scholarly journals Analysis of Hyperthermia Effect on Peripheral Blood Natural Killer Cell Cytotoxicity against SW-872 Cell Line

2015 ◽  
Vol 4 (3) ◽  
pp. 75-81
Author(s):  
Mohammad Reza Atashzar ◽  
Sajad Jalili
Keyword(s):  
Heat Treatment ◽  
Natural Killer Cell ◽  
Nk Cells ◽  
Natural Killer ◽  
Cell Line ◽  
Peripheral Blood ◽  
Killer Cell ◽  
Target Cells ◽  
Cell Cytotoxicity ◽  
Heat Treated

Background: Natural killer (NK) cells are a type of cytotoxic lymphocyte. It is revealed that hyperthermia which is used in cancer treatment, increases natural killer cell activity. In this study, our aim was to analyze effects of in-vitro hyperthermia on human Natural Killer cytotoxicity against SW-872 cell line. Materials and Methods: In this study, we used SW-872 liposarcoma cell line as a target cell. Peripheral blood NK cells were isolated from normal individuals by MACS. NK cells were treated at 39°C for 1hr .The expression of CD69 on the surface of NK cells was examined by flow cytometry at different time points including 0, 6 and 12 hrs. NK cell cytotoxicity was measured by LDH assay 12hrs after co-culture with SW-872. The results were compared to the conditions that both NK cells and SW-872 cells were heat treated at 39°C for 12hrs. Results: Our results revealed that cell killing activity of NK cells treated with heating alone was increased 6hrs after heat treatment at 39°C compared with heat-treated NK-target cells co-culture. While in heat-treated NK-target cells co-culture the maximum cytotoxicity was observed 12hrs after heat treatment at 39°C. Conclusion: These results showed that thermotherapy could be a feasible method to stimulate immune response against tumor cells. [GMJ.  2015;4(3):75-81]

scholarly journals Aggressive natural killer cell leukemia in an adult with establishment of an NK cell line

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 925-930 ◽  
Author(s):  
LA Fernandez ◽  
B Pope ◽  
C Lee ◽  
E Zayed
Keyword(s):  
Platelet Count ◽  
Nk Cells ◽  
Natural Killer ◽  
Cell Line ◽  
Peripheral Blood ◽  
Nk Cell ◽  
Killer Cell ◽  
Chromosome 1 ◽  
Cell Leukemia ◽  
Wbc Count

Abstract There have been many reports of cases in which chronic increases in the numbers of natural killer (NK) cells have been reported. Whether this is reactive or neoplastic in nature has been debated. We report the first case of an aggressive NK cell leukemia in an adult with establishment of an NK cell line. A 70-year-old man had two spontaneous episodes of jejunal perforation and one month later developed a severe febrile illness with moderate splenomegaly. Hemoglobin was 13.1 g/L, and WBC count was 1.8 X 10(9)/L with 2% large granular lymphocytes (LGLs). Platelet count was 143 X 10(9)/L; prothrombin time (PT) and partial thromboplastin time (PTT) were normal. Bone marrow was infiltrated with 25% to 30% LGLs; serum lysozyme was normal. Serum LDH was initially 1,191 U/L and rose to 6,408 (normal 240 to 525 U/L). Ten days later, the WBC count increased to 99.9 X 10(9)/L with 70% LGL cells; the PT and PTT increased, and the platelet count dropped. No bacterial or viral cause of fever was identified. The cells from peripheral blood were LGLs that stained positively for acid phosphatase. All of the LGLs reacted with a monoclonal antibody reactive with NK cells (LEU-11b). Functionally, the patient's peripheral blood mononuclear cells (PBMs) demonstrated 100 times more lytic activity against K562 tumor cell lines than did normal PBMs. The patient's PBMs were propagated in vitro. The cultured cells showed the morphological, cytochemical, immunological, and functional characteristics of NK cells. In addition, partial trisomy involving chromosome 1 q with duplication in regions of q21 through q31 was observed in all metaphases analyzed. The extra chromosome 1q with duplication in regions q21 through q31 was translocated to the p- terminal of chromosome 5. One percent to 5% of normal PBMs comprise NK cells; in most cases, leukemias arise from normal phenotypic counterparts. This case demonstrated that aggressive NK cell leukemia may occur in adults. In addition, the chromosomal abnormalities suggest that this is not a reactive process but a malignancy.

scholarly journals A research-driven approach to the identification of novel natural killer cell deficiencies affecting cytotoxic function

Blood ◽  
2020 ◽  
Vol 135 (9) ◽  
pp. 629-637
Author(s):  
Michael T. Lam ◽  
Emily M. Mace ◽  
Jordan S. Orange
Keyword(s):  
Natural Killer Cell ◽  
Nk Cells ◽  
Natural Killer ◽  
Nk Cell ◽  
Killer Cell ◽  
Cell Development ◽  
Impact Testing ◽  
Primary Immune Deficiency ◽  
Cell Cytotoxicity ◽  
Nk Cell Cytotoxicity

Abstract Natural killer cell deficiencies (NKDs) are an emerging phenotypic subtype of primary immune deficiency. NK cells provide a defense against virally infected cells using a variety of cytotoxic mechanisms, and patients who have defective NK cell development or function can present with atypical, recurrent, or severe herpesviral infections. The current pipeline for investigating NKDs involves the acquisition and clinical assessment of patients with a suspected NKD followed by subsequent in silico, in vitro, and in vivo laboratory research. Evaluation involves initially quantifying NK cells and measuring NK cell cytotoxicity and expression of certain NK cell receptors involved in NK cell development and function. Subsequent studies using genomic methods to identify the potential causative variant are conducted along with variant impact testing to make genotype-phenotype connections. Identification of novel genes contributing to the NKD phenotype can also be facilitated by applying the expanding knowledge of NK cell biology. In this review, we discuss how NKDs that affect NK cell cytotoxicity can be approached in the clinic and laboratory for the discovery of novel gene variants.

scholarly journals Triggering of human monocyte activation through CD69, a member of the natural killer cell gene complex family of signal transducing receptors.

