Differences In Immune Response Research Articles

Differential immune response between fundic and pyloric abomasal regions upon experimental ovine infection with Haemonchus contortus

The effect of experimental haemonchosis on the number of tissue eosinophils, plasma cells and lymphocyte subpopulations was evaluated in the fundic abomasal region, the pyloric abomasal region and the abomasal lymph node of Blackbelly lambs, which are resistant to infection, and Columbia lambs, which are susceptible to infection. An increase in the number of tissue eosinophils and CD4+ and WC1(+)γδ T-cells was observed in the pyloric abomasal region of Blackbelly lambs and correlated with lower worm burden and greater resistance to infection. Increases in IgA+ plasma cells from the pyloric abomasal region were observed in both infected groups, but there was no difference between the groups. Therefore, increases in IgA+ plasma cells did not explain the resistance observed. Infection caused a significant increase in tissue eosinophils in the abomasal lymph node of Blackbelly lambs and a decrease in the number of CD4+ T-cells in lambs of both breeds. CD8+ T-cells and IgG+ and IgM+ plasma cells were not associated with either infection or resistance. In this work, clear differences were observed in the numbers of CD4+ and WC1(+)γδ T-cells, tissue eosinophils and IgA+ plasma cells between the abomasal regions studied. These differences indicate that the immunological response is not homogenous in all abomasal mucosa and that evaluating the response from a single abomasal region may not be representative of the cellular response across the abomasum.

Host immune-related gene responses against highly pathogenic avian influenza virus infection in vitro differ among chicken cell lines established from different organs

Highly pathogenic avian influenza virus (HPAIV) induces acute disease in chickens causing high mortality and morbidity and is a major threat to poultry industries in Southeast Asian countries. The mechanisms of disease manifestation and host innate immune responses against HAPIV in chickens are not well understood. In this study, we examined virus replication and host gene expressions in four chicken cell lines in vitro to elucidate the impact of host innate immune responses against viral replication. It was demonstrated that viral replication efficiencies were different depending on the cell line. The viral replication appeared to be affected by the basal expression of IFN related genes. The expression of immune-related genes against the viral infection also varied in a cell line dependent manner. In non-immune derived cell lines, but not in immune derived cell lines, the expression of the CCL5 and CCL20 genes were induced by HPAIV infection. Reverse genetics HPAIV, with internal genes from avirulent avian influenza, reduced virus replication and affected immune-related gene expression in a cell line dependent manner. These results suggest the possibility that differential immune responses in different cell types in local tissues could modulate the consequences of HPAIV infection in chickens.

Differential immune response of rainbow trout ( Oncorhynchus mykiss) at early developmental stages (larvae and fry) against the bacterial pathogen Yersinia ruckeri

Innate immune factors play a crucial role in survival of young fish especially during early stages of life when adaptive immunity is not fully developed. In the present study, we investigated the immune response of rainbow trout ( Oncorhynchus mykiss) larvae and fry at an early stage of development. We exposed 17 and 87° days post hatch larvae and fry (152 and 1118 degree days post hatch; avg. wt. 70 and 770 mg, respectively) to the bacterial pathogen, Yersinia ruckeri for 4 h by bath challenge. Samples were taken at 4, 24, 72 and 96 h post exposure for qPCR and immunohistochemical analyses to elucidate the immune response mounted by these young fish. Larvae showed no mortality although infected larvae at 48 h post exposure showed hyperaemia in the mouth region and inflammation on the dorsal side of the body. Gene expression studies showed an up-regulation of iNOS and IL-22 in infected larvae 24 h post exposure but most of the investigated genes did not show any difference between infected and uninfected larvae. Immunohistochemical studies demonstrated a high expression of IgT molecules in gills and CD8 positive cells in thymus of both infected and uninfected larvae. Infection of rainbow trout fry with Y. ruckeri, in contrast, induced a cumulative mortality of 74%. A high expression of cytokines (IL-1β, TNF-α, IL-22, IL-8 and IL-10), acute phase proteins (SAA, hepcidin, transferrin and precerebellin), complement factors (C3, C5 and factor B), antimicrobial peptide (cathelicidin-2) and iNOS was found in infected fry when compared to the uninfected control. IgT molecules and mannose binding lectins in gills of both infected and uninfected fry were detected by immunohistochemistry. The study indicated that early life stages (yolk-sac larvae), merely up-regulate a few genes and suggests a limited capacity of larvae to mount an immune response by gene regulation at the transcriptional level. Based on the observed clearance of bacteria and lack of mortality it could be speculated that larvae may be covered by protective shield of different immune factors providing protection against broad range of pathogens. However, the increased susceptibility of older fry suggests that Y. ruckeri may utilize some of the immune elements to enter the naive fish. The up-regulation of iNOS and IL-22 in the infected larvae implicates an important role of these molecules in immune response at early developmental stages. A dense covering of surfaces of gill filaments by IgT antibody in the young fish suggest a role of this antibody as innate immune factor at early developmental stages.

