Mononuclear Cells Of Healthy Subjects Research Articles

Effects of vitamin K2 combined with methotrexate against mitogen-activated peripheral blood mononuclear cells of healthy subjects and rheumatoid arthritis patients.

Methotrexate (MTX) is used as anchor drug for patients with early and established rheumatoid arthritis (RA). Vitamin K2 administration was also reported to be associated with decreased disease activity in RA. Immunosuppressive pharmacodynamics of vitamin K2 combined with MTX was investigated. Mitogen-activated peripheral blood mononuclear cells (PBMCs) were used to evaluate immunosuppressive pharmacodynamics of drugs in vitro. Vitamin K2 alone dose-dependently suppressed T cell mitogen-activated proliferation of PBMCs of both healthy subjects and RA patients. 446.5 and 2232.5ng/mL vitamin K2 significantly decreased the IC50 values of MTX on the proliferation of PBMCs of RA patients, with little influences on the pharmacodynamics of MTX in the healthy PBMCs. 4465ng/mL vitamin K2 potentiated the pharmacodynamics of MTX in both RA patients and healthy PBMCs. The additional effects of vitamin K2 to potentiate the suppressive effects of MTX seemed not to be related to the regulation of CD4+ CD25+ T cells or CD4+ CD25+ Foxp3+ Treg cells. MTX alone at 100ng/mL significantly decreased the percentage of CD4+ T cells in PBMCs of healthy subjects (p<0.001) with a slight influence in that of RA patients (not significant) and the combination did not show synergistic inhibitory effect. Vitamin K2 alone tended to suppress the secretion of IL-17, IFN-γ, and TNF-α from the activated PBMCs of RA patients with smaller influences on the cytokine productions from healthy PBMCs. These additional effects of vitamin K2 were also observed in combination with MTX. The above information may partially elucidate the potentiation effects of vitamin K2 on the immunosuppressive efficacy of MTX.

Genetic variability of immune‐related lncRNAs: polymorphisms in LINC‐PINT and LY86‐AS1 are associated with pemphigus foliaceus susceptibility

Pemphigus foliaceus (PF) is an autoimmune blistering disease of the skin, clinically characterized by erosions and, histopathologically, by acantholysis. PF is endemic in the Brazilian Central-Western region. Numerous single nucleotide polymorphisms (SNPs) have been shown to affect the susceptibility for PF, including SNPs at long non-coding RNA (lncRNA) genes, which are known to participate in many physiological and pathogenic processes, such as autoimmunity. Here, we investigated whether the genetic variation of immune-related lncRNA genes affects the risk for endemic and sporadic forms of PF. We analysed 692 novel SNPs for PF from 135 immune-related lncRNA genes in 227 endemic PF patients and 194 controls. The SNPs were genotyped by Illumina microarray and analysed by applying logistic regression at additive model, with correction for sex and population structure. Six associated SNPs were also evaluated in an independent German cohort of 76 sporadic PF patients and 150 controls. Further, we measured the expression levels of two associated lncRNA genes (LINC-PINT and LY86-AS1) by quantitative PCR, stratified by genotypes, in peripheral blood mononuclear cells of healthy subjects. We found 27 SNPs in 11 lncRNA genes associated with endemic PF (p<.05 without overlapping with protein-coding genes). Among them, the LINC-PINT SNP rs10228040*A (OR=1.47, p=.012) was also associated with increased susceptibility for sporadic PF (OR=2.28, p=.002). Moreover, the A+ carriers of LY86-AS1*rs12192707 mark lowest LY86-AS1 RNA levels, which might be associated with a decreasing autoimmune response. Our results suggest a critical role of lncRNA variants in immunopathogenesis of both PF endemic and sporadic forms.

Absolute configuration of tetrandrine and isotetrandrine influences their anti-proliferation effects in human Tcells via different regulation of NF-κB.

