Mitogen-induced Blastogenesis Research Articles

Rapid Quantification of Mitogen-induced Blastogenesis in T Lymphocytes for Identifying Immunomodulatory Drugs

Lymphocyte proliferation in response to antigenic or mitogenic stimulation is a readily quantifiable phenomenon useful for testing immunomodulatory (i.e., immunosuppressive or immunostimulatory) chemical compounds and biologics. One of the earliest steps during mitogenesis is cell enlargement or blastogenic transformation, whereupon the cell volume increases before division. It is usually detectable in the first several hours of T-lymphocyte stimulation. Here, we describe a rapid method to quantify blastogenesis in T lymphocytes isolated from mouse spleens and human peripheral blood mononuclear cells (PBMCs) using an automated cell counter. Various commonly used proliferation assays for the most part are laborious and only reflect the overall population effect rather than individual cellular effects within a population. In contrast, the presented automated cell counter assay provides rapid, direct, and precise measurements of cell diameters that can be used for assessing the effectiveness of various mitogens and immunomodulatory drugs in vitro.

Rapid Quantification of Mitogen-induced Blastogenesis in T Lymphocytes for Identifying Immunomodulatory Drugs.

Lymphocyte proliferation in response to antigenic or mitogenic stimulation is a readily quantifiable phenomenon useful for testing immunomodulatory (i.e., immunosuppressive or immunostimulatory) chemical compounds and biologics. One of the earliest steps during mitogenesis is cell enlargement or blastogenic transformation, whereupon the cell volume increases before division. It is usually detectable in the first several hours of T-lymphocyte stimulation. Here, we describe a rapid method to quantify blastogenesis in T lymphocytes isolated from mouse spleens and human peripheral blood mononuclear cells (PBMCs) using an automated cell counter. Various commonly used proliferation assays for the most part are laborious and only reflect the overall population effect rather than individual cellular effects within a population. In contrast, the presented automated cell counter assay provides rapid, direct, and precise measurements of cell diameters that can be used for assessing the effectiveness of various mitogens and immunomodulatory drugs in vitro.

Suppressive potencies of calcineurin inhibitors against the mitogen-induced blastogenesis of peripheral-blood mononuclear cells of myasthenia gravis patients

Abstract The calcineurin inhibitors, tacrolimus and ciclosporin, are two useful immunosuppressive drugs for the treatment of myasthenia gravis (MG), for patients who have low responses to glucocorticoids. We have studied the suppressive potencies of tacrolimus and ciclosporin on concanavalin A-induced blastogenesis of peripheral-blood mononuclear cells (PBMCs) obtained from 38 MG patients and 26 healthy volunteers. Differences in the IC50 values of the two calcineurin inhibitors between the patients and the healthy subjects were evaluated. The median (range) IC50 values for tacrolimus and ciclosporin on the blastogenesis of PBMCs of MG patients were 0.06 (0.001–100) and 0.41 (0.09–83.0) ng mL−1, respectively. In contrast, the median (range) IC50 values of tacrolimus and ciclosporin on healthy PBMCs were 0.16 (0.001–0.33) and 5.59 (1.4–31.3), respectively, and thus ciclosporin potencies against PBMCs of MG patients were significantly higher than those against PBMCs of healthy subjects (P < 0.0001). The differences in tacrolimus IC50 values between the patients and healthy subjects were not significant. There was a correlation between ciclosporin IC50 values against the blastogenesis of PBMCs of MG patients and the duration of the disease (r = 0.35, P = 0.049). A significant correlation between the IC50 values of ciclosporin and those of prednisolone against the blastogenesis of PBMCs of MG patients was also observed (r = 0.56, P = 0.003). Furthermore, the ciclosporin IC50 values significantly correlated with the periods of glucocorticoid administration for MG treatment (r = 0.42, P = 0.038). Such correlations were not observed with the tacrolimus IC50 values. These results suggested that glucocorticoid administration had an influence on PBMC response to the suppressive efficacy of ciclosporin in MG.

