The latter event facilitated the dissociation of Bim from Bcl-2 without affecting Bim abundance in IL-15-treated CD8αα+ iIELs. Using an adoptive cell transfer approach, we found that either overexpression of Bcl-2 or removal

of Bim from CD8αα+ iIELs promoted their survival in Il15ra−/− mice. Taken together, IL-15 promotes CD8αα+ iIEL survival by both increasing Bcl-2 levels and dissociating Bim from Lapatinib Bcl-2 through activation of a Jak3-Jak1-PI3K-Akt-ERK1/2 pathway, which differs from a previously reported IL-15-induced survival signal. Intestinal intraepithelial lymphocytes (iIELs) are T cells located between the epithelial cells lining the intestinal lumen. In the small intestine of C57BL/6J (B6J) mice, approximately half the iIELs are conventional T cells, while the other half are CD4−CD8β−CD8α+ (CD8αα+) cells that consist of 30% TCRαβ+ (αβ) cells and 70% TCRγδ+ NSC 683864 clinical trial (γδ) cells. CD8αα+ iIELs are developmentally and functionally distinct from conventional T cells. Most CD8αα+ iIEL precursors go through a thymic stage of development, and complete maturation in the intestine [1-4]. Functionally, CD8αα+ iIELs

appear to assume an immune regulatory role in the gut mucosa, as implied by their production of immune suppressive cytokines, such as TGF-β and IL-10, and by their ability to inhibit colitis [5, 6]. IL-15 is a pleiotropic cytokine widely expressed with its exclusive high affinity receptor IL-15Rα, while IL-15Rβγ chains are the intermediate affinity receptors for both IL-15 and IL-2 and expressed mainly by hematopoietic cells [7-9]. IL-15 and IL-15Rα form a complex during synthesis

in the ER and exist as transmembrane and soluble forms [10]. Afatinib in vivo The transmembrane IL-15–IL-15Rα complex is “in trans presented” to the IL-15Rβγ on neighboring cells for usage [11]. This mode of IL-15 usage has been implied to control the homeostasis of several lymphoid lineages, including CD8αα+ iIELs [1, 12-14]. More than 90% of CD8αα+ iIELs are missing in Il15−/− [15], Il15ra−/− [16], and Il15rb−/− [17] mice. Bone marrow chimera studies indicate that parenchymal IL-15Rα is essential for the development and maintenance of CD8αα+ αβ and γδ iIELs in the intestine [2, 14]. IL-15 also sustains the survival of primary CD8αα+ αβ and γδ iIELs in vitro [2, 18, 19]. As specific expression of IL-15Rα in the intestinal epithelial cell (IEC) of Il15ra−/− mice restores CD8αα+ iIELs and their Bcl-2 level [1], Bcl-2 has been implicated in the prosurvival effect of the IL-15 system. However, overexpression of Bcl-2 only moderately restored CD8αα+ γδ iIELs in Il15−/− mice [20], suggesting that the increase in the level of Bcl-2 alone is not sufficient to account for the prosurvival effect of IL-15.

Median age of patients was

34 years (range 1–73) and 37% had less than 18 years. Acute leukaemia was the most common underlying haematological disease (68/84; 81%). The phase of treatment was as follows: first induction Pexidartinib cost in 21/84 (25%), consolidation phase in 18/84 (21%) and reinduction/salvage in 45/84 (54%). The main site of infection was lung with or without other sites. The principal fungal pathogens were as follows: Aspergillus sp. 68 cases (81%), Candida sp. six cases (8%), Zygomycetes four cases (5%) and Fusarium sp. four cases (5%). The most used combo was caspofungin+voriconazole 35/84 (42%), caspofungin + liposomal amphotericin B (L-AmB) 20/84 (24%) and L-AmB+voriconazole 15/84 (18%). The median duration of combo was 19 days (range 3–180). The overall response rate (ORR) was 73% (61/84 responders) without significant differences between the combo regimens. The most important factor that significantly influenced the response was granulocyte (PMN) recovery (P 0.009). Only one patient discontinued therapy (voriconazole-related neurotoxicity) and 22% experienced mild and reversible adverse

