Differences In Inflammatory Response Research Articles

P.1.d.002 - Differential microglial inflammatory responses in the spinal cord and brain towards chronic neuropathic pain in rats

P.1.d.002 - Differential microglial inflammatory responses in the spinal cord and brain towards chronic neuropathic pain in rats

Abstract PR04: Inflammatory responses within the lung tumor microenvironment correlate with oncogenic mutation and histologic subtype

Abstract Lung cancer is a heterogeneous disease comprised of multiple histologic subtypes that harbor disparate mutational landscapes. Small cell lung carcinoma (SCLC), in which RB1 and TP53 mutations are common, accounts for 10-20% of lung cancer diagnoses. Approximately half of non-small cell lung carcinoma cases are classified as lung adenocarcinomas (ADCA), in which KRAS, EGFR, and TP53 mutations are the predominant genetic drivers. Novel immunotherapeutic strategies have offered new hope for the management of ADCA and SCLC, but the clinical success of immunomodulatory agents will depend on a strong foundational knowledge of the cells that comprise the lung tumor microenvironment (TME) in these molecularly and histologically distinct diseases. We therefore sought to determine whether the host immune response to lung cancer is predicated, at least in part, by histologic and genetic differences, as such correlations would have important clinical ramifications. Using three mouse models of lung ADCA KrasLSL-G12D, KrasLSL-G12D;Trp53Fl/Fl, and EgfrL858R as well as the Rb1Fl/Fl;Trp53Fl/Fl model of SCLC, we show that SCLC and ADCA tumors exhibit unique immune cell composition. Lung specimens from tumor-bearing and control animals (n ≥ 5 per cohort) were harvested at multiple time points and subjected to histological assessment and comprehensive flow cytometric immunophenotyping. Tumor-associated inflammation was discernibly lower in SCLC compared to all ADCA models. Moreover, the leukocyte composition of SCLC was dominated by cells of the adaptive rather than innate arm of the host immune system, a finding subsequently validated in 2 human SCLC surgical specimens. We further identified key differences in leukocyte content of mouse ADCA that correlated with oncogenic mutation. Although Egfr-mutant cancers displayed robust myeloid cell recruitment, they failed to mount a CD8+ cellular immune response. In contrast, Kras-mutant tumors displayed significant expansion of multiple immune cell types, including CD8+ cells, regulatory T cells, IL17-producing lymphocytes, and myeloid cells. Although loss of Trp53 promoted malignancy, it had minimal effect on immune cell composition within the Kras TME. Pre-clinical in vivo investigations with immune checkpoint inhibitor compounds are currently being conducted to assess the importance of these distinct immune responses to tumor progression and survival. To determine whether the differential inflammatory responses observed in mice are clinically relevant, we conducted an immune profiling study of a large lung cancer patient cohort that included 55 ADCA specimens. Notably, patients with EGFR mutations (n = 5) exhibited fewer total CD8+ T cells, as well as decreased exhausted CD8+PD1+ and CD8+TIM3+ content, compared to all other ADCA cases, thereby substantiating the murine phenotype. Taken together, these findings establish a strong foundational knowledge of the immune cell contexture of lung ADCA and SCLC and suggest that molecular and histological traits shape the host immune response to cancer. This abstract is also presented as Poster A01. Citation Format: Stephanie E. Busch, Julia Kargl, Mark L. Hanke, Heather E. Metz, Kyoung-Hee Kim, A McGarry Houghton. Inflammatory responses within the lung tumor microenvironment correlate with oncogenic mutation and histologic subtype. [abstract]. In: Proceedings of the AACR Special Conference: Function of Tumor Microenvironment in Cancer Progression; 2016 Jan 7–10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2016;76(15 Suppl):Abstract nr PR04.