Suppressive potencies of calcineurin inhibitors against the mitogen-induced blastogenesis of peripheral-blood mononuclear cells of myasthenia gravis patients

The calcineurin inhibitors, tacrolimus and ciclosporin, are two useful immunosuppressive drugs for the treatment of myasthenia gravis (MG), for patients who have low responses to glucocorticoids. We have studied the suppressive potencies of tacrolimus and ciclosporin on concanavalin A-induced blastogenesis of peripheral-blood mononuclear cells (PBMCs) obtained from 38 MG patients and 26 healthy volunteers. Differences in the IC50 values of the two calcineurin inhibitors between the patients and the healthy subjects were evaluated. The median (range) IC50 values for tacrolimus and ciclosporin on the blastogenesis of PBMCs of MG patients were 0.06 (0.001-100) and 0.41 (0.09-83.0) ng mL(-1), respectively. In contrast, the median (range) IC50 values of tacrolimus and ciclosporin on healthy PBMCs were 0.16 (0.001-0.33) and 5.59 (1.4-31.3), respectively, and thus ciclosporin potencies against PBMCs of MG patients were significantly higher than those against PBMCs of healthy subjects (P < 0.0001). The differences in tacrolimus IC50 values between the patients and healthy subjects were not significant. There was a correlation between ciclosporin IC50 values against the blastogenesis of PBMCs of MG patients and the duration of the disease (r = 0.35, P = 0.049). A significant correlation between the IC50 values of ciclosporin and those of prednisolone against the blastogenesis of PBMCs of MG patients was also observed (r = 0.56, P = 0.003). Furthermore, the ciclosporin IC50 values significantly correlated with the periods of glucocorticoid administration for MG treatment (r = 0.42, P = 0.038). Such correlations were not observed with the tacrolimus IC50 values. These results suggested that glucocorticoid administration had an influence on PBMC response to the suppressive efficacy of ciclosporin in MG.

Cartilage-Hair Hypoplasia

Cartilage-hair hypoplasia (CHH) is a rare autosomal recessive metaphyseal chondrodysplasia seen primarily in people of Amish and Finnish origin. It is characterized by disproportionately short stature; limb deformities; fine, sparse, light-colored hair; and immunodeficiency. The aberrant gene was recently mapped to chromosome 9p21-p13 (the ribonuclease mitochondrial RNA processing [RMRP] gene). The immunologic deficiency in CHH affects primarily the T-cell system but may also involve humoral immunity. Patients with CHH are prone to certain viruses, including varicella, poliovirus, and vaccinia. They also have an increased risk of malignancies. A 39-year-old man with a history of dwarfism first presented with recurrent respiratory infections, allergies, and chronic cough. He experienced chronic sinusitis and recurrent pneumonias that required antibiotics and sinus surgery. Physical examination findings were notable for short stature (114 cm) with typical skeletal deformities; very fine, lightcolored hair; serous otitis media; expiratory rattles; and 2 warts on his left hand. Computed tomography of the chest revealed central bronchiectasis. Moderate lymphopenia was identified, with decreased counts in all subpopulations except natural killer cells. Diminished mitogen-induced blastogenesis was evident. Natural killer cell and neutrophil function were normal. Although CHH predominantly affects cellular immunity, our patient had a profound humoral deficit with remarkably low IgG2, IgG3, and IgG4 subclass levels despite a high-normal total IgG level. His response to pneumococcal polysaccharide was extremely limited. Genomic DNA analysis characterized the patient as compound heterozygotic, with 2 changes in the RMRP gene. Initial treatment with an alternating regimen of antibiotics resulted in improvement. After administration of intravenous immunoglobulin, antibiotics were discontinued with sustained clinical benefit.