events (hypokalaemia, ALT/AST increase and creatinine increase). The IFDs-attributable mortality was 17%. This study indicates that combo was both well tolerated and effective in haematological patients. The most used combo regimens were caspofungin + voriconazole (ORR find more 80%) and caspofungin + L-AmB (ORR 70%). The ORR was 73% and the mortality IFD related was 17%. PMN recovery during combo predicts a favourable outcome. Clinical Trials Registration: CYTH4 NCT00906633. “
“Hepatic fungal infection is a frequent complication in patients receiving intensive chemotherapy for acute leukaemia. Hepatic lesions may be detected

using computerised tomographic (CT) scans, but there is no standardised CT protocol for the diagnosis and follow-up of hepatic fungal infection. We therefore retrospectively analysed the number and the volume of hepatic fungal lesions in 24 CT of 20 consecutive patients treated for acute leukaemia during late-arterial and porto-venous phase. The mean number of lesions per patient was 31 (range: 3–105) in the late-arterial and 26 (3–81) in the porto-venous CT (P = 0.026). The mean total volume of all lesions was 6.45 ml in the late-arterial and 4.07 ml in the porto-venous CT representing a 1.6fold difference between the two CT scans (P = 0.008). The total volume of the lesions negatively correlated to the absolute contrast difference between liver parenchyma and liver vein (Pearson correlation, r = −0.62; P = 0.002).

Protective immunity against L. monocytogenes infection requires the coordinated action

of a diverse group of immune cells and cytokines (26, 27). Listeria monocytogenes infection led to increased relative spleen weights in the PC and LGG-fed groups, they did not increase in the JWS 833-fed group. Previous studies have reported that decreases in the relative weight of organs such as the spleen are indicative of increased host resistance. Administration of Lactobacillus plantarum reduced the spleen weight in L. monocytogenes-infected mice (29, 31). Meanwhile, the JWS 833-fed group had relatively heavier livers than the PC and LGG-fed groups. An earlier study by Tsai et al. showed a similar result in terms of increased liver weight (32). Rats Selleck Fulvestrant were fed with E. faecium TM39 for 4 weeks at a dose of 1 × 1012 cfu/kg. They found that E. faecium had no adverse effects in terms of changes in the relative weights of the heart, kidney and spleen weight in male or female Wistar rats; however, relative liver weights were higher in the female rats. Moreover, administration of Lactobacillus ingluviei in female BALB/c mice increased body and liver weights;

metabolic changes and amount of mRNA TNF-α was also significantly Selleck DMXAA increased (33). Puertollano et al. injected L. monocytegenes after oral administration of L. plantarum (29). According to them, liver weights were greater in the probiotic-fed than control group, although the difference between the two groups was not statistically significant. In our study, JWS 833-fed mice showed reduced spleen weights, suggesting protection from L. monocytogenes. JWS 833 induced higher serum concentrations

of NO and inflammatory cytokines after L. monocytogenes infection than did LGG. This immunomodulatory effect in JWS 833-fed mice correlated with increased survival rates and mean survival times after L. monocytogenes infection. The number of viable L. monocytogenes in the JWS 833-fed mouse livers was significantly lower than Resminostat in those of the control group. In our study we injected, the mice intravenously with L. monocytogenes. Most recent studies have also used i.v. injections to examine immune responses against L. monocytogenes infection in mice. L. monocytogenes is highly virulent in mice; however, JWS 833-fed mice infected with this bacterium i.v. were partially protected from this lethal infection. Since our goal was to determine whether JWS 833 protects mice from lethal infection with L. monocytogenes, we determined a lethal dose of L. monocytogenes based on published reports and our pilot experiments. Irons et al. (31) and Puertollano et al. (29) injected mice with a lethal dose of 106 cfu of L. monocytogenes; the infected mice died within 48–120 hrs. We carried out pilot experiments to determine the lethal dose of L. monocytogenes in BALB/c mice. We found that mice survived for 120 hr after an i.v. injection of 1.2 × 105 cfu/mouse.

