Interaction of IFN-γ with cholinergic agonists to modulate rat and human goblet cell function

The adenomatous polyposis coli ( APC) gene is a known tumor suppressor gene, and mice with mutations in Apc (ApcMin/+) spontaneously form multiple intestinal neoplasms. In this model of human colorectal cancer (CRC), it has been reported that CD4+ T-cell-derived interleukin 17 (IL-17) promotes intestinal tumor development, but it is not known if the Apc mutation actually directly alters T-cell function and subsequently tumor immunosurveillance. To investigate the ApcMin/+ mutation on T-cell function, flow cytometric, histochemical, and immunofluorescent studies on both wild-type (Apc+/+) and ApcMin/+ mice were performed. We identified decreased levels of interferon gamma (IFN-γ+)IL-17+ double-positive CD4+ cells in the mesenteric lymph nodes and Peyer’s patches of ApcMin/+ mice. In addition, altered levels of CD8+ cells, and changes in CD8+ production of IFN-γ and granzyme B were observed. These T-cell alterations did modify tumor immunosurveillance, as the adoptive transfer of splenocytes from ApcMin/+ animals into a chemically induced CRC model resulted in the inability to prevent epithelial dysplasia. These results suggest an altered T-cell balance in ApcMin/+ mice may disrupt intestinal homeostasis, consequently limiting intestinal tumor immunosurveillance.

Download Full-text

IL-12 gene transfer alters gut physiology and host immunity in nematode-infected mice

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.2001.281.1.g102 ◽

2001 ◽

Vol 281 (1) ◽

pp. G102-G110 ◽

Cited By ~ 23

Author(s):

Waliul I. Khan ◽

Patricia A. Blennerhassett ◽

Yikang Deng ◽

Jack Gauldie ◽

Bruce A. Vallance ◽

...

Keyword(s):

Immune Response ◽

Goblet Cell ◽

Recombinant Adenovirus ◽

Trichinella Spiralis ◽

Nematode Infection ◽

Interleukin 12 ◽

Cell Hyperplasia ◽

Goblet Cell Hyperplasia ◽

Worm Expulsion ◽

Ifn Γ

Immune responses elicited by nematode parasite infections are characterized by T helper 2 (Th2) cell induction. The immunologic basis for changes in intestinal physiology accompanying nematode infection is poorly understood. This study examined whether worm expulsion and associated goblet cell hyperplasia and muscle contractility share a similar immune basis by shifting the response from Th2 to Th1 using interleukin-12 (IL-12) overexpression. We used a single administration of recombinant adenovirus vector expressing IL-12 (Ad5IL-12) in Trichinella spiralis-infected mice. Ad5IL-12 administered 1 day after infection prolonged worm survival and inhibited infection-induced muscle hypercontractility and goblet cell hyperplasia. This was correlated with upregulated interferon-γ (IFN-γ) expression and downregulated IL-13 expression in the muscularis externa layer. We also observed increased IFN-γ production and decreased IL-4 and IL-13 production from in vitro stimulated spleen and mesenteric lymph node cells of infected Ad5IL-12-treated mice. These results indicate that transfer and overexpression of the IL-12 gene during Th2-based nematode infection shifts the immune response toward Th1 and delays worm expulsion. Moreover, the immune response shift abrogated the physiological responses to infection, attenuating both muscle hypercontractility and goblet cell hyperplasia. These findings strongly indicate that worm expulsion, muscle hypercontractility, and goblet cell hyperplasia share a common immunologic basis and may be causally linked.

Download Full-text

The tyrosine phosphatase Shp-2 confers resistance to colonic inflammation by driving goblet cell function and crypt regeneration

The Journal of Pathology ◽

10.1002/path.5177 ◽

2018 ◽

Vol 247 (1) ◽

pp. 135-146

Author(s):

Jessica Gagné-Sansfacon ◽

Ariane Langlois ◽

Marie-Josée Langlois ◽

Geneviève Coulombe ◽

Sarah Tremblay ◽

...

