scholarly journals Lymphokine-activated killer cells in rats. III. A simple method for the purification of large granular lymphocytes and their rapid expansion and conversion into lymphokine-activated killer cells.

1988 ◽  
Vol 167 (1) ◽  
pp. 15-29 ◽  
Author(s):  
N L Vujanovic ◽  
R B Herberman ◽  
A A Maghazachi ◽  
J C Hiserodt
Keyword(s):  
Nk Cells ◽  
Rapid Expansion ◽  
Mononuclear Leukocytes ◽  
Target Cells ◽  
Large Granular Lymphocytes ◽  
Lymphokine Activated Killer Cells ◽  
Simple Method ◽  
Killer Cells ◽  
Very High ◽  
Granular Lymphocytes

A simple method for the purification and rapid expansion of large granular lymphocytes into cells with efficient broad antitumor cytotoxicity after stimulation by human rIL-2 is described. Nylon-wool nonadherent splenic mononuclear leukocytes from Fischer 344 rats were cultured in medium containing 1,000 U/ml rIL-2. The initial response of a small subpopulation of cells (less than 2%) to rIL-2 was their adherence to the plastic surface. This response was noted as soon as 2 h after addition of rIL-2. 2-h rIL-2-activated plastic adherent lymphocytes were 90-98% LGL, expressed surface markers characteristic of rat NK cells (OX8 [CD8]+, asialo GM1, laminin+, OX19 [CD5]-, R1-3B3 [CD5]-, W3/25 [CD4]-, OX39 [CD25]-, Ia-, and Ig-), and expressed very high levels of cytotoxicity against YAC-1 target cells. In addition to the above markers, plastic-adherent LGLs obtained at 24, 48, or 72 h progressively expressed Ia surface antigens, but were not phagocytic and contained less than 1% monocytes/macrophages by morphology. When 24- or 48-h plastic-adherent LGL/NK cells were cultured over 3-4 d in rIL-2, the cells expanded between 30- and 100-fold, reaching densities between 2-3 X 10(6) cells/ml. These rapidly expanding LGL/NK cells also generated very high levels of LAK activity (including lysis of fresh NK-resistant solid tumor cells), expressed a phenotype characteristic of activated rat NK/LAK cells, and incorporated [3H]TdR into DNA. This technique not only provides a novel method for the purification of LGL/NK cells for in vitro studies but also provides a means for the rapid expansion of highly purified cells with high levels of broad antitumor (LAK) cytotoxicity.

scholarly journals Natural killer-mediated lysis of normal and malignant target cells, and its regulation by monocytes.

1985 ◽  
Vol 162 (2) ◽  
pp. 472-486 ◽  
Author(s):  
K Oshimi ◽  
Y Oshimi ◽  
M Satake ◽  
H Mizoguchi
Keyword(s):  
Nk Cells ◽  
Natural Killer ◽  
B Lymphocytes ◽  
Leukemia Cells ◽  
Target Cells ◽  
Large Granular Lymphocytes ◽  
Percoll Density Gradient ◽  
Granular Lymphocytes ◽  
Density Gradient Sedimentation

After depletion of monocytes, natural killer (NK) cells were partially purified from peripheral blood by Percoll density gradient sedimentation. The NK cells were then cultured for 1 d and assayed for their cytotoxicity against various types of normal and malignant target cells. All types of target cells tested were found to be susceptible to NK cells. The susceptible targets were autologous T and B lymphocytes, mitogen-induced T and B blasts, monocytes, large granular lymphocytes, autologous or allogeneic lymphoma and leukemia cells isolated from patients, and cultured cell lines, including those resistant to interferon-activated lymphocytes. Such a broad spectrum of cytotoxicity was demonstrated in 1 d of culture, and freshly prepared NK cells were not cytotoxic, or, if anything, were less cytotoxic. Monocytes and their supernatants, added throughout the course of culture, markedly inhibited the development of their cytotoxicity. These results may suggest that, although NK cells having ability to lyse autologous normal and malignant target cells are present in vivo, their lytic activity is regulated by coexisting monocytes.

scholarly journals Monoclonal antibody to a triggering structure expressed on rat natural killer cells and adherent lymphokine-activated killer cells.

