PHA-stimulated Peripheral Blood Lymphocytes Research Articles

Burkholderia cepacia in cystic fibrosis children and adolescents: overall survival and immune alterations.

In current literature there are only scarce data on the host inflammatory response during Burkholderia cepacia complex (Bcc) persistence. The primary objective of the present research was to carry out cross-sectional analyses of biomarkers and evaluate disease progression in cystic fibrosis (CF) patients with chronic Bcc infection and pathogen-free ones. The secondary aim was to assess prospectively overall survival of the study participants during up to 8 years of follow-up. The study included 116 paediatric patients with CF; 47 CF patients were chronically infected with Bcc, and 69 individuals were Bcc free. Plasma and sputum biomarkers (neutrophil elastase, MMP-8, MMP-9, MMP-12, IL-2, IL-4, IL-6, IL-8, IL-10, IL-18, IL-22, IL-23, IL-17, IFN-γ, TGFβ1, TNF-α) were analysed using commercially available kits. Besides, inhibitory effect of dexamethasone on proliferative response of PHA-stimulated peripheral blood lymphocytes had been assessed. Bcc infected patients did not differ from Bcc free ones in demographic and clinical parameters, but demonstrated an increased rate of glucose metabolism disturbances and survival disadvantage during prolong follow-up period. Biomarkers analyses revealed elevated TNF-α and reduced IL-17F levels in sputum samples of Bcc infected patients. These patients also demonstrated improvement of peripheral blood lymphocyte sensitivity to steroid treatment and reduction in plasma pro-inflammatory (IL-17F and IL-18) and anti-inflammatory (TGFβ1 and IL-10) cytokine concentrations. Reduction in IL-17F levels may have several important consequences including increase in steroid sensitivity and glycemic control disturbances. Further investigations are needed to clarify the role of IL-17 cytokines in CF complication development. Low plasma TGFβ1 and IL-10 levels in Bcc infected group may be a sign of subverted activity of regulatory T cells. Such immune alterations may be one of the factors contributing to the development of the cepacia syndrome.

Functional analysis of membrane-bound complement regulatory protein on T-cell immune response in ginbuna crucian carp.

Complements have long been considered to be a pivotal component in innate immunity. Recent researches, however, highlight novel roles of complements in T-cell-mediated adaptive immunity. Membrane-bound complement regulatory protein CD46, a costimulatory protein for T cells, is a key molecule for T-cell immunomodulation. Teleost CD46-like molecule, termed Tecrem, has been newly identified in common carp and shown to function as a complement regulator. However, it remains unclear whether Tecrem is involved in T-cell immune response. We investigated Tecrem function related to T-cell responses in ginbuna crucian carp. Ginbuna Tecrem (gTecrem) proteins were detected by immunoprecipitation using anti-common carp Tecrem monoclonal antibody (mAb) and were ubiquitously expressed on blood cells including CD8α(+) and CD4(+) lymphocytes. gTecrem expression on leucocyte surface was enhanced after stimulation with the T-cell mitogen, phytohaemagglutinin (PHA). Coculture with the anti-Tecrem mAb significantly inhibited the proliferative activity of PHA-stimulated peripheral blood lymphocytes, suggesting that cross-linking of Tecrems on T-cells interferes with a signal transduction pathway for T-cell activation. These findings indicate that Tecrem may act as a T-cell moderator and imply that the complement system in teleost, as well as mammals, plays an important role for linking adaptive and innate immunity.

Changes in Serum and Cell Homeostasis under Irradiation: Factors of Distant Action

Extracellular DNA-containing substances can be used as a marker of the balance between two processes: elimination of cells with critical number of lesions and repair of genetic material due to the work of adaptive systems. The idea of using this criterion was inspired by the results of measuring exDNA in blood plasma of chronically irradiated individuals. Here we used material obtained after gamma-irradiation (total dose 0.5 Gy) and subsequent culturing of PHA-stimulated human peripheral blood lymphocytes. The culture medium and exDNA isolated by various methods were treated with DNase and pronase. The concentration of free exDNA isolated by phenol extraction was an individual sign. The culture medium supernatant contained considerable amounts of exDNA in the form of nucleoprotein; cleavage of the protein component of this nucleoprotein considerably increased the size of fragments (from 1 to 20 kb) and reduced their resistance to DNase.