Changes in the Cytokine and Toll-Like Receptor Gene Expression Following Infection of Indigenous and Commercial Chickens With Infectious bursal disease virus

A comparative study of cytokine and toll-like receptor (TLR) mRNA expression in 3weeks old indigenous and commercial chickens infected with a very virulent strain of Infectious bursal disease virus (IBDV) was performed using a custom-made microarray chip. In uninfected indigenous chickens, the basal levels of interleukin (IL) 15 were lower and IL 16 was higher than their commercial counterparts. In the IBDV infected indigenous chickens, only IL16 gene expression was down regulated, while TLR3 expression was up regulated significantly. In the IBDV infected commercial chickens IL15, IL16 and TLR3 were down regulated. But, IL1-β, IL2, IL8, IL12, IL17, interferon (IFN)-α and β were significantly increased compared with the control. In IBDV infected indigenous chickens, IL15, IFN-γ, beta-defensin and TLR3 were up regulated compared to virus-infected commercial chickens. The results suggested that up regulation of TLR3, a ligand for double-stranded (ds) RNA probably could account for the possible clinical resistance in these birds. There was a 5.2 fold difference by quantitative real-time RT-PCR between indigenous and commercial chickens in TLR3 mRNA expression. Therefore, TLR3, a receptor for dsRNA could be a putative molecule that could play a role in differential innate and adaptive immune responses to IBDV in commercial and indigenous chickens.

Differential immune response of adipocytes to virulent and attenuated Mycobacterium tuberculosis

Mycobacterium tuberculosis ( M. tb) takes advantage of various cell types, allowing it to remain in the host for long periods. Because adipocytes have been proposed as niches for dormant M. tb in the latent state, understanding the interaction of virulent M. tb with adipocytes is important. We compared changes in cytokine secretion from 3T3-L1 murine adipocytes infected with virulent M. tb H37Rv (V- M. tb) and attenuated M. tb H37Ra (A- M. tb) strains. Both strains maintained non-replicating states within adipocytes until 10 days post-infection. Adipocytes infected with V- M. tb secreted lower levels of pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-12p40, IL-6, and IL-17, and lower levels of nitric oxide than those infected with A- M. tb. In contrast, the anti-inflammatory cytokines, IL-10 and IL-4, were markedly induced in V -M. tb-infected adipocytes versus those infected with A -M. tb at an early time point. Heat-killed or formalin-fixed bacteria induced lower levels of cytokines and no difference was observed between strains. Moreover, V- M. tb induced a high level of necrosis versus A -M. tb in conjunction with increased levels of LHD. These results suggest that V- M. tb regulates cytokine expression in its favor, increasing cytokines necessary for immune evasion and decreasing those required for protective immunity.

Differential acute phase immune responses by Angus and Romosinuano steers following an endotoxin challenge