Cartilage-Hair Hypoplasia

Cartilage-hair hypoplasia (CHH) is a rare autosomal recessive metaphyseal chondrodysplasia seen primarily in people of Amish and Finnish origin. It is characterized by disproportionately short stature; limb deformities; fine, sparse, light-colored hair; and immunodeficiency.1Makitie O Kaitila I Cartilage-hair hypoplasia—clinical manifestations in 108 Finnish patients.Eur J Pediatr. 1993; 152: 211-217Crossref PubMed Scopus (156) Google Scholar The aberrant gene was recently mapped to chromosome 9p21-p13 (the ribonuclease mitochondrial RNA processing [RMRP] gene).2Ridanpaa M van Eenennaam H Pelin K et al.Mutations in the RNA component of RNase MRP cause a pleiotropic human disease, cartilage-hair hypoplasia.Cell. 2001; 104: 195-203Abstract Full Text Full Text PDF PubMed Scopus (368) Google Scholar The immunologic deficiency in CHH affects primarily the T-cell system but may also involve humoral immunity.3Pierce GF Polmar SH Lymphocyte dysfunction in cartilage-hair hypoplasia: evidence for an intrinsic defect in cellular proliferation.J Immunol. 1982; 129: 570-575PubMed Google Scholar, 4Makitie O Kaitila I Savilahti E Deficiency of humoral immunity in cartilage-hair hypoplasia.J Pediatr. 2000; 137: 487-492Abstract Full Text Full Text PDF PubMed Scopus (49) Google Scholar Patients with CHH are prone to certain viruses, including varicella, poliovirus, and vaccinia. They also have an increased risk of malignancies.5Makitie O Kaitila I Savilahti E Susceptibility to infections and in vitro immune functions in cartilage-hair hypoplasia.Eur J Pediatr. 1998; 157: 816-820Crossref PubMed Scopus (68) Google Scholar A 39-year-old man with a history of dwarfism first presented with recurrent respiratory infections, allergies, and chronic cough. He experienced chronic sinusitis and recurrent pneumonias that required antibiotics and sinus surgery. Physical examination findings were notable for short stature (114 cm) with typical skeletal deformities; very fine, light-colored hair; serous otitis media; expiratory rattles; and 2 warts on his left hand. Computed tomography of the chest revealed central bronchiectasis. Moderate lymphopenia was identified, with decreased counts in all subpopulations except natural killer cells. Diminished mitogen-induced blastogenesis was evident. Natural killer cell and neutrophil function were normal. Although CHH predominantly affects cellular immunity, our patient had a profound humoral deficit with remarkably low IgG2, IgG3, and IgG4 subclass levels despite a high-normal total IgG level. His response to pneumococcal polysaccharide was extremely limited. Genomic DNA analysis characterized the patient as compound heterozygotic, with 2 changes in the RMRP gene. Initial treatment with an alternating regimen of antibiotics resulted in improvement. After administration of intravenous immunoglobulin, antibiotics were discontinued with sustained clinical benefit.

生体適合性を改良したチタン-ジルコニウム合金:ラット埋植試験におけるチタン-ジルコニウム合金の純チタンおよびジルコニウムと比較した組織反応性と感作性

Titanium-zirconium (Ti-Zr) binary alloy has better corrosion resistance and mechanical properties than commercially pure Ti. The present study was designed to determine the biocompatibility of Ti-Zr alloy by an implantation test in animal bodies in comparison with pure Ti, Zr, and chromium (Cr) implants. Sample specimens were placed in a subcutaneous position in rats for 8 months. No significant decreases in body weight, the weight of any organ, or the weight of any organ relative to body weight were found in the implant groups compared to a no-implant control group. On hematological examination, small differences in several parameters were found in some groups, but these changes were not attributable to the materials implanted. Mitogen-induced blastogenesis was observed in similar degrees among all implant groups. These results suggest that the implantation of test samples did not cause systemic toxicity or a decrease in immune activity. The fibrous capsule membranes around the Ti and Ti-Zr alloy implants were thinner than those around Cr implants. The numbers of macrophages, inflammatory cells, and other cells involved in immune responses in and around the fibrous capsules of the Cr- and Ti-implant groups were higher than those of the Ti-Zr alloy- and Zr-implant groups. The Ti-Zr alloy had the lowest total score of tissue responses among the materials tested. None of the animals from the Ti-, Zr-, and Ti-Zr alloy-implant groups exhibited a skin reaction following exposure to Ti or Zr salt solutions. These results indicate the Ti-Zr alloy has better biocompatibility than Ti for use as an artificial surgical implant.

