Effects of sarcolectin (SCL) on human peripheral blood mononuclear cells

Sarcolectin (SCL) is a tissue growth factor found in various human or animal tissues, functioning in balance with interferons (IFNs) that can inhibit growth and affect cell differentiation. Like somatotropin, SCL is found in the pituitary gland. In humans, the SCL gene is located on chromosome 12 (q12-q13) and expressed as a 55 kDa protein consisting of 469 amino-acids. After a single activation of peripheral blood mononuclear cells (PBMC) obtained from more than 30 individuals, highly significant cell proliferation was found to peak after 7 days in culture. The presence of adherent cells was necessary for cell proliferation. SCL induced over-expression of alpha-IL-2 receptor (CD25) leading to proliferation of CD3+/CD4+/CD45RO+ T cells. Thus in PBMC, SCL induced CD4+ T cell growth and expression of inflammatory cytokine genes, including TNF-alpha, IL-1beta, IL-6 and IL-8. IFNs are also produced following activation as a feedback response which is maintained for about 20 days.     

Effects of 10-T static magnetic field on human peripheral blood immune cells

The exposure of peripheral blood mononuclear cells (PBMCs) was performed in a 10-T static magnetic field. Without lymphocyte stimulation, there were no significant differences in the viability of the exposed and unexposed CD4(+) T cells, CD8(+) T cells, B cells, and natural killer (NK) cells. The expression of Th1 type chemokine receptor, CXCR3, and Th2 type receptor, CCR3, was unaltered after magnetic-field exposure. No differences were observed in the naive T cells and memory T-cell subclasses in either CD4(+) or CD8(+) T cells. In contrast to the unstimulated condition, the magnetic-field exposure reduced the viability of phytohemagglutinin (PHA)-activated T cells in both the CD4(+) and CD8(+) subclasses. In particular, the number of PHA-treated naive CD8(+) T cells (CD45RA(+)CD4(-)CD8(+)) was markedly decreased after the magnetic-field exposure, while PHA-treated memory CD8(+) cells (CD45RA(-)CD4(-)CD8(+)) were resistant to the exposure. The number of PHA-treated naive CD4(+) T cells (CD45RA(+)CD4(+)CD8(-)) and memory cells (CD45RA(-)CD4(+)CD8(-)) was markedly decreased to a similar degree. Thus the susceptibility of lymphocytes to the magnetic-field exposure differed among activated T-cell subtypes. The magnetic-field exposure significantly increased the death of PHA-stimulated lymphocytes by apoptosis. These results suggest that a strong static magnetic field has acute effects on immune cells during cell division, while the field exposure has a minimal effect on immune cells in a nondividing phase.     

Effects of exercise on gene expression in human peripheral blood mononuclear cells.

Exercise leads to increases in circulating levels of peripheral blood mononuclear cells (PBMCs) and to a simultaneous, seemingly paradoxical increase in both pro- and anti-inflammatory mediators. Whether this is paralleled by changes in gene expression within the circulating population of PBMCs is not fully understood. Fifteen healthy men (18-30 yr old) performed 30 min of constant work rate cycle ergometry (approximately 80% peak O2 uptake). Blood samples were obtained preexercise (Pre), end-exercise (End-Ex), and 60 min into recovery (Recovery), and gene expression was measured using microarray analysis (Affymetrix GeneChips). Significant differential gene expression was defined with a posterior probability of differential expression of 0.99 and a Bayesian P value of 0.005. Significant changes were observed from Pre to End-Ex in 311 genes, from End-Ex to Recovery in 552 genes, and from Pre to Recovery in 293 genes. Pre to End-Ex upregulation of PBMC genes related to stress and inflammation [e.g., heat shock protein 70 (3.70-fold) and dual-specificity phosphatase-1 (4.45-fold)] was followed by a return of these genes to baseline by Recovery. The gene for interleukin-1 receptor antagonist (an anti-inflammatory mediator) increased between End-Ex and Recovery (1.52-fold). Chemokine genes associated with inflammatory diseases [macrophage inflammatory protein-1alpha (1.84-fold) and -1beta (2.88-fold), and regulation-on-activation, normal T cell expressed and secreted (1.34-fold)] were upregulated but returned to baseline by Recovery. Exercise also upregulated growth and repair genes such as epiregulin (3.50-fold), platelet-derived growth factor (1.55-fold), and hypoxia-inducible factor-I (2.40-fold). A single bout of heavy exercise substantially alters PBMC gene expression characterized in many cases by a brisk activation and deactivation of genes associated with stress, inflammation, and tissue repair.     

