Although more experimental data are needed to

resolve thi

Although more experimental data are needed to

resolve this question, epidemiological data documenting resistance in HESN sex workers suggests that repeated mucosal exposure and the associated cell infiltrates do not result in higher infectivity, but rather a sustained resistance against HIV-1 infection [1]. Further research utilizing animal models of SIV mucosal exposure would be helpful in elucidating if pathogen-induced DC activation at the site of exposure is associated with recruitment of NK cell activity and protection from HIV-1 infection in spite of the recruitment of CD4+ T cell targets. Most anti-viral mechanisms are expected to act both at preventing infection during exposure and in reducing viral replication after infection. However, adaptive Proteasome inhibitor T cell responses may be more effective at control of viral replication after infection, as memory responses are probably amplified as CD8 T cell effectors only after infection is established. In contrast, NK cells remain an immune cell type associated with both resistance

to HIV-1 infection in HESN subjects and control over viral replication following infection. The case for the anti-viral capacity of NK subsets during infection is suggested by its loss of function in chronic infection. Progressive HIV disease is associated clearly with increasingly DNA Damage inhibitor impaired NK responses and the selective depletion of CD56dim Tobramycin NK cells during chronic HIV-1 infection [112–115]. The loss of CD56dim NK cells, the main circulating NK subset that mediates cytotoxicity, results in the enrichment of CD56null NK cells with decreased function [113,116–118]. HIV-1 replication also results in the altered expression of inhibitory and activating receptors on NK cells further impairing the lytic potential of the remaining NK pool [119–121]. Defects in the NK cell compartment have been hypothesized to

be part of the profound immunodeficiency observed during chronic HIV-1 infection and host susceptibility to opportunistic infections [122]. In contrast, NK frequency and IFN-γ production have been shown to be retained in HIV-1 long-term non-progressors [123]. HIV-1-infected elite controllers that suppress viral replication in the absence of anti-retroviral therapy also exhibit NK activity that is comparable to uninfected control donors [124]. Together, these results correlate an increasingly dysfunctional NK cell compartment after infection with loss in control over HIV-1 replication during chronic infection. Genetic studies of the KIR3DL1 locus in disease progression studies indicate that inheritance of KIR3DS1 and KIR3DL1high receptor alleles in conjunction with their HLA ligands can delay disease progression [87,125]. These genotypes are the same as those observed to be over-represented in a high-risk cohort of HESN i.v. drug users and HESN partners of HIV-1-infected subjects [17,28].

The overall score is the simple sum of the four symptom scores T

The overall score is the simple sum of the four symptom scores. Traditionally, a questionnaire has many items with the same minimum and maximum score (e.g. IPSS).27 However, with the OABSS, scales vary. For instance, the item “How often do you have a sudden desire to urinate, which is difficult to defer?” (urgency) ranges from 0 to 5. Scores for “How often do you leak urine because you cannot defer the sudden desire to urinate?” (urge incontinence) also range from 0 to 5. “How many times

do you typically wake up to urinate from sleeping at night until waking in the morning?” (nocturia) ranges from 0 to 3, while “How many times do you typically urinate from waking in the morning until sleeping?” (frequency) ranges from 0 to 2. Homma mentioned

that the relative weight among the four scores was determined on the basis of the maximal influence rate of the symptom in the epidemiologic survey.29 As Romidepsin concentration urgency is the core symptom of OAB, the design of OABSS is meant to show a clear separation between subjects with OAB and controls. One source of concern is that the OABSS was developed and validated using only Japanese patients. The authors did mention that cultural background may affect the psychometric properties of symptom questionnaires.28 Although different questionnaires are now available and validated for OAB, most of them are written in English. For non-English-speaking people, the questionnaires must be translated into the appropriate language. In 2006, Acquadro et al. translated the OABq into 14 languages.30 The process included six steps: (i) two forward translations; GS-1101 mouse (ii) comparison and reconciliation of the translations; (iii) back-translation; (iv) comparison of the source and back-translation; (v) review by one urologist or gynecologist; and (vi) a comprehension test, using patients. However, none of these versions was in traditional Chinese. In 2008, the president Tyrosine-protein kinase BLK of the Taiwan Continence Society (TCS), Professor Kuo, commenced linguistic validation and other elements of production of a Chinese version of the Homma OABSS. The process involved forward- and back-translation, and review by urologists and gynecologists

in expert meetings in Taiwan (hosted by Professor Kuo) and in Japan (hosted by Professor Homma). The validated OABSS in Traditional Chinese is now available (Appendix II) and posted on the official website of the TCS ( OAB is a symptom-based condition without physiological markers of disease activity. Appropriate tools are needed to assess patients with OAB. There is still no consensus for the evaluation of OAB. Patients may need to be assessed from different aspects, such as clinical symptoms, FVC, and multi-item questionnaires to obtain patient-reported outcomes to fully understand the condition in patients with OAB. On the other hand, a simple and effective symptom score is needed to meet the requirements of clinical work.

