[Present condition of knowledge as to lymphocytes (author's transl)].
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Biomedical subjects
Publications and source records attributed to G Astaldi.
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The effect of in vitro irradiation with increasing in logarythmic progress X-ray doses on lymphocyte viability and on T and B lymphocyte populations was studied in normal adults, patients with myasthenia gravis and in patients undergoing long-term steroid therapy. Decrease in numbers of lymphocytes carrying T or B lymphocyte surface markers was higher than viable cell loss. The decrease showed no linear correlation with X-ray doses applied, which might reflect the existence of radioresistant T and B lymphocytes. A higher so called early radiosensitivity of B lymphocytes was demonstrated. In patients with myasthenia gravis early radioresistance of T lymphocytes was detected. In patients undergoing long-term steroid therapy, an increase in numbers of cells lacking markers of any of lymphocyte populations was found in parallel with a decrease in T lymphocyte number which, in these patients, showed a higher radiosensitivity.
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In 24 men aged 32 to 58 years with precancerous states of the larynx, i.e., leukoplakia, papillomas and pachydermia the peripheral blood lymphocytes were cytochemically stained for N-acetyl-beta-glucosaminidase, beta-glucuronidase, acid phosphatase and glycogen; and the neutrophils were stained for alkaline phosphatase, myeloperoxidase and lipids. The results were expressed in terms of the absolute counts of reaction-positive cells and of the activity index score. The serum immunoglobulins IgG, IgA and IgM were also determined by Mancini's method. The results obtained were compared with those in 20 healthy men aged 20 to 30 years. The patients exhibited elevated numbers of N-acetyl-beta-glucosaminidase and beta-glucuronidase-positive lymphocytes. A characteristic feature was an increase in the absolute counts of lymphocytes with diffuse and granular-diffuse types of cytochemical reaction for all enzymes studied. The number of cells with the granular type of enzymatic reaction (intact enzyme-positive lysosomes) was significantly diminished. These cytochemical alterations were accompanied by a significant increase in the serum IgA level. These results are discussed with reference to the lymphoid system response to tissues of precancerous lesions of the larynx. So far as the neutrophils are concerned the patients exhibited significant intracellular deficiency of beta-glucuronidase and decrease in the lipid content as well as an elevated alkaline phosphatase activity. The possible significance of the beta-glucuronidase deficiency in neutrophils for the diminished cytotoxic response of these cells against the tumor and precancerous lesion cells is discussed.
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The Authors studied the effect of the human serum thymic factor on the peripheral-blood leukocytes from ALL patients, by testing the frequency of the E+cells by means of the e rosette assay before and after cell incubation with the above mentioned factor. In 3 out of 24 patients tested, the thymic factor incubation of peripheral-blood mononuclears increased the number of E+cells. The shift from R to E+cells caused by the human serum thymic factor occurred among null cells (E-, SM-Ig-). The possible value of the test in investigating whether a T0 leukemia might really exists is pointed out, as well as its prospective importance in monitoring the variations of the immature T cell counts in relationship to the patient's treatment and his disease course.
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In acute leukemia, the frequency of the E+ cells among the blood mononuclears before and after their incubation with human serum thymic factor was investigated by means of the E rosette assay. In 4 of 24 patients tested, the thymic factor incubation of their peripheral blood cells increased the number of the E+ cells among the mononuclears. Prospective importance of the test either in investigating whether a T0 leukemia might really exist, or in monitoring possible variations of the immature T cell counts in relationship to the patient's treatment and his disease course, are discussed.
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In 37 patients with acute leukemia and in 13 patients with other hematologic malignancies, E-rosette formation of peripheral blood mononuclear cells, before and after incubation with human serum thymic factor, was studied. This assay showed that incubation with thymic factor caused a clear-cut increase of E-rosette-forming cells in 8 of the 37 acute leukemia patients. Among the 13 patients with other hematologic malignancies, a similar effect was observed in two with pediatric Hodgkin's disease and in one adult with Waldenström's macroglobulinemia. These results support the contention that a significant percentage of the so-called "null" cells circulating in the blood of the above-mentioned patients were, in fact, immature T cells (T0 lymphocytes) that were shifted to a more mature stage (T1) by thymic factor.
Hyperthermia (42 degrees C) and peptichemio (PTC) applied simultaneously result in an increased killing effect and in a significant induction of single strand breaks (SSB) of DNA molecule. The enhanced killing effect was observed in all circumstances, whenever hyperthermia was applied before, during or after PTC treatment, but the most effective cell killing was obtained when hyperthermia and PTC were applied simultaneously. The results show that PTC concentrations used in these experiments do not induce SSB in DNA molecule during incubation for 2 hours at 37 degrees C. When the cells were exposed to PTC and hyperthermia the induction of SSB was observed. The number of SSB depends on the exposure time and PTC concentration. Cells with a greater number of SSB lost the capacity for repair during post-incubation at 42 degrees C.
