Cell electrophoresis is a useful method to measure electric charge on cell membrane and recognize precisely biological changes on the surface of cells. In the present paper, cell electrophoresis was used in an attempt to investigate various changes on the surface of human lymphocytes, which were separated from human peripheral blood by means of 3% gelatin sedimentation method, with the following results.
1. Cell electrophoretic mobility was 0.983±0.189μ/sec/V/cm in average of eighty one normal subjects. Human lymphocytes were shown to be heterogeneous electrophoretically.
2. Cell electrophoretic mobility of the human lymphocytes from patients with lymph tissue disorders was investigated. Electrophoresis of four cases of acute myelogenous leukemia (AML) did not show significant change in both electrophoretic mobility and cytopherogram. A case of AML in which 70% of cells in peripheral blood were leukemic, however, showed a cytopherographically homogeneous pattern. The lymphocytes from giant follicular lymphoma and reticulosarcoma did not show marked changes.
3. Cell electropheretic mobility of the lymphocytes treated with anti-human lymphocyte antibody (ALS) was investigated. Cytotoxic antibodies were obtained from the pregnant female sera as judged by the micro-droplet lymphocyte cytotoxicity test. Electrophoresis was carried out after the lymphocytes were incubated with ALS. Electrophoretic mobility of the lymphocyte treated with high titer ALS was lower than that treated with low titer ALS. It was shown that lowering of electrophoretic mobility was correlated with cytotoxic activity of ALS.
4. Cell electrophoretic mobility of the lymphocytes sensitized with the anti-homologous γ-globulin and various anti-immunoglobulin polypeptide chains was investigated.
Electrophoretic mobility of the lymphocytes treated with anti-human γ-globulin was lower. The phenomenon was significantly found in the lymphocytes treated with anti-μ, anti-κ, and anti-λ polypeptide chain sera, but not so markedly in those treated with anti-γ or anti-α chain sera. However, electrophoresis of the lymphocytes treated with anti-γ and anti-α sera showed that the delay phenomenon of mobility occurred on some of the cells. From the results of the experiments, it was suggested that large amounts of μ-chain, κ-chain and λ-chain and small amounts of γ-chain and α-chain are present on the surface of human lymphocytes.
The results were discussed with comprehensive survey of literatures on cell electrophoretic investigations of human lymphocytes.

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The supernatants of human peripheral blood lymphocytes stimulated with antigen and non specific mitogen influence electrophoretic mobility of sheep erythrocytes as indicator cells of lymphocyte sensitization. This test system has been used for application to clinical immunology.
Lymphocytes from human peripheral blood were obtained by centrifugation in Ficoll-Conray method. Then separation of T and non T lymphocytes was performed by the ordinary E-rosetting technique. Separated T and non T lymphocyte fractions were incubated with PHA, Con A and DNA obtained from calf thymus. To cultured supernatants the tanned sheep erythrocytes were added, and electrophoretic mobility of the sheep erythrocytes was measured in an analytical cell microelectrophoretic apparatus.
In the supernatant from normal T lymphocytes stimulated with PHA and Con A, decrease of electrophoretic mobility of the sheep erythrocytes was more remarkable than in that from normal non T lymphocytes. In the supernatants of the lymphocyte subpopulation from patients with SLE stimulated with Con A, delay of electrophoretic mobility of the sheep erythrocyte was observed in non T lymphocytes. When the lymphocytes from the patients with SLE were stimulated with DNA, electrophoretic mobility of the sheep erythrocyte showed change in the supernatants from T lymphocyte culture.

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Change of surface charge of the tanned sheep erythrocytes added with supernatants of human lymphocytes stimulated with antigens, mitogens or immunopotentiators was investigated. In supernatants of the sensitized lymphocytes stimulated with PPD, percentage slowing of electrophoretic mobility of sheep erythrocytes were increased. In normal lymphocytes cultured with PHA and Con A, there were high percentage slowings in electrophoretic mobility test, and in immunopotentiators such as OK-432, PSK, LVS, methyl B₁₂, poly-A:U, poly-I:C and NaIO₄, percentage slowings were not significantly changed.
Percentage slowing was reduced in RA lymphocytes stimulated with PHA, but not changed in SLE lymphocytes. In normal lymphocytes incubated with Lentinan, percentage slowing value was similar to the result obtained by Con A, but it was low in lymphocytes from the patients with collagen diseases and malignancies.
In SLE lymphocytes added with DNA from calf thymus and RA lymphocytes added with human IgG, electrophoretic mobility of sheep erythrocytes delayed.

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Antitumor effects and changes in electrophoretic mobility pattern of splenocytes and thymocytes were studied in sarcoma 180 or plasmacytoma X5563-bearing mice treated with Mitomycin C (MMC) and/or Krestin (PSK). The percentages of the inhibition of tumor growth were 80∼90% in the MMC-treated group and further improved in the MMC+PSK-treated group. In splenocyte electrophoretic pattern, intermediate mobility cells locating between high mobility cells (HMC: T cells) and low mobility cells (LMC: B cells) increased in tumor-bearer but recovered by treatment with antitumor drugs. On the other hand, in thymocyte electrophoretic pattern, LMC (immature thymocytes) which decreased by tumor-bearing were further reduced in MMC-treated group, but its reduction was suppressed by the combination with PSK. These results suggest that the combination therapy of MMC and PSK is more effective from the viewpoints of antitumor activity and antiinfectious activity because PSK restores immature thymocytes damaged by MMC.

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The use of automated devices in the last ten years have made biomedical applications of cell electrophoresis more attractive. We are using routinely the automated single cell electrophoresis microscope PARMOQUANT. Using the PARMOQUANT we applied this method to discriminate lymphocytes and study the interaction of substances with cells and synthetic particles. Electrophoretic histograms allowed the determination of changes in the proportion of lymphocyte populations after kidney transplantation, during dialysis treatment, open heart surgery and during pregnancy. Discrimination of leukemic cells on the basis of electrophoresis was used as an additional parameter in diagnosis. In a mouse tumor model histograms determination enabled evaluation of the in vivo effect of tumor necrosis factor on immune cells. Cell electrophoresis was shown to be suitable to detect the influence of antibodies, lectins and bacteria on the cell surface. Protein adsorption was studied on synthetic particle using cell electrophoresis. This method was applied in investigating the phenomena of blood interaction with biomaterials for use in artificial organs and to determine differences in the protein composition in serum or other body fluids connected with diseases. On the basis of this principle a test to detect hetrozygotes in cystic fibrosis is now in progress.

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