dublinienis isolates (r = 0 452; P = 0 046) However, the differe

dublinienis isolates (r = 0.452; P = 0.046). However, the difference in the effect elicited by nystatin on CSH did not have a positive relationship with the clampdown of adhesion to BEC (r = 0.127; P = 0.584)

and GT formation (r = 0.106; P = 0.658). C. dubliniensis is now well recognised as an opportunistic emerging pathogen associated with oral Doxorubicin mouse candidosis. Particular attention has been paid to studying candidal adhesion to BECs of the oral mucosa, as it is intimately associated with all forms of oral candidosis.[8, 9] In addition, GT, which marks the onset of hyphal growth, is a phenotypic characteristic associated with candidal adhesion. One reason for the pathogenic nature of C. dubliniensis may be its ability to transform from the blastospore or yeast phase to the mycelial or hyphal phase.[26] For instance, candidal hyphae are thigmotrophic Pirfenidone in nature and traverse along surface irregularities both in vivo and in vitro, thus helping in the

retention of the organism in hostile habitats such as the oral cavity.[12] In addition, the sheer physical size of the hyphal element poses a problem for the host phagocytic response.[11] Apart from the aforementioned biological phenotypic traits, the relative CSH of Candida is considered a non-biological physical force of critical importance pertaining to candidal adhesion. For instance, new Hazen and Hazen [27] have demonstrated that hydrophobic Candida are more virulent than their hydrophilic counterparts. Shibl et al., [28] and Ramadan et al., [29] have shown that the reduction in CSH following limited exposure to antimicrobials promoted increased ingestion of microbes by polymorphonuclear leukocyte (PMNL), thus increasing the susceptibility of the organisms to the killing effect of PMNL. Hydrophobic cells also exhibited greater adherence to epithelial cells and extracellular matrix proteins and decreased susceptibility to phagocytic killing.[30] In addition, it has been stated that

enhanced virulence of hydrophobic cells over hydrophilic cells may be due to the potential of hydrophobic cells to bind to various organs following clearance from the bloodstream.[30] Furthermore, to these adhesion-related traits, another form of measuring Candida virulence is with the PAFE, which measures the growth recovery capacity after a limited exposure to antifungal agents, where more virulent and resistant organisms will have low PAFE, whereas a susceptible and less virulent organism will have higher PAFEs.[18-20, 31] The PAFE, suppression of adhesion to BEC and almost complete abrogation of GT production by limited exposure to the polyene antifungal agent may be related to the mechanism of action of nystatin on the Candida cell wall. Polyenes bind to the sterol components in the cell wall of Candida and make it more permeable.

Finally, knowledge-driven gene expression-based predictors can be

Finally, knowledge-driven gene expression-based predictors can be translated into assays that are simpler and more robust than measurement of transcript abundance for many genes. Gene expression predictors have historically been limited by a lack of reproducibility between experiments [10, 25]. This is thought to be related to the high variance of individual gene measurements commonly seen in datasets of relatively few replicates. This variance results in discordance between lists of predictive genes even in high quality experiments. Using a larger set of genes rather than a small

number of genes may Trichostatin A provide some degree of robustness lacking in single gene level predictors. Indeed several platforms have now been developed [26, 27] that allow focused sets of genes to be profiled at high throughput and low cost. Moreover, because gene set based predictors Lumacaftor can identify not just predictive genes but predictive biological processes, this approach could overcome the limits of predicting clinical responses by measuring gene expression. For instance, our analysis shows that signatures associated with cellular proliferation are predictive of a protective antibody response. It would be relatively easy to translate

this to a flow-cytometry based assay of cellular proliferation in PBMCs using Ki67 staining, for example, that could rapidly be applied to many samples. In contrast, developing and validating a multigene predictive signature of unknown biological significance may prove to be more significantly more complex. Future studies will be required to determine how successfully biological processes discovered by gene set based approaches can