Natural compound tetrandrine was reported to inhibit the proliferation of Tcells by inhibiting activation of NF-κB. Chemically, isotetrandrine differs from tetrandrine only in the stereochemistry at the chiral centers. The present study aimed to compare their anti-proliferation effects on human Tcells with a focus on NF-κB. The IC50 values of tetrandrine against MOLT-4cells, MOLT-4/DNR cells, and concanavalin A-activated peripheral blood mononuclear cells of healthy subjects and dialysis patients were 4.43±0.22, 3.62±0.22, 1.91±0.22 and 3.03±0.28μM, respectively. Whereas, the IC50 values of isotetrandrine against the above immune cells were 2.19±0.27, 2.28±0.33, 1.29±0.14 and 1.55±0.26μM, respectively. The inhibitory effect of isotetrandrine against the proliferation of Tcells was stronger than that of tetrandrine significantly (p<0.05). Molecular mechanism investigation showed that 10μM of isotetrandrine largely decreased the expression of p-NF-κB and NF-κB in both MOLT-4 and MOLT-4/DNR Tcells (p<0.05), whereas 10μM of tetrandrine slightly inhibited the phosphorylation of p-NF-κB with little influence on the expression of NF-κB. Taken together, absolute configurations of tetrandrine and isotetrandrine are suggested to influence on their anti-proliferation effects in human Tcells via different regulation of NF-κB.

Brief Report: Differential Effects of Tenofovir, Abacavir, Emtricitabine, and Darunavir on Telomerase Activity In Vitro.

In vitro, tenofovir and abacavir induced a significant dose-dependent inhibition of telomerase activity at therapeutic concentrations in peripheral blood mononuclear cells of healthy subjects. Median inhibition of telomerase activity by tenofovir at 0.5 and 1 μM was 29% [Interquartile range (IQR) 29%-34%, P = 0.042] and 28% (IQR 28%-41%, P = 0.042), respectively. Abacavir inhibition was 12% (IQR 9%-13%, P = 0.043) at 3 μM and 14% (IQR 10%-29%, P = 0.043) at 10 μM. Tenofovir and abacavir did not change human telomerase reverse transcriptase (hTERT) levels or mRNA levels of other telomerase complex genes. Exposure to emtricitabine or darunavir did not affect telomerase activity, hTERT protein levels, or mRNA levels of telomerase/shelterin genes.

Host Th1/Th2 immune response to Taenia solium cyst antigens in relation to cyst burden of neurocysticercosis.

Neurocysticercosis (NCC), Taenia solium larval infection of the brain, is an important cause of acquired seizures in endemic countries, which relate to number, location and degenerating cysts in the brain. Multicyst infections are common in endemic countries although single-cyst infection prevails in India. Single-cyst infections in an endemic country suggest a role for host immunity limiting the infection. This study examined ex vivo CD4(+) T cells and in vitro Th1 and Th2 cytokine responses to T.solium cyst antigens of peripheral blood mononuclear cells of healthy subjects from endemic and nonendemic regions and of single- and multicyst-infected patients for association with cyst burden of NCC. T.solium cyst antigens elicited a Th1 cytokine response in healthy subjects of T.solium-endemic and T.solium-non-endemic regions and those with single-cyst infections and a Th2 cytokine response from subjects with multicyst neurocysticercosis. Multicyst neurocysticercosis subjects also exhibited low levels of effector memory CD4(+) T cells. Th1 cytokine response of T.solium exposure and low infectious loads may aid in limiting cyst number. Th2 cytokines and low effector T cells may enable multiple-cyst infections to establish and persist.

Genes and miRNA expression signatures in peripheral blood mononuclear cells in healthy subjects and patients with metabolic syndrome after acute intake of extra virgin olive oil

Extra virgin olive oil (EVOO) consumption has been associated with reduced cardiovascular risk but molecular mechanisms underlying its beneficial effects are not fully understood. Here we aimed to identify genes and miRNAs expression changes mediated by acute high- and low-polyphenols EVOO intake. Pre- and post-challenge gene and miRNAs expression analysis was performed on the peripheral blood mononuclear cells (PBMCs) of 12 healthy subjects and 12 patients with metabolic syndrome (MS) by using microarray and RT-qPCR. In healthy subjects, acute intake of EVOO rich in polyphenols was able to ameliorate glycaemia and insulin sensitivity, and to modulate the transcription of genes and miRNAs involved in metabolism, inflammation and cancer, switching PBMCs to a less deleterious inflammatory phenotype; weaker effects were observed in patients with MS as well as in healthy subjects following low-polyphenol EVOO challenge. Concluding, our study shows that acute high-polyphenols EVOO intake is able to modify the transcriptome of PBMCs through the modulation of different pathways associated with the pathophysiology of cardio-metabolic disease and cancer. These beneficial effects are maximized in healthy subjects, and by the use of EVOO cultivars rich in polyphenols. Nutrigenomic changes induced by EVOO thus legitimate the well-known beneficial effects of EVOO in promoting human health and, potentially, preventing the onset of cardiovascular disease and cancer.