The effect of calyx infusion of hibiscus sabdariffa l. On t-cells-mediated immune response in mitogen-induced blastogenesis of human lymphocytes in vitro

Free radical production and disturbance in the redox status may modulate the expression of a variety of immune and inflammatory molecules. Antioxidants could inhibit the proliferation of T lymphocytes induced by mitogens. The aim of this study was to investigate the effect of the infusion of dried calyx of Hibiscus sabdariffa L. on T-cell-mediated immune response. We examined the effect of the infusion on spontaneous and mitogen-induced blast transformation of human lymphocytes in vitro. Peripheral blood lymphocytes from heparinised venous blood of healthy subjects (n=17) were examined. Five and 10 mg ml-1 phytohemagglutinin (PHA) and 5 mg ml-1 pokeweed mitogen (PWM) were added. Depending on the concentration, calyx infusion of H. sabdariffa decreased mitogen-induced blastogenesis in normal subjects. It did not affect, however, the level of spontaneous proliferation of human lymphocytes in vitro. The infusion of hibisci flos may alleviate over-stimulated condition of the immune system in hypersensitivity ...

Improved Biocompatibility of Titanium&amp;ndash;Zirconium (Ti&amp;ndash;Zr) Alloy: Tissue Reaction and Sensitization to Ti&amp;ndash;Zr Alloy Compared with Pure Ti and Zr in Rat Implantation Study

Titanium–zirconium (Ti–Zr) binary alloy has better corrosion resistance and mechanical properties than commercially pure Ti. The present study was designed to determine the biocompatibility of Ti–Zr alloy by an implantation test in animal bodies in comparison with pure Ti, Zr, and chromium (Cr) implants as positive controls. Sample specimens were placed in a subcutaneous position in rats for 8 months. No significant decreases in body weight, the weight of any organ, or the weight of any organ relative to body weight were found in the implant groups compared to a no-implant control group. On hematological examination, small differences in several parameters were found in some groups, but these changes were not attributable to the materials implanted. Mitogen-induced blastogenesis was observed in similar degrees among all implant groups. These results suggest that the implantation of test samples did not cause systemic toxicity or a decrease in immune activity. The fibrous capsule membranes around the Ti and Ti–Zr alloy implants were thinner than those around Cr implants. The numbers of macrophages, inflammatory cells, and other cells involved in immune responses in and around the fibrous capsules of the Cr- and Ti-implant groups were higher than those of the Ti–Zr alloy- and Zr-implant groups. The Ti–Zr alloy had the lowest total score of tissue responses among the materials tested. None of the animals from the Ti-, Zr-, and Ti–Zr alloy-implant groups exhibited a skin reaction following exposure to Ti or Zr salt solutions. These results indicate the Ti–Zr alloy has better biocompatibility than Ti for use as an artificial surgical implant.

Fish oil diet affects on oxidative senescence of red blood cells linked to degeneration of spleen cells in mice

The effect of dietary polyunsaturated fatty acids and α-tocopherol supplementation on erythrocyte lipid peroxidation and immunocompetent cells in mice was studied comparatively using seven dietary oils (15% oil/diet, w/w) including fish oil rich in eicosapentaenoic acid (EPA, 20:5, n–3) and docosahexaenoic acid (DHA, 22:6, n–3). A 43% increase in spleen weight, about twice as many spleen cells and no change in the subpopulations of spleen cells, as well as a significant depression of mitogen-induced blastogenesis of both T and B cells in the spleen were observed in mice fed fish oil for 30 days in comparison with soybean oil diet-fed mice. In the fish oil diet-fed mice, membranous lipid hydroperoxide (hydroperoxides of phosphatidylcholine and phosphatidylethanolamine) accumulation as a marker of oxidative senescence in red blood cells (RBC) was 2.7–3.5 times higher than that in mice fed soybean oil, although there was no difference in the plasma phosphatidylcholine hydroperoxide concentration. In spite of the supplementation of α-tocopherol to up to 10 times the level in the basal diet, the degeneration of spleen cells and the stimulated oxidative senescence of RBC found by the fish oil feeding could not be prevented. The results suggest that oral intake of excess polyunsaturated fatty acids, i.e. EPA and DHA, in a fish oil diet can lead to acceleration of membrane lipid peroxidation resulting in RBC senescence linked to the lowering of immune response of spleen cells, and that supplementation of α-tocopherol as antioxidant does not always effectively prevent such oxidative degeneration as observed in spleen cells and RBC in vivo.