be deployed as simpler, more robust diagnostic tools. Gene set based predictors predicated on biological knowledge may therefore provide a sensitive, relevant, and robust analysis of the human immune response. We analyzed two existing datasets of gene expression profiles of PBMC Sorafenib in vivo from vaccinated subjects: raw Affymetrix array data for subjects vaccinated with YF-17D from Gene Expression Omnibus with the accession number GSE13486 [4], and raw Affymetrix array data from subjects vaccinated with influenza TIV with accession number GSE29619 [16]. The Genepattern module “CollapseDataset” was used to extract the expression values of genes from the raw data file and to map Affymetrix probes to gene symbols [28]. Then we applied quantile normalization and a log2 transformation. The final transformed data were used for the single sample GSEA projection (see below). For analysis of data from the influenza vaccinated subjects, gene expression fold change was calculated as the ratio of expression levels from PBMC profiles day 7 (postvaccine)/day 0 (prevaccine).

Irradiated splenocytes that were used as a source of APCs in our

Irradiated splenocytes that were used as a source of APCs in our experiments could

be treated with Ficoll–Hypaque and separated from the CD4+ T cells only after 1 day in cultures. In preparation for later experiments, Fig. 1(c) was included, showing that anergy could be demonstrated using beads instead of antigen to stimulate secondary cultures. In addition to proliferative unresponsiveness, Th1 cells stimulated with antigen in the presence of n-butyrate demonstrated a 37–77% decrease in IL-2 and a 26–55% decrease in interferon-γ secretion when stimulated in secondary culture with three different stimulation indices (Fig. 1d). Hence, n-butyrate-induced anergy Ku-0059436 concentration was demonstrated by a loss of both antigen-induced proliferation and cytokine production. It has

been reported previously that n-butyrate increased p21Cip1expression in antigen-stimulated Th1 cells.8 However, p21Cip1 is also induced in antigen-stimulated Th1 cells in the absence of n-butyrate. Consequently, the kinetics of p21Cip1 up-regulation was studied in antigen-stimulated Th1 cells in the presence and absence of n-butyrate during the 6-day primary cultures to compare the two groups for PD0332991 in vivo any possible difference in p21Cip1 expression. When antigen was added in the initiation of the primary culture (day 0), p21Cip1 was up-regulated in control Th1 cells by day 1, remained high on day 2, but decreased significantly by day 3 and was back to resting levels by day 5 (Fig. 2a). In contrast, when antigen was added on day 0 and n-butyrate was added on day 1, the p21Cip1 levels remained

elevated in anergic Th1 cells during the entire 6-day primary culture. p27Kip1 is another cdk inhibitor thought to play a role in T-cell anergy. As expected, p27Kip1 was high in resting Th1 cells. Its level decreased with the antigen stimulation and was later restored to resting levels in control Th1 cells by day 5 of the primary cultures. In contrast, p27Kip1 levels failed to be completely restored in Th1 cells incubated with antigen and n-butyrate in 6-day primary cultures (Fig. 2b). Hence, because p21Cip1 rather than p27Kip1 was high in the anergic Th1 cells at the end of the 6-day primary cultures, subsequent experiments OSBPL9 were focused on the role of p21Cip1 in maintaining proliferative unresponsiveness. The kinetics of other cell cycle proteins was also studied to assess their possible involvement in n-butyrate-induced T-cell anergy. No significant differences between the antigen-stimulated control and anergic Th1 cells were observed in the expression of cdk2, cdk4, cdk6, cyclin D2, cyclin D3 and cyclin E (Fig. 2b). In summary, the kinetics studies on cell cycle proteins revealed that the most detectable difference between anergic and control Th1 cells was the high level of p21Cip1 maintained throughout the primary cultures in the anergic Th1 cells. Localization of proteins such as p21Cip1 in the cell can have important functional consequences.

As shown in Fig  5, Flt3L gene expression was significantly incre

As shown in Fig. 5, Flt3L gene expression was significantly increased in MPPs from Fli-1∆CTA/∆CTA B6 mice compared with that cultured from wild-type B6 mice. The expressions of STAT3, Csf1 and Flt3