Fetal sex is associated with maternal stimulated cytokine production, but not serum cytokine levels, in human pregnancy

Some studies suggest that fetal sex plays a role in maternal physiological processes during pregnancy including glycemic control, blood pressure, and cortisol regulation. However, data examining fetal sex-specific differences in maternal immune parameters is lacking. In the current study, serum levels of interleukin(IL)-6, IL-8, and tumor necrosis factor(TNF)-α as well as LPS-stimulated production of IL-6, IL-8, TNF-α, and IL-1β by PBMCs incubated for 24h were assessed in early, mid, and late pregnancy among 80 women (46 with male and 34 with female fetuses). Linear mixed models showed that women carrying females versus males exhibited greater stimulated production of IL-6 at each timepoint (ps⩽0.03), TNF-α in early pregnancy (p=0.04), and IL-1β in mid- and late pregnancy (ps⩽0.05). Despite changes in serum levels of IL-8 (p=0.002) and TNF-α (p<0.0001) across pregnancy, no differences in any serum cytokines were observed in relation to fetal sex (ps>0.85). In conclusion, in pregnant women, those carrying female versus male fetuses exhibited greater stimulated cytokine production across pregnancy. Differential inflammatory responses could affect maternal health and fetal development. Fetal sex should be considered as a factor in studies of maternal inflammation. These findings have relevance both clinically and conceptually. For example, maternal asthma is exacerbated among women carrying female versus male fetuses. In addition, data on associations between fetal sex and maternal immune function among women with health conditions (e.g., preeclampsia) and adverse pregnancy outcomes (e.g., preterm birth) would be informative.

Elevated CXC chemokines in urine noninvasively discriminate OAB from UTI.

Overlapping symptoms of overactive bladder (OAB) and urinary tract infection (UTI) often complicate the diagnosis and contribute to overprescription of antibiotics. Inflammatory response is a shared characteristic of both UTI and OAB and here we hypothesized that molecular differences in inflammatory response seen in urine can help discriminate OAB from UTI. Subjects in the age range of (20-88 yr) of either sex were recruited for this urine analysis study. Urine specimens were available from 62 UTI patients with positive dipstick test before antibiotic treatment. Six of these patients also provided urine after completion of antibiotic treatment. Subjects in cohorts of OAB (n = 59) and asymptomatic controls (n = 26) were negative for dipstick test. Urinary chemokines were measured by MILLIPLEX MAP Human Cytokine/Chemokine Immunoassay and their association with UTI and OAB was determined by univariate and multivariate statistics. Significant elevation of CXCL-1, CXCL-8 (IL-8), and CXCL-10 together with reduced levels for a receptor antagonist of IL-1A (sIL-1RA) were seen in UTI relative to OAB and asymptomatic controls. Elevated CXCL-1 urine levels predicted UTI with odds ratio of 1.018 and showed a specificity of 80.77% and sensitivity of 59.68%. Postantibiotic treatment, reduction was seen in all CXC chemokines with a significant reduction for CXCL-10. Strong association of CXCL-1 and CXCL-10 for UTI over OAB indicates mechanistic differences in signaling pathways driving inflammation secondary of infection in UTI compared with a lack of infection in OAB. Urinary chemokines highlight molecular differences in the paracrine signaling driving the overlapping symptoms of UTI and OAB.

Surgical Trauma Caused by Different Abdominal Access Routes-Comparison of Open Surgical, Laparoscopic, and NOTES Transgastric Techniques in a Porcine Model.

Investigations indicate that natural orifice translumenal endoscopic surgery (NOTES) procedures induce a less pronounced postoperative inflammatory response than open or laparoscopic surgery, inflicting less trauma. In NOTES procedures, no skin incision is performed. We compare the inflammatory response added by the type of incision by measuring C-reactive protein (CRP) and tumors necrosis factor-alfa (TNF-α). Twenty-seven pigs were randomized to open surgical, laparoscopic, or transgastric NOTES abdominal access. After completion of the accesses, no surgery was performed. All accesses were left open for 40 minutes followed by closure, animals were survived for 7 days. Blood samples were drawn at the start of the accesses, at 20 and 40 minutes during the procedure, and at postoperative day (POD) 1, 3, and 7. Analyses of CRP and TNF-α were performed. CRP increased in all animals until POD1. This increase was greater in the open group (P = .006). No significant differences in CRP-levels were found at POD 1, 3, or 7. TNF-α showed a peak during the procedure, at 20 and 40 minutes, with normalization at POD1 for 1/3 of the open and laparoscopic animals, but not for the NOTES animals. Due to variations within the groups, no statistical difference was shown between them. At postmortem, 1/3 of the pigs in the laparoscopic and open groups had wound infections, while no NOTES animals showed infections. This study provides no statistically significant differences in inflammatory response after the different abdominal accesses. However, the lack of a TNF-α-peak in the NOTES group might indicate a less pronounced response, supporting the initial theories.

