Propagation of large numbers of T cells with natural killer cell markers

Schmidt‐Wolf, Ingo G.H.; Lefterova, Petja; Johnston, Valerie; Huhn, D.; Blume, Karl G.; Negrin, Robert S.

doi:10.1111/j.1365-2141.1994.tb08297.x

Cited by 110 publications

(77 citation statements)

References 14 publications

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“…30 In that report, the T cells that expanded in vitro partly coexpressed the NK marker CD56. Moreover, the expansion of CD3 þ CD56 þ T cells from normal donors, 31 patients with CML, 32 and patients undergoing autologous hematopoietic cell transplantation 33 has also been reported.…”

Section: Discussionmentioning

confidence: 99%

Expansion of natural killer (NK) and natural killer-like T (NKT)-cell populations derived from patients with B-chronic lymphocytic leukemia (B-CLL): a potential source for cellular immunotherapy

et al. 2003

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B-cell chronic lymphocytic leukemia (B-CLL) is the most common leukemia in the Western world. It is currently an incurable disease, making new treatment options such as immunotherapy desirable. Monoclonal antibodies (Mabs) to surface antigens of the tumor cell is one option. Administration of cytotoxic cells such as natural killer (NK) and natural killerlike T (NKT) cells expanded in vitro might be a useful treatment modality alone or in combination with MAbs. A limiting step in the development of successful cellular immunotherapy has been the availability of appropriate cytotoxic cells. Here, we report the feasibility of expanding populations of the human killer cells, CD3ÀCD56 þ NK and CD3 þ CD56 þ NKT cells, from peripheral blood mononuclear cells (PBMCs) of B-CLL patients. The influence of tumor B cells on the in vitro expansion of killer cells was assessed by depleting B cells from PBMCs by microbead separation before culture. The 21-day cultures from both B-cell-and non-B-cell-depleted PBMC showed a marked expansion of NK cells, and also of T cells, among which almost half had the NKT phenotype. Depletion of B cells before culture did not change the expansion rates of NK and NKT cells significantly. In patients with progressive B-CLL, NK cell expansion capacity was improved after fludarabine treatment when compared to samples obtained before treatment. Repeated samples of PBMCs from individual untreated patients with both indolent and progressive disease cultured under identical conditions gave similar NK cell expansion rates. Expanded killer cell populations had cytotoxic function against the NK-sensitive target K562 cell line and expressed high levels of Granzyme B. From our studies, we conclude that NK cells as well as NKT cells from the peripheral blood of B-CLL patients can be expanded, and that these cells have cytotoxic capacity.

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Section: Discussionmentioning

confidence: 99%

Expansion of natural killer (NK) and natural killer-like T (NKT)-cell populations derived from patients with B-chronic lymphocytic leukemia (B-CLL): a potential source for cellular immunotherapy

et al. 2003

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“…5,6 Briefly, nonadherent Ficoll -separated human peripheral blood mononuclear cells were prepared and grown in RPMI 1640 medium (Gibco BRL, Berlin, Germany ), consisting of 10% fetal calf serum (Gibco BRL ), 25 mM HEPES, 100 U /mL penicillin, and 100 g/ ml streptomycin. On day 0, 1000 U / mL human recombinant IFN -(Boehringer, Mannheim, Germany ) were added.…”

Section: Generation Of Nk -Like T Cellsmentioning

confidence: 99%

Transfection of dendritic cells (DCs) with the CIITA gene: increase in immunostimulatory activity of DCs

et al. 2001

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Dendritic cells ( DCs ) are the major antigen -presenting cells. They are able to present tumor antigens to immunologic effector cells. MHC class II molecules on DC surfaces play an important role in priming effector cells against tumor cells and their antigens. The transactivator CIITA ( MHC class II transactivator ) is a non ± DNA -binding transactivator, which regulates the expression of MHC class II, HLA -DM, and invariant chain and behaves as a master controller of constitutive and inducible MHC class II gene activation. Here, we transfected DCs with the CIITA gene using a novel transfection technique. The vector system consisted of a plasmid bound to an adenovirus via poly -L -lysine, which is covalently bound to a UV -irradiated adenovirus. After transfection, expression of MHC class II on DCs increased from 27% to 75% on day 2 after transfection. Transfected DCs were co -cultured with immunologic effector cells. Cytotoxicity of effector cells against tumor cells increased after co -culture with transfected DCs to 63% compared to 15% with effector cells co -cultured with irrelevantly transfected DCs ( P = .037 ) . This effect was dependent on the timing and period of coculture. In conclusion, transfection of DCs led to an increase in antitumoral immunostimulatory capacity of DCs. We can further conclude that DCs could be efficiently transfected with the CIITA gene. Transfection of DCs led to an increase in antitumoral immunostimulatory capacity of DCs and may have a major impact on immunotherapeutic protocols for patients with cancer.