Suppressive efficacies of antimicrobial agents against human peripheral-blood mononuclear cells stimulated with T cell mitogen and bacterial superantigen

The immunomodulatory efficacies of 12 antimicrobial agents clinically used were examined against T cell mitogen- or bacterial superantigen-induced proliferation of peripheral-blood mononuclear cells of healthy subjects. Minocycline (CAS 13614-98-7), rifampicin (CAS 13292-46-1), trimethoprim (CAS 738-70-5), and ribavirin (CAS 36791-04-5) significantly inhibited the proliferation of mitogen-stimulated mononuclear cells at 1-100 microg/ ml (p < 0.001), whereas ofloxacin (CAS 82419-36-1) enhanced the proliferation (p < 0.001). Ampicillin (CAS 69- 53-4), lincomycin (CAS 859-18-7), vancomycin (CAS 1404-93-9), sulfamethoxazole (CAS 723-46-6), fosfomycin (CAS 26016-99-9), colistin (CAS 1264-72-8), and polymyxin B (CAS 1405-20-5) showed no significant effect. Minocycline, rifampicin, trimethoprim, and ribavirin also inhibited the proliferation of superantigen-stimulated peripheral-blood mononuclear cells at 1-100 microg/ml (<0.001), whereas ofloxacin stimulated the proliferation (p < 0.001). Rifampicin and minocycline at 10-100 microg/ml significantly inhibited interleukin-2 production from mitogen- or superantigen-stimulated peripheral-blood mononuclear cells (p < 0.025). These results suggest that certain kinds of antimicrobial agents inhibited the proliferation of mitogen- and superantigen-stimulated human peripheral-blood mononuclear cells and suppressed interleukin-2 production from these cells. The ofloxacin effect is immunostimulative, while the drug did not influence the interleukin-2 production.

Benidipine increases the number and activity of in vitro human endothelial progenitor cells via NO-dependent pathway

Objective: Benidipine has been proved to exhibit protective effects on endothelial function. Enhanced number and activity of circulating endothelial progenitor cells (EPCs) are closely correlated with improved endothelial function. However, the effect of benidipine on EPCs and its possible mechanisms are unknown. Therefore, we investigated the effect of benidipine on the number and activity of in vitro human EPCs and its relationship with nitric oxide (NO) known to regulate EPCs. Methods: The peripheral blood mononuclear cells of healthy subjects were induced into EPCs.

Laminar shear stress-induced activation of Raf/MEK/ERK1/2 signaling pathway contributes to upregulation of t-PA expression in human endothelial progenitor cells

Objective: Shear stress has been proved to enhance the expression of tissue-type plasminogen activator (t-PA) in endothelial progenitor cells (EPCs), which may contribute to maintenance of vascular homeostasis. However, the underlying mechanism is not clear. Here, we investigated the possible role of mitogen-activated protein kinases (MAPK) and nuclear factor (NF-kB) signaling in shear stress-mediated upregulation of t-PA expression in endothelial progenitor cells (EPCs). Methods: The peripheral blood mononuclear cells of healthy subjects were induced into EPCs.

Laminar shear stress-induced activation of Raf/MEK/ERK1/2 signaling pathway contributes to upregulation of t-PA expression in human endothelial progenitor cells

BackgroundShear stress has been proved to enhance the expression of tissue-type plasminogen activator (t-PA) in endothelial progenitor cells (EPCs), which may contribute to maintenance of vascular homeostasis. However, the underlying...

An oat bran meal influences blood insulin levels and related gene sets in peripheral blood mononuclear cells of healthy subjects

The understanding of how fibre-rich meals regulate molecular events at a gene level is limited. This pilot study aimed to investigate changes in gene expression in peripheral blood mononuclear cells (PBMCs) from healthy subjects after consumption of an oat bran-rich meal. Fifteen subjects (8 men and 7 women, aged 20-28years) ingested meals with oat bran or a control meal after an overnight fast. Blood samples for analysis of postprandial glucose, insulin and triglyceride concentrations were taken during 3h, while PBMCs for microarray gene expression profiling from five men and five women were taken before and 2h after the meal. Analysis of transcriptome data was performed with linear mixed models to determine differentially expressed genes in response either to meal intake or meal content, and enrichment analysis was used to identify functional gene sets responding to meal intake and specifically to oat bran intake. Meal intake as such affected gene expression for genes mainly involved in metabolic stress; indicating increased inflammation due to the switch from fasting to fed state. The oat bran meal affected gene sets associated with a lower insulin level, compared with the control meal. The gene sets included genes involved in insulin secretion and β-cell development, but also protein synthesis and genes related to cancer diseases. The oat bran meal also significantly lowered postprandial blood insulin IAUC compared to control. Further studies are needed to compare these acute effects with the long-term health effects of oat bran.

