In this study, we explored the origins of 8-month-old infants’ me

In this study, we explored the origins of 8-month-old infants’ means-end action production using a cloth-pulling training paradigm. We examined whether highlighting the goal (toy) or the means (cloth) was more valuable for learning to perform a well-organized means-end action. Infants were given the opportunity to both practice cloth-pulling and view modeling of the action performed by an adult throughout the session. Infants saw either the same toy or the same cloth in successive trials, so that the goal or means were highlighted prior to modeling of the action. All infants improved throughout Ivacaftor chemical structure the session regardless

of which aspect of the event was highlighted. Beyond this general improvement, repetition of goals supported more rapid learning and more sustained learning than did repetition of means. These findings provide novel evidence that, at the origins of means-end action production, emphasizing the goal that structures an action facilitates the learning of new means-end actions. “
“Infants and their mothers

participated in a longitudinal study of the sequelae of infant goal-blockage responses. Tipifarnib research buy Four-month-old infants participated in a standard contingency learning and goal-blockage procedure during which anger and sad facial expressions to the blockage were coded. When infants were 12 and 20 months old, mothers completed a questionnaire about their children’s tantrums. Tantrum scores increased with age and boys tended to show more tantrum behavior than girls. Anger expressed to goal blockage at 4 months was unrelated to tantrum behavior. There was a gender by sad expression interaction. Girls who expressed sadness in response to the goal blockage had lower total tantrum scores than boys; otherwise

there was no difference. These results suggest that tantrums of infants who Parvulin display sad, not anger expression, in response to goal blockage, are differentially influenced by children’s gender. “
“The goal of this study was to examine developmental change in visual attention to dynamic visual and audiovisual stimuli in 3-, 6-, and 9-month-old infants. Infant look duration was measured during exposure to dynamic geometric patterns and Sesame Street video clips under three different stimulus modality conditions: unimodal visual, synchronous audiovisual, and asynchronous audiovisual. Infants looked longer toward Sesame Street stimuli than geometric patterns, and infants also looked longer during multimodal audiovisual (synchronous and asynchronous) presentations than during unimodal visual presentations. There was a three-way interaction of age, stimulus type, and stimulus modality. Significant differences were found within and between age groups related to stimulus modality (visual or audiovisual) while viewing Sesame Street clips. No significant interaction was found between age and stimulus type while infants viewed dynamic geometric patterns.

This inhibition is mainly mediated by LXRβ, as demonstrated by th

This inhibition is mainly mediated by LXRβ, as demonstrated by the fact that lymphoid hyperplasia and enhanced responses to antigenic challenge

have been observed in Lxrβ−/− mice, but not in Lxrα−/− mice [28]. Accordingly, IL-2- and IL-7-induced T-cell proliferation and cell cycle progression are inhibited upon LXR activation [29]. LXRs are also involved in Th17-cell differentiation, Selleck Veliparib as demonstrated by experiments in Lxrα−/−, Lxrβ−/−, and Lxrα−/−Lxrβ−/− mice, in which Th17 induction was found to be increased as compared with Th17 induction in WT mice [30]. In addition to LXR-dependent mechanisms, oxysterols regulate crucial innate and adaptive immune cell functions through the engagement of GPCRs. For example, the oxysterol 7α,25-OHC can bind and activate the GPCR Epstein–Barr virus-induced 2 (EBI2), which is upregulated on B cells and T cells under specific conditions [31, 32]. EBI2 is required for B-cell migration to intra- and extrafollicular sites of secondary lymphoid organs, where they then

differentiate into plasma cells FK506 concentration during T-cell-dependent Ab responses [31, 32]. The 7α,25-OHC–EBI2 axis is also involved in the homeostasis, localization, and function of a splenic CD4+ DC subset expressing EBI2. Specifically, 7α,25-OHC guides EBI2+CD4+ DCs to marginal-zone bridging channels [33], where CD4+ DCs interact with blood-borne Ags, thereby promoting T-cell-dependent Ab responses. Some oxysterols (such as 22R-HC, 27-HC, and 24S-HC) are also chemo-attractants for neutrophils, thereby inducing their recruitment within tumor microenvironment and promoting tumor growth [34]. This axis is independent of LXRs and requires the activation of the GPCR CXCR2 [34]. This unexpected activity of oxysterols amplifies the spectrum of biologic functions exerted by these molecules on immune cells and identifies new biologic fields of investigation of immune cells in different pathophysiologic conditions. Immune cells infiltrating the tumor microenvironment may be conditioned by a multitude of factors that are released by tumor cells [35].