Postmenopausal hormone replacement: effects on autonomic, neuroendocrine, and immune reactivity to brief psychological stressors.

Postmenopausal status increases some aspects of women's physiological responses to psychological stress; however, the influences of chronic hormone replacement with estrogen and progestogen on these responses are not known. We investigated possible effects of long-term estrogen replacement therapy (ERT), both with and without progestogen, on physiological reactivity to brief laboratory stressors. We studied three groups of postmenopausal women: 16 on estrogen alone, 14 on estrogen and progestogen, and 25 control participants receiving no replacement therapy. Cardiovascular, neuroendocrine, and immune data were collected at baseline and after speech and math tasks. In all groups, the stressors reduced vagal cardiac control (indexed by respiratory sinus arrhythmia); increased heart rate and plasma epinephrine, adrenocorticotropic hormone, and cortisol levels; and altered T lymphocyte response (measured by mitogen-induced cell proliferation), natural killer cell lysis, and circulating leukocyte subsets. Women on either type of ERT had higher total cortisol levels (reflecting an estrogen effect on cortisol binding globulin) and greater mitogen-induced blastogenesis across measurement periods than controls. They also showed greater vagal withdrawal and less decline in mitogen-stimulated blastogenesis in response to the stressors. Combined estrogen and progestogen was associated with higher epinephrine and lower circulating total lymphocytes, T cells, and CD4+ T cells across measurement periods, and with intermediate levels of vagal withdrawal in response to the stressors. Long-term ERT was associated with enhanced parasympathetic responsiveness to stress, suggesting possible reduced demand for potentially detrimental sympathetic activation; and with higher overall levels and smaller stress-induced reductions of mitogen-stimulated blastogenesis, suggesting up-regulated T cell function.

Possible association between immunocompetence and enhanced risk of multicentric carcinogenesis in both the stomach and other organs in patients with gastric cancer

In a retrospective study, we analyzed the association between the immunocompetence and multicentric carcinogenesis in both the stomach and organs other than the stomach in patients with single gastric cancer (SGC) and those with multiple gastric cancers (MGC). The incidence of primary cancers in other organs in patients with MGC (4/17, 23.5%) was significantly higher than that with SGC (15/202, 7.4%). Our analysis of the immunocompetence revealed a trend towards decreased activity of NK cells and of mitogen-induced blastogenesis in patients with MGC as compared to those with SGC. In patients with SGC, patients with primary cancers in other organs had significantly lower levels of both PHA-induced blastogenesis and CD4/CDs, and revealed a greater trend towards decreased activity of both NK cells and Con A-induced blastogenesis than patients without other primary cancers. These findings may suggest that immunocompetence plays a role in the enhanced multicentric carcinogenesis not only in the stomach but also in cancers of other organs.

Immunomodulatory activity of blood serum from chromium-supplemented periparturient dairy cows

Our previous research showed enhanced immune responses, including mitogen-induced blastogenesis of peripheral blood mononuclear cells from feedlot calves and periparturient dairy cows supplemented with dietary chromium (Cr). The objectives of the present study were to test whether blood sera from Cr-supplemented periparturient cows contained immunomodulatory activity for mitogen-stimulated peripheral blood mononuclear cells and, if so, to determine if this activity was explicable by differences in blood profiles of some glucose-regulating hormones (insulin, cortisol, growth hormone, insulin-like growth factor I, and tumor necrosis factor-α) between Cr-supplemented and unsupplemented (control) animals. Blood sera from ten unsupplemented cows and nine Cr-supplemented cows (0.5 ppm day −1) were collected weekly from 2 weeks before to 6 weeks after parturition, and were used to supplement (1, 10, and 20% vol/vol) culture medium supporting concanavalin A (Con A) stimulated mononuclear cells enriched from blood of four nulliparous donor cows. Hormone concentrations were determined using radioimmunoassays. Con A-induced blastogenesis was enhanced when 1, 10, and 20% sera from Cr-supplemented cows was added to the mononuclear cell cultures, and this was particularly evident around parturition. Conversely, peripartum sera from unsupplemented cows depressed Con A-induced blastogenesis. Except for a marginal rise in blood cortisol 2–4 weeks after parturition, no significant effects of Cr supplementation on other hormones (insulin, growth hormone, insulin-like growth factor I, tumor necrosis factor-α) were observed. These observations suggest that factors in peripheral blood serum from Cr-supplemented cows, other than absolute concentrations of the glucose-regulating hormones studied, modulate Con A-induced blastogenesis of mononuclear leukocytes.

