In vivo localization of lymphocytes labelled with low concentrations of Hoechst 33342

Loeffler, D.; Ratner, Stuart

doi:10.1016/0022-1759(89)90385-2

Cited by 22 publications

(17 citation statements)

References 13 publications

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“…Analysis of serial sections indicated that approximately 30% of the rhodamine-labelled cells are visible in more than one section. The data given in Tables 3 and 4 can therefore be expected to overestimate the true number of rhodamine-labelled cells per organ by approximately 30%, Since H33342 stains only the cell nucleus [18], fewer cells would be visible in more than one section and the overestimation of the number of H33342-1abelled cells per organ because of this phenomenon would be rauch less pronounced.…”

Section: Methodsmentioning

confidence: 93%

Tissue distribution of adoptively transferred adherent lymphokine-activated killer cells assessed by different cell labels

Basse

Herberman

Hokland

et al. 1992

Cancer Immunol Immunother

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Assessment of the tissue distribution of adoptively transferred adherent lymphokine-activated killer A-LAK) cells by use of 51Cr indicated that these effector cells, after an initial phase in the lungs, distributed in high numbers to liver and spleen (30% and 10% of injected dose, respectively). However, when this experiment was repeated with 125IdUrd as cell label, fewer than 2% and 0.5% of the injected cells distributed into liver and spleen respectively. To analyse this discrepancy, we compared the tissue distribution of 51Cr- and 125IdUrd-labelled A-LAK cells with that indicated by alternative direct visual methods for identification of the injected cells, such as fluorescent dyes (rhodamine and H33342) or immunohistochemical staining of asialo-GM1-positive cells. The number of i.v. injected A-LAK cells found in the liver by all visual methods ranged from 1% to 5% of the injected dose, supporting the data obtained with 125IdUrd, whereas 25%-30% of the 51Cr label was consistently found in this organ. Autoradiography of the liver 24 h after i.v. injection of 51Cr-labelled cells revealed a background activity that was four- to fivefold higher than the control level, indicating substantial non-specific accumulation in the liver of 51Cr released from A-LAK cells. We conclude that 51Cr cannot be reliably used in investigations of cell traffic to the liver because of non-specific accumulation of the 51Cr label, particularly in this organ. In contrast, labelling with 125IdUrd or rhodamine and immunohistochemical staining of asialo-GM1-positive cells appear to be reliable and essentially equivalent methods for investigations of the fate of adoptively transferred A-LAK cells. Using these methods, we found that only few A-LAK cells redistribute to the liver upon i.v., i.e. systemic, injection, whereas 40%-50% of locally (intraportally) injected A-LAK cells remain in the liver for at least 24 h.

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Section: Methodsmentioning

confidence: 93%

Tissue distribution of adoptively transferred adherent lymphokine-activated killer cells assessed by different cell labels

Basse

Herberman

Hokland

et al. 1992

Cancer Immunol Immunother

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“…However, undoubtedly the greatest problem associated with the use of H33342 is that it can inhibit lymphocyte proliferation, presumably due to its ability to bind to cellular DNA. A number of groups have reported this difficulty, 9 , 10 , 24 although it can be overcome if cells are labelled with concentrations of H33342 that are subsaturating for cellular DNA 24 . There is one report that H33342 labelling reduces the motility and migration pattern of lymphocytes, 24 although numerous studies in my laboratory over many years have failed to observe such an effect.…”

Section: Dna‐binding Fluorescent Dyesmentioning

confidence: 99%

Fluorescent dyes for lymphocyte migration and proliferation studies

1999

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Summary Fluorescent dyes are increasingly being exploited to track lymphocyte migration and proliferation. The present paper reviews the properties and performance of some 14 different fluorescent dyes that have been used during the last 20 years to monitor lymphocyte migration. Of the 14 dyes discussed, two stand out as being the most versatile in terms of long-term tracking of lymphocytes and their ability to quantify lymphocyte proliferation. They are the intracellular covalent coupling dye carboxyfluorescein diacetate succinimidyl ester (CFSE) and the membrane inserting dye PKH26. Both dyes have the advantage that they can be used to track cell division, both in vitro and in vivo, due to the progressive halving of the fluorescence intensity of the dyes in cells after each division. However, CFSE appears to have the edge over PKH26 based on homogeneity of lymphocyte staining and cost. Two other fluorescent dyes, although not suitable for lymphocyte proliferation studies, are valuable tracking dyes for short-term (up to 3 day) lymphocyte migration experiments, namely the DNA-binding dye Hoechst 33342 and the cytoplasmic dye calcein. In the future it is highly likely that additional fluorescent dyes, with different spectral properties to CFSE, will become available, as well as membrane inserting fluorescent dyes that more homogeneously label lymphocytes than PKH26.

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“…These cells drill deep into the interior of the tumor and accumulate in it due to attractants secreted from the tumor tissue and the high locomotive ability of activated immune cells (Ratner and Heppner 1986). Indeed during the avascular stage, tumor development can be effectively eliminated by tumor-infiltrating cytotoxic lymphocytes (TLs) (Loeffler and Ratner 1989). …”

Section: Introductionmentioning

confidence: 99%

A mathematical model of pre-diagnostic glioma growth

Sturrock

Hao

Schwartzbaum

et al. 2015

Journal of Theoretical Biology

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Due to their location, the malignant gliomas of the brain in humans are very difficult to treat in advanced stages. Blood-based biomarkers for glioma are needed for more accurate evaluation of treatment response as well as early diagnosis. However, biomarker research in primary brain tumors is challenging given their relative rarity and genetic diversity. It is further complicated by variations in the permeability of the blood brain barrier that affects the amount of marker released into the bloodstream. Inspired by recent temporal data indicating a possible decrease in serum glucose levels in patients with gliomas yet to be diagnosed, we present an ordinary differential equation model to capture early stage glioma growth. The model contains glioma-glucose-immune interactions and poses a potential mechanism by which this glucose drop can be explained. We present numerical simulations, parameter sensitivity analysis, linear stability analysis and a numerical experiment whereby we show how a dormant glioma can become malignant.

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In vivo localization of lymphocytes labelled with low concentrations of Hoechst 33342

Cited by 22 publications

References 13 publications

Tissue distribution of adoptively transferred adherent lymphokine-activated killer cells assessed by different cell labels

Tissue distribution of adoptively transferred adherent lymphokine-activated killer cells assessed by different cell labels

Fluorescent dyes for lymphocyte migration and proliferation studies

A mathematical model of pre-diagnostic glioma growth

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