Lymphocyte gates were set manually according to forward-scatter (FSC) and side-scatter (SSC), and subpopulations were subsequently determined. T cells within the lymphocyte gate were identified selleck screening library as either CD4+CD8- events (T helper cells) or CD4-CD8+ events (T cytotoxic cells). Natural killer (NK)

cells and B cells were approximated within the lymphocyte gate as CD56+ and CD19+ events, respectively. To determine the percentage of total monocytes/macrophages, the total live events were first gated and CD14+ events were then plotted versus SSC. Activated monocytes/macrophages were subsequently determined as CD16+ events within the CD14+ population. Therefore the results, reported as BEZ235 cost CD14+CD16+, represent the percentage of CD14+ cells expressing CD16, not double-positive events within the total live population. Plasma levels of the following interleukins IL-1β, IL-6, IL-8, IL-10 and tumour necrosis factor (TNF)-α were determined using the Milliplex™ MAP high sensitivity human cytokine kit with sensitivities of (0·06, 0·10, 0·11, 0·15 and 0·05 pg/ml), respectively (Millipore Corp. Billerica, MA, USA). The plates were read on a Luminex-200 fluorescent analytical test instrument (Luminex Corp., Austin, TX, USA). All assays were performed in duplicate according to the manufacturers’ instructions. For parametric variables, statistical significance between groups

was determined by t-test or analysis of variance (anova) using the Tukey–Kramer post-hoc multiple comparison test. The Kruskall–Wallis test was used to compare gender differences between groups. Correlations between parameters were determined using Pearson’s correlation. For non-parametric variables, correlations were determined by Spearman’s rho. The data was considered significantly different if P < 0·05. Calculations were accomplished with the aid of statistical data analysis software (spss version 17; SPSS Inc., Chicago, IL, USA). A total of 46 subjects (25 CRPS, 21 controls) were recruited for this study. The number of subjects in each group, their age, gender, body mass index (BMI), as

well as the duration of disease and NRS pain score for the CRPS group are tabulated in Table 1. There Anidulafungin (LY303366) were no significant differences in age, gender or BMI (P > 0·05) between the CRPS and control groups. For the CRPS subjects, the location of the initial injury, most prominent signs and symptoms, their overall pain score, the medications they were taking at the time the blood was sampled and other conditions with which the subjects were afflicted are listed in Appendix I. Eighteen of the 25 CRPS subjects had quantitative thermal tests performed as part of their clinical evaluation. None of the subjects demonstrated low thresholds (hypersensitivity) to cold or warm stimuli. The majority (10 of 18) had cold and heat thresholds within the normal range.

However, comparing the two patient groups regarding alloimmune and infectious history, we found no difference (data not shown). Remarkably,

we did not find a correlation between either severity of time to rejection and donor-specific CD8 precursor frequency, implying that other factors predominate in this respect. This could be due to differences in drug metabolism, concomitant with viral infections after transplantation that went unnoticed or the presence of Tregs that somehow delays the alloimmune response. Several groups have shown the IFN-γ ELISPOT assay to be a sensitive assay in predicting cellular alloreactivity pre- and post-transplantation. We Atezolizumab therefore compared the results of this Maraviroc order assay with the results of the MLC–CFSE assay [4,26]. Indeed, the number of IFN-γ-producing cells as detected by ELISPOT was increased significantly in rejectors compared to non-rejectors. In addition, we found a correlation between the number of IFN-γ-producing cells detected by ELISPOT and the dsp CD8 pf. This indicates that the CD8+ allospecific T cells are the most important IFN-γ-producing cells in the ELISPOT assay. However, in

the relatively small populations studied, there was a great overlap between rejectors and non-rejectors both in the ELISPOT assay and the MLC–CFSE assay. Because the difference in precursor frequency between rejectors and non-rejectors could not be explained by a difference in number of HLA-mismatches only, we measured the strength of alloreactive T cell activation by examining the difference in common-γ chain receptor expression after allostimulation. Importantly, we observed a significantly lower frequency of IL-7Rα expressing alloreactive

CD8+ T cells after both donor-specific and third-party see more stimulation in rejectors compared to non-rejectors. A higher pretransplant number of alloreactive IL-7Ra- CD8+ cells could cause this increase in pf. Indeed, we found a fair correlation between dsp CD8pf and the percentage of alloreactive IL-7Rα- CD8+ T cells. An explanation for the difference in percentage of IL-7Rα+ CD8+ T cells between the two patient groups may be a genetic polymorphism that influences the down modulation of IL-7Rα surface expression induced after T cell receptor (TCR) signalling or IL-7 binding [26,30,31]. In line with this, there are known polymorphisms associated with rejection after bone marrow transplantation as well as polymorphisms associated with increased immune activation playing a role in multiple sclerosis [32–34]. The finding of a low proliferative recall response to alloantigens of sorted IL-7Rα- CD8+ T cells is consistent with data from murine and human anti-viral responses [31,35]. These cells resemble the chronic antigen-addicted memory cells as described by Wherry et al. [36].