Activation of human keratinocytes by Leishmania spp.: divergent effects of Leishmania infantum versus Leishmania major.

Abstract All Leishmania infections are initiated in skin, but clinical manifestations vary greatly with infecting species. Leishmania major (Lm) causes localized cutaneous lesions, whereas L. infantum (Li) causes potentially fatal disseminated disease without skin pathology. Early responses at the skin infection site influence adaptive responses to Leishmania, yet little is known of the role of keratinocytes. We hypothesized that Leishmania induce keratinocytes to produce factors influencing the immune response. Incubation of Li with primary or immortalized human keratinocytes caused a significant increase in pro-inflammatory cytokine transcripts il6, il8, tnfa, and il1b measured by RT-qPCR. However, keratinocytes exposed to five distinct Lm isolates did not induce these transcripts, highlighting a species-specific difference in inflammatory response. Similar to live parasites, Li-derived exosomes induced more il8 mRNA compared to control (p&amp;lt;0.01) or Lm-derived exosomes (p&amp;lt;0.05). Western blotting confirmed NFkB-p65 activation in keratinocytes exposed to Li but not Lm. To examine whether soluble keratinocyte factors influence nearby immune cells, Li-infected human monocytes were cocultured with keratinocytes across a transwell membrane. Soluble products of Li-exposed keratinocytes improved monocyte control of parasite replication compared with unexposed controls (p&amp;lt;0.01). However, culture with Lm-exposed keratinocytes across the transwell did not affect monocyte Lm infection. These data suggest that (1) activated keratinocytes may increase monocyte leishmanicidal activity and (2) keratinocytes support an early inflammatory environment, uniquely tailored to each Leishmania species, at the infection site.

Vitamin D increases the human endosomal TLR response against Mycobacterium tuberculosis nucleic acids

Abstract Mycobacterium tuberculosis (Mtb) infects macrophages where it avoids elimination by interfering with host defense mechanisms. Type I IFNs are produced by activation of endosomal TLRs after recognition of nucleic acids. TLR importance is underlined by the fact that the TLR-adaptor molecule MyD88 pathway in macrophages is important in confining Mtb within phagolysosomes, mycobacterial clearance, and an adequate innate and adaptive immune response against Mtb. Vitamin D (VD) has a positive effect in macrophage activation, phagocytosis, restriction of Mtb growth, and killing of the mycobacteria. We aimed to characterize the endosomal TLR-signaling of human macrophages, as they are appear crucial in determining the fate of the disease, and explore if variations in Type I IFN responses correlate with differences in Mtb nucleic acids derived from different Mtb genotypes. Stimulation with DNA from Mtb of different lineages elicited differential NF-κB and IRF inflammatory responses in THP-1 derived macrophages. We also studied the effect of VD on these responses, as it is known to activate macrophages and enhance their antimycobacterial activity. Pretreatment with VD increased the response to DNA from HN878 and virulent strain H37Rv. It also enhanced the response to RNA from M. smegmatis and attenuated M. bovis strain BCG. No previous study has evaluated Type I or Type II IFNs response differences between different Mtb strains. This is of great importance as type I IFNs appear to strongly inhibit macrophages responsiveness to IFN-γ, and are associated with active TB. Understanding VD-mediated activation occurring at the pulmonary alveoli will impact the design of novel therapeutic and successful immunization strategies.