Effect of some nickel compounds on red blood cell characteristics

The effect of certain inorganic and coordinated nickel compounds on the resistance to different destructive substances, rheological properties, and functional activity of healthy human red blood cells (RBC), was investigated. It is shown that nickel compounds affect the erythrocyte membrane lipid bilayer, as well as membrane proteins to various extents, depending on the type of compounds used. In general, the acceleration of erythrocyte aging was observed to be more pronounced in young erythrocytes. The observed results suggest that nickel compounds decrease water permeability across erythrocyte membranes. Almost all the investigated nickel compounds decrease erythrocyte thermostability, deformability, and the rate of O₂ release by erythrocytes.     

硼对大白菜生长发育及若干生理生化指标的影响

研究了不同水平的硼对大白菜生长发育及若干生理生化指标的影响。结果表明,缺硼明显抑制大白菜植株生长、抽薹和开花,导致花粉活力下降、叶绿素含量降低、可溶性糖含量增加、膜脂过氧化产物丙二醛(MDA)含量增加、超氧化物歧化酶(SOD)活性下降、过氧化物酶(POD)活性上升,最终导致植株代谢紊乱,生长发育受抑制。     

Cytotoxicity of human peripheral blood monocytes

Native tumoricidal activity of human peripheral blood mononuclear cells was examined before and after their separation by counterflow centrifugation elutriation (CCE). Tumoricidal activity was found in the subpopulation of small mononuclear cells but not within the relatively pure subpopulation of large monocytes. Addition of lymphokine and/or lipopolysaccharide demonstrated that large monocytes were resistant to activation for tumor killing, in contrast to small mononuclear cells. However, cryopreservation or simply exposure to dimethyl sulfoxide (DMSO) rendered the large monocytes sensitive to activating agents without altering their unstimulated tumoricidal activity. Cryopreservation was not detrimental to small or large monocytes either in number or tumoricidal function but did decrease the number of large granular lymphocytes (LGL). The small mononuclear cell fraction was enriched for small monocytes to 80% by combining CCE with Percoll gradient separation. HNK-1 mouse monoclonal antibody against human LGL was used with complement to remove virtually all LGL from cryopreserved cells as judged by morphology and tumoricidal activity against K-562 human lymphoblastoid cells. Such treatment actually augmented rather than suppressed tumoricidal activity against P-815 mastocytoma cells. Therefore, we conclude that small monocytes but not large monocytes possess native tumoricidal activity distinct from that attributed to LGL or natural killer lymphocytes. Further, small monocytes are readily activated for tumor killing and can be cryopreserved without loss of tumoricidal activity.     

Effects of deoxyribonucleosides and cell-stimulation on frequencies of ultraviolet-B-induced micronuclei in human peripheral blood lymphocytes

In the present study the involvement of deoxyribonucleotides (dNTPs) in the clastogenicity of ultraviolet-B (UVB) in unstimulated peripheral blood lymphocytes (G(0)-PBLs) was investigated. This was studied by analyzing the frequency of UVB-induced micronuclei (MN), either after adding a cocktail of the four deoxyribonucleosides to the PBLs immediately after exposure to UVB, or by stimulating the cells before exposure. In total, PBLs obtained from two different donors were investigated. For both donors, it could be demonstrated that addition of deoxyribonucleosides to UVB-irradiated G(0)-PBLs resulted in a significant reduction of the clastogenic effect of UVB. A gradual reduction of the clastogenic effect of UVB could also be realized by irradiating PBLs that were progressively more stimulated with the lectin PHA before exposure. The latter finding is explained by upregulation of intracellular pool sizes of dNTPs in stimulated PBLs.     

