For example, epithelial cells from the upper tract of postmenopau

For example, epithelial cells from the upper tract of postmenopausal women lack the capacity to secrete antimicrobials compared to pre-menopausal women.13 When planning studies of response to microbicides or vaccination, investigators should decide whether to include menopausal women or whether

to control for menopausal status in analyses. Pregnancy may increase the risk of HIV acquisition and is associated with marked hormonal and immunologic changes. A large, rigorous study carried out in Rakai, Ku-0059436 purchase Uganda, found that women were at significantly increased risk of HIV acquisition during pregnancy. Data from a community cohort with longitudinal data were analyzed for the incidence rate of HIV during pregnancy and lactation, and compared to the incidence rate during periods of non-pregnancy and non-lactation. The incidence rate was 2.3

per 100 person years in pregnancy when compared to 1.1 per 100 person years in non-pregnant and lactating women. This study was rigorous because sexual behavior was recorded Luminespib mouse as part of a community, epidemiologic study. This difference in incidence rates resulted in an incident rate ratio of HIV acquisition in pregnancy of 2.16 (95% CI 1.39–3.37) after adjusting for age, marital status, education, multiple sex partners, genital ulcer disease, and condom use.14 Data remain conflicting, however, regarding the risk of HIV infection in pregnancy. Other studies also carried out in Africa failed to confirm the findings in the Rakai study.15,16 The ability of the mother’s body to tolerate a fetus that is not genetically identical

to her has long been a topic of immunologic interest. While there are immunologic changes that occur at the Isotretinoin maternal–fetal interface to allow the mother to tolerate her semi-allogeneic fetus, there are also major components of the lower genital tract that play an important role in immunity and modification of these may not be beneficial to the mother. The concentration of some antimicrobial peptides thought to be important in anti-HIV activity is frequently altered in pregnancy. In normal pregnancy, secretory leukocyte protease inhibitor concentrations are significantly greater than in the non-pregnant state, particularly in the cervical mucous.17 Kutteh and Franklin18 followed 36 pregnant women through pregnancy and found increasing concentrations of IL-1β, a pro-inflammatory cytokine during the course of pregnancy. Donders et al. performed a small, prospective cohort study examining the changes in cytokine concentrations of 30 women during normal pregnancy. They found that, compared to non-pregnant women, pregnant women were less likely to have detectable IL-6 and IL-8 and that the concentrations of these molecules dipped during the second trimester. The concentrations then returned to pre-pregnancy levels in the third trimester.

As shown in Fig  3(b) both m-S100A9 and LPS stimulated NO

As shown in Fig. 3(b) both m-S100A9 and LPS stimulated NO selleck production, again with LPS as the more potent inducer. These results further supported the pro-inflammatory activity of S100A9. Our next step was to determine whether h-S100A9 would exert its effects on NF-κB activation through the same or a different

signalling pathway than LPS. Hence, we pre-incubated THP-1 cells with selected inhibitors to block key steps in the main pathway involved in NF-κB activation and then stimulated the cells and measured TNF-α secretion. Figure 4 shows that BAY11-7082, which reduces IκBα phosphorylation,[31] effectively blocked both the LPS-induced and h-S100A9-induced response. Further, PD98059 and SB203580, which are inhibitors of MEK1[33] and p38,[32] respectively, strongly inhibited the TNF-α response triggered both by LPS and h-S100A9, suggesting that mitogen-activated protein kinase proteins were involved both in the LPS and h-S100A9-induced signalling pathways. The inhibitor of proteasome activity MG132,[34] which blocks IκBα degradation, inhibited TNF-α responses almost completely, suggesting that IκBα could be involved in the h-S100A9 signalling pathway. For all the inhibitors tested, we could observe more than 50% inhibition of LPS-mediated