1989 ◽  
Vol 169 (4) ◽  
pp. 1373-1389 ◽  
Author(s):  
W H Chambers ◽  
N L Vujanovic ◽  
A B DeLeo ◽  
M W Olszowy ◽  
R B Herberman ◽  
...  
Keyword(s):  
Nk Cells ◽  
Cell Function ◽  
Nk Cell ◽  
Cytolytic Activity ◽  
Lak Cells ◽  
Target Cells ◽  
Lymphokine Activated Killer Cells ◽  
Killer Cells ◽  
And Function ◽  
Lak Activity

To study the cellular structures involved in NK and lymphokine-activated killer (LAK) cell function, we have produced a panel of mAbs that modulate the cytolytic function of a population of cells with LAK activity that derive from large granular lymphocyte (LGL)/NK cells (adherent LAK [A-LAK] cells). In this report, we describe an mAb (3.2.3; IgG1k) that recognizes a triggering structure that is expressed on rat LGL/NK cells and A-LAK cells. This epitope is also expressed on polymorphonuclear leukocytes (PMN). The expression of the epitope identified by mAb 3.2.3 increased progressively on A-LAK cells after culture in the presence of rIL-2. mAb 3.2.3 enhanced the cytolytic activity of NK and A-LAK cells against FcR+ target cells, but not FcR- target cells. However, this effect was not induced by F(ab')2 fragments of 3.2.3. This antibody also induced the release of N-alpha-benzyloxycarbonyl-L-lysine thiobenzy esteresterase by A-LAK cells. These data suggest that the epitope identified by mAb 3.2.3 is on a triggering structure expressed on rat NK cells and A-LAK cells. The expression of the epitope recognized by mAb 3.2.3 on LGL/NK cells and PMN suggests that this structure may be analogous to that identified by the anti-CD16 (-FcR) mAbs. However, the molecule immunoprecipitated by mAb 3.2.3 was a 60-kD dimer composed of two 30-kD chains. These data suggest that mAb 3.2.3 recognizes a unique triggering structure. As mAb 3.2.3 is the first antibody recognizing a determinant with functional significance, selectively expressed on both rat NK cells and A-LAK cells, it will be a useful tool for the study of NK cell ontogeny and function, and the development of cells with LAK activity from the NK cell compartment.

scholarly journals Thy-1+ and Thy-1- natural killer cells. Only Thy-1- natural killer cells suppress dendritic cells.

1986 ◽  
Vol 163 (4) ◽  
pp. 1012-1017 ◽  
Author(s):  
P D Shah ◽  
J Keij ◽  
S M Gilbertson ◽  
D A Rowley
Keyword(s):  
Natural Killer Cells ◽  
Nk Cells ◽  
Natural Killer ◽  
Poly I:C ◽  
Target Cells ◽  
Killer Cells ◽  
Different Populations ◽  
High Cytotoxicity ◽  
Granular Lymphocytes

Cells enriched for NK activity (poly I:C induced, x-ray resistant, and nonadherent), include two phenotypically and functionally different populations. Both populations of NK cells are AGM1+, Ly-1.1-, Ly-2.1-, Ia-, and have the morphology of large granular lymphocytes. One population, however, is Thy-1+ while the second population is Thy-1-. Thy-1+ NK cells lyse YAC-1 and P815 target cells; Thy-1- NK cells lyse YAC-1 but not P815 target cells. The FACS was used to obtain homogeneous populations of Thy-1+ and Thy-1- NK cells, which retain high cytotoxicity. While Thy-1- NK cells suppress the antibody response in vitro by suppressing or eliminating DC, Thy-1+ NK cells do not suppress antibody responses in vitro.

scholarly journals Battle of Thermopylae: 300 Spartans (natural killer cells plus obinutuzumab) versus the immortal warriors (chronic lymphocytic leukemia cells) of Xerxes’ army