Streptococcus uberis ST439 and ST475 induce differential inflammatory responses in a mouse intramammary infection model

Streptococcus uberis causing mastitis is a growing challenge to the dairy industry. Molecular, epidemiological and population structure studies have revealed clonal diversity among the infecting strains. In this study, mouse intramammary infection model was used to uncover the host immune response to two epidemiologically important live strains of S. uberis (SU1and SU2) obtained from subclinical case of mastitis possessing specific and unique multi locus sequence types (ST), pulsed field gel electrophoresis (PFGE) pulsotypes and virulence profiles. Temporal (2h, 4h, 8h, 12h, 24h and 48h) expression of key inflammatory mediators (IL2, IL4, IL6, IL12, TNFα, IFNγ, GMCSF, TLR2, TLR4, TLR9, TLR11, TLR12, CD14, IL1β, RANTES, Lactoferrin, and CXCl1) by reverse transcription and probe-based quantitative real-time PCR showed relative mRNA levels higher (p<0.05) in response to SU2 compared with SU1 with 24h PI serving as a critical point for the deviating behavior (SU1 versus SU2). Further employing the predicted biological processes under the influence of this pool of tested genes, the delineation of gene regulatory networks suggested SU1−favoring its persistence in the host environment; in contrast, SU2−which elevated gene expression indicating towards pathogen clearance or immune surveillance. This study suggested how these unique strains could manipulate the host immune response to influence the severity of mastitis; our results expand the available information on host pathogen interaction and provide a firm foundation needing further investigations to gain control over this pathogen.

FasL expression in cardiomyocytes activates the ERK1/2 pathway, leading to dilated cardiomyopathy and advanced heart failure.

Increase in the apoptotic molecule Fas ligand (FasL) in serum and cardiomyocytes has been shown to be associated with progressive dilated cardiomyopathy (DCM) and congestive heart failure (CHF) in humans. However, the underlying mechanism(s) of FasL-related deterioration of heart function remain obscure. The aim of the present study is to determine roles of myocardial FasL in the activation of alternative pathways such as extracellular-signal-regulated kinase 1/2 (ERK1/2), inflammation or fibrosis and to identify effective treatments of progressive DCM and advanced CHF. Transgenic mice with cardiomyocyte-specific overexpression of FasL were investigated and treated with an ERK1/2 inhibitor (U-0126), losartan (los), prednisolone (pred) or placebo. Morpho-histological and molecular studies were subsequently performed. FasL mice showed significantly higher mortality compared with wild-type (WT) littermates due to DCM and advanced CHF. Prominent perivascular and interstitial fibrosis, increased interleukin secretion and diffuse CD3-positive cell infiltration were evident in FasL hearts. Up-regulation of the short form of Fas-associated death domain (FADD)-like interleukin 1β-converting enzyme (FLICE) inhibitory protein (s-FLIP), RIP (receptor-interacting protein) and ERK1/2 and down-regulation of transforming growth factor beta 1 (TGFβ1) and nuclear factor-κB (NF-κB) was determined in the myocardium, whereas expression of ERK1/2, periostin (Postn) and osteopontin increased in cardiac fibroblasts. U-0126 and los increased CHF survival by 75% compared with pred and placebo groups. U-0126 had both anti-fibrotic and anti-apoptotic effects, whereas los reduced fibrosis only. Myocardial FasL expression in mice activates differential robust fibrotic, apoptotic and inflammatory responses via ERK1/2 in cardiomyocytes and cardiac fibroblasts inducing DCM and CHF. Blocking the ERK1/2 pathway prevented progression of FasL-induced DCM and CHF by reducing fibrosis, inflammation and apoptosis in the myocardium.

Histological evaluation of mineral trioxide aggregate and enamel matrix derivative combination in direct pulp capping: An in vivo study.

Aim:The aim of this study is to evaluate the response of human pulp tissue to mineral trioxide aggregate (MTA), Emdogain (EMD), and combination of MTA/EMD.Materials and Methods:This study was performed on sixty intact first and second premolars of human maxillary and mandibular teeth. A standard pulpal exposure was done on all the teeth and was divided into three groups of twenty teeth each and was capped with MTA, EMD, and MTA/EMD combination. The final restoration was done with resin-modified glass ionomer cement. The teeth were then extracted on the 15th or 45th day and histological evaluation done.Results:Differences in inflammatory response and thickness of dentin bridge formation of the exposed pulp to the three different groups were statistically evaluated using Chi-square and Mann–Whitney tests and were found to be significant. No significant difference was found between MTA/EMD and MTA in terms of calcified bridge formation and pulp inflammatory response to the capping materials.Conclusions:MTA and MTA/EMD combination produced a better quality hard tissue response compared with the use of EMD.

