Cancer-Specific Density Changes in Lymphocytes After Stimulation With Phytohæmagglutinin

Pritchard, J A; Seaman, J.E.; Evans, I.H.; James, K.; Sutherland, W.H.; Deeley, T. J.; Kerby, I.J.; Paterson, Ian C.; Davies, B H

doi:10.1016/s0140-6736(78)92039-1

Cited by 25 publications

(4 citation statements)

References 7 publications

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“…It follows that decreases in P of 10-50% after PHA or tumour-antigen stimulations, as observed in the SCM measurements (Cercek et al, 1974;Cercek & Cereek, 1975, 1976, 1977, 1978aTakaku et al, 1977;Nishifuku et al, 1977;Hashimoto et al, 1978, Kreutzmann et al, 1978Pritchard & Sutherland, 1978;Stewart et (1974), was considered carefully, and comparisons made to exclude prestimulation. In fact the much greater pH stability of the HEPES-buffered medium allowed us to avoid the pH-related variation in fluorescent intensity (Visser et al, 1979) which would have caused pHrelated variations in P. We question the problem of impurities in acetone.…”

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confidence: 99%

Comments on “Response of Human Lymphocytes to PHA and Tumour-Associated Antigens as Detected by Fluorescence Polarization”

Preece¹,

Light²,

Balding³

1980

Br J Cancer

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confidence: 99%

Comments on “Response of Human Lymphocytes to PHA and Tumour-Associated Antigens as Detected by Fluorescence Polarization”

Preece¹,

Light²,

Balding³

1980

Br J Cancer

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“…The polarization changes observed depend on the experimental conditions used during the isolation of these lymphocytes (2-7). We have examined the leukocyte isolation procedure used by Price' and find their isolation conditions to be different from those used by us (2,3) and other laboratories which confirmed the SCM-test (4)(5)(6)(7)(8)(9). Furthermore, the intracellular fluorescein emission polarization spectrum recorded on these leukocytes ( Fig.…”

Section: Dear Sirmentioning

confidence: 88%

Response to "Comments on the Fluorescein Excitation and Emission Polarization Spectra in Living Cells"

Cercek¹,

Cercek²

1979

Biophysical Journal

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“…Second, the detection of early-stage cancers is limited by its relatively low sensitivity. However, these difficulties may possibly be solved if the mechanism of this phenomenon is clarified from the physical and biological viewpoints.Various mechanisms to explain this fluorescence depolarization in living cells have been proposed so far, (6,8,13,(17)(18)(19)21,23). such as free rotational Brownian motion of the dye molecules in the cytoplasm (2,241, restricted motion of the dye molecules bound to proteins (20), and the mixture of these two (16,251.…”

mentioning

confidence: 99%

“…Various mechanisms to explain this fluorescence depolarization in living cells have been proposed so far, (6,8,13,(17)(18)(19)21,23). such as free rotational Brownian motion of the dye molecules in the cytoplasm (2,241, restricted motion of the dye molecules bound to proteins (20), and the mixture of these two (16,251.…”

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confidence: 99%

Spectroscopic properties of fluorescein in living lymphocytes

et al. 1987

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Detailed studies have been performed on various spectroscopic properties such as time dependence and excitation wavelength dependence of the fluorescence anisotropy for fluorescein molecules introduced into rat thymus lymphocytes. Experimental results have been found to be well interpreted in terms of the coexistence of two types of dye molecules, i.e., free and bound molecules. The fluorescence spectrum of only the bound molecules has been obtained from the difference in the time-resolved spectra of fluorescence with two polarization directions. The time gate has been set at a sufficiently late time after the excitation, so that the polarization memories of the free molecules are lost. The spectrum thus determined agrees very well with that calculated from the spectral data in the stationary condition. From the above results, we come to the conclusion that the main factors which determine the fluorescence anisotropy inside the cell are the fraction and the anisotropy of the bound dye molecules. Finally, we discuss how these factors are related to biological quantities.Key terms: Neoplasms, spectrometry, fluorescence, lasers, fluorescence polarization Recently, several experiments have been reported in which human cancers are detected by the measurement of fluorescence depolarization in lymphocytes. This method is based on the fact that the degree of polarization of fluorescence from fluorescein introduced into living cells changes according to antigenic stimulation of lymphocytes. Cercek and Cercek (3) examined this property in peripheral lymphocytes from various cancer patients and found that the ratio of the change in the degree of polarization by the stimulation with nonspecific mitogen PHA (phytohemagglutinin) to that with socalled cancer basic proteins, gives a good measure for the purpose of the diagnosis of human cancers. This finding has been confirmed by many researchers Although this method provides an easy way of a medical checkup for human cancers, there exist clear limitations to the wide applicability of this method at the present time. First, a very complicated pretreatment method for this assay lowers the reproducibility of the experiment. Second, the detection of early-stage cancers is limited by its relatively low sensitivity. However, these difficulties may possibly be solved if the mechanism of this phenomenon is clarified from the physical and biological viewpoints.Various mechanisms to explain this fluorescence depolarization in living cells have been proposed so far, (6,8,13,(17)(18)(19)21,23). such as free rotational Brownian motion of the dye molecules in the cytoplasm (2,241, restricted motion of the dye molecules bound to proteins (20), and the mixture of these two (16,251. However, no model has been accepted generally yet, and also, the quantitative evaluation of the degree of polarization has not been made. To determine the physical mechanism of this fluorescence depoIarization, we have measured the time dependence of the fluorescence anisotropy of flourescein introduced into rat th...

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Cancer-Specific Density Changes in Lymphocytes After Stimulation With Phytohæmagglutinin

Cited by 25 publications

References 7 publications

Comments on “Response of Human Lymphocytes to PHA and Tumour-Associated Antigens as Detected by Fluorescence Polarization”

Comments on “Response of Human Lymphocytes to PHA and Tumour-Associated Antigens as Detected by Fluorescence Polarization”

Response to "Comments on the Fluorescein Excitation and Emission Polarization Spectra in Living Cells"

Spectroscopic properties of fluorescein in living lymphocytes

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