and h-S100A9-mediated TNF-α secretion. The above-mentioned inhibitors did not significantly affect cell viability (see Supplementary material, Fig. S2a). Taken together, these data indicate that LPS and h-S100A9 exerted their pro-inflammatory effects through basically the same signalling pathway to activate NF-κB. To further confirm the activation of NF-κB by human and mouse S100A9, we monitored IκBα degradation. IαBκ 3-MA mw is activated via phosphorylation by IKK proteins upon proper cellular stimulation. In this way, IκBα is targeted for proteasomal degradation and NF-κB subunits are able to interact and form the mature NF-κB dimers.[35] As human S100A9 was less potent than LPS in promoting cytokine secretion, we expected to find

that h-S100A9 provoked a weaker IκBα degradation. Surprisingly, Western blot analysis revealed the opposite. Hence, h-S100A9-mediated stimulation of THP-1 XBlue cells effectively reduced the IκBα level already after 15 min and it remained reduced for up to 60 min after stimulation. The LPS-induced degradation was significant only at 60 min of Tolmetin stimulation and in this case there was only a slight IκBα degradation (Fig. 5a). These results further confirmed that h-S100A9 activated the NF-κB transcription factor. Most importantly, the kinetics of the h-S100A9-induced NF-κB activation was more rapid, even though it led to a weaker cytokine response. In contrast, LPS provoked delayed and weaker NF-κB activation but a more potent and sustained cytokine response. These results were in agreement with the pro-inflammatory role of h-S100A9 but in apparent contrast with Fig. 1, which showed that h-S100A9 promoted NF-κB activity in a comparable way to LPS.

Interestingly, however, in spite of higher

Interestingly, however, in spite of higher MAPK Inhibitor Library concentration parasitaemia, iNOS-inhibited birds did not pay a higher cost of infection because

haematocrit values were similar for iNOS-inhibited and control birds. This result parallels those reported for Plasmodium chabaudi-infected mice and suggests that the cost of higher parasitaemia in iNOS-inhibited birds might be compensated by a reduced cost of immunopathology. Overall, these results also point towards a possible trade-off between resistance and tolerance. As mentioned above, the control of the acute proliferation of asexual malaria parasites relies on several inflammatory effectors. Up-regulating the inflammatory response however adds a potential immunopathology toll to the overall cost of infection. Breaking down immunological tolerance therefore constitutes a possible mechanism underpinning a physiological trade-off between resistance and tolerance. A pending important question is now how parasites do adapt to hosts depending on the defence strategy (resistance vs. tolerance) and the possible trade-off between strategies. Again insight into the possible evolutionary trajectory followed by parasites experiencing particular immune environments comes from studies on rodent malaria, where Plasmodium chabaudi serially passaged in vaccinated mice

evolved to become a more serious threat to their host [59]. The reason for increased virulence of parasites evolving in vaccinated host lies on the relaxed cost of virulence. GS-1101 clinical trial Vaccinated hosts are protected from infection-induced mortality but they still contribute to parasite transmission [60]. Therefore, rapidly growing parasites are favoured Amine dehydrogenase in vaccinated hosts and can

be highly pathogenic in nonvaccinated hosts. Evidence in support to parasite evolution as a function of host immunity [61] also comes from a recent study involving Plasmodium relictum-infected canaries. Cornet et al. [62] assessed the infection dynamics and the cost of infection in canaries facing two diets. Birds enjoying a protein- and vitamin-enriched food were better able to control parasite growth (they had lower parasitaemia, and peak parasitaemia was reached earlier than for control, nonsupplemented hosts). Protein and vitamins are important environmental determinants of immune competence as shown in several organisms, including humans [63, 64]. Therefore, reduced parasitaemia in food-supplemented birds is consistent with an improved resistance. Nevertheless, food-supplemented birds also paid the highest per-parasite cost of infection (Figure 2a). In a follow-up experiment, parasites grown in food-supplemented and control hosts were inoculated in another group of hosts following a fully factorial design (parasites grown in food-supplemented hosts passaged in food-supplemented and in control hosts; parasites grown in control hosts passaged in food-supplemented and in control hosts) [62].