Abstract 2148: Thalidomide and liposomal doxorubicin inhibit differentiation and function of human osteoclasts

Abstract Background: Thalidomide inhibits angiogenesis and growth of multiple myeloma. Liposomal doxorubicin is mainly absorbed and degraded by monocyte-macrophage lineage in vivo, implicating a possible targeting effect. Given that osteoclasts (OCs) are derived from precursors of monocyte-macrophage lineage, we hypothesized that thalidomide, liposomal doxorubicin and their combination may affect osteoclastogenesis. Materials and Methods: We isolated CD14+ monocytes from peripheral blood mononuclear cells of healthy subjects and generated OCs under stimulation with macrophage-colony forming factor and receptor activator of NK-κB ligand. Cell viability, surface CD51/61 expression, and tartrate-resistant acid phosphatase (TRAP) activity were assessed by using MTT, flow cytometry and immunohistochemical staining, respectively. Bone resorption activity of OCs was examined. Results: The combination of thalidomide (purchased from TTY Biopharm, Taiwan) and liposomal doxorubicin (purchased from TTY Biopharm, Taiwan) profoundly inhibited the amount of harvested viable OCs. The expression of osteoclast-specific surface antigen CD51/61 was markedly inhibited by each drug and their combination. Specifically, the amount of multinucleated TRAP-positive cells characteristics of OCs and bone resorption activity of OCs were suppressed by each drug and, the mostly, their combination. Conclusion: Thalidomide, liposomal doxorubicin and their combination, beyond currently clinical indications, might be effective regimen to inhibit human osteoclastogenesis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2148. doi:10.1158/1538-7445.AM2011-2148

Hydrochlorothiazide increases interleukin-1 beta (IL-1β) secretion by peripheral blood mononuclear cells in healthy subjects

Recent study shows close relationship between hypertension and inflammation. The concentration of inflammatory mediators is increased in patients with essential hypertension. Angiotensin II (Ang II) may contribute to inflammatory process. Previous studies showed that individuals with essential hypertension had increased interleukin-1beta (IL-1β) secretion by peripheral blood mononuclear cells (PBMCs) and also valsartan and simvastatin reduced this inflammatory marker. In this study, the effect of hydrochlorothiazide on IL-1β secretion by PBMCs in healthy subjects was investigated. PBMCs in healthy subjects were isolated by gradient centrifugation. After incubation with Ang II and hydrochlorothiazide, IL-1 β concentrations in supernatant from PBMCs were measured by enzymelinked immunosorbent assay (ELISA). When compared with the control group, hydrochlorothiazide (10

Abstract 1661: Effect of thalidomide and liposomal doxorubicin on human osteoclastogenesis

Abstract Background: Thalidomide has been reported capable of inhibiting angiogenesis and growth of multiple myeloma. Liposomal doxorubicin is known to be mainly absorbed and degraded in reticuloendothelial system, especially the monocyte-macrophage lineage. Since osteoclasts (OCs) are derived from precursors of monocyte-macrophage lineage, we aimed to examine the effect of thalidomide, liposomal doxorubicin and their combination on human osteoclastogenesis. Materials and Methods: We isolated CD14+ cells from peripheral blood mononuclear cells of healthy subjects and generated OCs under stimulation with macrophage-colony forming factor and receptor activator of NK-κB ligand. Cell viability, surface CD51/61 expression, and tartrate-resistant acid phosphatase (TRAP) activity were assessed by using MTT, flow cytometry and immunohistochemical staining, respectively. Results: In comparison with control differentiated OCs, combination of thalidomide (purchased from TTY Biopharm, Taiwan) and liposomal doxorubicin (purchased from TTY Biopharm, Taiwan) profoundly inhibited the amount of harvested viable OCs. By treatment with each drug alone, thalidomide increased and liposomal doxorubicin decreased the amount of viable OCs. The expression of osteoclast-specific surface antigen CD51/61 was greatly inhibited by each drug and their combination. Furthermore, the amount of multinucleated TRAP-positive cells characteristics of OCs, was suppressed by each drug and, the mostly, their combination. Elucidation of cellular and molecular mechanisms of action has been undergoing. Conclusion: Thalidomide, liposomal doxorubicin and their combination may effectively inhibit human osteoclastogenesis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1661.