Keyword(s):

Goblet Cell ◽

Cell Function ◽

Tyrosine Phosphatase ◽

Colonic Inflammation

Download Full-text

Clinical significance of insulin peptide-specific interferon-γ-related immune responses in ketosis-prone type 2 diabetes

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/clinem/dgab912 ◽

2021 ◽

Author(s):

Atsushi Satomura ◽

Yoichi Oikawa ◽

Akifumi Haisa ◽

Seiya Suzuki ◽

Shunpei Nakanishi ◽

...

Keyword(s):

Type 2 Diabetes ◽

Immune Responses ◽

Elispot Assay ◽

Mononuclear Cells ◽

Cell Function ◽

Β Cell ◽

Peripheral Blood Mononuclear ◽

Β Cell Function ◽

Ifn Γ

Abstract Context Unprovoked A−β+ ketosis-prone type 2 diabetes (KPD) is characterized by the sudden onset of diabetic ketosis/ketoacidosis (DK/DKA) without precipitating factors, negative anti-islet autoantibodies (“A−”), and preservation of β-cell function (“β+”) after recovery from DKA. Although this phenotype often appears with acute hyperglycemia and DK/DKA just like acute-onset type 1 diabetes (AT1D), the involvement of anti-islet immune responses remains unknown. Objective We sought to clarify the immunological role of insulin-associated molecules in unprovoked A−β+ KPD. Methods In this cross-sectional study, blood samples from 75 participants (42 with AT1D and 33 with KPD) were evaluated for interferon (IFN)-γ-secreting peripheral blood mononuclear cells (PBMCs) reactive to four insulin B-chain amino acid 9–23-related peptides (B:9–23rPep) using an enzyme-linked immunospot (ELISpot) assay. Results Overall, 36.4% (12/33) of KPD participants showed positive IFN-γ ELISpot assay results; the positivity rate in KPD was similar to that in AT1D (38.1%; 16/42) and significantly higher than the previously reported rate in type 2 diabetes (8%; 2/25; P < 0.0167). Moreover, B:9–23rPep-specific IFN-γ-producing PBMC frequency was negatively correlated with age and ad lib serum C-peptide levels in all KPD participants and positively correlated with HbA1c level in KPD participants with positive IFN-γ ELISpot results. Conclusions These findings suggest the involvement of B:9–23rPep-specific IFN-γ-related immunoreactivity in the pathophysiology of some unprovoked A−β+ KPD. Moreover, increased immunoreactivity may reflect transiently decreased β-cell function and increased disease activity at the onset of DK/DKA, thereby playing a key role in DK/DKA development in this KPD phenotype.

Download Full-text

Regulation of Macrophage Activation and Polarization by HCC-Derived Exosomal lncRNA TUC339

International Journal of Molecular Sciences ◽

10.3390/ijms19102958 ◽

2018 ◽

Vol 19 (10) ◽

pp. 2958 ◽

Cited By ~ 41

Author(s):

Xue Li ◽

Yi Lei ◽

Miao Wu ◽

Nan Li

Keyword(s):