2019 ◽  
Vol 5 (10) ◽  
pp. FSO425
Author(s):  
Ricardo García-Muñoz ◽  
María-Josefa Nájera ◽  
Jesús Feliu ◽  
Judith Antón-Remírez ◽  
Enrique Ramalle-Gómara ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Nk Cells ◽  
Natural Killer ◽  
Peripheral Blood ◽  
Nk Cell ◽  
Killer Cell ◽  
Lymphocytic Leukemia ◽  
Leukemic Cells ◽  
Lymphokine Activated Killer Cells ◽  
Killer Cells

Aim: To analyze the effects of subcutaneous or intravenous rituximab + lymphokine-activated killer cells, obinutuzumab or ibrutinib on natural killer (NK) cell levels in chronic lymphocytic leukemia and follicular lymphoma patients. Patients & methods: The distribution of peripheral blood NK cells of 31 patients was analyzed by flow cytometry. Results: We detected a decrease of NK cells in peripheral blood below normal range after obinutuzumab treatment. During maintenance treatment with subcutaneous rituximab, an NK cell reduction was less pronounced than after intravenous rituximab treatment, despite lymphokine-activated killer cell infusions. Conclusion: After one dose of obinutuzumab, each NK cell in peripheral blood destroys 25 leukemic cells.

scholarly journals Malignant clonal expansion of large granular lymphocytes with a Leu- 11+, Leu-7- surface phenotype: in vitro responsiveness of malignant cells to recombinant human interleukin 2

Blood ◽  
1986 ◽  
Vol 68 (5) ◽  
pp. 1065-1073 ◽  
Author(s):  
S Koizumi ◽  
H Seki ◽  
T Tachinami ◽  
M Taniguchi ◽  
A Matsuda ◽  
...  
Keyword(s):  
Interleukin 2 ◽  
Receptor Expression ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Target Cells ◽  
Transferase Activity ◽  
Large Granular Lymphocytes ◽  
Surface Phenotype ◽  
Granular Lymphocytes ◽  
Leu 7

Abstract A 14-year-old Japanese female with neutropenia showed malignant proliferation of the large granular lymphocytes (LGLs). These LGLs were E rosette+ and Fc(IgG) receptor+ and therefore are referred to as T gamma lymphocytes. They were also Leu-11+ and OKT11+; however, they were clearly negative for Leu-7, OKT3, OKT8, OKM1, and HNK-1 antigens as well as for terminal deoxynucleotidyl transferase activity. Karyotype analysis revealed 47, XXX. The LGLs showed no rearrangement of T cell receptor C beta genes. The natural killer (NK) cell activity against K562 target cells was low, but was significantly augmented after stimulation by recombinant human interleukin 2 (IL 2) in contrast to minimal NK boosting by recombinant human gamma-interferon (gamma- IFN). Such a unique responsive ability to lymphokines was quite similar to that noted in fetal and cord blood cells. These LGLs also demonstrated a considerable increase in antibody-dependent cell- mediated cytotoxicity (ADCC) and lymphokine-activated killer (LAK) activity after a short incubation with IL 2. Although in a resting stage they showed no IL 2 receptor expression as examined by anti-Tac antibody, Tac antigen appeared after IL 2 treatment followed by a marked increase in 3H-thymidine incorporation and a remarkable production of gamma-IFN. To investigate the mechanism of neutropenia, in vitro IL 2-stimulated coculture studies of these cells with normal bone marrow cells were performed. Colony formation of myeloid progenitors (CFU-C) was significantly suppressed. In addition, the conditioned medium from IL 2-stimulated LGLs indicated a remarkable suppression of CFU-C. These results suggest that these LGLs with a Leu- 11+, Leu-7- surface phenotype might belong to a unique subset of pre-NK cells that are functionally and phenotypically similar to those represented at any early stage of human ontogeny and that they strongly express Tac antigen under the influence of IL 2 administration, followed by remarkable cell proliferation and gamma-IFN production.