Malignant fibrous histiocytoma two years after autologous stem cell transplant for Hodgkin lymphoma: Evidence for genomic instability

Secondary malignancies (SMs) in Hodgkin lymphoma (HL) are thought to be related to exposure to alkalating agents, topoisomerase II inhibitors and ionizing radiation, and tend to occur a decade after initial therapy. We report a 14 year old autistic male, who developed malignant fibrous histiocytoma (MFH) two years after autologous stem cell transplantation for advanced stage HL. The MFH and post-surgical reactive tissues exhibited multiple clonal abnormalities. In addition, PHA-stimulated peripheral blood lymphocytes showed increased frequency of non-clonal chromosomal aberrations. The potential role of genomic instability in early onset of SM in our patient is discussed.

The Effect of Mesenchymal Stem Cells on Preventing Graft-Versus-Host Disease After Allogeneic Hematopoietic Stem Cell Transplantation—the Effect of Mesenchymal Stem Cells on Active T Lymphocytes(in vitro)

AbstractAbstract 5182 ObjectiveTo study the immune regulatory effects of human bone marrow Mesenchymal stem cells(MSCs) on active T lymphocytes in vitro and to explore the new strategy to prevent Graft-Versus-Host Disease(GVHD) in allogeneic hematopoietic stem cell transplantation(allo-HSCT). Methods1. Mononuclear cells from human peripheral blood cells were isolated and cultured in the presence of phytohemagglutinin (PHA) (final concentration was 10μ g/ml) for different time (24hours, 36hours, 48hours, 72hours). 2. The ability of T lymphocyte proliferation and activation was measured by 3H-Thydimine incorporation. 3. MSCs from human bone marrow cells were isolated and cultured.The purity of MSCs were identified with morphological characterization by microphotograph and the phenotypes were tested by flow cytometry (FCM). 4.MSCs were planted in 6-well plates (2×104/well for group B,4×104/well for group C and 8×104/well for group D) and cocultured for 72 hours with T cells activated as the control group (group A). CD3+CD4+, CD3+CD8+, CD4+CD25+ and CD4+CD152+ expressed on T cells were analyzed by FCM after coculture with MSCs for 72 hours respectively.T lymphocyte proliferation after cocultured with MSCs was evaluated by 3H-Thydimine incorporation. The expressions of IL-2, sIL-2R and TGF-β1 proteins in these four groups was detected by ELISA. Results1. The ability of T lymphocyte proliferation in the same PHA concentration increased with the time changing. Its ability was strongest when culturing for 48 hours (P<0.01)and had no difference between 48 hours and 72 hours (P>0.05). 2. The phenotypes of MSCs were measured by FCM:they were positive on the expression of CD44, CD105, CD29 and FIK1 and negtive on the expression of CD33, CD34, CD45 and HLA-DR. 3. MSCs inhibited T lymphocyte proliferation and the inhibitory effect depended on the amount of MSCs:CD3+CD8+, CD4+CD25+ and CD4+CD152+ T cells cocultured with bone marrow MSCs (group B, group C, group D) increased obviously and a significant decrease of CD3+CD4+ T cell proliferation as compared with control group (T lymphocytes uncocultured with MSCs) (P<0.01), and there were no difference between group C and group D (P>0.05). In group B, C and D, IL-2 and sIL-2R levels were lower than control group (P<0.01) and TGF-β1 level was higher obviously than group A (P<0.01). Every of IL-2, sIL-2R and TGF-β1 expressions has difference during group B, C and D (P<0.05). Conclusions1. Mitogen PHA could increase proliferation function of T lymphocytes and this function was strongest when cultured for 48 hours in same concentration of PHA. 2. MSCs inhibit T lymphocyte proliferation and perform their immunosuppressive function by up-regulation of CD3+CD8+ T cells, CD4+CD25+ Tcells and CD4+CD152+ Tcells. MSCs decreased IL-2 and sIL-2R levels and increased TGF-β1 level in PHA-stimulated peripheral blood lymphocytes. 3. MSCs play a critical immune negative regulatory effect on preventing GVHD. MSCs pretreatment may be useful in the prevention of GVHD in allo-HSCT and provides a new strategy to induce transplantation tolerance. Disclosures:No relevant conflicts of interest to declare.