Among these factors, we have recently found that LXR ligands are released by human and mouse tumors [36]. The biochemical characterization of tumor-conditioned media from the mouse lymphoma RMA highlighted the presence of two main oxysterol species, namely 22R-HC and 27-HC. These results were in agreement with the expression of Cyp11a1 and Cyp27a1 transcripts by RMA tumor cells, two enzymes responsible for the generation of 22R-HC and 27-HC, respectively [34]. Once produced, oxysterols can activate LXRs in different subsets of immune cells infiltrating the tumor microenvironment. A related critical issue concerns the activation of LXRα and LXRβ isoforms under conditions where both isoforms may be activated.

Pim1 binds to the aminoterminal

Pim1 binds to the aminoterminal TAM Receptor inhibitor transactivation domain of Myc and is

thereby recruited to its target genes, where it mediates histone H3S10 phosphorylation at the Myc-binding site in these loci. In its co-activating role with Myc, Pim1 is required for the expression of one-fifth of all Myc-target genes, a majority of them encoding transcription factors and cell cycle- as well as apoptosis-controlling genes 22. Expression of Pim1 is upregulated upon CD40 signaling in mature B cells 23. Pim1 has been shown to enhance 24 or decrease 25 cell survival, depending on the cellular context. Furthermore, Pim1 is involved in the transition from G2 to M-phase during cell cycle 26. In the present study, we examined the effects of the proto-oncogenes Myc and Pim1 on proliferation and survival of B cells along the pathway of B-cell differentiation from pre-BI cells to the mature, antigen-sensitive stages in vitro and in vivo. Inducible overexpression of the proto-oncogenes Pim1 and Myc was achieved by using the doxycycline-inducible

TetON expression system. cDNAs of Myc, Pim1 and Egfp (control) were integrated into self-inactivating (SIN) retroviral vectors under PF-562271 clinical trial the control of a doxycycline-inducible promoter (Supporting Information Fig. 1A). Two fetal liver-derived pre-BI cell lines were transduced with a vector containing the improved reverse transactivator rtTA-M2 (27, Supporting Information Fig. 1B) and several cell lines thereof were generated. These were then transduced with the EGFP control vector. Concentrations of 1–3 μg/mL doxycycline-induced EGFP expression in transduced pre-BI cells to maximal levels within 1–2 days (Supporting Information Fig. 1C) in 30–60% of the cells, depending on the cell line. The cell lines with the highest inducible potential were used for subsequent transductions with the vectors containing inducible Pim1 and Myc genes. Inducible expression Dichloromethane dehalogenase of

both proto-oncogenes was confirmed on RNA levels in pre-BI cells (Fig. 1A and B) and mature cells (see later), for Myc also on protein expression level (Fig. 1C). The effect of doxycycline-induced expression of Pim1 or Myc alone, as well as Pim1 together with Myc on cell cycle progression of pre-BI cells was tested by staining the pre-B cells with propidium iodide for their DNA content 2 days after removal of IL-7 and, hence, 2 days after the start of differentiation of these pre-BI cells. The results of these analyses show that Pim1 does not influence entry into cell cycle, while overexpression of Myc alone as well as Myc and Pim1 together increase entry into cell cycle approximately to the same extent (Fig. 1D, and Supporting Information Fig. 1D for gating strategy).

The role of CMV infection in acute rejection after renal transpla

The role of CMV infection in acute rejection after renal transplantation remains controversial; several studies have suggested that it can lead to allograft

rejection [6, 7]. Because investigation of strategies for preventing CMV RXDX-106 nmr replication and acute rejection is of ongoing interest [8], we have concentrated on this matter in our series of our studies. Cytomegalovirus, a member of the herpesvirus family, has a large genome which encodes over 65 unique glycoproteins [9]. It is well known that some of the glycoproteins encoded by CMV induce strong immune responses, as do other viral components. Among the glycoproteins gB, one of the most abundant envelope components, is essential for viral replication and considered one of the major target molecules for neutralizing antibodies as well as for cellular immune response [10]. Three linear antibody-binding sites have been described: it is well Fostamatinib clinical trial known that the AD2 site