Our primary objective of this experiment was to evaluate potential genetic differences between two diverse Bos taurus breeds [Angus (AG) and Romosinuano (RO)] in response to an endotoxin challenge. Eighteen steers (n = 9 steers/breed; 299.4 ± 5.2 kg BW) were acclimated to environmentally controlled chambers maintained at thermoneutrality (19.7 °C) and then fitted with indwelling jugular catheters and rectal temperature (RT) recording devices 1 d before the endotoxin challenge. The next day, blood samples were collected at 30-min intervals from −2 to 8 h, and RT was measured continuously at 1-min intervals throughout the study. At time 0, all steers received an intravenous bolus injection of lipopolysaccharide (LPS; 2.5 μg/kg BW). Serum samples were stored at −80 °C until analyzed for cortisol, proinflammatory cytokines [tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), IL-6, and interferon gamma (IFN-γ)], and acute phase proteins (serum amyloid A, acid soluble protein, ceruloplasmin, and α-acid glycoprotein). Rectal temperatures increased in both breeds within 1 h after LPS, with RO producing a greater increase in RT than AG steers ( P < 0.001). Serum cortisol and TNF-α increased ( P < 0.01) in both breeds within 1 h after the LPS challenge. For cortisol, an overall breed effect ( P < 0.02) was detected, such that AG steers had a higher cortisol response than RO steers. A breed × time interaction ( P < 0.01) was observed for TNF-α, such that the response was delayed and extended in the RO steers compared with the AG steers. At 2 and 2.5 h after LPS, TNF-α concentrations were greater ( P < 0.03) in RO steers than in AG steers. For IL-1β, a breed × time interaction ( P < 0.04) was also observed. At 3 h after LPS, IL-1β concentrations were greater ( P < 0.01) in RO steers than in AG steers. Serum IL-6 and IFN-γ increased ( P < 0.01) in a similar manner in both groups after the LPS challenge. These data show differences in the innate immune response between two diverse Bos taurus breeds which may provide insight about differences observed in productivity, heat tolerance, disease resistance, and longevity among cattle breeds.

Immunological and pathological responses of red deer resistant or susceptible genotypes, to experimental challenge with Mycobacterium avium subsp. paratuberculosis

This study aimed to monitor the clinical, immunological and pathological changes in red deer for 49 weeks after experimental oral challenge with Mycobacterium avium subsp. paratuberculosis (MAP) and to assess the heritability of resistance in the offspring of two red stags. Eighteen young deer, which were bred from unselected hinds and sired by two stags resistant (R) or susceptible (S) to paratuberculosis, were challenged with MAP and monitored for 49 weeks. Biopsy samples of the jejunal lymph node were collected at Weeks 4 and 13 and at necropsy after euthanasia of clinically affected animals or when electively killed at Week 49. Three animals (two S and one R) developed clinical disease and were euthanised. The nine S offspring had significantly more severe lesions than the nine R offspring (Mantel–Haenszel Chi-square P = 0.017). The average Lesion Severity Score (LSS) of R offspring was 5.9 (mild), and 7/9 had no or very mild lesions. In contrast, the LSS of S offspring averaged 11.7 (severe), and 7/9 had severe lesions. Most of the resistant, but not the susceptible, animals showed evidence of resolving lesions and a reduction in the number of MAP between 13 and 49 weeks after challenge. One R offspring appeared to completely cure itself, and progressed from mild culture-positive paratuberculosis lesions at Week 13 to having no signs of disease or infection 36 weeks later. This study showed significant heritable resistance/susceptibility to paratuberculosis and key differences in immunological responses in the first 3 months after challenge, indicating different paths to relative success or failure to control MAP. In general, R deer had higher IFN-γ levels, low antibody titres and fewer MAP, while S deer had lower IFN-γ levels, higher antibody and more MAP.

Differential Innate Immune Responses to Low or High Dose Oral SIV Challenge in Rhesus Macaques

Mucosal transmission of HIV predominately occurs during sexual intercourse or breast-feeding and generally results in a successful infection from just one or few founder virions. Here we assessed the impact of viral inoculum size on both viral and immune events within two groups of Rhesus macaques that were non-traumatically, orally inoculated with either multiple low (1000 to 4000 TCID(50)) or high (100,000 TCID(50)) doses of SIV. In agreement with previous studies, more diverse SIV variants were observed in macaques following infection with high dose oral SIV compared to a low dose challenge. In peripheral blood cells, the immune gene transcript levels of CXCL9, IFNγ, TNFα and IL10 remained similar to uninfected macaques. In contrast, OAS and CXCL10 were upregulated following SIV infection in both the high and low dosed macaques, with a more rapid kinetics (detectable by 7 days) following the high SIV dose challenge. In peripheral lymph nodes, an increase in CXCL10 was observed irrespective of viral dose while CXCL9 and OAS were differentially regulated in the two SIV dosed groups. Magnetic bead sorting of CD3+, CD14+ and CD3- /CD14- cells from peripheral blood identified the increase in OAS expression primarily within CD14+ monocytes, whereas the CXCL10 expression was primarily in CD3+ T cells. These findings provide insights into the impact of SIV challenge dose on viral and innate immune factors, which has the potential to inform future SIV/HIV vaccine efficacy trials in which vaccinated hosts have the potential to be infected with a range of viral challenge doses.