Macrophage Activation ◽

Cell Function ◽

Regulatory Function ◽

Receptor Interaction ◽

Loss Of Function ◽

Critical Function ◽

Over Expression ◽

Ifn Γ ◽

Non Coding Rnas

Exosomes released by cells can serve as vehicles for delivery of biological materials and signals. Long non-coding RNAs (lncRNAs) are non-coding RNAs longer than 200 nt, which roles are increasingly appreciated in various biological content. Tumor-derived exosomal lncRNAs have been implicated as signaling mediators to orchestrate cell function among neighbor tumor cells. However, the role of tumor-derived lncRNAs in cross-talk with environmental macrophages has yet to be explored. In this paper, we demonstrated that hepatocellular carcinoma (HCC) cells–derived exosomes contain elevated levels of lncRNA TUC339 and that HCC-derived exosomes could be taken up by THP-1 cells. In seeking to dissect the biological function of tumor secreting TUC339 in macrophages, we applied loss-of-function and gain-of-function strategies. We observed increased pro-inflammatory cytokine production, increased co-stimulatory molecule expression, and enhanced phagocytosis upon suppression of TUC339 by siRNA in THP-1 cells, and the opposite effect upon over-expression of this lncRNA, which indicates that TUC339 was involved in the regulation of macrophage activation. Moreover, we detected an elevated level of TUC339 in M(IL-4) macrophages as compared to M(IFN-γ + LPS) macrophages and a down-regulation of TUC339 expression during M(IL-4)-to-M(IFN-γ + LPS) repolarization and vice versa. Furthermore, suppression of TUC339 in macrophages diminished the expression of M(IL-4) markers upon IL-4 treatment while overexpression of TUC339 in macrophages enhanced M(IL-4) markers upon IFN-γ + LPS treatment, which suggests a critical function of TUC339 in the regulation of macrophage M1/M2 polarization. Lastly, using microarray analysis, we identified cytokine-cytokine receptor interaction, CXCR chemokine receptor binding, Toll-like receptor signaling, FcγR-mediated phagocytosis, regulation of the actin cytoskeleton, and cell proliferation are related with TUC339 function in macrophages. Our results provide evidence for a novel regulatory function of tumor-derived exosomal lncRNA TUC339 in environmental macrophages and shed light on the complicated interactions between tumor and immune cells through exosomal lncRNAs.

Download Full-text

Effects and interaction of dietary electrolyte balance and citric acid on the intestinal function of weaned piglets

Journal of Animal Science ◽

10.1093/jas/skaa106 ◽

2020 ◽

Vol 98 (5) ◽

Author(s):

Qingqing Deng ◽

Yirui Shao ◽

Qiye Wang ◽

Jianzhong Li ◽

Yali Li ◽

...

Keyword(s):

Citric Acid ◽

Mrna Expression ◽

Goblet Cell ◽

Microbial Community Composition ◽

Electrolyte Balance ◽

Intestinal Function ◽

Weaned Piglets ◽

Ifn Γ ◽

Relative Abundances ◽

Relative Mrna Expression

Abstract Fifty-six piglets (6.26 ± 0.64 kg BW) were weaned at 21 d and randomly assigned to one of the eight dietary treatments with seven replicate pens for a 14-d experimental period. The eight experimental diets were prepared via a 2 × 4 factorial arrangement with citric acid (CA; 0% and 0.3%) and dietary electrolyte balance (dEB, Na + K − Cl mEq/kg of the diet; −50, 100, 250, and 400 mEq/kg). Varying dEB values were obtained by altering the contents of calcium chloride and sodium bicarbonate. An interaction (P < 0.05) between dEB and CA in diarrhea score and the number of goblet cell in jejunum were observed. Ileum pH significantly decreased in weaned piglets fed 250 mEq/kg dEB diet compared with those fed −50 and 400 mEq/kg dEB diets (P < 0.05). Supplementation of 0.3% CA decreased the number of goblet cell in the ileal crypt (P < 0.05) and the relative mRNA expression of cystic fibrosis transmembrane conductance regulator, tumor necrosis factor-α, interferon-γ (IFN-γ), interleukin-1β (IL-1β), interleukin-10 (IL-10), zona occludens-1, and Claudin-1 (P < 0.05). Increasing dEB values increased the number of goblet cells in the jejunal crypt (P < 0.05). A 250-mEq/kg dEB diet decreased the relative mRNA expression of IFN-γ, IL-1β, and IL-10 (P < 0.05) than 100-mEq/kg dEB diet. The interaction between dEB and CA on the relative abundances of Cyanobacteria and Saccharibacteria was observed (P < 0.05). Supplementation of 0.3% CA increased relative abundances of and Streptococcus hyointestinalis. Piglets fed 250-mEq/kg diet increased relative abundances of Firmicutes and Lactobacillus rennini, and decreased the relative abundance of Proteobacteria, Veillonella, Actinobacillus minor, and Escherichia–Shigella.In conclusion, supplementation of 0.3% CA resulted in differential expression of inflammatory cytokines, ion transporters, and tight junction proteins, and changes in the microbial community composition. A 250-mEq/kg dEB diet reduced gastrointestinal pH and promoted the enrichment of beneficial microbes in the gut microbiota, thereby suppressing inflammation and harmful bacteria. However, the addition of CA to diets with different dEB values did not promote intestinal function in weaned piglets.