berghei infection. Increased expression of ECM components was observed in thymi from infected mice. In contrast, down-regulated surface expression of fibronectin and laminin receptors was observed in thymocytes from these animals. Moreover, in thymi from infected mice there was increased CXCL12 and CXCR4, and a decreased expression of CCL25 and CCR9. An altered thymocyte migration towards ECM elements

and chemokines was seen when the thymi from infected mice were analysed. Evaluation of ex vivo migration patterns of CD4/CD8-defined thymocyte subpopulations revealed that double-negative (DN), and CD4+ and CD8+ single-positive (SP) cells from P. berghei-infected mice have higher migratory responses compared with controls. Interestingly, selleck chemicals llc increased numbers of DN and SP subpopulations were found in the spleens of infected mice. Overall, we show that the thymic atrophy observed in P. berghei-infected mice is accompanied by thymic microenvironmental changes that comprise altered expression of thymocyte

migration-related C646 molecular weight molecules of the ECM and chemokine protein families, which in turn can alter the thymocyte migration pattern. These thymic disturbances may have consequences for the control of the immune response against this protozoan. The immune response during malaria is highly complex; this is partially the result of the intricate molecular structure of Plasmodium sp., the aetiological agent of the disease. This protozoan stimulates multifaceted immune responses, including antibodies, natural killer (NK) and NKT cells, and CD4+ and

CD8+ T cells.1,2 The immune response to the intraerythrocytic stages of the parasite has been better characterized by the use of murine experimental models. In this stage the CD4+ T helper type 1 response is essential for the development of the next events of the immune response in experimental malaria.3,4 We previously reported that the thymus gland is also a target organ in Plasmodium berghei infection: Suplatast tosilate there is atrophy with depletion of CD4+ CD8+ double-positive (DP) thymocytes, and histological alterations with loss of delimitation between the cortical and medullar regions. Moreover, we detected the intrathymic presence of parasites.5 The thymus is a primary lymphoid organ, responsible for the differentiation of T lymphocytes, including the shaping of an appropriate T-cell repertoire. This process is controlled by the cells and molecules of the thymic microenvironment, a tri-dimensional network essentially formed by epithelial cells, together with small numbers of dendritic cells, macrophages and fibroblasts.

The objective of this study was to assess whether peptidoglycan (PGN) derived from Gram-positive bacteria induces trophoblast stem (TS) cell death or alters TS cell cytokine

production. Method of study  Toll-like receptor (TLR) transcript expression was assessed by RT-PCR. Protein expression was determined by confocal microscopy or flow cytometry. 7-Aminoactinomycin D (7-AAD) staining was used to assess TS cell death. Morphological features of cell death were evaluated by transmission electron microscopy. The presence of cleaved caspase-3 and high mobility group box 1 (HMGB1) protein was examined by Western blot. Cytokine levels Decitabine ic50 in cell supernatants were determined using a mouse cytokine 23-plex panel. Results  Toll-like receptor 2 and TLR4 protein was expressed from the 1-cell stage through the blastocyst stage of murine embryo development. Murine TS cells expressed TLR2 and TLR6 but not TLR1 or TLR4 RNA. Only TLR2 protein was detected at the plasma membrane of TS cells.

PGN induced TS cell death by a caspase-3-independent mechanism. The cell death pathway induced by PGN was morphologically consistent with necrosis. Finally, PGN induced HMGB1 release Mdm2 antagonist and increased MIP-1β secretion while inhibiting the constitutive release of RANTES. Conclusion  Peptidoglycan-induced TS cell necrosis and the subsequent selleckchem release of HMGB1 and MIP-1β may regulate an infection-induced inflammatory response at the maternal–fetal interface and thus may play a role in the pathogenesis of infection-associated pregnancy complications. “
“A good understanding of the immunological correlates of protective immunity is an important requirement for the development of effective vaccines against malaria. However,

this concern has received little attention even in the face of two decades of intensive vaccine research. Here, we review the immune response to blood-stage malaria, with a particular focus on the type of vaccine most likely to induce the kind of response required to give strong protection against infection. Malaria still causes serious illness and many deaths in some of the poorest countries in the world. Over 200–300 million new cases are reported each year with 1·2 million deaths, mainly of young children [1]. There is still no vaccine that confers strong protective immunity to infection. Gaps in our understanding both of putative vaccine antigens and of the nature of antimalarial immunity have held back the development of a protective vaccine. While some immunity is acquired to infection after several years of repeated exposure to malarial infection, it is never complete. Such partial immunity or naturally acquired immunity that does develop, in an age and exposure related manner, involves both antibody and cell-mediated immune responses.