I epitope of gB is conserved in CMV isolates and is the major epitope for neutralization [9, 11, 12]. The antibody-binding site on AD2 is located between a.a. 28 and 84 of gB [9, 11]. gB is also a target for CMV-specific T-cell immunity. Although little is known about any association between gB AD2 and CMV-specific T-cells, Elkington et al. isolated CD4+ cytotoxic T lymphocytes [13], which recognize epitopes from CMV gB in association with HLA-DR7 and DR11 antigens. In addition to gB, gH has Racecadotril been used to identify preexisting strain-specific

antibodies [14, 15]. Previously, we found that reinfection of seropositive recipients with a different type of CMV is also associated with acute rejection and CMV disease in renal transplant patients [15]. A study which reevaluated the previous study has also indicated that the absence of antibodies against gB in transplantation recipients is a good indicator of CMV disease [16]. In this study, we investigated whether, in addition to CMV disease, antibodies against gB AD2 contribute to prediction of acute rejection in renal transplantation in D + R+ setting, irrespective of gH serological matching. This study investigated 77 CMV seropositive renal transplant recipients whose donors were also CMV seropositive (D + /R+ setting) and in whom antibodies against amino-terminal regions of CMV-gH had been detected; these recipients were enrolled at Fukushima Medical University and Tokyo Women’s Medical University and have been described previously [15]. All study recipients had received hemodialysis treatment before transplantation and had received living-related renal transplants. This study was approved by the Institutional Ethics Committee and written informed consent was obtained from all subjects. All serum specimens were obtained before transplantation. To detect antibody against CMV gB AD2 site I, which is located between a.a.

Members of the S100 family of calcium-binding proteins play essen

Members of the S100 family of calcium-binding proteins play essential roles in epithelial tissues and participate in a wide range of cellular processes, including transcription, proliferation and differentiation.19,20 S100A8, S100A9 and S100A12 are specifically linked to innate immune functions Selleck Ivacaftor by their expression in cells of myeloid origin.21 S100A8 and S100A9

are found in granulocytes, monocytes and the early differentiation stages of macrophages. Their expression can also be induced in keratinocytes and epithelial cells under inflammatory conditions. In contrast, S100A12 is restricted more to granulocytes.22 S100 proteins are related to pro-inflammatory mechanisms and a significant over-expression can be found at sites of inflammation.23 These proteins have been shown to exert their pro-inflammatory activity through receptor for advanced glycation end products (RAGE).24 Interestingly, Gebhardt et al. have demonstrated that RAGE-deficient Idasanutlin mice are resistant to DMBA (7,12-dimethyl-benz[a]anthracene)/TPA (12-O-Tetradecanoylphorbol-13-Acetate)-induced skin carcinogenesis and exhibit a severe defect in sustaining inflammation during the promotion phase, indicating a pivotal role for S100/RAGE in promoting inflammation-induced carcinogenesis.25 S100A8 and S100A9 are reportedly up-regulated in many cancers, including

colon cancer,26 and have been implicated in the regulation of tumour cell proliferation and metastasis. Murine MDSC have been shown to secrete S100A8/A9 proteins and

blocking of the binding of S100A8/A9 to MDSC reduces MDSC levels in blood.27 Multiple suppressor functions still remain as the major hallmark of MDSC. NOS2 and arginase-1 are both strongly expressed in MDSC and have been shown to be responsible for immune suppression. Because S100 is an intracellular protein we could not use this marker for direct isolation of cells followed by functional analysis. Instead, we were able to demonstrate that CD14+ S100A9high but not CD14+ S100A9low cells expressed NOS2 in response to lipopolysaccharide and interferon-γ stimulation, suggesting that S100A9 can specifically identify MDSC and distinguish them from CD14+ HLA-DR+ monocytes. It should be noted that S100 family members are all intracellular Montelukast Sodium proteins, which makes it impossible to use this marker to isolate MDSC and use them in functional studies. Therefore, we were able to provide indirect data indicating that CD14+ S100A9high cells are MDSC. In addition, our data clearly demonstrated a better discrimination between MDSC and non-MDSC when MDSC in whole blood were analysed for S100A9 expression. Therefore, we would suggest using this marker when whole blood is available for the analysis of MDSC. In summary, we describe S100A9, as a new marker in MDSC that can be used to identify CD14+ MDSC. S100A9 can be used instead of or in combination with HLA-DR staining.