Download Full-text

2028 Discovery and evaluation of FOXP3 dimerization inhibitors

Journal of Clinical and Translational Science ◽

10.1017/cts.2018.65 ◽

2018 ◽

Vol 2 (S1) ◽

pp. 10-10

Author(s):

Ravyn Thompson ◽

Cara Coleman ◽

Nathan G. Dolloff

Keyword(s):

T Cell ◽

Cell Function ◽

Cell Activity ◽

Cell Responses ◽

Treg Suppression ◽

Study Population ◽

Ifn Γ ◽

Treg Phenotype ◽

Dimerization Inhibitors

OBJECTIVES/SPECIFIC AIMS: Immuno-oncology (IO) strategies are promising new approaches for the treatment of a variety of malignancies, including multiple myeloma (MM). Regulatory T cells (Tregs), which suppress effector T cell function, are a limitation to durable IO responses. The transcription factor FOXP3 is critical for the mature Treg phenotype. FOXP3 homodimerization is required for DNA binding and transcriptional activity, and mutations mapping to the dimerization region are associated with IPEX syndrome, resulting in dysfunctional Tregs in humans. We therefore hypothesize that inhibitors of FOXP3 dimerization will repress Treg suppression and enhance the anti-MM activity of IO. METHODS/STUDY POPULATION: To discover FOXP3 dimerization inhibitors, we are modeling FOXP3 homodimerization in vitro. Currently, we are optimizing an ALPHA screen and an ELISA-based dimerization assay using recombinant full length and truncated versions of FOXP3 to discover peptidomimetics that inhibit homodimerization. Induced Tregs expanded from human PBMCs will be treated with lead biologics and functional assays will be performed. RESULTS/ANTICIPATED RESULTS: Here we demonstrate Treg suppression of T cell proliferation and IFN-γ secretion after 5 days of co-culture under basal conditions. Additionally, we developed a MM/T cell co-culture system to measure anti-MM T cell responses and show decreased anti-MM T cell activity in the presence of Tregs. We expect to exploit the assays outlined here to demonstrate defective Treg suppression when FOXP3 dimerization is inhibited. DISCUSSION/SIGNIFICANCE OF IMPACT: These studies support drug discovery efforts that will ultimately improve IO therapies for patients with MM.

Download Full-text

The Interleukin-12–Mediated Pathway of Immune Events Is Dysfunctional in Human Immunodeficiency Virus–Infected Individuals

Blood ◽

10.1182/blood.v94.3.1003.415a15_1003_1011 ◽

1999 ◽

Vol 94 (3) ◽

pp. 1003-1011 ◽

Cited By ~ 68

Author(s):

Jason D. Marshall ◽

Jihed Chehimi ◽

Giorgia Gri ◽

Jay R. Kostman ◽

Luis J. Montaner ◽

...

Keyword(s):

Immune Deficiency ◽

Human Immunodeficiency Virus ◽

Nk Cells ◽

Mononuclear Cells ◽

Cell Function ◽

Interleukin 12 ◽

Accessory Cell ◽

Mrna Accumulation ◽

Immunodeficiency Virus ◽

Ifn Γ

Interleukin-12 (IL-12) is a potentially critical factor in the immune response against human immunodeficiency virus (HIV) because it is important for regulating proliferation and interferon-γ (IFN-γ) production by T cells and natural killer (NK) cells, antigen presentation and accessory cell function by macrophages and dendritic cells, and cytolytic activities of cytotoxic T-lymphocyte cells and NK cells, which are all functions known to be dysfunctional in patients with acquired immune deficiency syndrome. Peripheral blood mononuclear cells (PBMC) from HIV-infected patients have been previously shown to be deficient in the ability to produce IL-12 in response to the bacterial pathogen Staphylococcus aureus Cowan. In this study, impaired IL-12 production in cells from PBMC of HIV-infected patients compared with healthy donors was observed across a broad panel of stimuli derived from infectious pathogens with or without priming with cytokines such as IFN-γ and IL-4, which amplify the IL-12 induction signal. Analysis of p40 and p35 mRNA accumulation showed that reductions in both subunits contribute to the lower IL-12 secretion of cells from HIV-infected individuals. PBMC from HIV-infected donors also failed to upregulate the IL-12 receptor β2 chain (IL-12Rβ2) in response to mitogenic stimuli. The expression of the IL-12Rβ2 gene could, however, be restored by in vitro exposure to rIL-12. Thus, it is possible that a primary IL-12 defect may lead to secondary deficiencies in expression of the genes for IL-12Rβ2 and IFN-γ, thus amplifying immune deficiency during HIV infection.