1994 ◽  
Vol 180 (5) ◽  
pp. 1999-2004 ◽  
Author(s):  
R De Maria ◽  
M G Cifone ◽  
R Trotta ◽  
M R Rippo ◽  
C Festuccia ◽  
...  
Keyword(s):  
Natural Killer Cell ◽  
Natural Killer ◽  
Cell Line ◽  
Peripheral Blood ◽  
Killer Cell ◽  
Cross Linking ◽  
Gene Complex ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Cell Gene

The expression and function of CD69, a member of the natural killer cell gene complex family of signal transducing receptors, was investigated on human monocytes. CD69 was found expressed on all peripheral blood monocytes, as a 28- and 32-kD disulfide-linked dimer. Molecular cross-linking of CD69 receptors induced extracellular Ca2+ influx, as revealed by flow cytometry. CD69 cross-linking resulted also in phospholipase A2 activation, as detected by in vivo arachidonic acid release measurement from intact cells and by direct in vitro measurement of enzymatic activity using radiolabeled phosphatidylcholine vesicles. Prostaglandin E 2 alpha, 6-keto-prostaglandin F 1 alpha, and leukotriene B4 were detected by radioimmunoassay in supernatants from CD69-stimulated monocytes, suggesting the activation of both cyclooxygenase and lipoxygenase pathways after CD69 stimulation. CD69 cross-linking, moreover, was able to induce strong nitric oxide (NO) production from monocytes, as detected by accumulation of NO oxydixed derivatives, and cyclic GMP. It is important to note that NO generation was responsible for CD69-mediated increase in spontaneous cytotoxicity against L929 murine transformed fibroblast cell line and induction of redirected cytotoxicity towards P815 FcRII+ murine mastocytoma cell line. These data indicate that CD69 can act as a potent stimulatory molecule on the surface of human peripheral blood monocytes.

Proteomic Analysis for the Identification of Disease Biomarkers and Therapeutic Targets in Natural Killer Cell Lymphoma.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1774-1774
Author(s):  
Hojung Lee ◽  
Evan Shereck ◽  
Mitchell S. Cairo ◽  
Seiler E Seiler ◽  
Venkatesha Basrur ◽  
...  
Keyword(s):  
Natural Killer Cell ◽  
Western Blot ◽  
Nk Cells ◽  
Natural Killer ◽  
Cell Line ◽  
Blot Analysis ◽  
Proteomic Analysis ◽  
Cell Lymphoma ◽  
Killer Cell ◽  
Differentially Expressed Proteins

Abstract Natural killer cell lymphoma (NKL) is a highly aggressive form of hematolymphoid malignancy that arises from natural killer cells, a critical component of the innate immune system. Early diagnosis of the tumor is critical however there are no biomarkers which are useful currently for the diagnosis and monitoring of patients with NKLs. We hypothesized that comprehensive analysis of differentially expressed proteins by NKL and normal NK cells would be a rational strategy for the discovery of NKL biomarkers and lead to better understanding of pathogenetic mechanisms of NKL. We used total cell lysates obtained from highly enriched peripheral blood NK cells (CD3-, CD56dim) and an Epstein-Barr virus negative, IL-2 dependent cell line (NK-92MI) for quantitative proteomic analysis. Isotopecoded affinity tags (ICATTM)-labeling followed by liquid chromatography-tandem mass spectrometry analyses (LC-MS/MS) identified 114 differentially expressed proteins in NKL compared to the normal NK cells. Out of these, 80 were upregulated 1.5-fold or greater and 34 were downregulated 1.5-fold or greater in NKL. Proteins within diverse cellular functional categories including cell signaling, transcriptional regulation, DNA metabolism, and immune response were differentially expression. Furthermore, known proteins relevant to the function of NK cells such as perforin, granzyme and defensins were downregulated in the NKL cell line. We selected eight proteins to validate the changes identified by ICAT-LC-MS/MS using Western blot analysis. HSP90, PCNA, alpha-enolase, GSTP1, galectin-1, and PARK7 were upregulated, and lactoferrin and annexin A1 were downregulated by western blot analysis demonstrating high concordance with the results of ICAT-LC-MS/MS analysis. We validated the functional significance of HSP90 as a potential therapeutic target in NKL by investigating the effect of inhibitors of HSP90 (geldanamycin) on the survival of an NKL cell line. Inhibition of HSP90 led to significant decrease in cell viability and apoptotic cell death as confirmed by western blotting demonstration of PARP-1 induction and detection of its cleavage products. This study represents the first global quantitative proteomic cataloguing of proteins expressed by NKL and normal NK cells, and reveals potential candidate proteins that may be useful for the diagnosis and function as therapeutic targets in patients with NKL.

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