On day 6, fresh medium containing GM-CSF, IL-4, IL-1β, IL-6,

On day 6, fresh medium containing GM-CSF, IL-4, IL-1β, IL-6,

PGE2, and TNF was added to the culture. After additional 48 h of culture, nonadherent cells were harvested and used as APCs. INK 128 research buy Purified CD4+, CD8+ and DN T cells (1×105/well) from donor A were cocultured with allogeneic mature DC (2.5×104/well) from donor B or with anti-CD3/CD28-coated beads (2.5×104/well; Dynabeads CD3/CD28, Invitrogen) in 96-well U-bottom plates in complete medium supplemented with 3% TCGF. T cells were restimulated weekly with fresh allogeneic DC. Viability and purity of the T cells were monitored by flow cytometry. Further purification via magnetic separation was performed if purity decreased to lower than 95%. T cells were used for functional assays 6 days after last stimulation. Cells were stained with fluorescein isothiocyanate (FITC)-conjugated anti-IFN-γ, anti-CD4, anti-CD8, anti-TCR-γδ, phycoerythrin (PE)-conjugated anti-CD25, anti-CD45RO, anti-TCR-, and allophycocyanin-conjugated anti-CD38, anti-CD45RA, anti-CTLA4 monoclonal antibodies (mAb) (all from BD Biosciences, Heidelberg, Germany). Isotype control mAb, FITC-labeled annexin V, and 7AAD were purchased from BD. Foxp3 stains were performed

with allophycocyanin-conjugated anti-Foxp3 mAb and the respective control from eBioscience (San Diego, USA). For intracellular IFN-γ staining, activated CD4+ T cells were cocultured with DC and DN T cells in the presence Roxadustat molecular weight of monensin (GolgiStop, BD) for 5 h. After washing, cells were stained for surface markers, fixed and permeabilized (Cytofix/Cytoperm kit, BD), and then stained for intracellular cytokines. Flow cytometry was performed on a FACSCanto II (BD); cell sorting was accomplished on a MoFlo (Beckman Coulter). ZD1839 datasheet Data were analyzed with FlowJo software (Treestar, Ashland, OR, USA). CFSE (Sigma, Munich, Germany) labeled CD4+ and CD8+ T cells (5×104/well) from donor A were stimulated in 96-well U-bottom plates with allogeneic DC (2.5×104/well) from donor B, anti-CD3/CD28

beads (2.5×104/well, Invitrogen/Dynal, Oslo, Norway), or plate-bound anti-CD3 (0.25 μg/well, Orthoclone OKT3, Janssen-Cilag) in complete medium in the presence or absence of DN T cells or CD4+CD25+ Tregs (5×104/well). Anti-CD2/CD3/CD28 loaded particles (Treg Suppression Inspector, Miltenyi Biotec) were used according to the manufactures instructions. After 5–6 days of culture, cells were harvested and stained with anti-CD4, anti-CD8, anti-TCR-αβ, and anti-CD25 mAb. Proliferation of cells was determined by flow cytometry. For blocking experiments, mAb to IL-10 (10 μg/mL JES3-19F1; BD), TGF-β (10 μg/mL 1D11; R&D Systems), Fas (10 μg/mL ZB4; Biomol), or isotype-matched controls were added to the MLR. To block TCR-signaling and protein translocation, DN T cells were incubated with Lck-inhibitor II (100 μM; Calbiochem, Darmstadt, Germany) or with monensin (GolgiStop, according to the manufacture’s protocol; BD) for 3 h, and then used as suppressor cells in the MLR.

