Endogenous natural killer cells do not play a role in antitumor effects induced by interleukin-2 in a syngeneic rat colon tumor model

Hagenaars, Martin; Ensink, N. G.; Eggermont, Alexander M.M.; Velde, E. A. van der; Velde, C.J.H. van de; Fleuren, Gert Jan; Kuppen, Peter J. K.

doi:10.1007/pl00006674

Cited by 2 publications

(2 citation statements)

References 32 publications

(32 reference statements)

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“…Cells were then prepared for flow cytometry as described. 45 Briefly, the isolated NK cells and blood lymphocyte fractions were adjusted to a concentration of 5 ϫ 10 5 cells/ml in FACS buffer consisting of PBS enriched with 1% bovine serum albumin (BSA). The cells were incubated for 30 min at 4°C with 100 l of 3.2.3 IgG1 mAb, washed and incubated for 20 min in the dark at 4°C with 100 l of the FITC-conjugated donkey antimouse mAb (Jackson Immuno Research Laboratories, West Grove, PA).…”

Section: Assessment Of Nk Cells By Flow Cytometry and Immunohistochemmentioning

confidence: 99%

Interactions between rat colon carcinoma cells and Kupffer cells during the onset of hepatic metastasis

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et al. 2004

Intl Journal of Cancer

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Liver sinusoids harbor populations of 2 important types of immunocompetent cells, Kupffer cells (KCsColon cancer is one of the most common cancers in the Western world. 1 Metastases to the liver, a common event in cancer patients, is an important cause of death, and the rate of metastasis generally determines survival time. 2,3 The growth and development of metastasis involve a cascade of tumor-host cell interactions.The first vascular bed encountered by blood-borne tumor cells from the gastrointestinal tract is in the liver, in the sinusoids of which they encounter 2 immunocompetent cells, Kupffer cells (KCs) and natural killer (NK) cells. 4,5 KCs represent the largest macrophage population in the body. 5 They move along sinusoids, scavenge degenerated cells, macromolecules and microorganisms and function as antigen-presenting cells. 4,6,7 By virtue of these capacities and their privileged location within the circulation, KCs form the first line of defense against disseminating colorectal tumor cells. 8 Their role in controlling hepatic metastasis in vivo has been demonstrated by their capacity to kill tumor cells in vitro. 9 -13 It has been shown that binding to tumor cells is necessary for effective killing in vitro, 14 -16 which is accomplished by a variety of mechanisms, including phagocytosis and the release of proteinases, tumor necrosis factor and oxygen metabolites. 4,17,18 The role of KCs in controlling metastasis has been demonstrated at late stages of metastasis (3 weeks); metastasis is enhanced by selectively eliminating KCs and reduced by stimulating them. 8,19 -22 Whether or not this effect can also be observed during the early stages of metastasis has not been reported yet.Liver 32 and the receptor-mediated pathway (members of the TNF family). 33 Both KCs and liver NK cells belong to the innate immune system and are often observed in close contact with each other. 24,27 It is supposed that these cells affect the growth of liver metastases only if they interact with incoming tumor cells shortly after their arrival in the liver (ϳ 24 hr) because later on the natural host defenses may be incapable of limiting metastasis due to the rapid proliferation of the cancer cell population, angiogenesis and weakened host defenses. Synergism between KCs and NK cells in the elimination of tumor cells has been postulated in vivo 34 and illustrated in vitro. 12 However, most studies describing the in situ KC-tumor cell interactions focused on events occurring 2-3 weeks after tumor cell inoculation, 20,22,34,35 and only a few studies examined the early stages of hepatic metastases (Ͻ 24 hr). 36,37 Routine microscopic methods are not optimal for studying early hepatic metastasis because they require thin sections, low sampling volumes, complicated preparation methods and immunohistochemical procedures. 38 In contrast, confocal laser scanning microscopy (CLSM) enables the study of large sample volumes and sections 50 -100 m thick, facilitating investigation of rare cellular events such as the occurrence of tumor...

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Section: Assessment Of Nk Cells By Flow Cytometry and Immunohistochemmentioning

confidence: 99%

Interactions between rat colon carcinoma cells and Kupffer cells during the onset of hepatic metastasis

Timmers

Vekemans

Vermijlen³

et al. 2004

Intl Journal of Cancer

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“…These slides were first pretreated with 0.3% H 2 O 2 in methanol and then incubated with normal goat serum. They were incubated overnight at 4°C with ALB and G6PC antibodies (1:200 dilution) for HCs [22], DES, and VIM for HSCs [23], LYZ and ED2 for KCs [24], CK18 and GGT for BECs [25], CD14 and ET-1 for SECs [26,27], CD8 and CD56 for PCs [28,29], CD86 and CD103 for DCs [30], OC2 and OV6 for OCs [31]. Biotinized secondary antibodies (1:2000 dilution) was added and allowed to react at 37°C for 60 min.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Reference Gene Selection for Real-Time RT-PCR in Eight Kinds of Rat Regenerating Hepatic Cells

Wang

2010

Mol Biotechnol

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Liver regeneration (LR) is a process during which the liver recovers its mass and function after damage due to various causes such as partial hepatectomy (PH). It involves a sequence of well-orchestrated changes in physiological and biochemical activities, especially in the gene expression profile in a variety of liver cells. In order to produce reliable gene expression of target genes in eight kinds of rat hepatic cells during LR, the determination of internal control housekeeping genes (HKGs) is required. Eight kinds of hepatic cells were first isolated from liver tissue with high purity and activity. Then quantitative real-time reverse transcription (RT)-PCR was applied to detect expression changes of six commonly used HKGs (18SrRNA, B2M, ACTB, UBC, GAPDH, and HK1) in eight types of hepatic cells isolated from regenerating liver at 0, 2, 6, 12, 24, 30, 36, 72, 120, and 168 h after PH. The amplification of the HKGs was statistically analyzed by using geNorm algorithm. Using this method, 18SrRNA-UBC, ACTB-HK1, ACTB-GADPH, B2M-ACTB, 18SrRNA-UBC, B2M-UBC, B2M-ACTB, and B2M-UBC were found to be the two most stable reference genes for rat regenerating hepatocytes, hepatic stellate cells, Kupffer cells, biliary epithelial cells, sinusoidal endothelial cells, pit cells, dendritic cells, and oval cells, respectively, regardless of the stages of LR. In conclusion, this study has laid a good foundation for investigating gene expression of target genes in different types of hepatic cells during LR.

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Endogenous natural killer cells do not play a role in antitumor effects induced by interleukin-2 in a syngeneic rat colon tumor model

Cited by 2 publications

References 32 publications

Interactions between rat colon carcinoma cells and Kupffer cells during the onset of hepatic metastasis

Interactions between rat colon carcinoma cells and Kupffer cells during the onset of hepatic metastasis

Reference Gene Selection for Real-Time RT-PCR in Eight Kinds of Rat Regenerating Hepatic Cells

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