• IMMUNE NETWORK
• /
• v.22 no.2
• /
• pp.20.1-20.9
• /
• 2022

Despite the high prevalence of chronic dermatitis and the accompanied intractable itch, therapeutics that specifically target itching have low efficacy. Increasing evidence suggests that TLRs contribute to immune activation and neural sensitization; however, their roles in chronic itch remain elusive. Here, we show that the RBL-2H3 mast cell line expresses TLR4 and that treatment with a TLR4 antagonist opposes the LPS dependent increase in mRNA levels of Th2 and innate cytokines. The pathological role of TLR4 activation in itching was studied in neonate rats that developed chronic itch due to neuronal damage after receiving subcutaneous capsaicin injections. Treatment with a TLR4 antagonist protected these rats with chronic itch against scratching behavior and chronic dermatitis. TLR4 antagonist treatment also restored the density of cutaneous nerve fibers and inhibited the histopathological changes that are associated with mast cell activation after capsaicin injection. Additionally, the expression of IL-1β, IL-4, IL-5, IL-10, and IL-13 mRNA in the lesional skin decreased after TLR4 antagonist treatment. Based on these data, we propose that inhibiting TLR4 alleviated itch in a rat model of chronic relapsing itch, and the reduction in the itch was associated with TLR4 signaling in mast cells and nerve fibers.

• IMMUNE NETWORK
• /
• v.20 no.1
• /
• pp.5.1-5.15
• /
• 2020

The γδ T cells are unconventional lymphocytes that function in both innate and adaptive immune responses against various intracellular and infectious stresses. The γδ T cells can be exploited as cancer-killing effector cells since γδ TCRs recognize MHC-like molecules and growth factor receptors that are upregulated in cancer cells, and γδ T cells can differentiate into cytotoxic effector cells. However, γδ T cells may also promote tumor progression by secreting IL-17 or other cytokines. Therefore, it is essential to understand how the differentiation and homeostasis of γδ T cells are regulated and whether distinct γδ T cell subsets have different functions. Human γδ T cells are classified into Vδ2 and non-Vδ2 γδ T cells. The majority of Vδ2 γδ T cells are Vγ9δ2 T cells that recognize pyrophosphorylated isoprenoids generated by the dysregulated mevalonate pathway. In contrast, Vδ1 T cells expand from initially diverse TCR repertoire in patients with infectious diseases and cancers. The ligands of Vδ1 T cells are diverse and include the growth factor receptors such as endothelial protein C receptor. Both Vδ1 and Vδ2 γδ T cells are implicated to have immunotherapeutic potentials for cancers, but the detailed elucidation of the distinct characteristics of 2 populations will be required to enhance the immunotherapeutic potential of γδ T cells. Here, we summarize recent progress regarding cancer immunology of human γδ T cells, including their development, heterogeneity, and plasticity, the putative mechanisms underlying ligand recognition and activation, and their dual effects on tumor progression in the tumor microenvironment.

• IMMUNE NETWORK
• /
• v.20 no.6
• /
• pp.49.1-49.15
• /
• 2020

C-C chemokine receptor type 5 (CCR5) regulates the trafficking of various immune cells to sites of infection. In this study, we showed that expression of CCR5 and its ligands was rapidly increased in the kidney after systemic Candida albicans infection, and infected CCR5^-/- mice exhibited increased mortality and morbidity, indicating that CCR5 contributes to an effective defense mechanism against systemic C. albicans infection. The susceptibility of CCR5^-/- mice to C. albicans infection was due to impaired fungal clearance, which in turn resulted in exacerbated renal inflammation and damage. CCR5-mediated recruitment of NK cells to the kidney in response to C. albicans infection was necessary for the anti-microbial activity of neutrophils, the main fungicidal effector cells. Mechanistically, C. albicans induced expression of IL-23 by CD11c⁺ dendritic cells (DCs). IL-23 in turn augmented the fungicidal activity of neutrophils through GM-CSF production by NK cells. As GM-CSF potentiated production of IL-23 in response to C. albicans, a positive feedback loop formed between NK cells and DCs seemed to function as an amplification point for host defense. Taken together, our results suggest that CCR5-mediated recruitment of NK cells to the site of fungal infection is an important step that underlies innate resistance to systemic C. albicans infection.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.57 no.3
• /
• pp.239-252
• /
• 2024