, 1986; Walden & Kim, 2005) This suggests that infants are visua

, 1986; Walden & Kim, 2005). This suggests that infants are visually referencing the adult with the appropriate advice and information pertaining to the visual stimulus or event at hand, rather than seeking emotional or physical comfort. The observed increase in vocalizations accompanying the greater number of manual gestures toward the impossible cube may also be interpreted as the preverbal infants’ means of communicating their interest in such a novel and unusual visual display. Recent work examining the spectral frequency of infants’ babbling and utterances has shown that vocalizations may serve as a communicative mechanism

co-occurring with pointing and reaching gestures, which together may convey meaning among preverbal infants (Bernardis et al., 2008). In addition to referencing the two adults in the Adriamycin cost test room, infants may have been trying to communicate learn more their interest or curiosity in the depicted images. Interestingly, we also observed mouthing in some of the infants as an exploratory behavior that occurred

only with the impossible cube display. In addition to haptic exploration, infants between the ages of 6 and 9 months also rely on their mouths as a primary means of exploring the distinct features of objects, such as texture and shape (Ruff, 1984), although this particular behavior tends to wane by the end of the first year as infants expand their repertoire of manual exploration skills (McCall, 1974; Ruff, 1984). In addition to the increased manual exploration efforts among these infants, some also employed mouthing as a final means of determining what the object might be. In our study, infants were more persistent in focusing their exploration and reaching activity on the impossible cube, and this was directly affected by the perception of the incompatible depth relations in the display. Other researchers have also shown that these types of manual exploration activities are purposeful

in ascertaining features, properties, Selleckchem Rucaparib and functions of surfaces and objects, rather than random, haphazard, and indiscriminate motions (Bourgeois et al., 2005; Palmer, 1989; Ruff, 1984). As infants’ fine motor skills improve toward the end of the first year, there is progressive increase in coordinated action and haptic exploration of objects, which simultaneously complements and enhances visual and other sensory input (McCall, 1974; Palmer, 1989). Indeed, the manual action system was directly affected by the depiction of an impossible object. We observed differences in a variety of “whole body” behaviors ranging from more persistent manual gestures to increased social referencing, mouthing, and vocalizations toward the picture of an impossible cube.

As shown in Fig 4, TREM-2-deficient DCs had more I-AbhighCD86hig

As shown in Fig. 4, TREM-2-deficient DCs had more I-AbhighCD86high mature cells than WT DCs after CpG DNA and Zymosan stimulation. Importantly, the maturation level of TREM-2-deficient DCs was very similar to that of DAP12-deficient DCs, suggesting that TREM-2 signaling is mediated by DAP12 in BMDCs. We also compared TREM-2-deficient DCs to those deficient in both DAP12 and FcRγ. Similar to what we found for cytokine production, TREM-2-deficient DCs showed less CpG DNA- and Zymosan-induced maturation than DAP12/FcRγ-deficient DCs. Interestingly,

whereas WT, DAP12-deficient and TREM-2-deficient DCs had a similar amount of maturation in the absence of stimulus, DCs lacking both DAP12 and FcRγ consistently had less learn more basal maturation even though they had the highest amount of stimulus-induced click here maturation (Fig. 4B). In conclusion, these results show that TREM-2/DAP12 signaling negatively regulates DC TLR responses. It has been reported that Siglec-H is involved in the negative regulation of type I IFN responses through DAP12 signaling in plasmacytoid DCs (pDCs) 20, 21.

Though TREM-2 is not expressed in pDCs (Ito and Hamerman, unpublished data), we hypothesized that TREM-2 may inhibit type I IFN production in conventional DCs, such as BMDCs. We assessed IFN-α4 and IFN-β expression by qRT-PCR in WT and TREM-2-deficient BMDCs after CpG DNA stimulation. Expression of mRNAs encoding both type I IFNs analyzed were higher in TREM-2-deficient BMDCs compared with WT BMDCs at 2 and