Constitutional chromosomal aberration, t(4;12) (q23;p11), in a patient with Ph1 positive chronic myelocytic leukemia

A 54-year-old man with Ph1 positive chronic myelocytic leukemia and two balanced reciprocal translocations in bone marrow cells is described. One was the typical translocation between chromosomes No. 9 and 22, t(9;22), while the other was a constitutional translocation between chromosomes No. 4 and 12, t(4;12)(q23;p11), which was also present in all PHA-stimulated peripheral blood lymphocytes. Two brothers, one sister, one son, and one daughter also had the same constitutional translocation between chromosomes No. 4 and 12. None of them had signs of hematological disorder. The follow-up investigation of the family may clarify whether this constitutional translocation involves increased risk of developing CML.

Timing of apoptosis onset depends on cell cycle progression in peripheral blood lymphocytes and lymphocytic leukemia cells

Apoptosis results in cell death within 10 min after initiation by Bcl-2 family proteins and mitochondria; however, cells enter the apoptotic pathway at different elapsed times after being triggered. Intrinsic factors related to chemical or physical cell damage can initiate apoptosis at a specific cell cycle phase; it is not clear whether cells insulted via an extrinsic pathway also die at a specific cell cycle phase, or how apoptosis is related to cell cycle progression in cells. To illustrate the kinetic changes of apoptosis during cell cycle progression, we examined both intrinsically and extrinsically induced apoptosis in MOLT-4 and Jurkat lymphocytic leukemia cells and in cultured peripheral blood lymphocytes (PBLs) using a recently modified annexin V and propidium iodide method, which detects cell cycle-specific apoptosis. Apoptosis predominantly occurred at a specific cell cycle phase. Leukemia cells were sensitive to induction by both intrinsic (X-rays, UV light, camptothecin, arsenic trioxide, and the traditional Chinese medicine Jinke, which is an extract of Auricularia auricula) and extrinsic factors (via Fas and TNF receptor pathways). The phase at which leukemia cells entered apoptosis depended on the nature of the insult (X-ray or UV, G1-phase; camptothecin, S-phase; arsenic, G1/S phases; Jinke, G1/S phases; and TNF or Fas ligand, G1/S phases), whereas PBLs did not exhibit such insult-dependent differences. PHA-stimulated PBLs entered apoptosis, and additional cells were recruited following additional insults. Unstimulated PBLs remained unresponsive to apoptosis, and proliferating cells became insensitive to the insults after the cell cycle checkpoint was abolished by caffeine. Confluent or starving PBLs were also unresponsive to apoptotic triggers. Thus, apoptotic cell death is a cell cycle event with most, if not all, apoptosis being initiated during a particular cell cycle phase, and changes in the cell cycle result in changes in the apoptotic pattern and schedule. The coordination of apoptosis and proliferation in cells offers a mechanism for the integration of both cell cycle and apoptotic signals.

CDNA analysis of gene expression associated with DNA-dependent protein kinase activity.