A randomized controlled trial on the effects of goal-directed therapy on the inflammatory response open abdominal aortic aneurysm repair.

IntroductionGoal-directed therapy (GDT) has been shown in numerous studies to decrease perioperative morbidity and mortality. The mechanism of benefit of GDT, however, has not been clearly elucidated. Targeted resuscitation of the vascular endothelium with GDT might alter the postoperative inflammatory response and be responsible for the decreased complications with this therapy.MethodsThis trial was registered at ClinicalTrials.gov as NCT01681251. Forty patients undergoing elective open repair of their abdominal aortic aneurysm, 18 years of age and older, were randomized to an interventional arm with GDT targeting stroke volume variation with an arterial pulse contour cardiac output monitor, or control, where fluid therapy was administered at the discretion of the attending anesthesiologist. We measured levels of several inflammatory cytokines (C-reactive protein, Pentraxin 3, suppressor of tumorgenicity--2, interleukin-1 receptor antagonist, and tumor necrosis factor receptor-III) preoperatively and at several postoperative time points to determine if there was a difference in inflammatory response. We also assessed each group for a composite of postoperative complications.ResultsTwenty patients were randomized to GDT and twenty were randomized to control. Length of stay was not different between groups. Intervention patients received less crystalloid and more colloid. At the end of the study, intervention patients had a higher cardiac index (3.4 ± 0.5 vs. 2.5 ± 0.7 l/minute per m2, p < 0.01) and stroke volume index (50.1 ± 7.4 vs. 38.1 ± 9.8 ml/m2, p < 0.01) than controls. There were significantly fewer complications in the intervention than control group (28 vs. 12, p = 0.02). The length of hospital and ICU stay did not differ between groups. There was no difference in the levels of inflammatory cytokines between groups.ConclusionsDespite being associated with fewer complications and improved hemodynamics, there was no difference in the inflammatory response of patients treated with GDT. This suggests that the clinical benefit of GDT occurs in spite of a similar inflammatory burden. Further work needs to be performed to delineate the mechanism of benefit of GDT.Trial registrationClinicalTrials.gov Identifier: NCT01681251. Registered 18 May 2011.

Progression of inflammation during immunodeficient mouse skeletal muscle regeneration.

The skeletal muscle injury triggers the inflammatory response which is crucial for damaged muscle fiber degradation and satellite cell activation. Immunodeficient mice are often used as a model to study the myogenic potential of transplanted human stem cells. Therefore, it is crucial to elucidate whether such model truly reflects processes occurring under physiological conditions. To answer this question we compared skeletal muscle regeneration of BALB/c, i.e. animals producing all types of inflammatory cells, and SCID mice. Results of our study documented that initial stages of muscles regeneration in both strains of mice were comparable. However, lower number of mononucleated cells was noticed in regenerating SCID mouse muscles. Significant differences in the number of CD14-/CD45+ and CD14+/CD45+ cells between BALB/c and SCID muscles were also observed. In addition, we found important differences in M1 and M2 macrophage levels of BALB/c and SCID mouse muscles identified by CD68 and CD163 markers. Thus, our data show that differences in inflammatory response during muscle regeneration, were not translated into significant modifications in muscle regeneration.

Brain large artery inflammation associated with HIV and large artery remodeling.