6 h after stimulation (Fig. 5A and B). As expected, TREM-2-deficient BMDCs also expressed more mRNA encoding IL-12 p40 (il12b) at 2 and 6 h after CpG DNA treatment than WT BMDCs (Fig. 5C). Intriguingly, IRF7 expression was not changed between WT and TREM-2-deficient BMDCs (Fig. 5D). IRF7 is induced by type I IFN stimulation and plays a major role in the positive feedback regulation of type I IFN expression 22, 23. We also measured IFN-β secretion after 16 h of CpG DNA stimulation by ELISA. TREM-2-deficient BMDCs secreted significantly more IFN-β protein than WT BMDCs after CpG DNA stimulation (Fig. 5E). These results suggest that increased type I IFN response in TREM-2-deficient Aldol condensation DCs was due to lack of TREM-2/DAP12 signaling at the primary TLR response phase. In conclusion, these results demonstrate that TREM-2 negatively regulates DC production of type I IFN in addition to IL-12 p70 and TNF in response to TLR ligation. Because TREM-2-deficient BMDCs matured more efficiently than WT BMDCs, we investigated whether the antigen-presenting activity of TREM-2-deficient DCs was higher than that of WT DCs. We co-cultured OVA peptide-pulsed BMDCs in the presence of high (100 nM) and low (25 nM) doses of CpG DNA with CFSE-labeled OT-II TCR transgenic CD4+ T cells for 72 h and detected CFSE dilution of CD4+ T cells by flow cytometry (Fig. 6A).

Developing B cells in the bone marrow express CD25 during the pre

Developing B cells in the bone marrow express CD25 during the pre-B-cell stage [8, 9] but the function of CD25 on these immature B cells is largely unknown as they do not proliferate in response to IL-2 this website [9]. CD25+ B cells in the periphery are today believed to be activated B cells; however, most of these studies are performed in vitro [10] and very little is known about the expression of CD25

on B cells after activation in vivo. The CD25+ B-cell population consists of about 1% of the whole B-cell population in a naïve mouse spleen and previous studies have revealed considerable phenotypical difference between the CD25+ B cells in bone marrow and those present in secondary lymphoid organs [2]. While CD25+ B cells isolated from bone marrow displayed an immature phenotype, CD25+ B cells isolated from secondary lymphoid organs display a more mature and activated phenotype when compared with CD25− B cells characterized by higher expression

of surface IgA and IgG as well as a higher expression of the costimulatory molecules CD80 and CD86 [2]. In addition, we have shown that human circulating CD25+ B cells display different phenotypic and functional properties when compared with the CD25− B cells. CD25+ B cells performed significantly better as antigen-presenting cells in allogeneic mixed lymphocyte reaction (MLR) and B cell–specific blocking of the CD25 expression led to abrogation of the

MLR. CD25+ B cells also expressed significantly higher levels of surface immunoglobulin but lacked the ability to secrete them [3]. Overall, the human CD25+ B cells display a more mature phenotype and seem belong to the Vildagliptin memory B-cell population [4]. The aim of this study has been to analyse the functional properties of CD25+ B cells in mice with respect to immunoglobulin and cytokine production, antigen presentation, migration and homing. Our results clearly show that CD25+ B cells are highly differentiated and might belong to the memory B-cell subset. Mouse strains.  Naval Medical Research Institute (NMRI) and C57BL/6 female mice were used. C57Bl/6 mice were used only in the mixed lymphocyte reaction experiments. Permission from the local animal research ethics committee, in accordance with national animal welfare legislation, was obtained for all the mice experiments. B-cell isolation.  Spleens were passed through a 70-μm nylon mesh (BD Bioscience, Erembodegem, Belgium) into a Petri dish containing 10 ml phosphate-buffered saline (PBS). Cell suspension was centrifuged; the pellet resuspended in NH4Cl solution (0,83%, pH 7.29) and kept on ice for 7 min to lyse erythrocytes, followed by two washing steps in cold PBS. The cells were counted and incubated with optimal concentration of Fc-block (2.4G2; BD Bioscience) for 8 min at room temperature to avoid unspecific binding via Fc-receptor interaction.