Differential immune responses of the green neon shrimp ( Neocaridina denticulate) to dipropyl phthalate

This study set out to understand the sublethal effect of xenobiotic phthalate esters (PAEs) on the relationship between the susceptibility of shrimp to bacterial infection and the immune response of the shrimp. Neocaridina denticulate were exposed to different concentrations of the PAE dipropyl phthalate (DPrP), and mortality and six immune parameters were measured on days 1, 3, 5, and 10 after exposure. On days 1 and 3 after exposure, shrimp exposed to 0, 1, 5, 10, and 50 mg/L of DPrP and challenged with Aeromonas veronii experienced 14% and 16%, 16% and 16%, 18% and 18%, 34% and 24%, and 38% and 26% mortality, respectively. On day 1, five immune parameters (acid phosphatase, AcP; β-glucuronidase, β-Glu; phenoloxidase, PO; superoxide dismutase, SOD; and haemocyanin mRNA) were significantly altered in the all of the groups treated with DPrP compared to the untreated shrimp and were elevated in the 10 mg/L- and 50 mg/L-treated groups. Beta–Glu activity and haemocyanin mRNA levels were significantly increased in a dose-dependent manner. These results suggest that the increased susceptibility of N. denticulate exposed to DPrP is short-term and may be related to the increased expression of DPrP-induced immune mediators.

Differential Cell Proliferation, Apoptosis, and Immune Response in Healthy and Evaporative-Type Dry Eye Conjunctival Epithelia

To assess cell viability and cell cycle kinetics of the conjunctival epithelium and to identify intraepithelial lymphocyte (IEL) subsets in patients with evaporative-type dry eye disease (ev-DED) caused by meibomian gland dysfunction and in healthy subjects. The effect of topical treatment and correlations between clinical symptoms and signs and epithelial and immune variables were also determined. Inferior fornix and tarsal conjunctival cells were collected by brush cytology (BC) from patients with mild to moderate ev-DED (n = 23) before and after 2 months of treatment that included lid hygiene, artificial tears, and a 3-week course of topical unpreserved steroids. Healthy subjects (n = 17) served as controls. Two symptom questionnaires were self-answered, and multiple DED-related clinical tests were performed. Epithelial or immune lineage, IEL subtypes, cell viability, apoptosis, and cell cycle stage of the BC-recovered cells were determined by flow cytometry. Conjunctival cell viability was dramatically decreased in ev-DED patients compared with controls. For both groups, two different cell populations, differentiated by cell size and complexity, were present in the apoptosis assay. After 2 months of treatment, 87% of subjects subjectively improved, CD8 cells increased, and CD4 cells and the CD4/CD8 ratio significantly decreased. The pretreatment and posttreatment proliferative capacity of the conjunctival epithelium was significantly lower in ev-DED patients than in healthy controls. The viability and proliferative capacity of ev-DED patient conjunctival cells were reduced, suggesting a potential role for these parameters as disease biomarkers.

Single-Cell Mass Cytometry of Differential Immune and Drug Responses Across a Human Hematopoietic Continuum

Flow cytometry is an essential tool for dissecting the functional complexity of hematopoiesis. We used single-cell "mass cytometry" to examine healthy human bone marrow, measuring 34 parameters simultaneously in single cells (binding of 31 antibodies, viability, DNA content, and relative cell size). The signaling behavior of cell subsets spanning a defined hematopoietic hierarchy was monitored with 18 simultaneous markers of functional signaling states perturbed by a set of ex vivo stimuli and inhibitors. The data set allowed for an algorithmically driven assembly of related cell types defined by surface antigen expression, providing a superimposable map of cell signaling responses in combination with drug inhibition. Visualized in this manner, the analysis revealed previously unappreciated instances of both precise signaling responses that were bounded within conventionally defined cell subsets and more continuous phosphorylation responses that crossed cell population boundaries in unexpected manners yet tracked closely with cellular phenotype. Collectively, such single-cell analyses provide system-wide views of immune signaling in healthy human hematopoiesis, against which drug action and disease can be compared for mechanistic studies and pharmacologic intervention.