Bromohydrin pyrophosphate enhances antibody-dependent cell-mediated cytotoxicity induced by therapeutic antibodies

In human blood, 1% to 5% of lymphocytes are gammadelta T cells; they mostly express the gammadelta T-cell receptor (TCR)Vgamma9, recognize nonpeptide phosphoantigens (PAgs) produced by microbes and tumor cells, and mediate different modes of lytic activities directed against tumor target cells. Antibody-dependent cell-mediated cytotoxicity (ADCC) mediated by cytolytic lymphoid cells is essential for the clinical activity of anticancer monoclonal antibodies (mAbs), but whether PAgs affect ADCC by gammadelta T cells is unknown. Here we report that, in association with the CD20(+)-specific mAb rituximab (RTX), the synthetic PAg bromohydrin pyrophosphate (BrHPP) increased TCRVgamma9(+) cell binding to CD20(+) lymphoma cells in vitro. This combination activated phospho-ZAP70 and phospho-ERK1/2 signaling in TCRVgamma9(+) cells and strongly enhanced their ADCC activity. We obtained similar results with BrHPP in the context of the mAbs alemtuzumab and trastuzumab. Furthermore, BrHPP enhanced RTX-mediated depletion of CD20(+) cells in vitro from peripheral blood mononuclear cells of healthy subjects and enhanced ADCC by gammadelta T cells from patients with chronic lymphocytic leukemia. In cynomolgus macaques, a regimen combining RTX, BrHPP, and IL2 activated TCRVgamma9(+) lymphocytes and enhanced B-cell depletion from blood and lymph nodes. Thus, the combination with BrHPP PAg is able to improve the efficacy of cancer immunotherapy by therapeutic mAbs.

Drug Metabolizing Enzymes and Transporters mRNA in Peripheral Blood Mononuclear Cells of Healthy Subjects: Biological Variations and Importance of Preanalytical Steps

Quantification in peripheral blood mononuclear cells of mRNA of drug metabolizing enzymes or drug targets could give interesting, new information in the field of pharmacogenomics and molecular mechanisms. However, for the interpretation of these data, it is necessary to know mRNA biological variations. In this review, we propose a strategy based on the production and interpretation of clinical chemistry reference values. We discuss the concept of reference values; the necessity to master pre-analytical variations of CYP and ABC transporters; the choice of the analytical methods and of the reference genes; and finally the biological variations themselves. In particular, we focus on the importance of considering homogeneity for age, sex, degree of adiposity, tobacco and alcohol intake, food habits, and drug consumption, including their inductive effects, at the phase of subject recruitment. All this information is useful to define the partition and exclusion factors to obtain mRNA reference limits.

The Effect of Repeated Exposures to Low-Dose UV Radiation on the Apoptosis of Peripheral Blood Mononuclear Cells

To investigate whether repeated exposures to low-dose UV or solar-simulated radiation induce apoptosis of peripheral blood mononuclear cells. Cohort study in a healthy population. Departments of Dermatology and Hematology, Medical University of Lodz, Lodz, Poland. Ninety-eight healthy volunteers were divided into the following 4 groups: group A, whole-body irradiated with a 0.7-minimal erythema dose (MED) of UV-B daily on 10 consecutive days followed by a single dose of 3 MEDs on a small body area 24 hours later; group B, whole body irradiated with 120 J/m(2) solar-simulated radiation (10 consecutive days, then single-dose UV-B exposure of 3 MEDs on a small body area after 24 hours); group C, irradiated with a single UV-B dose of 3 MEDs on a small body area; and group D, irradiated with a single UV-B dose of 4 MEDs on a small body area. Apoptosis of peripheral blood mononuclear cells as well as expression of several apoptosis-regulating proteins in response to irradiation. Ten daily whole-body suberythemal exposures to UV-B or solar-simulated radiation enhanced the apoptosis of peripheral blood mononuclear cells, while the single erythemal doses on the small body area, either on their own or following the repeated exposures, did not increase the level significantly. Increase in Bax and p73 protein expression and down-regulation of Mcl-1 and Bcl-2 correlated with enhanced apoptosis. Repeated suberythemal UV exposures enhance the apoptosis of peripheral blood mononuclear cells in healthy subjects.