were higher in MPPS from Fli-1∆CTA/∆CTA B6 mice compared with that cultured from wild-type B6 mice, though the difference was not statistically significant (Fig. 5). To assess whether Fli-1 directly or indirectly regulates the expression of Flt3L, we analysed the promoter region of the Flt3L gene. There are 15 putative Fli-1 binding sites in the promoter region of the mouse Flt3L gene. We designed 15 pairs of primers to cover these sites, and a ChIP assay was performed to examine if Fli-1 binds to the promoter of Flt3L. The primers used are listed in Table 1. We examined the expression of Fli-1 and Flt3L in MS1 endothelial cell lines by RT-PCR and found that both Fli-1 and Flt3L are expressed in the cell line (data not shown). After immunoprecipitation by a Fli-1-specific antibody with cross-linked protein/DNA complexes from MS1 cell lines, two Fli-1 sites were significantly enriched with specific Fli-1 antibodies as detected by PCR amplification and compared with normal rabbit IgG controls (Fig. 6). These results clearly indicate Fli-1 can directly bind to the promoter of the Flt3L gene and probably regulate the expression of Flt3L. Fli-1 transcription factor regulates the differentiation

and development of haematopoietic lineages, especially megakaryocytic and erythrocytic lineages.[28-30] We previously demonstrated that Fli-1 modulates B-cell development and is implicated in autoimmune Y-27632 supplier disease.[22, 26, 27, 31] We report here that Fli-1 also plays an important role in mononuclear phagocyte

development. We found that Fli-1∆CTA/∆CTA mice had significantly increased populations of HSCs and CDPs in BM compared with wild-type littermates (Fig. 1). Therefore, Fli-1 is likely to play an important role in regulating HSC and CDP development. Expression of Fli-1 clearly affects the HSC population and lack of the CTA domain in Fli-1 resulted in the increase of the HSC population. Previous studies have demonstrated that expression of Fli-1 affects development and differentiation Montelukast Sodium of megakaryocytes, erythrocytes, neutrophils and monocytes in Fli-1-deficient or Fli-1 heterozygous mice.[28, 29] Complete Fli-1 deficiency in HSCs resulted in a decrease in neutrophilic granulocyte and monocyte populations in mice.[29] In this report, we used Fli-1ΔCTA/ΔCTA mice with expression of a truncated Fli-1 protein, lacking the C-terminal transcriptional activation domain.[24] Cell proportion and absolute cell number of pDCs, cDCs, pre-cDCs and macrophages in the spleen from Fli-1∆CTA/∆CTA mice were significantly increased when compared with wild-type littermates (Fig. 2). The splenic cDC population can be subdivided into three groups according to their surface markers.

2B) The altered response to anti-IgM could arise from a decrease

2B). The altered response to anti-IgM could arise from a decreased FO/MZ ratio, since BCR engagement causes proliferation of FO cells but apoptosis of MZ cells 14, 15. However, reduced anti-IgM-mediated proliferation was also observed in B cells from Foxo1f/fCd21Cre mice in which no changes in FO/MZ ratios were reported 10, and is consistent with the presence of a prominent IgM− B-cell population (Supporting Information Fig. 1C). Measuring live cell number using a metabolic dye conversion (MTS) assay confirmed the finding of impaired anti-IgM response in Foxo1f/fCd19Cre B cells (Supporting Information Fig. 2A). The LPS response in Foxo1f/fCd19Cre B cells was increased when measured using MTS assays (Supporting

buy Dabrafenib Information Fig. 2A), but not using the CFSE assay (Fig. 2A). This might indicate that LPS-stimulated B cells have altered metabolism when Foxo1 is absent, leading to increased MTS conversion despite equivalent cell number. TGF-β is a cytokine with potent anti-proliferative effects in lymphocytes 16. TGF-β signaling activates Smad transcription factors, which in several cellular systems cooperate with Foxo proteins to activate target promoters 17, 18. Furthermore,

the TGF-β/Smad signaling axis regulates MZ B-cell development 19. Although we obtained evidence for functional cooperation of Foxo1 and Smad transcription factors in B cells (Supporting Information Fig. 2B and C), Foxo1 was not required for TGF-β-mediated suppression of B-cell proliferation triggered by anti-IgM or LPS (Supporting Information Fig. 2A). CD62L mRNA was consistently reduced about threefold in Foxo1f/fCd19Cre B cells (Fig. 2C), indicating that lower CD62L protein expression on the surface of these cells is at least partly due to reduced steady-state mRNA levels, resulting from altered