Arsenic and Infectious Disease: A Potential Factor in Morbidity among Bangladeshi Children

Vol. 119, No. 5 NewsOpen AccessArsenic and Infectious Disease: A Potential Factor in Morbidity among Bangladeshi Children Angela Spivey Angela Spivey Search for more papers by this author Published:1 May 2011https://doi.org/10.1289/ehp.119-a218aCited by:6AboutSectionsPDF ToolsDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InReddit Lower respiratory tract infection (LRTI) and diarrhea are two of the most common causes of morbidity and mortality in children under 5 years old, especially in low-income countries. A new prospective cohort study of the link between these types of infections and arsenic exposure revealed a dose-dependent increase in LRTI and diarrhea in relation to maternal arsenic exposure [EHP119(5):719–724; Rahman et al.].Earlier studies linked prenatal arsenic exposure to increased risk of infant mortality, and infectious disease has been suggested as a potential underlying cause in these deaths. No epidemiologic studies have been conducted to support that explanation, but there is evidence from a few animal and human studies that arsenic may cause immunosuppression.The current study included 1,552 live-born infants of women enrolled during 2002–2004 in Matlab, Bangladesh. Arsenic exposure was assessed by measuring inorganic arsenic in maternal urine samples collected at gestational weeks 8 and 30. After birth, information on symptoms of LRTI and diarrhea in infants was collected at monthly home visits in which mothers recalled symptoms that had occurred over the previous 7 days.The estimated relative risk of LRTI and severe LRTI increased by 69% and 54%, respectively, in the participants whose mothers had urinary arsenic concentrations in the highest quintile (262–977 μg/L), compared with offspring of mothers whose exposure was in the lowest quintile (less than 39 μg/L). The relative risk of diarrhea increased 20% for the highest-exposure group compared with the lowest-exposure group.The authors observed that relative risks of LRTI and diarrhea increased irrespective of sociodemographic factors or nutritional status of the women. However, further evaluation is needed in a larger sample to evaluate possible effect modification, because the risks appeared more pronounced in low social strata.Strengths of the study include the large sample size, the objective measure of arsenic exposure, and the followup of infants for one full year, which the authors say would reduce any influence of seasons on infection rates. Potential limitations include lack of measurements of infant exposure to arsenic, a lack of information about other potentially toxic substances in water and food, and reliance on mothers’ reports of disease symptoms and signs. Given the millions of people worldwide who drink well water with elevated arsenic concentrations, the study results could have serious public health implications and, taken together with previous studies showing health effects from this exposure, emphasize the need to reduce arsenic exposure via drinking water.More than 50 million people in Bangladesh are believed to be chronically exposed to drinking water with arsenic concentrations exceeding the WHO standard of 10 μg/L.FiguresReferencesRelatedDetailsCited by Zheng K, Zeng Z, Tian Q, Huang J, Zhong Q and Huo X (2023) Epidemiological evidence for the effect of environmental heavy metal exposure on the immune system in children, Science of The Total Environment, 10.1016/j.scitotenv.2023.161691, (161691), Online publication date: 1-Jan-2023. Taylor S, Terkildsen M, McQuilty R, Lee D, Wing-Simpson A and Gray R (2022) Non-essential heavy metals and protective effects of selenium against mercury toxicity in endangered Australian sea lion (Neophoca cinerea) pups with hookworm disease, Environment International, 10.1016/j.envint.2022.107521, 169, (107521), Online publication date: 1-Nov-2022. Chatterjee S, Chetia M, Voronina A and Gupta D (2017) Prospects of Combating Arsenic: Physico-chemical Aspects Arsenic Contamination in the Environment, 10.1007/978-3-319-54356-7_5, (103-121), . Chatterjee S, Datta S and Gupta D (2017) Studies on Arsenic and Human Health Arsenic Contamination in the Environment, 10.1007/978-3-319-54356-7_3, (37-66), . Choudhury S, Gupta P, Ghosh S, Mukherjee S, Chakraborty P, Chatterji U and Chattopadhyay S (2016) Arsenic-induced dose-dependent modulation of the NF-κB/IL-6 axis in thymocytes triggers differential immune responses, Toxicology, 10.1016/j.tox.2016.06.005, 357-358, (85-96), Online publication date: 1-May-2016. Naujokas M, Anderson B, Ahsan H, Aposhian H, Graziano J, Thompson C and Suk W (2013) The Broad Scope of Health Effects from Chronic Arsenic Exposure: Update on a Worldwide Public Health Problem, Environmental Health Perspectives, 121:3, (295-302), Online publication date: 1-Mar-2013. Vol. 119, No. 5 May 2011Metrics About Article Metrics Publication History Originally published1 May 2011Published in print1 May 2011 Financial disclosuresPDF download License information EHP is an open-access journal published with support from the National Institute of Environmental Health Sciences, National Institutes of Health. All content is public domain unless otherwise noted. Note to readers with disabilities EHP strives to ensure that all journal content is accessible to all readers. However, some figures and Supplemental Material published in EHP articles may not conform to 508 standards due to the complexity of the information being presented. If you need assistance accessing journal content, please contact [email protected]. Our staff will work with you to assess and meet your accessibility needs within 3 working days.