To test the hypothesis that brain arteries from HIV+ cases have a greater degree of inflammation than brain arteries from HIV- cases, and that inflammation is associated with brain arterial remodeling. Case-control study, cross-sectional. Brain arteries from 162 autopsy cases (84 with HIV) were systematically analyzed for thickness of the intima, media, and adventitia, and atherosclerosis and dolichoectasia. Inflammation was assessed with CD68 immunohistochemistry, and measured with a semiquantitative score reflecting the number and location (i.e., arterial layer) of activated macrophages infiltrating the arterial wall. Latent varicella zoster virus (VZV) was assessed with anti-VZV gene 63 product immunohistochemistry. Demographic and clinical variables were available in all cases, and longitudinal data about CD4 cell counts were available among cases with HIV. Multilevel generalized linear models were used to test the association between inflammation and HIV. Arteries from HIV+ cases had a higher inflammation score (B = 0.36, P = 0.05) compared with arteries from HIV- cases, although the association was attenuated after controlling for demographic variables, vascular risk factors, and latent VZV (B = 0.20, P = 0.18). Although intimal inflammation was similar in cases with and without HIV, adventitial inflammation was associated with HIV. Intimal inflammation was associated with intracranial atherosclerosis independent of HIV status, but adventitial inflammation was associated with HIV-associated dolichoectasia in arteries with a thin media. Adventitial inflammation is associated with HIV and dolichoectasia independent of intracranial atherosclerosis. This suggests that differential inflammatory responses may play a role in intracranial atherosclerosis and dolichoectasia.

Differential microglial inflammatory responses in the spinal cord and brain towards the chronic neuropathic pain in rats

Accumulating evidences suggest the involvement of immune cells, especially microglia, in development of the central nervous system and persistence of neuropathic pain after peripheral nerve injuries in adulthood, but the exact roles that microglia play are unclear. In this study, by utilizing an efficient and reproducible multicolor flow cytometry-based approach, we quantitatively evaluated both microgliosis and microglial activation in the lumbar spinal cord (SC), amygdala and prefrontal cortex (PFC) of adult male Hannover-Wistar rats in response to a chronic neuropathic pain induced by ligation of the tibial and common peroneal nerves (known as the spared nerve injury (SNI) model). When comparing the SNI and sham-operated rats, we observed increased microgliosis in the ipsi-lateral lumbar SC on both post-operative day (POD) 7 and 21, and a milder increase in the contralateral side of SC on POD21. Regarding microglial activation, we unexpectedly found increased numbers of the anti-inflammatory type of microglia (CD172a+MHCII-/+) in the ipsilateral lumbar SC but an opposite change in the contra-lateral PFC on POD21. Moreover, these inflammatory responses were accompanied by the mechanical allodynia and anxiety-related behaviors of the SNI rats. Our results suggest that microglia in the SC and brain are differentially activated by the spinal nerve injury, and that microglial responses in the SC do not necessarily develop into a proinflammatory prototype as many literature suggested.

Age-dependent alterations in the inflammatory response to pulmonary challenge.

The aging lung is increasingly susceptible to infectious disease. Changes in pulmonary physiology and function are common in older populations, and in those older than 60 years, pneumonia is the major cause of infectious death. Understanding age-related changes in the innate and adaptive immune systems, and how they affect both pulmonary and systemic responses to pulmonary challenge are critical to the development of novel therapeutic strategies for the treatment of the elderly patient. In this observational study, we examined age-associated differences in inflammatory responses to pulmonary challenge with cell wall components from Gram-positive bacteria. Thus, male Sprague–Dawley rats, aged 6 months or greater than 18 months (approximating humans of 20 and 55–65 years), were challenged, intratracheally, with lipoteichoic acid and peptidoglycan. Cellular and cytokine evaluations were performed on both bronchoalveolar lavage fluid (BAL) and plasma, 24 h post-challenge. The plasma concentration of free thyroxine, a marker of severity in non-thyroidal illness, was also evaluated. The older animals had an increased chemotactic gradient in favor of the airspaces, which was associated with a greater accumulation of neutrophils and protein. Furthermore, macrophage migration inhibitory factor (MIF), an inflammatory mediator and putative biomarker in acute lung injury, was increased in both the plasma and BAL of the older, but not young animals. Conversely, plasma free thyroxine, a natural inhibitor of MIF, was decreased in the older animals. These findings identify age-associated inflammatory/metabolic changes following pulmonary challenge that it may be possible to manipulate to improve outcome in the older, critically ill patient.