transcription and/or RNA processing. Foxo1 also controls expression of the Sell gene encoding CD62L in T cells 20–22. Another Foxo target gene, Klf2, regulates CD62L expression in T cells and might be a link between Foxo1 and CD62L 20, 21, 23–25. Klf2 Tolmetin mRNA expression was also significantly reduced in Foxo1-deficient B cells, though less prominently than the reduction in Sell mRNA (Fig. 2C). Previously, we identified Ccng2, Rbl2 and Klf4 as Foxo target genes in B cells 26, 27. By various criteria, including reporter assays, electrophoretic mobility shift assays and chromatin immunoprecipitation, these genes were regulated similarly by Foxo1 and Foxo3a 26, 27. RNA measurements using quantitative real-time PCR showed that none of these genes were differentially expressed in Foxo1-deficient B cells (Fig. 2C), further suggesting that Foxo1 and Foxo3a have redundant functions at these target promoters. The increased population of MZ B cells in Foxo1f/fCd19Cre mice was intriguing, since Foxo factors are turned off by the PI3K/AKT pathway and the opposite phenotype occurs in mice lacking PI3K genes 28–30.

Our results show that HIV-specific

CD8+ T cells contribut

Our results show that HIV-specific

CD8+ T cells contribute significantly to IL-10 production in the peripheral blood and that this subset modulates monocyte activation. Constitutive IL-10 gene transcription was reported to be upregulated in multiple cell subsets among PBMCs in chronically HIV-infected individuals but there is uncertainty as to whether this is universally reflected in increased spontaneous or antigen-driven cytokine production [7]. We therefore analysed the Obeticholic Acid cell line fractions of IL-10-producing cells among circulating CD4+ T cells, CD8+ T cells, CD19+ B cells and CD14+ monocytes ex vivo, after stimulation with either 0.05% DMSO or HIV-1 gag peptides, in three subject

groups: patients who were antiretroviral (ART) naïve (n = 31, median viral load – 17 964 copies/mL) or fully suppressed on ART for >1 year (n = 20) and HIV-uninfected healthy controls (n = 5). Study participants’ characteristics are described in Table 1. The gating strategy used to identify IL-10-producing cells is shown in Supporting Information Fig. 1. Constitutive IL-10 release (0.05% DMSO control) was detected in all cell subsets analysed; the proportion of IL-10-producing cells was highest among CD19+ B cells and CD14+ Caspases apoptosis monocytes in all three groups but there were no significant differences among the groups for each cell subset analysed, suggesting that constitutive IL-10 expression was not increased at the protein level in this patient cohort

(Supporting Information Fig. 2). By contrast, we observed significant IL-10 secretion in response to HIV-1 gag stimulation, predominantly in CD8+ T cells. These IL-10+ CD8+ T cells were rare but reproducibly detected in ART-naïve viraemic individuals, at a median frequency of 0.01% (range 0–0.13%, tenfold greater than the 0.05% DMSO control). Frequencies among ART-treated and uninfected most subjects were <0.001 and 0%, respectively (p < 0.01, Fig. 1A and B). Although the proportion of IL-10+ cells was lower among CD8+ T cells than monocytes, CD8+ T cells were the major contributors to IL-10 production in response to HIV-1 gag, due to the higher absolute numbers of CD8+ T cells in the peripheral blood (Fig. 1C). Phenotypic analysis of HIV-specific IL-10+ CD8+ T cells revealed that the majority were CD25- and FoxP3-negative and a substantial minority expressed CXCR3, a ligand for inflammatory chemokines that promotes migration to sites of inflammation and differentiation towards an effector phenotype [15] (Fig. 1D). We also investigated the expression of the gut-homing integrin alpha-4/beta-7 on HIV-specific IL-10+ CD8+ T cells, since IL-10 expression is upregulated in gut-associated lymphoid tissue (GALT) during acute HIV-1 infection [16].

However, renal biopsies have not revealed adequate information fo

However, renal biopsies have not revealed adequate information for predicting prognosis. Thus, this retrospective study was conducted of diabetic nephropathy to obtain prognostic information from histopathologic findings. Methods: The subjects were 28 diabetic nephropathy patients confirmed by renal biopsy who were seen between August 2007 and December 2012. Histopathological and clinical findings with renal outcomes were studied. The histopathological scores were determined according to Tervaert et al.: glomerular lesions (score 0–20)

were based on degree of mesangial expansion, GBM thickness, exudative lesion, nodular sclerosis, mesangiolysis, polar vasculosis, global sclerosis, segmental sclerosis, ischemic PDGFR inhibitor sclerosis, and hypertrophy; interstitial and tubular lesions (score 0–6) were based on degree of interstitial fibrosis, tubular atrophy, and interstitial inflammation; and vascular lesions (score 0–5) were based on degree of arteriolar hyalinosis and arteriosclerosis. Renal dysfunction was defined as doubling of serum creatinine