DNA-dependent protein kinase (DNA-PK) is thought to play a pivotal role in DNA double-strand break repair. We recently demonstrated the association of DNA-PK activity in peripheral blood lymphocytes (PBL) with the incidence of chromosomal aberrations and the risk of cancer. In this study, we applied cDNA array technology to find the expression of genes which are associated with DNA-PK activity in PBLs with various levels of DNA-PK activity. Most genes correlated with DNA-PK activity involved cell cycle regulation. Moreover, the transcription factor E2F1, which plays an important role in cell cycle progression, exhibited strong correlation with the DNA-PK activity and Rbp130, which is considered a negative regulator of E2F, showed inverse correlation with DNA-PK activity. In silico promoter analyses showed the presence of at least one E2F binding site in the promoter regions of Ku70, Ku86, DNA-PKcs and genes associated with DNA-PK activity. In order to examine the relationship among the E2F1 expression, the expression of genes related with DNA-PK activity, and DNA-PK activity, we activated PMLs by PHA to progress the cell cycle. After PHA activation of PML, the expression of E2F1 and DNA-PK activity increased. The expression of most genes in PHA-stimulated PBLs had a similar relationship with DNA-PK activity to that without PHA stimulation. These results indicate that the E2F transcription factor may regulate the concerted expression of genes related with DNA-PK activity.

Oncolytic Measles Virus Selectively Targets CD46 Overexpression on Myeloma Cells.

Multiple myeloma remains incurable and novel therapeutics are urgently needed. We have shown that live attenuated Edmonston strain measles virus (MV-Edm) is potently oncolytic in multiple myeloma xenograft models and can efficiently infect CD138 sorted primary myeloma cells inducing extensive intercellular fusion which leads to cell death (Blood, 2001, v98, p2002). In contrast, the virus causes minimal damage to normal PHA-stimulated peripheral blood lymphocytes, fibroblasts and smooth muscle cells. To facilitate non-invasive imaging of sites of viral gene expression, we introduced the human thyroidal sodium iodide symporter (NIS) gene into the viral genome (MV-NIS). MV-NIS infected myeloma cells efficiently concentrate radioiodine which provides a basis for noninvasive imaging and destruction of infected tumors using beta emitters such as 131I (Blood, 2004, v103, p1641). Clinical translation of MV-NIS in patients with refractory myeloma is planned. However, the mechanism underlying the tumor selectivity of MV-Edm has not previously been defined. MV-Edm uses either of two cellular receptors for cell entry and cell fusion; CD46, a regulator of complement activation, is expressed on all nucleated cells, wheras SLAM (signaling lymphocyte activation molecule) is expressed only on activated B lymphocytes, T lymphocytes, and monocytes, and is not present on myeloma plasma cells. Using a panel of CHO cell transfectants, we demonstrated that MV-Edm -mediated cell killing increases exponentially above a threshold CD46 receptor density (Can Res, 2004, v64, p4919). We subsequently obtained bone marrow aspirates from 18 myeloma patients and performed multi-parameter flow cytometry to determine the relative expression levels of CD46 on the myeloma plasma cells versus the rest of the hematopoietic cells in the bone marrow. The myeloma cells expressed significantly higher levels (5-fold) of CD46 receptors (MFI=294.9 + 204.4, means ± SD) on their cell surfaces compared to hematopoietic cells of the erythroid, myeloid and lymphoid lineages (MFI=58.6 ± 9.0). Analysis of the myeloma array data generated by the Shaughnessy lab indicated that, in comparison to plasma cells from healthy volunteers (2997 ± 1053, n=30, means ± SD), primary myeloma cells express 2-fold elevated levels of CD46 mRNA (6592 ± 4029, n=72, p=0.001, Wilcoxon signed rank test). Taken together, these findings provide strong support for the hypothesis that selective destruction of myeloma cells by oncolytic measles viruses is a direct consequence of their overexpression of the cellular receptor CD46.

DNA repair capacity after γ-irradiation and expression profiles of DNA repair genes in resting and proliferating human peripheral blood lymphocytes