Cellular Interactions of Murine Immune Cells Exposed to Live Mycobacterium intracellulare, Its Whole Lipid Extract, and Its Serovar-Specific Glycopeptidolipid

In this study we examined some of the immunological responses to Mycobacterium intracellulare and its lipid components. Our results indicate that infection with M. intracellulare can increase the expression of adhesion molecules, ICAM-1 and LFA-1, only at the site of injection (peritoneum). There was no change in the expression of these adhesion molecules in the lymphoid organs (thymus and spleen). Significant increases in the adhesion molecules were observed in the spleen cells incubated with the lipid derived from mycobacteria in the presence of concanavalin (Con A) compared to the Con A alone. The expression of the Thy 1.2 and Lyt-2 markers was not affected by the bacteria or their lipids. The results indicate a marked increase in the mitogenic response by the infected spleen cells removed at an early day. The blastogenic study also indicated that the lipids can reduce the mitogen-induced blastogenesis of spleen cells removed from M. intracellular- and saline-injected mice; moreover, they suggested that the spleen cells removed from Listeria monocytogens-infected mice can also be affected by mycobacterial lipids. This indicates a nonspecific effect by these lipids. The results that the immunological response was contingent upon prior exposure of the mice to M. intracellulare, and also was dependent on whether the cells came from the peritoneal cavity or lymphoid organs.

Natural flavonoids and lignans are potent cytostatic agents against human leukemic HL-60 cells

Anti leukemic-cell efficacy of 28 naturally occurring and synthetic flavonoids and 11 naturally occurring lignans on human promyelocytic leukemic cell line HL-60 were examined using MTT assay methods. Differences between anti cell-proliferative activity and cytotoxicity of these compounds were compared with thos of 4 clinical anti-cancer agents. Eight of the 28 flavonoids and 4 of the 11 lignans showed considerable suppressive effects on HL-60 cell growth with IC 50s ranging from 10–940 ng/ml. Among these compounds, genistein, honokiol, machilin A, matairesinol, and arctigenin had the strongest effects with IC 50s less than 100 ng/ml, which were almost equivalent to the effects of current anti-cancer agents. The flavonoid genistein and the lignans, however, showed little or no cytotoxicity against HL-60 cells as assessed by dye exclusion tests (LC 50s>2,900ng/ml), whereas the regular anti-cancer agents had potent cytotoxicity. All of the flavonoids and lignans, except for machilin A and arctigenin, were less effective against growth of human T lymphocytic leukemia cell line MOLT- 4. In addition, the flavonoid and the lignans showed little or no inhibiting activity on mitogen-induced blastogenesis of human peripheral-blood lymphocytes. The lignans and genistein were strongly suppressive against incorporations of [ 3H]thymidine, [ 3H]uridine, and [ 3H]leucine into HL-60 cells. These results showed that some of the naturally occurring flavonoids and lignans inhibited HL-60 cell growth with a non-toxic mechanism, possibly via cessation of DNA, RNA, and/or protein synthesis of the leukemic cells.

Variability in chromosome aberrations, sister chromatid exchanges, and mitogen-induced blastogenesis in peripheral lymphocytes from control individuals.