concentration, chronic hemodialysis initiation or renal transplantation. Results: Renal survival rates contrasting the low and high score groups in each of the three types of lesions were studied by Kaplan-Meier analysis. The mean survival periods of the low and high score groups for the glomerular Palbociclib (p = 0.24) and vascular (p = 0.22) lesions were not different. However, renal survival rates of 19 and 8 months for the low and high score groups (p = 0.010) respectively, in the interstitial and tubular lesions were significant. Conclusion: Interstitial and tubular lesions were a significant predictor for renal prognosis in diabetic nephropathy. Inasmuch as the histopathology of the glomerulus is known to provide important information of renal disease, our study indicates that significant prognostic information

may be associated with interstitial and tubular changes. MENG XIANJIE1, SHEN SHANMEI2, WAN YIGANG2, LUO XUNYANG2, ZHANG LE2, CHEN HAOLI1, SHI XIMIAO1, HUANG YANRU1, MAO ZHIMIN1 1Department of Graduate School, Nanjing University of Chinese Medicine; 2Nanjing MRIP Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School Introduction: Diabetic nephropathy (DN), as a common health problem worldwide, is a dominant cause of end-stage renal disease (ESRD). Therefore, noninvasive detections and dynamic managements in clinic are of major importance of preventing progression from DN to ESRD. The early diagnosis of DN has focused on measurement of urinary albumin (UAlb) excretion rate, but UAlb is actually not overall and sensitive marker for DN patients with inchoate and latent injuries in glomeruli and renal tubules.

The LPS from M huakuii (lines 1 and 2) migrated as three cluster

The LPS from M. huakuii (lines 1 and 2) migrated as three clusters of bands: a very intensively stained R-form LPS, an S-form, and an SR-form. A. lipoferum LPS (lines 3 and 4) was separated into two main fractions: the first one representing an R-form and

the second one, high molecular weight material. Those complete LPS molecules contained approximately 20 repeating units in the Cobimetinib mw O-chain, as calculated by comparison with the standard Salmonella LPS (line 7 and 14) (see also: 36). B. japonicum and B. yuanmingense LPSs (lines 5, 6 and 8, 9, respectively) were represented by complete molecules (S-form), mainly with short O-chains. The R fraction (containing only lipid A and core) was scarcely visible on the gel. In contrast, B. elkanii LPS (lines 12 and 13) occurred mainly as an R or SR form accompanied by a small amount of a

ladder-like S-form containing up to 20 repeating units. LPS from B. liaoningense (lines 10 and 11) was represented mainly by an SR-form, though a small amount of the R- and the S-forms was also present. The endotoxic properties of rhizobial LPSs were measured as their ability to gelate Limulus amebocyte lysate. For the LPSs from B. japonicum and B. yuanmingense, gelation was observed at a concentration of 0.1 μg/mL, whereas for the LPSs of B. elkanii, B. sp. (Lupinus), and B. liaoningense, the minimum LPS dose required for a positive reaction was ten times smaller (0.01 μg/mL). The LPSs from M. huakuii and A. lipoferum exhibited significantly greater endotoxic activity and gelated the amebocyte lysate at a concentration of 0.1 ng/mL. For the Florfenicol standard LPS preparations (Salmonella and E. coli), a positive reaction was observed at a concentration of 0.01 ng/mL. Production of NO was determined in cultures of THP-1 cells which were stimulated with 1 μg/mL LPS preparations for 24 hr (Fig. 3). A significant

amount of NO release was observed only for the standard LPS of Salmonella enterica bv Typhimurium (more than 300% of negative control). The amount of NO production by cells incubated with the B. sp. (Lupinus), B. elkanii, B. japonicum, M. huakuii, and A. lipoferum LPSs was just over half as much as that for Salmonella endotoxin, and exceeded by 50 to 100% the amount of spontaneous NO production by cells in the control sample. A statistically significant difference in NO production in comparison with the negative control (Student’s t-test, P value <0.05) was noted for B. sp. (Lupinus), B. japonicum, and M. huakuii. Production of the cytokines TNF, IL-1β, and IL-6 was determined in cultures of THP-1 cells stimulated with two LPS concentrations, 0.01 and 1 μg/mL (Fig. 4). At an LPS dose of 0.01 μg/mL, the Bradyrhizobium and the Azospirillum strains induced production of very small amounts of the cytokines. In the case of the two interleukins (IL-1β and IL-6), the measured amounts were within the same range as for the control sample (spontaneous activity of THP-1 cells) and the differences were not statistically significant.