune response. In addition, 1h dasatinib exposure in vivo improved the cytotoxicity of NK-cells. These data support the dual mode of action of dasatinib: potent BCR-ABL1 inhibition in leukemic cells is accompanied by enhancement of cellular immunity, which likely have implications in be the long term control of Ph+ leukemia. Disclosures: Porkka: Bristol-Myers Squibb: Honoraria, Research Funding; Novartis: Honoraria, Research Funding. Mustjoki:Bristol-Myers Squibb: Honoraria; Novartis: Honoraria.

Enhancement of the host immune responses in cutaneous T-cell lymphoma by CpG oligodeoxynucleotides and IL-15

Blood ◽  
2004 ◽  
Vol 104 (13) ◽  
pp. 4142-4149 ◽  
Author(s):  
Maria Wysocka ◽  
Bernice M. Benoit ◽  
Sarah Newton ◽  
Livio Azzoni ◽  
Luis J. Montaner ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Healthy Volunteers ◽  
Nk Cells ◽  
Cd8 T Cells ◽  
Cell Lymphoma ◽  
T Cell Lymphoma ◽  
Cell Mediated Immunity ◽  
Cutaneous T Cell Lymphoma ◽  
Cpg Oligodeoxynucleotides

Abstract Patients with advanced cutaneous T-cell lymphoma (CTCL) exhibit profound defects in cell-mediated immunity. Host immune functions appear to play an integral role in mediating disease-controlling responses in CTCL, therefore we investigated the effects of synthetic oligode-oxynucleotides with CpG motifs (CpG ODN), which have been recognized as immune stimulatory by virtue of activation of dendritic cells (DCs) following binding to Toll-like receptor (TLR) 9. Peripheral blood mononuclear cells (PBMCs) from patients with advanced CTCL (erythroderma with circulating malignant T cells) and healthy volunteers were cultured with either CpG-A or CpG-B ODN. Patients' PBMCs exhibited marked induction of interferon-α (IFN-α) release following culture with CpG-A. Similarly significant activation of NK cells and CD8 T cells occurred as assessed by up-modulation of CD69 expression and by natural killer lytic activity. Nevertheless, the PBMCs of patients exhibited blunted responses to CpG-A compared to healthy volunteers. In such cases, IL-15 was capable of producing levels of NK activation that were superior to CpG-A, while the combined effects of CpG-A plus IL-15 induced maximal activation of NK cells and further enhanced activation of CD8 T cells. These findings have important implications for the potential enhancement of antitumor immunity among patients with advanced CTCL.

scholarly journals Clinical and immunological features of the combined course of pertussis and rhinovirus infection in children

2021 ◽  
Vol 66 (5) ◽  
pp. 82-87
Author(s):  
O. P. Popova ◽  
M. S. Blyakher ◽  
I. M. Fedorova ◽  
S. I. Koteleva ◽  
I. V. Kapustin ◽  
...  
Keyword(s):  
Nk Cells ◽  
Research Methods ◽  
Interferon Gamma ◽  
Interferon Alpha ◽  
Respiratory Infections ◽  
Cytokine Profile ◽  
Low Level ◽  
Alpha Production ◽  
Rhinovirus Infection ◽  
Complicated Course

The association of pertussis with various respiratory infections in children is the leading factor determining the complicated course and unfavorable outcome of the disease.Objective. To analyze clinical and immunological features of the combined course of pertussis and rhinovirus infection.Children characteristics and research methods. The authors observed 20 patients: 10 (50%) children were under the age of 1 year, 5 (25%) children of 1–3 years old, 3 (15%) children of 4–6 years, 2 (10%) children of 7 -14 years old.Results. The rhinovirus infection developed mainly at 1-2 weeks of illness in 18 (90%) children. Bronchitis developed in 11 (55%) children, pneumonia – in 4 (20%). Special attention was drawn to the low content of NK cells in 82.4% of patients when assessing the subpopulation composition of lymphocytes. The cytokine profile was characterized by a low level of interferon-gamma and interferon-alpha production – in 94.4 and 61.1% of patients, respectively.Conclusion. The combination of pertussis and rhinovirus infection in children contributes to the uneven course of the disease, the frequent development of bronchopulmonary complications.

The NK and NKT Cell Compartment Is Suppressed in CML Patients before and after Imatinib Treatment.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4667-4667
Author(s):  
Christiane I.-U. Chen ◽  
Holden Maecker ◽  
Peter P. Lee
Keyword(s):  
T Cells ◽  
Nk Cells ◽  
Imatinib Mesylate ◽  
Cd8 T Cells ◽  
Lymphocyte Subsets ◽  
Nkt Cells ◽  
Healthy Controls ◽  
Nkt Cell ◽  
Cell Counts ◽  
Before And After

Abstract In CML patients, lymphocyte subsets have been shown to be disturbed. NK cells as well as CD8+ T cells have been shown to be significantly lower in patients with relapse after BMT than without relapse (Jiang et al., 1996). After successful treatment (IFN- α and BMT/SCT), MHC-restricted CD8+ T cells as well as MHC-unrestricted cytotoxic NK and lymphokine activated killer (LAK) cells play an important role in controlling the disease (Meseri et al., 1993, de Castro et al., 2003, Molldrem et al., 2000), which can lead not only to hematological, but also cytogenetic responses. The development of imatinib mesylate, a selective inhibitor of the bcr/abl tyrosine kinase, as a first-line therapy enormously enhanced treatment options for CML patients, and has led to hematologic, and occasionally cytogenetic, responses at various stages of disease. To explore, if CML patients, who develop clinical responses with imatinib mesylate treatment, have normalization of their lymphocyte subsets with restoration of CD8+ and NK cells, we studied peripheral blood T cell, B cell, and NK cell subpopulations in patients in chronic or accelerated phase before and after treatment with imatinib mesylate. In 5 CML patients, 8-color flow cytometry was applied to investigate the lymphocyte subsets using anti-CD3, anti-CD4, anti-CD8, anti-CD19, anti-CD56, anti-CD25, anti-CD27, and anti-CD45RA antibodies. 5 healthy donors served as controls. There was no significant difference in CD19+ B cells or CD3+ T cells as well as its CD4+ and CD8+ subsets between CML patients and healthy controls. T regulatory cells (CD4+CD25+) were also similar in patients in comparison to the healthy controls (0.6% – 3% vs 2.3%). In contrast, CD3–CD56+ NK cells (4% vs 14%, p&lt;0.05) as well as NK T cells (CD3+CD56+ 1% vs 14%, p&lt;0.02; CD8+CD56+ 2% vs 17%, p&lt;0.03) were profoundly decreased in CML patients compared to the healthy controls prior to treatment with imatinib mesylate. Importantly, these abnormalities persisted even during treatment and following hematological responses (range 10 months – 35 months) (CD3-CD56+ 4–8%, CD3+CD56+ 0.5–5.5%, CD8+CD56+ 2.7–7%). On further analysis, these NK and NKT cells were mostly effector (CD45RA+CD27−, 50–80%) or memory (CD45RA-CD27+, 10–30%) cells. These results suggest, that CML not only disturbs the myeloid and the lymphoid compartment before treatment, but persistently impairs the MHC-unrestricted cytotoxic NK cells and NKT cells after treatment with imatinib mesylate, even in patients in hematological (and cytogenetic) remission with normal white cell counts (2.0 – 8.0/nl). Further investigations are required to better understand the mechanism of the prolonged and specific NK and NKT cell suppression in imatinib mesylate treated patients, and its impact on the course of the disease.

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