Once the effect of the intervention on an outcome is calculated w

Once the effect of the intervention on an outcome is calculated within each trial (either the

RR or MD), the next step is to combine these treatment effects for each outcome together to calculate an overall RR (dichotomous variable) or MD (continuous variable) between two treatments (meta-analysis). Combining results from individual studies is not simply achieved by treating all studies equally and averaging their data. Instead, the studies are combined using a weighted average. The contribution of a trial to the overall effect size (weight) depends on its variance (the certainty of the trial’s effect size). Studies with smaller estimates of variance (greater precision) and/or with more events, make a larger contribution to the overall effect estimate of an intervention.14 Figure 2 shows a graphical representation (known as PF-6463922 purchase the forest plot) commonly used in systematic reviews to summarize data from a systematic review of haemoglobin targets in patients with CKD.1 In this example, studies are pooled to examine the risk of mortality using human recombinant erythropoietin to treat anaemia (higher haemoglobin vs lower haemoglobin level) in people with CKD.1 In this forest plot: 1 The left hand column shows the eight included randomized, MAPK Inhibitor Library cell line controlled trials that have mortality

data available for analysis. In this figure they are in chronological order. What happens if the meta-analysis is trying to combine apples with oranges? In other words, does the systematic review aggregate

poor-quality trials that possess a substantial risk of bias, together with higher-quality trials? Such inclusion of low-quality trials may provide an unreliable conclusion about treatment efficacy or toxicity. To explore the possibility that a meta-analysis includes trials of lower quality and provides a less precise estimate of treatment effect, the reader of a systematic review might assess whether the authors have conducted a formal assessment of method quality Methamphetamine for each included trial. Specifically, a systematic review should report an assessment of each domain considered to be indicative of study quality. These are: 1 Allocation concealment (‘selection bias’): Allocation concealment is adequate when the trial investigators cannot determine the treatment group to which a patient has been assigned. Knowledge of treatment allocation may lead to exaggerated treatment effects. It has been shown through systematic review of meta-analyses that the estimate of effect summarized by meta-analysis may be substantially more beneficial to the intervention when the trial conduct of included studies does not follow these principles, and particularly when allocation concealment is inadequate.

The growth and migration of cultured cells were quantified by usi

The growth and migration of cultured cells were quantified by using CL-Quant software to analyze time-lapse images in a Nikon BioStation CT. The real-time images of cell migration were monitored for 2 days. And also NRK-49F cells were stimulated with S1P after the addition of FTY720 (S1P 1, 3, 4, 5 agonist), or DMS (sphingosine kinase inhibitor) were evaluated. Results: S1P stimulated fibrosis of NRK-49F cells in a dose- and time-dependent manner as

previously observed, and induced morphological changes (elongation of the cell shape with spindle-like extension, increased migration) of the NRK-49F cells. Migration of NRK49F cells was accelerated and increase in a-SMA, COL1, COL4, TIMP1 and PAI1 expressions and decrease in E-cadherin expression were observed by addition of S1P. Antagonist and siRNA transfection to NRK-49F cells of Sphingosine 1-phosphate receptor-3 (S1PR-3) attenuated cell

growth and migration, Sirolimus manufacturer in addition, the expression of fibrotic markers was also diminished by antagonist and siRNA transfection to NRK-49F cells of S1PR-3. And also in the presence of FTY720 and DMS, fibrosis and migration induced by S1P were suppressed. Conclusion: These results suggest that activation of S1P signaling mediated by S1PR-3 results in chronic pathological fibrosis, such as in chronic kidney disease (CKD). LEUNG JOSEPH C K, CHAN LORETTA Y Y, LIM AI ING, WONG DICKSON W L, LAI KAR NENG, TANG SYDNEY C W The University of Hong Kong, Hong Kong Introduction: Protein overload check details induces apoptosis in proximal tubule epithelial cells (PTEC). We have recently shown that bone morphogenetic protein-7 (BMP-7) up-regulates the expression of cellular inhibitor of apoptosis 1 (cIAP1) and represses albumin-induced chemokines synthesis in kidney tubular epithelial cells (PTEC). In this study, we examined the effect of BMP-7 on albumin-induced apoptosis in PTEC. Methods: An in vitro PTEC culture model of albumin overload was used to examine the roles of BMP-7 on albumin-induced apoptosis. Either with Montelukast Sodium or without incubation with recombinant