Ketogenic Diet Impairs FGF21 Signaling and Promotes Differential Inflammatory Responses in the Liver and White Adipose Tissue

Background/HypothesisBeside its beneficial effects on weight loss, ketogenic diet (KD) causes dyslipidemia, a pro-inflammatory state involved in the development of hepatic steatosis, glucose intolerance and insulin resistance, although the latter is still being debated. Additionally, KD is known to increase fibroblast growth factor 21 (FGF21) plasma levels. However, FGF21 cannot initiate its beneficial actions on metabolism in these conditions. We therefore hypothesized and tested in the present study that KD may impair FGF21 signaling.Methods/ResultsUsing indirect calorimetry, we found that KD-fed mice exhibited higher energy expenditure than regular chow (RC)-fed mice associated with increased Ucp1 levels in white adipose tissue (WAT), along with increased plasma FGF21 levels. We then assessed the effect of KD on FGF21 signaling in both the liver and WAT. We found that Fgfr4 and Klb (β-klotho) were downregulated in the liver, while Fgfr1 was downregulated in WAT of KD-fed mice. Because inflammation could be one of the mechanisms linking KD to impaired FGF21 signaling, we measured the expression levels of inflammatory markers and macrophage accumulation in WAT and liver and found an increased inflammation and macrophage accumulation in the liver, but surprisingly, a reduction of inflammation in WAT.We also showed that KD enhances lipid accumulation in the liver, which may explain hepatic inflammation and impaired Fgfr4 and Klb expression. In contrast, import of lipids from the circulation was significantly reduced in WAT of KD-fed mice, as suggested by a downregulation of Lpl and Cd36. This was further associated with reduced inflammation in WAT.ConclusionAltogether, these results indicate that KD could be beneficial for a given tissue but deleterious for another.

Early Life Lung Antioxidant Levels and Response to Ozone: Influence of Sex and Maturation in Wistar Rats

Epidemiologic studies of air pollution effects on respiratory health report significant modification by sex. Studies of children suggest stronger effects among boys in early life and girls in later childhood. In adults, particularly the elderly, studies report stronger effects among women. Differential inflammatory or antioxidant responses are purported. Herein we examined sex or maturation influences in prepubertal Wistar rats [a strain prone to ozone (O3)‐induced lung injury/inflammation] on (A) lung antioxidant and related enzyme levels and (B) antioxidant response to O3. Female (F) and male (M) 14, 21, and 28 d (pre‐, at, post‐weaning) pups were exposed to air, 0.5 or 1 ppm O3 x 2h. In air controls, no sex‐based differences in lung antioxidant levels were observed. From 14‐28 d, weight increased (F: 1.7‐2.7‐fold; M: 1.7‐3.1‐fold); with M&gt;F at 28 d. Likewise, lung uric acid increased (1.1‐1.2‐fold), as did ascorbic acid (1.4‐2.7‐fold), glutathione (GSH) peroxidase (1.0‐3.5‐fold), GSH transferase (1.0‐2.9‐fold), GSH reductase (1.2‐4.2‐fold), and G‐6‐PDH (10‐fold) (per gwet wt). Conversely, SOD declined 30‐42%. Lung GSH was relatively stable. At 0h, 1 ppm O3‐exposed F 14 d pups had decreased GSH and related enzyme activity. At 21 d, exposure was without effect on GSH; and males showed O3‐induced increases in uric acid. At 28 d, no antioxidant changes were apparent. Together, data suggest health risk due to early life pollutant exposure is greatest in very immature females. With maturity, especially post‐weaning, rat lung antioxidant reserve increases, thus reducing susceptibility to pollutant‐induced oxidative stress. (Abstract does not reflect USEPA policy)

C-reactive protein response to influenza vaccination as a model of mild inflammatory stimulation in the Philippines