A C9 related channel forming protein in the cytoplasmic granules of human large granular lymphocytes

1985 ◽  
Vol 5 (12) ◽  
pp. 1093-1100 ◽  
Author(s):  
Leora S. Zalman ◽  
Mary A. Brothers ◽  
Hans J. Müller-Eberhard
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Target Cells ◽  
Cellular Cytotoxicity ◽  
Related Protein ◽  
Large Granular Lymphocytes ◽  
Cytoplasmic Granules ◽  
Granular Lymphocytes ◽  
Precipitation Test

Large granular lymphocytes (LGL) from human blood main-tained in culture for 2 to 6 weeks with IL-2 were found positive in the K562 cell killing assay. The cytoplasmic granules of the LGL were isolated, lysed and the soluble proteins were passed over a Sepharose-anti-C9 column. The retained protein was eluted with NaC1 and found to consist by SDS polyacrylamide gel electrophoresis of essentially one component with a molecular weight of approximately 70 000. The protein did not give a positive precipitation test with anti-human C9 by Ouchterlony analysis, but it reacted reproducibly with anti-human C9 by Western blot analysis. By ELISA the cross reaction with human C9 was less than 1%. The C9 related lymphocyte protein lacked C9 hemolytic activity, but it formed functional pores in liposomes in presence of Ca++. These results suggest that the cytoplasmic granules of human LGL that are capable of killing NK target cells contain C9 related protein which is involved in the cellular cytotoxicity reaction.

Malignant clonal expansion of large granular lymphocytes with a Leu- 11+, Leu-7- surface phenotype: in vitro responsiveness of malignant cells to recombinant human interleukin 2

Blood ◽  
1986 ◽  
Vol 68 (5) ◽  
pp. 1065-1073 ◽  
Author(s):  
S Koizumi ◽  
H Seki ◽  
T Tachinami ◽  
M Taniguchi ◽  
A Matsuda ◽  
...  
Keyword(s):  
Interleukin 2 ◽  
Receptor Expression ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Target Cells ◽  
Transferase Activity ◽  
Large Granular Lymphocytes ◽  
Surface Phenotype ◽  
Granular Lymphocytes ◽  
Leu 7

A 14-year-old Japanese female with neutropenia showed malignant proliferation of the large granular lymphocytes (LGLs). These LGLs were E rosette+ and Fc(IgG) receptor+ and therefore are referred to as T gamma lymphocytes. They were also Leu-11+ and OKT11+; however, they were clearly negative for Leu-7, OKT3, OKT8, OKM1, and HNK-1 antigens as well as for terminal deoxynucleotidyl transferase activity. Karyotype analysis revealed 47, XXX. The LGLs showed no rearrangement of T cell receptor C beta genes. The natural killer (NK) cell activity against K562 target cells was low, but was significantly augmented after stimulation by recombinant human interleukin 2 (IL 2) in contrast to minimal NK boosting by recombinant human gamma-interferon (gamma- IFN). Such a unique responsive ability to lymphokines was quite similar to that noted in fetal and cord blood cells. These LGLs also demonstrated a considerable increase in antibody-dependent cell- mediated cytotoxicity (ADCC) and lymphokine-activated killer (LAK) activity after a short incubation with IL 2. Although in a resting stage they showed no IL 2 receptor expression as examined by anti-Tac antibody, Tac antigen appeared after IL 2 treatment followed by a marked increase in 3H-thymidine incorporation and a remarkable production of gamma-IFN. To investigate the mechanism of neutropenia, in vitro IL 2-stimulated coculture studies of these cells with normal bone marrow cells were performed. Colony formation of myeloid progenitors (CFU-C) was significantly suppressed. In addition, the conditioned medium from IL 2-stimulated LGLs indicated a remarkable suppression of CFU-C. These results suggest that these LGLs with a Leu- 11+, Leu-7- surface phenotype might belong to a unique subset of pre-NK cells that are functionally and phenotypically similar to those represented at any early stage of human ontogeny and that they strongly express Tac antigen under the influence of IL 2 administration, followed by remarkable cell proliferation and gamma-IFN production.

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