Thirty thousands of sorted CD19+ CD25+ or CD19+ CD25− B cells were resuspended in KRG buffer (Krebs-Ringer phosphate buffer) selleck kinase inhibitor with Ca2+, containing 0,1% BSA (Sigma-Aldrich) in a final volume of 30 μl and were placed on the upper well in duplicates. Cells were migrated towards different concentration of CXCL13 (50, 100 and 500 ng/ml), KRG buffer containing 0.1% BSA as a negative control added to the lower wells in a final volume of 30 μl. To determine if the migration was random

(chemokinesis) or directed (chemotaxis), 500 ng/ml of CXCL13 was added to both the upper and lower chamber followed by addition of cells to the upper chamber. Cells were incubated in a humidified atmosphere containing 5% CO2 at 37° for 12 h, thereafter the upper cell suspensions was removed, and the plates with the net were centrifuged at 350 g at 4° for 10 min. The net was discarded followed by an addition of 2 μl trypan blue together with 28 μl formaldehyde (4%). Migrated HIF inhibitor cells were manually enumerated using a microscope. Expression of homing receptors.  For flow cytometry analyses, 106 spleen cells were placed in 96-well plates and pelleted (3 min, 300 g, 4 °C). To avoid unspecific binding via Fc-receptor interactions, cells were incubated with Fc-block (2.4G2; BD Bioscience) for 8 min at room temperature. All antibodies were diluted in FACS-buffer (PBS containing, 1% FCS, 0.1% sodium azide and 0.5 mm EDTA). The antibodies used were directly conjugated with phycoerythrin

(PE), Pacific blue (PB) and peridinin chlorophyll protein (PerCp). Antibodies used were anti-CD25 (PC61), anti-α4β7 (DATK32), anti-CD62L (MEL-14), anti-CXCR5 (2G8) cAMP purchased from BD Bioscience and anti-CD19 (1D3), anti-CXCR4 (2B11) purchased from eBioscience, (San Diego, CA, USA). Cells were stained as previously described, and gating of cells was performed using fluorochrome minus one settings

[13]. All data in the study are presented as levels above the background. Proliferation assay.  Triplicates of sorted CD19+ CD25+ or CD19+ CD25− B cells at a concentration of 2.5 × 105/ml were plated in a volume of 100 μl in round-bottomed 96-well plates and stimulated with either 3 μm CpG-PS, 5 μg/ml E-coli LPS or 0.5 μg/ml of Pam3Cys in a humidified atmosphere containing 5% CO2 at 37° for 48 h and pulsed with 1 μCi 3H-thymidine (Amersham Pharmacia Biotech) for additional 8 h. The cells were harvested onto glass fibre filters (Walluc Oy) and dried, where after incorporated 3H-thymidine was measured using a β-scintillation counter. Statistics.  All statistical analyses have been performed using the Prism software (GraphPad software version 4.0b; La Jolla, CA, USA), and Wilcoxon matched paired test was used when comparing CD25+ to CD25− B-cell subpopulations and Kurskal–Wallis test followed by Dunn’s test for multiple comparisons when comparing more than two cell populations. P < 0.05 was considered as significant. B cells were sorted in to two highly purified populations (>98.

An alternative

mechanism whereby neutrophils eliminate Leishmania parasites was proposed very recently, and involves the generation of neutrophil extracellular traps, which are webs composed of chromatin and granular proteins 34. However the most likely mechanism is that TLR-9-expressing neutrophils become activated by CpG DNA and increase (i) their ability to activate macrophages (ii) their phagocytic and killing capacity 35. We will study changes in neutrophil activation by the Lm/CpG vaccine in future studies. In summary, the present study suggests that IL-17 may become an important modulator of Leishmania infection. Elucidating the mechanisms involved Selleck RXDX-106 in Th17 generation and those that undermine T-cell lineage crossregulation