Obese mice have increased morbidity and mortality compared to non-obese mice during infection with the 2009 pandemic H1N1 influenza virus

Please cite this paper as: Easterbrook et al. (2011) Obese mice have increased morbidity and mortality compared to non‐obese mice during infection with the 2009 pandemic H1N1 influenza virus. Influenza and Other Respiratory Viruses 5(6), 418–425. Background Obesity has been identified as an independent risk factor for severe or fatal infection with 2009 pandemic H1N1 influenza (2009 pH1N1), but was not previously recognized for previous pandemic or seasonal influenza infections. Objectives Our aim was to evaluate the role of obesity as an independent risk factor for severity of infection with 2009 pH1N1, seasonal H1N1, or a pathogenic H1N1 influenza virus. Methods Diet‐induced obese (DIO) and their non‐obese, age‐matched control counterparts were inoculated with a 2009 pH1N1, A/California/04/2009 (CA/09), current seasonal H1N1, A/NY/312/2001 (NY312), or highly pathogenic 1918‐like H1N1, A/Iowa/Swine/1931 (Sw31), virus. Results Following inoculation with CA/09, DIO mice had higher mortality (80%) than control mice (0%) and lost more weight during infection. No effect of obesity on morbidity and mortality was observed during NY312 or Sw31 infection. Influenza antigen distribution in the alveolar regions of the lungs was more pronounced in DIO than control mice during CA/09 infection at 3 days post‐inoculation (dpi), despite similar virus titers. During CA/09 infection, localized interferon‐β and proinflammatory cytokine protein responses in the lungs were significantly lower in DIO than control mice. Conversely, serum cytokine concentrations were elevated in DIO, but not control mice following infection with CA/09. The effect of obesity on differential immune responses was abrogated during NY312 or Sw31 infection. Conclusions Together, these data support epidemiologic reports that obesity may be a risk factor for severe 2009 pandemic H1N1 influenza infection, but the role of obesity in seasonal or highly virulent pandemic influenza infection remains unclear.

Abstract SY14-01: 34+ parameter single-cell mass cytometry of differential immune and drug responses across healthy human hematopoiesis and related malignancies