Download Full-text

Contribution of Natural Killer Cells to Inhibition of Angiogenesis by Interleukin-12

Blood ◽

10.1182/blood.v93.5.1612 ◽

1999 ◽

Vol 93 (5) ◽

pp. 1612-1621 ◽

Cited By ~ 127

Author(s):

Lei Yao ◽

Cecilia Sgadari ◽

Keizo Furuke ◽

Eda T. Bloom ◽

Julie Teruya-Feldstein ◽

...

Keyword(s):

Endothelial Cells ◽

Nk Cells ◽

Natural Killer ◽

Cell Function ◽

Nk Cell ◽

Primary Cultures ◽

Interleukin 12 ◽

Ifn Γ

Abstract Interleukin-12 (IL-12) inhibits angiogenesis in vivo by inducing interferon-γ (IFN-γ) and other downstream mediators. Here, we report that neutralization of natural killer (NK) cell function with antibodies to either asialo GM1 or NK 1.1 reversed IL-12 inhibition of basic fibroblast growth factor (bFGF)-induced angiogenesis in athymic mice. By immunohistochemistry, those sites where bFGF-induced neovascularization was inhibited by IL-12 displayed accumulation of NK cells and the presence of IP-10–positive cells. Based on expression of the cytolytic mediators perforin and granzyme B, the NK cells were locally activated. Experimental Burkitt lymphomas treated locally with IL-12 displayed tumor tissue necrosis, vascular damage, and NK-cell infiltration surrounding small vessels. After activation in vitro with IL-12, NK cells from nude mice became strongly cytotoxic for primary cultures of syngeneic aortic endothelial cells. Cytotoxicity was neutralized by antibodies to IFN-γ. These results document that NK cells are required mediators of angiogenesis inhibition by IL-12, and provide evidence that NK-cell cytotoxicity of endothelial cells is a potential mechanism by which IL-12 can suppress neovascularization.

Download Full-text

The Level of Viral Antigen Presented by Hepatocytes Influences CD8 T-Cell Function

Journal of Virology ◽

10.1128/jvi.02415-06 ◽

2007 ◽

Vol 81 (6) ◽

pp. 2940-2949 ◽

Cited By ~ 58

Author(s):

Adam J. Gehring ◽

Dianxing Sun ◽

Patrick T. F. Kennedy ◽

Esther Nolte-'t Hoen ◽

Seng Gee Lim ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cd8 T Cells ◽

Viral Antigen ◽

Cell Function ◽

Cd8 T Cell ◽

T Cell Function ◽

Tnf Α ◽

Antiviral Function ◽

Ifn Γ

ABSTRACT CD8 T cells exert their antiviral function through cytokines and lysis of infected cells. Because hepatocytes are susceptible to noncytolytic mechanisms of viral clearance, CD8 T-cell antiviral efficiency against hepatotropic viruses has been linked to their capacity to produce gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α). On the other hand, intrahepatic cytokine production triggers the recruitment of mononuclear cells, which sustain acute and chronic liver damage. Using virus-specific CD8 T cells and human hepatocytes, we analyzed the modulation of virus-specific CD8 T-cell function after recognition peptide-pulsed or virally infected hepatocytes. We observed that hepatocyte antigen presentation was generally inefficient, and the quantity of viral antigen strongly influenced CD8 T-cell antiviral function. High levels of hepatitis B virus production induced robust IFN-γ and TNF-α production in virus-specific CD8 T cells, while limiting amounts of viral antigen, both in hepatocyte-like cells and naturally infected human hepatocytes, preferentially stimulated CD8 T-cell degranulation. Our data document a mechanism where virus-specific CD8 T-cell function is influenced by the quantity of virus produced within hepatocytes.

Download Full-text