will not only clarify the flexibility of T-cell differentiation, but may also shed insight into the pathogenesis of disease. Furthermore, understanding these phenomena will be critical for the design of immunotherapy that seeks to disrupt PLX4032 in vitro lineage-specific T-cell responses and may suggest ways to manipulate the balance between pathogenic and regulatory lymphocytes for the restoration of homeostasis. Six to eight wk old C57BL/6 and IL-17R−/− (C57BL/6 background) mice were purchased from Taconic (Germantown, NY). All mice were maintained in the Baker Institute Animal Care Facility under pathogen-free conditions. L. major clone V1 (MHOM/IL/80/Friedlin) promastigotes were grown at 26°C in medium 199 supplemented as described in 11. Infective-stage promastigotes of L. major were isolated

from stationary cultures (4–5 day-old) by Ficoll enrichment 36. Mice were vaccinated intradermally in both ears with 104L. major alone or in combination with 50 μg CpG DNA (5′ TCC ATG ACG TTC CTG ACG TT-3′, IDT, Coralville, IA) using a 27 1/2 G needle in a volume of 10 μL 10. Single cell suspensions from the ear dermis were obtained and processed as in Amobarbital 12. Briefly, the ear sheets were separated and deposited in DMEM containing Liberase CI enzyme blend (0.5 mg/mL) for 60 min at 37°C. The sheets were then cut and dissociated using a tissue homogenizer. For parasite titrations, a fraction of the homogenates were serially diluted in a 96-well flat bottom microtiter plate containing biphasic medium prepared using 50 μL Novy-MacNeal-Nicolle (NNN) medium containing 20% of defibrinated rabbit blood. The number of viable parasites in each sample was estimated from the highest dilution at which promastigotes could be grown out after 7 days of incubation at 26°C. For the analysis of the relative abundance of cell populations in the ears, single cell suspensions were generated as described above. In most experiments, ears were pooled to obtain enough cells for flow cytometry and microscopy assays. This will be indicated in each figure. Differential counts were performed manually on Giemsa-stained cytocentrifuge preparations.

Our data further suggest that the production of ROS and NO is linked. Since transcriptional selleck chemical regulation of iNOS is altered, this linkage is most likely at the level of the signaling pathways and ultimately NFκB associated. It remains unclear whether this effect is mediated by the ROS molecules themselves, the changes in vesicular pH, or another mechanism; however, our data are supported by findings in which the anti-inflammatory regulator Nrf2 was found to be defective

in CGD 42. This raises the possibility that increased iNOS transcription in CGD upon GlyAg stimulation could be a result of an inability to shut down the initial GlyAg-mediated TLR2-dependent signal 19 to activate iNOS synthesis in the first place. The difference between WT and CGD responses to an actual antigen like PSA from B. fragilis provides an ideal model system to explore the relationship between the control of ROS and NO production. Taken together, our findings suggested that NO in macrophages, but not neutrophils, is the primary mediator of hyperresponsiveness to GlyAg in CGD. Our adoptive transfer experimental selleck compound data further suggest that the loss of ROS in the T-cell population, which has been linked to a switch between T effector and T regulatory cells 43, does not explain the enhanced GlyAg response. These interpretations were confirmed

in vivo using iNOS inhibition which completely prevented abscess formation in 6 of 14 animals while significantly reducing the abscess severity in the remaining mice. Since 1400W

did not appear to increase the risk of bacterial sepsis, this strategy may represent a new pathway of treatment for CGD patients, although far more stringent testing with more invasive organisms would be needed to confirm these initial findings. In contrast to the CGD T-cell studies in which non-specific anti-CD3/anti-CD28 stimulation of T cells was used 44, 45, our findings suggest a novel pathway responsible for CGD-associated recurring abscess formation that is centered upon professional APCs, increased GlyAg processing, tuclazepam and antigen-mediated T-cell activation. This pathway can be specifically targeted through inhibition of iNOS activity in vivo, resulting in attenuation of CGD-associated immune pathology arising from bacterial infection. This approach could significantly improve treatment outcomes for CGD patients through increasing antibiotic efficacy and reducing the need for surgical drainage of abscesses. WT (C57BL/6J, stock 000664) and X-linked gp91phox-deficient CGD (B6. 129S6-Cybbtm1Din/J, stock 002365) breeders were purchased from Jackson Labs and colonies were housed at CWRU Animal Resource Center. Experiments were performed in accordance with the guidelines of the National Institutes of Health (NIH) and protocols approved by the Institutional Animal Care and Use Committee. All experimental mice were at least 12 wk old.