human BMP-7, apoptosis in cultured PTEC exposed to albumin (5 mg/ml for 3 days) was determined using an in situ cell death detection kit and quantitated with a fluorometric TUNEL assay. Expression of genes and proteins of TNF-α, the pro-apoptotic bax, bad and anti-apoptotic bcl-xL, c-FLIP, were determined by quantitative RT-PCR, western blotting or ELISA. Caspase-8 activity was determined using a luminescent assay. Activation of the NF-kB p65 and p50 subunits in PTEC were quantitated by ELISA-based transcriptional factor assays and western blotting. Results: In cultured human PTECs, albumin significantly up-regulated the gene and protein expression of bax, bad but down-regulated bcl-xL and c-FLIP. Addition of BMP-7 further amplified these increased apoptotic and reduced anti-apoptotic proteins expression elicited by albumin.

Experimental infection  Mycobacterium avium strain 2447 (smooth t

Experimental infection. Mycobacterium avium strain 2447 (smooth transparent variant kindly provided by Dr F. Portaels from the Institute of Tropical Medicine, Antwerp, Belgium) was grown in Middlebrook 7H9 medium containing 0.05% Tween 80 at 37 °C until mid-log phase of growth. Bacteria were harvested by centrifugation and resuspended in saline containing 0.05% Tween 80. The bacterial suspensions were briefly sonicated with a Branson

sonifier to disrupt bacterial clumps, diluted, and stored in aliquots at −70 °C until use. Intravenous infection with M. avium was performed through the tail lateral vein with 106 CFU per animal. At specific time-points (4, 8 and 20 weeks post infection), the organ bacterial load was determined as previously described [21]. Briefly, mice were anaesthetized and killed with isoflurane (Abbott, IL, USA). The organs were removed in aseptic conditions, homogenized, and serial dilutions were click here prepared in distilled sterile water with 0.05% Tween 80 and plated onto Middlebrook 7H10 agar medium. The numbers of CFU were

counted after 1 week of incubation at 37 °C. Flow cytometry.  Single cell suspensions were prepared from the spleen and the thymus of each mouse. Spleen erythrocytes were lysed with a haemolytic solution (155 mm NH4Cl, 10 mm KHCO3, Sunitinib nmr pH 7.2). For each staining, 5 × 105 cells from each organ were incubated with a specific set of antibodies for 20 min at 4 °C. Cell surface markers were analysed using anti-CD25 APC or Pe (clone PC61), anti-CD11b PE (clone M1/70), anti-CD3 FITC, PE or APC (clone 145-2C11), anti-CD4 FITC or PECy5 (clone RM4-5), anti-CD62L FITC (clone MEL-14), anti-CD44 PE (clone IM7), anti-CD19 FITC (clone 6D5), anti-NK-1.1 FITC (clone PK136), anti-CD8 FITC, APC or APCCy7 (clone 53-6.7) and anti-Ly-6G/Ly-6C PECy5 (Gr-1;

clone RB6-8C5) (all from Biolegend, San Diego, CA, USA). Cells were Niclosamide fixed with 2% formaldehyde after staining. The analysis of the cell populations was based on the acquisition of 30,000 events using CellQuest software on a FACscalibur flow cytometer or a FACSAria cell sorter (Becton Dickinson, NJ, USA). Data analysis was performed using FlowJo software (Tree Star, Inc, Ashland, OR, USA). Detection of IFN-γ in serum samples.  Mice were anaesthetized with isoflurane (Abbott, IL, USA), and retro-orbital bleeding was performed before killing. Blood was allowed to clot and serum was collected after centrifugation and frozen at −80 °C until use. Quantification of IFN-γ was done by a two-side sandwich ELISA using anti-IFN-γ-specific affinity-purified mAbs (R4-6A2 as capture and biotinylated AN-18 as detecting mAbs), and the standard curves were generated with known amounts of IFN-γ (Peprotech, Rocky Hill, NJ, USA). The sensitivity of the assay was 20 pg/ml. Statistical analysis.  All data are presented as means + SD.