Effects of cocaine and morphine on IgG production by human peripheral blood lymphocytes in vitro

Elevated serum levels of IgG are amongst the immunological abnormalities exhibited by intravenous drug addicts. We therefore addressed the hypothesis that cocaine and morphine (the major metabolite of heroin) exert a direct effect on human B cell function in vitro. Human peripheral blood mononuclear cells from normal individuals were incubated for 7 days with the T cell-dependent B cell activator pokeweed mitogen (PWM) and serial dilutions of either cocaine or morphine. At the end of this time total IgG was measured by use of a sandwich ELISA incorporating a biotin-labelled affinity-purified anti-IgG and streptavidin peroxidase. At concentrations relevant to those found in plasma, morphine and cocaine did not affect PWM-stimulated IgG synthesis in vitro. We suggest that these drugs of abuse do not directly influence human B cells, but in vivo exert immune modulatory effects via indirect mechanisms.     

Cryopreserving human peripheral blood progenitor cells

High-dose chemotherapy followed by autologous peripheral blood progenitor cell (PBPC) transplantation is used in the treatment of chemosensitive malignancies. Cryopreservation of PBPC in 10% dimethyl sulfoxide (DMSO) has been the standard procedure in most institutions. Infusion of PBPC cryopreserved with DMSO can be associated with toxic reactions such as vomiting, cardiac dysfunction, anaphylaxia and acute renal failure. The grade of toxicity experienced by patients is related to the amount of DMSO present in the PBPC. Cryopreservation with lower DMSO concentrations would be expected to reduce the toxicity. In recent studies done with PBPC cells cryopreserved with 5%, 4% and 2% DMSO, using 10% DMSO as a reference control, CD34+ cells were investigated for preservation of viability, apoptosis, and necrosis. Also preservation of mature colony-forming (CFU) cells, specifically mature myeloid, erythroid progenitors, CFU-megakaryocytes and long-term culture-initiating cells (LTC-ICs) were investigated, using 5% and 10% DMSO as cryoprotectant. All samples were frozen in a rate-controlled programmed freezer and stored in the vapor phase of liquid nitrogen until used. Conclusion: 5% DMSO is the optimal concentration for cryopreserving human PBPC in vitro. Consequently, some hospitals have started using 5% DMSO as cryoprotectant for the autologous PBPC as a standard procedure.     

Effects of long-term cryopreservation on peripheral blood progenitor cells

Background aimsThe long-term stability of cryopreserved peripheral blood progenitor cells is an important issue for patients experiencing disease relapse. However, there is no consensus on how to evaluate the long-term effects of cryopreservation. We describe the effect of cryopreservation on viability and progenitor colony activity from 87 individual samples processed at the Scripps Green Hospital Stem Cell Processing Center (La Jolla, CA, USA).MethodsWe randomly selected 87 peripheral blood hematopoietic stem cell (PBHSC) samples from 60 patients and evaluated the effect of cryopreservation on sample viability and red and white cell colony activity after < 24 h and 7, 10 and 15 years of cryopreservation. Viability was assayed via trypan blue dye exclusion and activity was measured following 14 days of culture.ResultsAn age at collection older than 50 years may result in suboptimal activity and viability following long-term cryopreservation, while gender and disease status had no effect. Cryopreservation did not significantly affect white or red cell activity following 10 years of cryopreservation. However, for samples stored longer than 10 years, viability and activity significantly decreased. We noted a positive association between higher pre-cryopreservation %CD34 count and colony activity.ConclusionsCryopreservation of peripheral blood progenitor cells for up to 10 years results in no loss of clonogenic capacity, as determined by culture activity, although longer durations of storage may affect activity. Until validated methods are developed, cryopreserved grafts should be evaluated based on pre-freeze CD34⁺ cell counts as assayed by flow cytometry, and post-thaw sample evaluation should be reserved for patients identified as poor mobilizers.     