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“…In previous reports, we presented a protocol generating large numbers of efficient cytotoxic effector cells. 25,26 These effector cells possess greater cytotoxicity and higher proliferation rate than lymphokine -activated killer cells. 39,40 Interestingly, two tumor cell lines transfected with wild -type p53 showed an increased sensitivity against CIK cells.…”

Section: Discussionmentioning

confidence: 99%

“…25,26 Most interestingly, CIK cells have been shown to eradicate established human lymphoma cells in a severe combined immunodeficient mouse xenograft model in vivo. 25 This part of the study was aimed at increasing the sensitivity of lymphoma cells to CIK cells.…”

Section: Discussionmentioning

confidence: 99%

“…25,26 In brief, nonadherent Ficoll -separated human peripheral blood mononuclear cells were prepared and grown in RPMI 1640 medium ( Gibco BRL, Berlin, Germany ), consisting of 10% fetal calf serum ( Gibco BRL ), 25 mM HEPES, 100 U / mL penicillin, and 100 g /mL streptomycin. On day 0, 1000 U /mL human recombinant interferon-( Boehringer Mannheim, Germany ) was added.…”

Section: Generation Of Nk -Like T Cellsmentioning

confidence: 99%

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Effects of adenoviral wild-type p53 gene transfer in p53 -mutated lymphoma cells

et al. 2001

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The present study assessed the role of adenoviral vector ± mediated wild -type p53 gene transfer in B lymphoma cells. Deficiency of p53 -mediated cell death is common in human cancer contributing to both tumorigenesis and chemoresistance. Lymphoma cells are being considered as suitable targets for gene therapy protocols. Recently, we reported an adenoviral protocol leading to highly efficient gene transfer to B lymphoma cells. All lymphoma cell lines ( n = 5 ) tested here showed mutations in the p53 gene locus. The aim of this work was to transduce lymphoma cells with the wild -type p53 gene. Using this protocol, 88% of Raji, 75% of Daudi, and 45% of OCI -Ly8 -LAM53 cells were transfected with the reporter gene green fluorescent protein at a multiplicity of infection of 200. The expression of green fluorescent protein in CA46 and BL41 cells was 27% and 42%, respectively. At this multiplicity of infection, growth characteristics of lymphoma cell lines were not changed significantly. In contrast, cells transduced with wild -type p53 gene showed an inhibition of proliferation as well as an increase in apoptosis. Cell loss by apoptosis after p53 gene transfer was up to 40% as compared to transduction with an irrelevant vector. In addition, we determined the effects of DNA damage produced by the DNA topoisomerase II inhibitor etoposide on wild -type p53 transfected lymphoma cells. In Ad -p53 ± transfected Raji cells, treatment with the drug resulted in a marked increase of cell loss in comparison to Ad --Gal ± transfected cells ( 45% vs. 77% ). Interestingly, performing cytotoxicity studies, we could show an increased sensitivity of Raji and Daudi cells against immunological effector cells. In conclusion, transduction of wild -type p53 into lymphoma cells expressing mutated p53 was efficient and led to inhibition of proliferation and increase in apoptotic rate in some cell lines dependent on p53 mutation. This protocol should have an impact on the use of lymphoma cells in cancer gene therapy protocols.

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Propagation of large numbers of T cells with natural killer cell markers

Cited by 110 publications

References 14 publications

Expansion of natural killer (NK) and natural killer-like T (NKT)-cell populations derived from patients with B-chronic lymphocytic leukemia (B-CLL): a potential source for cellular immunotherapy

Expansion of natural killer (NK) and natural killer-like T (NKT)-cell populations derived from patients with B-chronic lymphocytic leukemia (B-CLL): a potential source for cellular immunotherapy

Transfection of dendritic cells (DCs) with the CIITA gene: increase in immunostimulatory activity of DCs

Effects of adenoviral wild-type p53 gene transfer in p53 -mutated lymphoma cells

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