Effect of Bifidobacterium bifidum DSM 20082 Cytoplasmic Fraction on Human Immune Cells

The objective of the present study was to determine the effect of the soluble cytoplasmic fraction from Bifidobacterium bifidum DSM 20082 (Bb) lysate on peripheral blood T cells. In peripheral blood mononuclear cells of healthy subjects, cytotoxic activity, proliferation, apoptosis, and up-regulation of CD8 or CD4 molecules in T cells were examined. When peripheral blood mononuclear cells were stimulated with Bb lysate, the main effect was observed in CD8+ cells as a significant increase of CD8 molecules in a dose-dependent manner, and this behavior was observed at 24, 48, and 72 h after stimulation; in contrast, stimulation with Bb lysate showed no effect on the up-regulation of CD4 molecules in T helper cells. Further Bb lysate did not induce proliferation activity in either CD8+ or CD4+ cells. Bb lysate induced activation of CD8+ cytotoxic activity against autologous monocytes. Around 80% of the cells stimulated with Bb lysate were positive to peanut agglutinin (PNA), suggesting that the stimulated CD8+ cells corresponded to activated/effector cellular populations. When apoptosis was determined, there were no differences between stimulated and non-stimulated cells. Our results indicate that Bb lysate is able to increase cytotoxic activity of peripheral CD8+ cells, without affecting lymphocyte survival.

Expression of the TRAIL receptors in blood mononuclear cells in leukemia

TRAIL receptors are differentially expressed on restricted subpopulations of normal blood cells. In the present study, we investigated the utility of individual TRAIL receptors in evaluating the presence of circulating tumor cells in blood. Patients with chronic myeloid leukemia (CML) carrying the t(9;22) translocation were compared with patients in whom no translocation was detected, with patients with multiple myeloma and with a group of healthy individuals. TRAIL receptor expression was analyzed by RT-PCR in blood mononuclear cells. Blood mononuclear cells of healthy subjects expressed the TRAIL-R1 and TRAIL-R2 death receptors and the TRAIL-R4 decoy receptor while the other decoy receptor TRAIL-R3 was not detectable. This normal expression pattern was also observed in all cases with multiple myeloma and in almost all patients without translocation (42/43; 97.7%). However, in 24/56 (42.9%) of the translocation-positive patients, the expression pattern was completely different. In this group the TRAIL-R4 receptor alone or in combination with TRAIL-R1 disappeared from blood mononuclear cells, while the TRAIL-R2 was expressed at normal level, indicating that the loss of expression is specific for the TRAIL-R4 and TRAIL-R1. This expression pattern was also confirmed by real-time PCR. The differences between the translocation-positive and -negative groups for the TRAIL-R4 and TRAIL-R1 expression were highly significant (p=0.0001 and p=0.0004, respectively). However, the differential expression pattern did not correlate with the number of leukemic cells. Our results suggest a correlation between the presence of leukemic cells in circulation and the differential expression pattern of TRAIL receptors in blood mononuclear cells.

Polyomavirus BK non‐coding control region rearrangements in health and disease

BK virus is an increasingly recognized pathogen in transplanted patients. DNA sequencing of this virus shows considerable genomic variability. To understand the clinical significance of rearrangements in the non-coding control region (NCCR) of BK virus (BKV), we report a meta-analysis of 507 sequences, including 40 sequences generated in our own laboratory, for associations between rearrangements and disease, tissue tropism, geographic origin, and viral genotype. NCCR rearrangements were less frequent in (a) asymptomatic BKV viruria compared to patients viral nephropathy (1.7% vs. 22.5%), and (b) viral genotype 1 compared to other genotypes (2.4% vs. 11.2%). Rearrangements were commoner in malignancy (78.6%), and Norwegians (45.7%), and less common in East Indians (0%), and Japanese (4.3%). A surprising number of rearranged sequences were reported from mononuclear cells of healthy subjects, whereas most plasma sequences were archetypal. This difference could not be related to potential recombinase activity in lymphocytes, as consensus recombination signal sequences could not be found in the NCCR region. NCCR rearrangements are neither required nor a sufficient condition to produce clinical disease. BKV nephropathy and hemorrhagic cystitis are not associated with any unique NCCR configuration or nucleotide sequence.