Confidence in results from monitoring genetic end points in environmentally or occupationally exposed individuals can be improved with knowledge of the normal variability of changes in genetic end points in the general population. Confounding effects can be determined, and study interpretation can be improved by correlation of this variability with various lifestyle factors such as sex and age, smoking and drinking habits, viral infections, exposure to diagnostic X-rays, etc. Eight blood samples were taken from each of 24 male and 24 female volunteers over a period of 2 years. Questionnaires pertaining to lifestyle were completed at the time of each sampling. Whole blood was cultured and slides prepared for chromosome aberration (CA) or sister chromatid exchange (SCE) analysis. Separated mononuclear cells were cultured with a range of phytohemagglutinin concentrations, and the maximum level of mitogen-induced blastogenesis was determined by measurement of [3H]thymidine uptake. There was a significant effect of both year and season of sampling for all three end points. Because there was no consistent pattern in 2 successive years, effects were thought to be independent of season. No significant effects in any of the three end points were found with respect to sex or age nor any of the other lifestyle factors, although SCE frequency and mitogen-induced blastogenesis were nearly always higher in females than in males. These results point to the need for concurrent sampling of controls with exposed populations.

Variability in Chromosome Aberrations, Sister Chromatid Exchanges, and Mitogen-Induced Blastogenesis in Peripheral Lymphocytes from Control Individuals

Variability in Chromosome Aberrations, Sister Chromatid Exchanges, and Mitogen-Induced Blastogenesis in Peripheral Lymphocytes from Control Individuals

In vitro and ex vivo effect of tiaprofenic acid on human peripheral blood mononuclear cells

The effect of the non-steroidal anti-inflammatory drug (NSAID) tiaprofenic acid on different human immune parameters was investigated in vitro or following in vivo administration in healthy adult volunteers. Results from the in vitro study demonstrated an increased mitogen-induced blastogenesis and interleukin 2 (IL-2) production together with a reduced polyclonal immunoglobulin (Ig) secretion in the presence of the drug. Results from the ex vivo study showed that treatment with tiaprofenic acid had no significant effects on the immune parameters investigated, i.e. unstimulated and mitogen-induced proliferation and IL-2 production, spontaneous and stimulated Ig synthesis, lymphocyte subpopulations, serum Ig and complement levels.

Effects of exercise stress on various immune functions in horses

Summary Chemotactic locomotion and luminol-dependent chemiluminescence of neutrophils, mitogen-induced lymphocyte blastogenesis, serum cortisol concentration, immunoglobulin quantification, and leukocyte counts were determined to evaluate the effect of a single strenuous exercise in horses. Increased serum cortisol concentration (P &lt; 0.01) and an increased neutrophil-to-lymphocyte ratio (P &lt; 0.05) indicated that horses had been stressed. The chemotactic index and peak chemiluminescence production decreased significantly (P &lt; 0.05 and P &lt; 0.01, respectively) 1 day after exercise. Mitogen-induced blastogenesis of lymphocytes and serum immunoglobulin values remained unchanged in response to exercise. Results of this study indicated that a single bout of exercise may transiently impair neutrophil antimicrobial functions and nonspecific defense mechanisms, but not specific immunity in horses.

Chromosome aberrations (CA), sister-chromatid exchanges (SCE) and mitogen-induced blastogenesis in cultured peripheral lymphocytes from 48 control individuals sampled 8 times over 2 years

Confidence in the measurement of positive effects determined by monitoring of environmentally or occupationally exposed individuals can be enhanced by a knowledge of the normal variability in these endpoints in the general population. Confounding effects can be determined and study interpretation improved by correlation of this variability with various lifestyle factors such as sex and age of donor, smoking and drinking habits, viral infections, exposure to diagnostic X-rays, etc.8 blood samples were taken from each of 24 male and 24 female volunteers over a period of 2 years. Questionnaires pertaining to lifestyle were completed at the time of each sampling. Whole blood was cultured and slides prepared for CA or SCE analysis. Separated mononuclear cells were cultured with a range of phytohaemagglutinin concentrations and the maximum level of mitogen-induced blastogenesis was determined by measurement of [3H]thymidine uptake.There was a significant effect of both year and season of sampling for all 3 endpoints. No significant effects in any of the 3 endpoints were found with respect to sex or age of donor nor any of the other lifestyle factors, although SCE frequency and mitogen-induced blastogenesis were nearly always higher in females than males. These results point to the need for concurrent sampling of controls with exposed populations.