Abstract Fluorescence flow cytometry is an essential tool for dissecting the functional complexity of hematopoiesis. Using this, intracellular assays of signaling systems can be correlated to functional subsets of cells in complex populations based on active kinase states or other signaling junctions. Such correlations could be important to distinguish changes in signaling status that arise in rare cell subsets during environmental activation or pathogenesis. Historically, fluorescence spectral overlap has stymied the use of an increasing number of phenotypic parameters required to delineate an array of rare cell populations, thus limiting the application of this single-cell approach in studying complex diseases. Using a next-generation single-cell ‘mass cytometry’ platform we avoid these caveats, quantifying 34 parameters simultaneously with no spectral overlap (e.g., 31 antibodies, viability, DNA content and relative cell size). In healthy human blood and bone marrow, we identified cell subsets spanning the hematopoietic hierarchy using a panel of up to 31 surface markers. Additionally, 18 simultaneous functional markers and a battery of ex vivo stimuli and inhibitors were used to interrogate signaling. A novel algorithm for unsupervised high-dimensional clustering of single-cell data revealed a continuum of phenotypically contiguous subpopulations. This continuum was demarcated by surface marker expression, but paralleled by gradual changes in intracellular signaling architecture. These results have exposed unappreciated layers of human hematopoietic organization, and provide an opportunity to reevaluate diseases and pharmacological therapeutics as specific perturbations to this inherent order. Working from a 34-parameter template of normal hematopoiesis, we now are now focused on further dissecting the biology and improving risk stratification in hematopoietic malignancies. The application of these multidimensional approaches to human acute lymphoid leukemia (ALL) and acute myeloid leukemia (AML) samples will be presented. The simultaneous measurement of surface marker, cell cycle, apoptosis, and intracellular signaling parameters has revealed substantial heterogeneity in the blast populations of these diseases. This information is used to provide a high-content view of in vitro drug activity in blast subpopulations. Leveraging the wide variability between human individuals, we have integrated single-cell data from multiple individuals to construct a system-level correlation map of seemingly independent immune compartments. Specifically, we reframe the healthy immune system as a homeostatic network of immune parameters including cell frequency, surface marker intensity, and intracellular signaling capacity. Chronic diseases such as rheumatoid arthritis and prolonged bacterial infection fundamentally disrupt this balance, causing measurable and pathologically relevant shifts in the normal homeostatic balance. Taken together, these early successes point toward a future of actionable clinical information based on single-cell assays for diverse hematological malignancies and autoimmune disorders. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr SY14-01. doi:10.1158/1538-7445.AM2011-SY14-01

Differential immune responses mediated by adenovirus- and lentivirus-transduced DCs in a HER-2/neu overexpressing tumor model

Recent investigations have demonstrated that adenoviral and lentiviral vectors encoding HER-2 can be utilized in cancer immunotherapy. However, it is not known whether both viral systems elicit a similar immune response. Here, we compare the immune response in mice induced by dendritic cells (DCs) infected with either recombinant adenovirus or lentivirus encoding rat HER-2 (rHER-2). Both vaccine types yielded similar control of tumor growth, but we found clear differences in their immune responses 10 days after DC immunization. Adenovirus rHER-2-transduced DCs elicited locally and systemically high frequencies of CD4+ and CD8+ T cells, while lentivirus rHER-2-transduced DCs predominantly led to CD4+ T-cell infiltration at the tumor site. Splenocytes from mice immunized with lentivirus rHER-2-transduced DCs secreted higher levels of interferon (IFN)-γ, mainly by CD4+ T cells, following stimulation by RM-1-mHER-2 tumors. In contrast, the adenovirus vaccinated group exhibited CD4+ and CD8+ T cells that both contributed to IFN-γ production. Besides an established cellular immune response, the rHER-2/DC vaccine elicited a significant humoral response that was highest in the adenovirus group. DC subsets and regulatory T cells in the spleen were also differentially modulated in the two vaccine systems. Finally, adoptive transfer of splenocytes from both groups of immunized mice strongly inhibited in vivo tumor growth. Our results suggest that not only the target antigen but also the virus system may determine the nature and magnitude of antitumor immunity by DC vaccination.