In seven patients with polycythemia vera, the agar colony growth of bone marrow total nucleated cell suspensions and of the cell fractions obtained with an albumin discontinuous density gradient were studied. In one patient, the density distribution of colony-forming units in culture (CFUc) before and after alkylating treatment was evaluated and cell differentials on smears obtained from each density fraction were determined. A high percentage of low density CFUc compared with the total CFUc population and with that in the normal control subjects in the same density fractions was observed.
The Authors studied the effect of the human serum thymic factor on the peripheral blood leukocytes from ALL patients, by testing the frequency of the E+ cells by means of the E rosette assay before and after cell incubation with this factor. In four out of twenty-four patients tested, the thymic factor incubation of peripheral blood mononuclears increased the number of E+ cells. The shift from E- to E+ cells caused by the human serum thymic factor occurred among null cells (E-, SM-Ig-). The possible value of the test in investigating whether a T0 leukemia might really exists is pointed out, as well as its prospective importance in monitoring the variations of the immature T cell counts in relationship to the patient's treatment and his disease course.
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First, lymphocyte transplantation into total body-irradiated rats is discussed. The effect on spleen colony formation caused by the transplantation of untreated lymphocytes, as well as of lymphocytes previously incubated with PHA, or with PHA plus L-asparaginase, or with lymphokines, was studied. Then the effect of the urinary colony-stimulating factor in vitro, and the in vitro feeder-layer activity of leucocytes on colony formation of human and mice bone marrow cells in haematological diseases is dealth with. The injection of rat lymphocytes previously incubated for 24 hours with PHA resulted in a higher number and a larger size of colonies in the spleen of the recipient rats. Lymphocytes preincubated with lymphokines gave rise to the formation of spleen colonies which were larger than those developing after the injection of untreated lymphocytes. When the lymphocytes were previously incubated with PHA plus L-asparaginase, PHA failed to stimulate colony formation in the spleen. The phenomenon is explained by assuming that PHA, as an aspecific stimulator of cell division, initiated the division of CFUs, thus the CFUs content of the preincubated samples increased, resulting in an increase in the number of colonies formed after the transplantation of lymphocytes pretreated with PHA. Another possible explanation is that CFUs division, or their spleen take is enhanced by the immunocompetent lymphocytes activated by PHA, either directly or via soluble mediators produced or released by immunocompetent lymphocytes such as lymphokines. The study of colony-forming cells and colony-stimulating activity in primary myelofibrosis (PM) showed an increase in the number of circulating CFUc in this condition, and an abnormal density of these cells reaching a peak below 1.062. The lowering of CSA in the first two peripheral blood gradient fractions agrees with the observation in the same fractions of a high percentage of CFUc at the expense of the CSC population. Thus, double cell population seems to exist in PM. One is greatly abnormal with a low specific density and high plating efficiency, whereas the other population is almost normal, showing a higher specific density and a lower plating efficiency.
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Since four years, it has been possible to classify most cases of Chronic Lymphocytic Leukemia (CLL) as given to the expansion of abnormal B-lymphocytes. On the contrary, the T-cell class is apparently normal and the T cell extent in CLL-peripheral blood can be even greater than normal when taken as absolute value. The clonal nature theory of B-lymphocytes in CLL is substantiated by the fact that, in general, in every patient only one Ig light chain determinant is present. Again, when serum Ig monoclonality is present in CLL, it appears idiotypically identical to the Ig shown by the lymphocytes of the same patient. Among the most important B-lymphocyte abnormalities in CLL, there are: (a) fluorescence of surface Ig is usually less intense than that in normal subjects, and fluorescence intensity may vary not only from patient to patient, but also from cell to cell in the same patient; (b) the Fc-receptor can be lacking; (c) the C3b-receptor is not always present, or it is from 2 to 20 folds less frequent than the C3d-receptor, whereas normal human lymphocytes do not show any outstanding differences between the number of EAC rosette-forming cells either when tested with mouse complement (C3d-receptor) or with human complement (C3b-receptor); (d) the traffic capacity of peripheral-blood B-lymphocytes in CLL is quite defective. All the above-mentioned data, taken as a whole, suggest that CLL is in general given by the expansion of an abnormal clone of cells of B origin, arrested in their maturative development, non-responsive to the mitogen stimulation, accumulating in the peripheral-blood for a traffic deficiency.