BackgroundC-reactive protein (CRP) is increasingly measured as a marker of systemic inflammation that predicts elevated risk for cardiovascular disease. Influenza vaccination is a mild pro-inflammatory stimulus, and the CRP response to vaccination may provide additional information on individual differences in inflammatory response and risk for disease. AimTo document the pattern of CRP response to influenza vaccination among a large sample of older women in the Philippines. The Philippines exemplifies current global trends toward increasing rates of overweight/obesity, but also maintains relatively high rates of infectious disease. The secondary aim of the study is to investigate the impact of infectious symptoms on the pattern of response to vaccination. MethodsA community-based sample of 934 women (mean age=55.4 years) received the influenza vaccine. CRP was assessed at baseline and 72h post-vaccination. Descriptive, non-parametric, and parametric analyses were implemented to assess the magnitude of CRP response, and to investigate whether responses were associated with baseline CRP or the presence of infectious symptoms prior to vaccination. ResultsInfluenza vaccination resulted in a statistically significant CRP response of 0.35mg/L (p<0.001), representing a 30.2% increase from baseline. For individuals with symptoms of infectious disease at baseline, the CRP response was smaller (12.9%) and not statistically significant (p=0.77). Lower CRP at baseline was associated with larger CRP response to vaccination in the entire sample, and among participants without recent symptoms of infection. ConclusionsInfluenza vaccination produces a mild CRP response in the Philippines. This study extends prior research in US and European populations validating influenza vaccination as an in vivo model for investigating the dynamics of inflammation, but also raises potential complications in settings where rates of infectious disease are elevated.

Association of the Infant Gastrointestinal Microbiome with Nocturnal Symptoms in Children with Asthma

E S D A Y 887 Human Rhinovirus Species Induce Differential Antiviral and Inflammatory Responses in Peripheral Blood Mononuclear Cells Alalia W. Berry, MD, Megan Yanny, Mark DeVries, Yury Bochkov, PhD, Victoria Rajamanickam, James E. Gern, MD FAAAAI, Robert F. Lemanske, Jr, MD, FAAAAI, Daniel J. Jackson, MD; University of Wisconsin School of Medicine and Public Health, University of Wisconsin, University of Wisconsin School of Medicine and Public Health, Madison, WI, University of Wisconsin Hospitals and Clinics, Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, WI. RATIONALE: Human rhinoviruses (RV) are a common cause of asthma exacerbations in children. RV-C causes more severe respiratory illnesses than RV-A or RV-B. However, the mechanism(s) underlying this observation remain unclear. Prior studies have shown that children with allergic asthma have impaired antiviral responses in peripheral blood mononuclear cells (PBMCs). We hypothesize that RV-C would induce differential antiviral responses in PBMCs when compared to RV-A and RV-B. METHODS: PBMCs were isolated from peripheral blood samples of 40 children ages 13-14 years enrolled in the Childhood Origins of ASThma (COAST study). PBMCs were incubated with RV-A36, RV-B52, RV-C15 or medium alone for 22 hours. Supernatants were frozen at -80 and interferons (IFN-alpha2, IFN-gamma, IFN-lambda1) were measured using multiplex ELISA. Interferon levels induced by the different RV species were compared. RESULTS: Stimulation with RV-A led to significantly greater production of IFN-alpha2 and IFN-lambda1 compared to RV-B, RV-C, or control [(median IFN-alpha2: RV-A 1000 pg/ml, RV-B 4 pg/ml, RV-C <3.2 pg/ml, control <3.2 pg/ml), p<0.0001; (median IFN-lambda1: RV-A 142 pg/ml, RV-B <12.2 pg/ml, RV-C <12.2 pg/ml, control <12.2 pg/ml, p<0.0001]. All RV species increased production of IFN-gamma compared to control, with RV-C leading to the least IFN-gamma [(median IFN-gamma: RV-A 55 pg/ml, RV-B 15 pg/ml, RV-C 6 pg/ml, control <3.2 pg/ml), p<0.0001)]. CONCLUSIONS: PBMCs stimulated with RV-C15 secreted significantly less interferon compared to cells stimulated with other RV species. RV-C may evade the antiviral response of mononuclear cells in the airway, which could contribute to greater illness severity.

Human Rhinovirus Species Induce Differential Antiviral and Inflammatory Responses in Peripheral Blood Mononuclear Cells

Human Rhinovirus Species Induce Differential Antiviral and Inflammatory Responses in Peripheral Blood Mononuclear Cells