Interleukin-1 Receptor–Associated Kinase 3 Gene Associates with Susceptibility to Acute Lung Injury

Sepsis is the most common cause of acute lung injury (ALI), leading to organ dysfunction and death in critically ill patients. Previous studies associated variants of interleukin-1 receptor-associated kinase genes (IRAKs) with differential immune responses to pathogens and with outcomes during sepsis, and revealed that increased expression levels of the IRAK3 gene were correlated with poor outcomes during sepsis. Here we explored whether common variants of the IRAK3 gene were associated with susceptibility to, and outcomes of, severe sepsis. After our discovery of polymorphism, we genotyped a subset of seven single-nucleotide polymorphisms (SNPs) in 336 population-based control subjects and 214 patients with severe sepsis, collected as part of a prospective study of adults from a Spanish network of intensive care units. Whereas IRAK3 SNPs were not associated with susceptibility to severe sepsis, rs10506481 showed a significant association with the development of ALI among patients with sepsis (P = 0.007). The association remained significant after adjusting for multiple comparisons, population stratification, and clinical variables (odds ratio, 2.50; 95% confidence interval, 1.15-5.47; P = 0.021). By imputation, we revealed three additional SNPs independently associated with ALI (P < 0.01). One of these (rs1732887) predicted the disruption of a putative human-mouse conserved transcription factor binding site, and demonstrated functional effects in vitro (P = 0.017). Despite the need for replication in independent studies, our data suggest that common SNPs in the IRAK3 gene may be determinants of sepsis-induced ALI.

Differential cellular immune response in inductive and effectors sites after oral administration of quercetin

Differential cellular immune response in inductive and effectors sites after oral administration of quercetin

Selective colonization with Helicobacter bilis or Escherichia coli induces differential host immune responses following an inflammatory trigger.

Selective colonization with Helicobacter bilis or Escherichia coli induces differential host immune responses following an inflammatory trigger.

Glioma Indian scenario: Is there a human leucocyte antigen association?

Background:The central nervous system tumors are a rare neoplasm with little knowledge with Human Leukocyte Antigen (HLA) involvement. Primary brain tumors are cancers that originate in brain classified according to their appearance under a microscope as low grade (grade I and II) with diffuse astrocytomas, pliocytic astrocytomas, oligodendrogliomas, gangliogliomas, and mixed gliomas as common subtypes and high grade (grade III and IV).Materials and Methods:HLA associations in common glioma are reported from other parts of the world. The normal cancer treatment is surgery, followed by radiotherapy, and chemotherapy; nowadays immunotherapy is advised. HLA distribution in a Glioma patient was done based on serology and molecular techniques. The immune response gene studies have implicated the HLA allele association in most of the common diseases from India. Considerable variations are noted in HLA association with cancers; hence, we have summarized the HLA involvement in Glioma with respect to the literature.Results:HLA A*030101, A*310102, B*350101, B*4406, Cw*040101, Cw*070101, DRB1*070101, and DRB1*1001.Conclusion:Ethnic diversity and HLA polymorphism precipitate differential immune response genes involved in variable disease manifestations. Therefore, caste-specific HLA allelic specificity needs to be identified, which may help in early identification of the associated HLA allele and establishing clinical practices among glioma patients.

Tissue expression of steroid hormone receptors is associated with differential immune responsiveness

Glucocorticoids have been used as treatments against a number of diseases, especially autoimmune/inflammatory conditions in which the immune system is overactive. These treatments have varying degrees of responsiveness among individuals and in different tissues (including brain); therefore, it is important to determine what could account for these differences. In this study, we evaluated expression of stress hormone receptors in immune cells from lymphoid and non-lymphoid tissues (including brain) as a possible explanation. We analyzed leukocytes (CD45 +) in kidney, liver, spleen, and thymus tissues from healthy mice for expression of the receptor for stress hormone (glucocorticoid—GR) as well as other steroid hormones (androgen—AR, progesterone—PR) and found that all tissues expressed these steroid hormone receptors but with varying patterns. To determine whether tissue-specific differences were related to immune cell composition, we examined steroid hormone receptor expression in T lymphocytes from each of these tissues and found similar patterns of expression in these cells regardless of tissue source. Because glucocorticoids can also impact brain function, we further examined expression of the stress hormone receptor in brain tissue and found GR expressed in immune cells at this site. In order to investigate the potential impact in an area of neuropathology, we utilized a mouse model of West Nile Virus (WNV). We observed pathological changes in brains of WNV-infected animals and T lymphocytes in the areas of inflammation; however, these cells did not express GR. These data indicate that tissue-specific differences in steroid hormone receptor expression by immune cells could determine responsiveness to steroid hormone treatment.