DNA repair plays an important role in maintaining genomic integrity, and deficiencies in repair function are known to promote cancer development. Several studies have used the individual capacity to repair DNA damage in peripheral blood lymphocytes (PBLs) as a cancer risk marker. As the cell’s ability to remove DNA damage may be correlated with proliferative activity, it is an important question whether quiescent or dividing cells should be used in such studies. The aim of our study was to compare DNA repair capacity and expression profiles of 70 known DNA repair genes, both in resting and phytohemagglutinin (PHA) stimulated human PBLs. Using the comet assay, γ-radiation-induced DNA damage and repair in lymphocytes was analyzed. No difference, neither in the rate of radiation-induced DNA damage nor in DNA repair capacity between PHA-stimulated and non-stimulated PBLs was observed. Stimulated cells, however, showed significantly elevated values for background damage. Transcriptional profiles of repair genes were analyzed using cDNA arrays. Hybridization experiments were performed with mRNA isolated from both unstimulated and PHA-stimulated PBLs. More than 70% of all evaluated genes had constant expression levels. Twelve genes responded with a more than two-fold increase of transcripts to the mitogenic stimulus. Most of the up-regulated repair enzymes are also known to play a role in DNA replication. In conclusion, the data presented here suggest that all repair proteins needed for the repair of γ-irradiation induced DNA-damage, that can be detected by the alkaline comet assay, are already present in G 0 cells at sufficient amounts and do not need to be induced once lymphocytes are stimulated to start cycling. Our results thus do not support a general increase in DNA repair activity of PBLs by PHA stimulation, and the use of stimulated PBLs in molecular epidemiological studies on DNA repair of γ-irradiation induced DNA damage seems not to be mandatory.

Replication status in leukocytes of treated and untreated patients with polycythemia vera and essential thrombocytosis

The replication status of malignant cells is usually asynchronous. However, to date the pattern of replication has not been studied in myeloproliferative disorders nor has the effect of chemotherapy been systematically evaluated. Therefore, we used fluorescence in situ hybridization to interphase nuclei in PHA-stimulated peripheral blood lymphocytes to examine replication timing of three alleles associated with the malignant process. The study group comprised hydroxyurea treated and untreated patients with essential thrombocytosis (ET) or polycythemia vera (PV). A significantly higher rate of the asynchronous pattern of replication in both treated and untreated patients was found as compared to healthy controls. The highest rate of asynchronous replication was observed in untreated patients. Also, the frequency of the two doublets pattern was significantly higher in the untreated group compared to the treated patients and to the control groups. In conclusion, patients with PV and ET have a higher rate of asynchronous pattern of replication. A possible correlation between disease activity and the pattern of replication is suggested. The effect of hydroxyurea on the pattern of replication is variable.

Systemic therapy of myeloma xenografts by an attenuated measles virus

Conditionally replicating viruses are promising agents for the treatment of malignancy. Here it is shown that the live attenuated Edmonston-B vaccine strain of measles virus (MV-Edm) replicates selectively in human myeloma cells and has potent antitumor activity. In vitro, replication of MV-Edm was restricted in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocytes (PBLs) but proceeded efficiently in a panel of 6 myeloma cell lines-ARH-77, RPMI 8226, JJN-3, MM1, KAS-6/1, and KMS-11-and in primary myeloma cells isolated by CD138 sorting from the bone marrow aspirates of 6 patients. MV-Edm infection induced potent cytopathic effects in these myeloma cells, resulting in the formation of multinucleated syncytia that eventually became nonviable. In contrast, syncytial formation in PHA-stimulated PBLs was minimal after MV-Edm infection. In vivo, MV-Edm was antitumorigenic and inhibited the establishment of myeloma cells as xenografts in immunocompromised mice. When injected directly into ARH-77 myeloma xenografts in the mice, MV-Edm caused complete regression of these xenografts. MV-Edm administered intravenously into the tail veins of mice also showed significant antineoplastic activity against established RPMI 8226 and ARH-77 xenografts. In particular, the ARH-77 myeloma xenografts were exquisitely sensitive to MV-Edm therapy, and tumors in all mice regressed completely. In light of its selectivity for myeloma cells and its potent antineoplastic activity against myeloma xenografts in vivo, MV-Edm merits further development for the treatment of multiple myeloma.