trachomatis inclusions (Coers et al , 2008) This localization wa

trachomatis inclusions (Coers et al., 2008). This localization was observed only with C. trachomatis, while C. muridarum seems to have evolved mechanisms that prevent the accumulation of GTPases in the chlamydial inclusion, a possible immune evasion strategy (Coers et al., 2008). Although most of the assessed pathways seem to help the host cell in bacterial clearance, there is evidence that Chlamydiales also use TLRs to establish a replication-friendly environment. Chlamydia pneumoniae raises ATP levels through activation of the TLR2/Myd88 pathway. This behavior is crucial

because Chlamydiales are unable to produce ATP (Yaraei et al., 2005). MIP-2 and KC are two chemokines expressed upon Myd88 BI 6727 clinical trial activation. In infected mice, these chemokines attract polymorphonuclear neutrophils to the lungs. Chlamydia pneumoniae is thought to use these cells to spread Target Selective Inhibitor Library throughout the lungs (Rodriguez et al., 2005). Immune cells can therefore be used as vehicles to reach new tissues instead of fighting the infection. Interaction of Chlamydiales with TLRs is of particular interest because they control inflammation that can become chronic or, if uncontrolled, cause damage. For example,

TLR2 recognition of bacterial PAMPs was linked to trophoblast apoptosis (Abrahams et al., 2004), which could provoke preterm delivery. Similarly, exposure to chlamydial Hsp60 (CHsp60) induces apoptosis in trophoblasts. Trophoblast TLR4 recognized CHsp60 and, through an unknown signaling pathway, induced several downstream caspases (Equils et al., 2006). Development of atherosclerosis was reduced in TLR2-deficient mice infected with C. pneumoniae. Without the TLRs, the level of circulating cytokines

was reduced and less dendritic cells were activated these (Naiki et al., 2008). Thus, different yet unknown chlamydial antigens seem to induce such a strong response that they cause severe damage to the surrounding tissue. Downstream of PRRs, there are not only cytokines and their receptors but also several enzymes that synthesize microbicidal molecules. ROS are strong microbicidals produced by macrophages, dendritic cells and neutrophils. Most of them are produced by NADPH oxidase (Nox), a multiproteic transmembrane complex. This family of genes is found only in multicellular organisms, with few exceptions (reviewed in Bedard & Krause, 2007). There are three different classes of NADPH oxidases (reviewed in Bedard et al., 2007). In most mammals, all seven genes are found, while rodents lack Nox5. The Nox present in phagocytic cells is Nox2. It is not clear whether other members of the Nox family are also specifically induced upon infection of phagocytic cells. Chronic granulomatous disease is a severe and debilitating disease found in individuals with mutations in components of the Nox2 complex.

2a) Previous reports on DS have suggested that limited thymic ou

2a). Previous reports on DS have suggested that limited thymic output would lead to decreased function and immunosenescence in peripheral immune cells.[14] To assess lymphocyte functional capacity in Ts65Dn mice, whole splenocytes were stimulated with immobilized anti-CD3 antibody (at 0.5 or 5 μg/ml) for 48 and 72 hr in vitro. Proliferation of CFSE-labelled splenocytes was measured in TCR+ T cells by flow cytometry (representative flow data shown in Fig. 2b,c) as described in the ‘Materials and methods’. There was a significant decrease in the percentage of TCR+ cells that had undergone

at least one division in cells from Ts65Dn mice compared with euploid Navitoclax in vitro controls after 48 hr (Fig. 2d) and 72 hr (Fig. 2e). There was also a significant decrease in the percentage of TCR+ cells that had undergone more than three divisions after 72 hr (Fig. 2f) in cells from Ts65Dn mice. Changes in proliferation were not the result of cell death because the percentage of viable cells was not different between euploid and Ts65Dn cells at both time-points (not shown). Hence, although the proportions

of peripheral immune cells in Ts65Dn mice are relatively unchanged, there are significant defects in the function of peripheral T cells that suggest a senescent phenotype in this mouse model of DS. As a possible mechanism for thymic alterations in Ts65Dn mice, IL-7Rα was assessed because it plays a non-redundant role in thymic development,