Effects of rare earth elements on telomerase activity and apoptosis of human peripheral blood mononuclear cells

To study the effects of rare earth exposure on human telomerase and apoptosis of mononuclear cells from human peripheral blood (PBMNCs). The blood contents of 15 rare earth elements, including La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, and Y, were measured by inductively coupled plasma-mass spectrometry. Telomeric repeat amplification protocol assay and flow cytometer analysis were carried out to analyze the telomerase activity and apoptosis of PBMNCs, respectively. The total content of rare earth elements in the blood showed significant differences between the exposed group and the control group. The rare earth exposure increased the telomerase activity and the percentages of cells in the S-phase and the G₂/M phase in PBMNCs, but it had no effect on the apoptotic rate of PBMNCs. Under the exposure to lower concentrations of rare earth elements, the telomerase activity of PBMNCs in the exposed group was higher than that of the control group, and there was no effect on the apoptotic rate of PBMNCs, but promoted the diploid DNA replication and increased the percentages of G₂/M- and S-phase cells.     

Lectin-dependent cytotoxicity of human peripheral blood lymphocytes

The evaluative technique of lymphocyte cytolytic activity in human peripheral blood has been designed. The target cells, lectin (Con A) concentration and incubation time for measuring cytolytic activity of lymphocytes pre-purified from adherent cells have been selected. The mean values of lectin-dependent cytotoxicity in peripheral blood of 50 healthy donors are presented.     

Effects of PM10 in human peripheral blood monocytes and J774 macrophages

Background

Asthma is characterized by type 2 T-helper cell (Th2) inflammation, goblet cell hyperplasia, airway hyperreactivity, and airway fibrosis. Monocyte chemoattractant protein-1 (MCP-1 or CCL2) and its receptor, CCR2, have been shown to play important roles in the development of Th2 inflammation. CCR2-deficient mice have been found to have altered inflammatory and physiologic responses in some models of experimental allergic asthma, but the role of CCR2 in contributing to inflammation and airway hyperreactivity appears to vary considerably between models. Furthermore, MCP-1-deficient mice have not previously been studied in models of experimental allergic asthma.

Methods

To test whether MCP-1 and CCR2 are each required for the development of experimental allergic asthma, we applied an Aspergillus antigen-induced model of Th2 cytokine-driven allergic asthma associated with airway fibrosis to mice deficient in either MCP-1 or CCR2. Previous studies with live Aspergillus conidia instilled into the lung revealed that MCP-1 and CCR2 play a role in anti-fungal responses; in contrast, we used a non-viable Aspergillus antigen preparation known to induce a robust eosinophilic inflammatory response.

Results

We found that wild-type C57BL/6 mice developed eosinophilic airway inflammation, goblet cell hyperplasia, airway hyperreactivity, elevations in serum IgE, and airway fibrosis in response to airway challenge with Aspergillus antigen. Surprisingly, mice deficient in either MCP-1 or CCR2 had responses to Aspergillus antigen similar to those seen in wild-type mice, including production of Th2 cytokines.

Conclusion

We conclude that robust Th2-mediated lung pathology can occur even in the complete absence of MCP-1 or CCR2.     

Fibrinolytic activity of human peripheral blood granulocytes

The fibrinolytic activity of human peripheral blood leukocytes was studied by plating the cells on 125I-fibrin coated dishes. The separation of the three major leukocyte types allowed to demonstrate that most of the activity was produced by granulocytes. The rate of fibrinolysin was found to be linear with incubation time and cell number in the range of 1-4 X 10(5) cells/ml. Since little activity was found in absence of exogenous plasminogen, it was concluded that the cell fibrinolytic activity depended mostly upon the release of plasminogen activator. Plasmatic and granulocytic activators obtained from the same amount of blood were found to be of similar level suggesting a possible clinical implication of the cellular activity in the thrombolytic system.