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PubMed1993

Methods for the analysis of cellular kinetics in PHA-stimulated blood lymphocytes using BrdU incorporation. A comparative study.

The cellular cycle (cc) span was measured by using differential sister-chromatid staining (DSCS) and applying the following methods: cellular cycle time (cct) according to the graphic method of Dutrillaux and Fosse (G-cct); the analytical equation (A-cct) proposed in the present paper, and the average generation time (AGT) suggested by Ivett and Tice. The mean values obtained by the three methods were 12.5, 12.7, and 19.5 h, respectively. A-cct is the more precise method, since the equation of the analytical procedure allows the utilization of numerical data, and when the graphical method is used, the values plotted in a graph may vary according to the employed scale. Cct is the choice over AGT because the first evaluates actively dividing cells and only considers those at M2 or M3. It will be useful to study cell proliferation kinetics in genetic pathological conditions and to investigate with accuracy the effect of cytostatic and cytotoxic drugs.

Adolescent↗
PubMed1994

Influence of PHA stimulation on the cytotoxicity and mutagenicity of X-rays and ethylnitrosourea in human peripheral blood T-lymphocytes.

We investigated the effects of mitogenic stimulation on the cytotoxicity and mutagenicity of X-rays and ethylnitrosourea (ENU) in human peripheral blood lymphocytes using a cloning technique. Resistance to 6-thioguanine (TG) served as the genetic marker. Day 0 (unstimulated) lymphocytes were about two times more radiosensitive than day 3 (stimulated) lymphocytes to the cytotoxicity when compared for the D0 value (0.72 Gy vs. 1.54 Gy), and about five times more radiosensitive to its mutagenicity when compared for the frequency of TG-resistant cells following exposure to 4 Gy of X-rays (25.5 x 10(-6) vs. 126.0 x 10(-6). On the other hand, day 3 (stimulated) lymphocytes were about three times more sensitive to ENU with a D37 value of 1.03 mM compared with 2.82 mM for day 0 (unstimulated) lymphocytes, but as sensitive as day 0 lymphocytes to its mutagenic effect. These results indicate that the sensitivity of lymphocytes for cytotoxicity and mutagenicity is modified by mitogen stimulation, when lymphocytes are exposed to carcinogens or mutagens in vitro.

Cell Survival↗
PubMed1985

The decline in murine splenic PHA and LPS responsiveness following serial transplantation of young and old bone marrow cells.

An analysis was made of the ability of bone marrow cells derived from young (6 months) or old (28 months) murine (A X C57BL/6) donors to differentiate to B and T lymphocytes following serial bone marrow cell transplantations into groups of young syngeneic lethally irradiated recipients. Both number and mitogenic responsiveness of splenic B and T lymphocytes were recorded for recipients of young or old bone marrow cells 9 months following each serial transplant. The data suggests that lymphopoietic stem cells senescence in a manner analogous to that predicted by the clonal succession model of ageing.

Aging↗
PubMed1996

The novel immunomodulator, Linomide, stimulates interleukin-2-induced human natural killer (NK) cell and PHA-stimulated T cell proliferation from normal donors.

Donor-derived cell-mediated immunotherapy has been shown to be an effective tool for reinduction of remission in chronic myeloid leukaemia (CML) patients who have relapsed post-bone marrow transplantation (BMT). Linomide, quinoline-3-carboxamine (LS 2616), is a new immunomodulator shown to increase the number of NK precursors in mice in addition to upregulating the quantity of CD56(+), CD3(-) and CD16(+) NK cells in the peripheral blood of patients following autologous BMT (ABMT). We investigated the in vitro effects of Linomide on NK activity of normal human donors. Large granular lymphocytes (LGLs) and NK cells were incubated overnight with Linomide (0.02-4.8 mg/ml), recombinant human interleukin-2 (IL-2, 75 IU/ml), or a combination of both. Linomide, at 0.02-0.3 mg/ml, augmented IL-2-induced proliferation of LGLs and NK cells in an inversely proportional manner. In contrast, Linomide at 0.6-4.8 mg/ml inhibited IL-2-induced proliferation of LGLs and NK cells in a dose-dependent manner. Linomide was able to potentiate phytohemaglutinin-induced CD3(+) cell proliferation. In addition, supernatants derived from Linomide treated CD3(+) T cells were able to mimic the direct stimulatory effect of Linomide on activated NK cell proliferation. These supernatants were found to have low levels of tissue necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) and therefore Linomide stimulation of NK and T cell proliferation may be due to its inhibitory effect on the secretion of these cytokines by activated CD3(+) T cells. Linomide had no effect on cytotoxicity nor on the phenotypic expression of resting and IL-2-activated LGLs or NK cells. In view of our results, Linomide could possibly play a potential role in adoptive cell-mediated immunotherapy post-BMT.

Adjuvants, Immunologic↗
PubMed1995

Chromosome aberrations, spontaneous SCE, and growth kinetics in PHA-stimulated lymphocytes of five cases with Sézary syndrome.

Cytogenetic studies of five patients with Sézary syndrome (SS) revealed clonal chromosome aberrations in all cases. In one patient, a del(8)(p21) was the sole abnormality, whereas the remaining cases had karyotypes with multiple chromosome changes. In three SS cases with hypodiploid chromosome numbers, structural rearrangements affecting regions 10q22-24 and 12p11-13, and aberrations leading to loss of material from 17p were found concurrently. Bands 14q11 and 14q32 were involved in structural rearrangements in one case each. Our results and review of 51 published previously SS cases that were analyzed with banding techniques indicate that the chromosomes most frequently involved in structural changes were chromosomes 1 and 2 (in 43% of cases), 6 (in 38%), 17 (in 34%), 14 (in 27%), 11 (in 25%), 13 (in 21%), and 9 (in 20%). In particular, the breakpoints tended to aggregate at 1p11, 1p36, 2p11-24, 6q, 9q, 11q, 13q11-14, 14q11, 14q32, and in the pericentric region of chromosome 17. The most common numerical change was loss of chromosome 10, detected in 32% of SS cases. In our studies of three SS cases, sister chromatid exchange frequencies were significantly higher in comparison to the normal control. Cell cycle kinetics analysis revealed that the cell cycle time in the malignant cells was significantly longer than in lymphocytes of normal individuals.

Adult↗