promoting proliferation and survival of immature, DN thymocytes.[18] Consistent with our previous observations selleck in bone marrow lymphoid progenitors,[6] the percentage of specific thymocyte subsets that were IL-7Rα+ was decreased in the lineage-negative (total DN thymocytes), DN2 and DN3 populations (Fig. 3a). The absolute number DAPT cell line of cells expressing the IL-7Rα chain was significantly decreased in the Lin− and all the DN thymocyte populations (Fig. 3b). Interleukin-7Rα is normally down-regulated in DP thymocytes and re-expressed in positively selected CD4 and CD8 SP thymocytes.[15] In contrast to the data in DN thymocytes, when mature DP and SP thymocytes were analysed neither the percentage of IL-7Rα+ cells (Fig. S1c) nor the number of positive cells (not shown) was decreased. To measure whether changes in IL-7Rα were associated with altered cell proliferation in the thymus, mice were injected with BrdU for 2 days and incorporation was assessed ex vivo. Consistent with those populations having decreased IL-7Rα expression, lower percentages of BrdU+ cells were found in the DN2 and DN3 populations (Fig. 3c), and significantly fewer BrdU+ cells were detected in the DN2, DN3 and DN4 populations of Ts65Dn mice in comparison to euploid mice (Fig. 3d), indicating defects in thymocyte proliferation.

When logistic regression was used to carry out association analys

When logistic regression was used to carry out association analysis after modelling the SNP effects as additive, dominant or recessive, SNP6 (rs7749390, located on the splice site of the exon/intron of ifngr1 gene) showed a significant difference in co-dominant (OR: 1.86, 95% CI: 1.04–3.31) and log-additive (OR: 1.35, Navitoclax concentration 95% CI: 1.02–1.80) models, and P-values were <0.05 after adjustment for sex (Table 4). The log-additive model was accepted as the best inheritance model because of the smaller Akaike information criterion (AIC) value

(565.6). The other SNP showed no association with tuberculosis in any of the five inheritance models (all P > 0.05, data not shown). Pairwise LD between the three SNP of the ifng gene and the other four SNP of ifngr1 was calculated for the cases and controls in the Chinese Han population. The three SNP of the ifng gene had no association with tuberculosis (data not shown). D′ and r for all possible pairs of the SNP in the ifngr1 gene are shown in Table 5. We found strong LD (D′ > 0.75) between the following pairs of the markers in the ifngr1 gene: SNP4/SNP5 (D′ = 0.941), SNP4/SNP6 (D′ = 0.830) and SNP5/SNP6 PD-0332991 ic50 (D′ = 0.998). Therefore, we constructed SNP4/SNP5/SNP6/SNP7 as haplotype blocks in the ifngr1 gene (because the distance was about 141 bp between SNP6 and

SNP7, SNP7 was list to the haplotype analysis). The frequencies of the estimated haplotypes are presented in Table 6. The association analysis of the haplotypes with tuberculosis is shown in Table 6. The haplotype of SNP4/SNP5/SNP6/SNP7 showed significant association with the disease (P = 0.00079). The Cyclin-dependent kinase 3 C-A-A-TT haplotype was observed more frequently in the cases than in the controls (OR: 3.96, 95% CI: 1.90–8.21). The association analysis of haplotypes was adjusted also by sex. In China, tuberculosis is still prevalent with about 5 million cases every year. As only about 10% of the population that

is infected by M. tuberculosis will develop clinical tuberculosis, differences in host immunity and genetic factors may account for the development of tuberculosis after infection [3]. In this study, we tested the hypothesis that the ifng and ifngr1 genes play a role in the pathogenesis of tuberculosis. Seven SNP in these two genes were selected as the gene markers for association analysis. It is accepted generally that IFN-γ plays a pivotal role in the pathogenesis of tuberculosis [7]. Abnormality of the ifng gene is considered as one of the causative factors [8]. An initial study of the association of the ifng gene with tuberculosis indicated that allele A of the +874 A/T in the first intron region was a susceptibility factor for M. tuberculosis infection, both by population- and family-based analysis.