Restricted IgG1 subclass of anti-Yo antibodies in paraneoplastic cerebellar degeneration

Paraneoplastic cerebellar degeneration (PCD) occurs as a non-metastatic manifestation of cancer in a small proportion of patients with certain breast or gynaecological tumours, and is characterised by widespread Purkinje cell loss. Antibodies against a Purkinje cell cytoplasmic antigen, called Yo, that is expressed by the tumours, are present in the majority of these patients, but the pathogenic role of the antibodies is not clear. To characterise further the immune response in these cases, 13 anti-Yo positive sera were tested for IgG subclasses by immunohistochemistry and western blotting and, in four cases, PHA-stimulated cytokine secretion by peripheral blood lymphocytes was measured. Surprisingly, anti-Yo antibodies were entirely restricted to the IgG1 subclass, whereas antibodies against the small cell cancer-associated antigen, Hu, were found in all four IgG subclasses. There was a trend towards raised IgG1 levels in the total IgG of the anti-Yo positive patients and, in two, PHA-stimulated peripheral blood lymphocytes secreted raised levels of IFN-γ. By contrast, in the other two cases tested, raised levels of IL-4 were secreted. Patients with PCD associated with anti-Yo antibodies appear to have strong immune responses that are polarised with respect to the IgG subclass and Th cytokine profiles.

Modification in the inherent mode of allelic replication in lymphocytes of patients suffering from renal cell carcinoma: A novel genetic alteration associated with malignancy

Using fluorescence in situ hybridization (FISH) to interphase nuclei, we examined the replication timing of 1 allele relative to its counterpart in PHA-stimulated peripheral blood lymphocytes of normal subjects and patients suffering from a solid tumor (renal cell carcinoma). In the FISH assay, an unreplicated DNA sequence is identified by a single dot-like hybridization signal, whereas a replicated region gives rise to a duplicated, bipartite signal. Accordingly, lymphocytes of normal individuals show 2 patterns of allelic replication: (i) synchronized replication of allelic counterparts, as exemplified by the biallelically expressed loci TP53 and D21S55; and (ii) non-synchronized replication of allelic partners, as exemplified by the early and late replicating alleles of GABRB3, an imprinted locus subjected to monoallelic expression. However, when present in lymphocytes of the cancer patients, all 3 loci change their replication mode: alleles of TP53 and D21S55 become asynchronous, whereas the early replicating allele of GABRB3 delays replication, leading to relaxation in the imprinted mode of replication. Based on the tight relationship between temporal order of allelic replication and allelic mode of expression, the modified order of allelic replication observed in nonmalignant cells of individuals diagnosed with cancer represents a novel genetic alteration associated with malignancy. This alteration detected by simple cytogenetic means, applied to peripheral blood lymphocytes, offers a potential test for cancer identification. Genes Chromosomes Cancer 27:270-277, 2000.

Effect of parathyroid adenoma excision on interleukin-6 (IL-6) and IL-2 receptor levels

1,25-Dihydroxycholecalciferol [1,25(OH)2D], besides its role in calcium and phosphorus homeostasis, is also an important immunoregulatory molecule. Plasma levels of this hormone may be normal or elevated in patients with primary hyperparathyroidism. 1,25(OH)2D has been reported to inhibit production of the cytokines interleukin-2 (IL-2) and IL-6. In the present study, we examined the effect of parathyroid adenoma excision on serum IL-2 receptor (IL-2R) levels and the release and production of IL-2R and IL-6 by peripheral blood lymphocytes (each measurement was performed twice). Ten patients (5 females and 5 males aged 45 to 78 years) with primary hyperparathyroidism were enrolled in the study. The diagnosis of primary hyperparathyroidism was based on the presence of asymptomatic hypercalcemia, hypophosphatemia, and elevated serum intact PTH levels. Three weeks after removal of the parathyroid adenoma, there was a significant increase in the serum level of IL-2R, as well as the PHA-stimulated peripheral blood lymphocyte production of IL-6 and release of IL-2R. The results indicate that the removal of a parathyroid adenoma in patients with primary hyperparathyroidism causes a significant increase in IL-2R and IL-6 levels. The mechanism by which hyperparathyroidism may affect these cytokines and how they seem related to the levels of vitamin D is discussed.

The impact of the new fish technologies on the cytogenetics of haematological malignancies.

The impact of the new fish technologies on the cytogenetics of haematological malignancies.