The current studies are limited by the fact that the measure of p

The current studies are limited by the fact that the measure of performance, infant looking time, has had only modest success as a measure of individual differences (e.g., Frick & Richards, 2001). It has been used primarily as a group measure that yields a categorical outcome in which performance is either above chance or not different from chance. Our studies therefore suggest that a gender difference in mental rotation ability exists, but may not be especially sensitive to revealing the magnitude of this difference. However, the recent work of Krogh, Moore, and Johnson (2013) suggests that progress may be achieved by examining

individual differences in posthabituation Selleck ACP-196 looking times as a measure of mental rotation performance and correlating them with other measures. Krogh et al. eye-tracked 5-month-olds while they performed a mental rotation task and observed this website that males allotted more visual attention to the top of the stimulus and that higher levels of this top-bias were associated with successful performance; by contrast, female visual attention was distributed more evenly throughout the stimulus and with no relation to performance. Additionally, Krogh et al. reported

a positive relation in females, but not in males, between mental rotation performance and prior tactile manipulation with the three-dimensional object to be presented in the looking time task. Taken together, the findings of Krogh et al. suggest that there may be different determinants of performance for male and female infants in mental rotation tasks. An additional possibility worth exploring in future work is whether Dehydratase males and females might be only quantitatively different, rather than qualitatively different, in their mental rotation abilities. Such a possibility might be manifested if females were found capable of performing at an above-chance level in the mental rotation task, but just needed more time to complete it. It is additionally worth

noting that the procedure used in our second study suggests that the gender difference exists between 3 and 10 months, but is not well-suited to determining whether this difference increases during that time window as has been reported for the time period between childhood and adulthood (Geiser, Lehmann, & Eid, 2008). Additional studies could examine whether a sex difference in infant mental rotation changes in magnitude over time. This research was supported by NIH Grant HD-46526. The authors thank Scott P. Johnson and an anonymous reviewer for helpful comments on the initial submission, and Paige Valeski and Laurie A. Yarzab for their assistance. “
“An abundance of experience with own-race faces and limited to no experience with other-race faces has been associated with better recognition memory for own-race faces in infants, children, and adults.

Mixtures of opsonized Candida in mouse autologous serum (10%) wer

Mixtures of opsonized Candida in mouse autologous serum (10%) were added to 0.2 mL of macrophage suspension. The mixture was incubated for 30 min at 37°C. The percentage of phagocytosis was expressed as the percentage of phagocytosing macrophages in 200 cells counted using an optical microscope (15). Alveolar and peritoneal macrophages SB525334 cost monolayers were prepared as described above. In order to determinate the influence of lactobacilli on the capacity of macrophages to produce cytokines, alveolar and peritoneal macrophages were challenged in vitro with heat-killed C.

albicans AV4 at a concentration of 107 cells/mL. After incubation at 37°C in 5% CO2, the supernatant was recovered and kept frozen until cytokine analyses.

IL-1β and TNF-α were determined using the corresponding mouse ELISA kits Vemurafenib (R & D Systems). In order to evaluate the influence of lactobacilli treatments on the immune response against C. albicans in vivo, challenges with pathogenic C. albicans AV4 were performed. Yeast cells were grown in Sabouraud broth at 37°C until the log phase was reached. The pathogens were harvested by centrifugation at 3600 g for 10 min at 4°C and washed three times with sterile PBS. Intraperitoneal challenge with C. albicans AV4 was performed on the day after the end of each Lactobacillus treatment (third or sixth days). The mice were challenged with injections of 200 μL of an inoculum containing 108 cells. For yeast cell counts in blood, liver and spleen, mice were killed on day 2 post-infection. The livers and spleens were excised, weighed and homogenized in 5 mL of sterile peptone water. The homogenates were diluted appropriately, plated in duplicate on Sabouraud agar and MRIP incubated at 37°C. C. albicans colonies were counted and the results expressed as log10 CFU/g of organ or mL of blood. Intranasal challenge

with C. albicans AV4 was performed on the day after the end of each Lactobacillus treatment (third or sixth days). The mice were challenged nasally with the pathogen by dripping 25 μL of an inoculum containing 107 cells into each nostril. To facilitate migration of the inoculum to the alveoli, the mice were held in a head-up vertical position for 2 min. For yeast cell counts in lung and blood, mice were killed on day 2 post-infection. The lungs were excised, weighed and homogenized in 5 mL of sterile peptone water. The homogenates were diluted appropriately, plated in duplicate on Sabouraud agar and incubated at 37°C. The C. albicans colonies were counted and the results expressed as log10 CFU/g of organ or ml of blood. In order to evaluate innate immune responses after challenges, the concentrations of TNF-α and IFN-γ and the number of leukocytes and neutrophils were determined in BAL and peritoneal fluid according to techniques described in a previous report (15).