Estrogenic xenobiotics affect the intracellular activation signal in mitogen-induced human peripheral blood lymphocytes : immunotoxicological impact

The present study was an attempt to elucidate the effect of estrogenic xenobiotics on the proliferation of mitogen-stimulated human peripheral blood lymphocyte (PBL). Our findings follow : (a) the proliferation of PBL in response to phytohemagglutinin (PHA) was mediated by protein kinase C activity, but estrogenic xenobiotics had a strong inhibitory effect on protein kinase C activity of PHA-stimulated PBL ; (b) cytoplasmic extracts from PHA-stimulated PBL greatly activated DNA replication, but estrogenic xenobiotics had a strong inhibitory effect on these activities. The results suggest that the cytoplasmic signal-generating system in mitogen-treated PBL is inhibited by estrogenic xenobiotics, and that the defect occurs at all stages in the sequence of events leading to DNA synthesis and cell proliferation.

Improved DNA-repair parameters in PHA-stimulated peripheral blood lymphocytes of human subjects with low body mass index

Clinically healthy subjects of the Indian population were divided into three age groups: young, 8–14 years; adult, 20–35 years; old, ≥55 years and were further classified based on body mass index (BMI) as normal BMI (NBMI) ≥20 and low BMI (LBMI) between 16 and 18, respectively. The ability of the peripheral blood lymphocytes from these subjects to respond to PHA stimulation in vitro and DNA-repair parameters, thereafter as a function of BMI and aging, were studied. The DNA-repair markers like unscheduled DNA synthesis (UDS), activities of DNA polymerase β and of two endodeoxy-ribonucleases, (UV- and AP-DNases) were assessed under different conditions. The LBMI group, considered to be going through chronic but mild undernutrition, showed higher repair capacity and exhibited no appreciable age-dependent decline in DNA-repair potential as was seen in normal subjects. These results correlate well with those seen in unstimulated human lymphocytes and also confirm the observations made earlier in experimental animals, where dietary restriction was shown to have beneficial effects on DNA-repair capacity.

Sex hormones affect the intracellular activation signal in mitogen-stimulated human blood lymphocytes

The present study was an attempt to elucidate the effect of sex steroids (estradiol-17β and 5α-dihydrotestosterone) on the mitogen-stimulated human peripheral blood lymphocytes (PBL) proliferation. Our findings were as follows: (a) the proliferation of PBL in response to phytohemagglutinin (PHA) was mediated by protein kinase C (PKC) activity, but a strong inhibitory effect of sex steroids on the PKC activity of PHA-stimulated PBL was observed; (b) cytoplasmic extracts of PHA-stimulated PBL showed high activity as an activator of DNA replication (ADR) as well as increased levels of the mitosis stimulator p34cdc2 kinase, and the steroid hormones studied all significantly reduced these activities; and (c) these effects of sex steroids are similar to those found with hydrocortisone in parallel experiments. The results suggest that the cytoplasmic signal-generating system stimulating DNA synthesis and mitosis is inhibited in PHA-stimulated PBLs by several steroid hormones. The inhibition affects different factors stimulating DNA replication and cell proliferation.

Effects of prenatal morphine exposure on behavioral responses to LPS in rats

Phytohemagglutinin (PHA) and concanavalin A (Con A)-induced blastogenesis of peripheral blood lymphocytes was examined in heat-stressed pre- and postpartal sheep. The peak responses of lymphocytes to PHA and Con A in heat-stressed sheep revealed significant reduction before and after parturition compared with those in the corresponding control animals kept under thermoneutral conditions. Furthermore, the effect of serum from control or heat-stressed sheep on PHA-induced lymphocyte blastogenesis was examined. Supplementation of serum from heat-stressed sheep significantly suppressed the blastogenesis of lymphocytes obtained from healthy sheep, bovine, and human donors. Unlike dexamethasone, heat-stressed sheep serum did not inhibit IL-2 production by PHA-stimulated human peripheral blood lymphocytes. These results indicate that the immunosuppression of heat-stressed sheep is in part mediated by serum factor(s) that can modulate T-cell function in a species nonspecific manner.