Recombination activating genes (RAGs) play a crucial role in initiating V(D)J recombination, which is essential for developing adaptive immunity in vertebrates. In this study, we cloned and characterized RAG1/2 cDNA from the marine medaka Oryzias dancena (OdRAG1/2) and investigated their mRNA expression patterns during ontogenetic developmental stages. The OdRAG1 and OdRAG2 cDNA contained open reading frames (ORFs) encoding proteins containing 1,078 and 531 amino acids, respectively. Multiple sequence alignment and phylogenetic analysis revealed that OdRAG1 and OdRAG2 are highly conserved with their corresponding orthologs, featuring distinct core and non-core regions. Notably, expression analysis showed that, in contrast to other fish RAGs studied, OdRAG1/2 expression peaked at 0 days post-hatching (DPH). Additionally, for the expression of T and B cell differentiation markers, CD3γ and CD20, also peaked at 0 DPH. Collectively, adaptive immunity in O. dancena potentially begins during embryonic development, which is critical for V(D)J recombination and essential immune component development, suggesting the early ontogenetic stage interactions between innate and adaptive immunity.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.57 no.4
• /
• pp.355-363
• /
• 2024

Although aquaculture shrimp production has increased, disease mortality has also increased. Acute hepatopancreatic necrosis disease (AHPND) is an emerging disease that significantly reduces production. In this study, we have investigated the potential of medicinal herbs in enhancing the immune system of shrimp. A mixture of medicinal herb extracts (Han In-jin, Artemisia iwayomogi kitamura and Cham Dang-gwi, Angelicae gigas Nakai) was absorbed into pellets and fed to whiteleg shrimp Litopenaeus vannamei for 12 weeks. The weight gain, immune activity, and disease resistance effects of the shirmp were examined. The growth performance of shrimp fed the mixture of medicinal herbal extracts was higher than that of the control group. The prophenoloxidase (proPO) activity was found to be significantly higher at 8 weeks in the 0.1% medicinal herb extract fed group. The cumulative mortality when artificially infected with AHPND pathogens commonly decreased in the 8 weeks fed group when compared with the control group. Furthermore, the RPS (relative percentage survival) was improved. This study confirmed that a mixture of medicinal herbal extracts has a positive effect on growth performance, innate immunity and disease resistance in shrimp. The optimum concentration of the medicinal herb extract for shrimp feed was 0.1% .

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.36 no.6
• /
• pp.481-489
• /
• 2010

Introduction: TLR-5, a member of the toll-like receptor (TLR) family, is a element of the type I transmembrane receptors, which are characterized by an intracellular signaling domain homolog to the interleukin-1 receptor. These receptors recognize microbial components, particularly bacterial flagellin. All-trans retinoic acid (atRA, tretinoin), a natural metabolite of vitamin A, acts as a growth and differentiation factor in many tissues, and is also needed for immune functions. In this study, THP-1 human macrophage-monocytes were used to examine the mechanisms by which atRA regulated the expression of TLR-5. Because the molecular mechanism underlying this regulation at the transcriptional level is also unclear, this study examined which putative transcription factors are responsible for TLR-5 expression by atRA in immune cells. Materials and Methods: This study examined whether atRA induces the expression of TLR-5 in THP-1 cells using reverse transcription-polymerase chain reaction (RT-PCR), and which transcription factors are involved in regulating the TLR-5 promoter in RAW264.7 cells using a reporter assay system. Western blot analysis was used to determine which signal pathway is involved in the expression of TLR-5 in atRA-treated THP-1 cells. Results: atRA at a concentration of 10 nM greatly induced the expression of TLR-5 in THP-1 cells. Human TLR-5 promoter contains three Sp-1/GC binding sites around -50 bp and two NF-kB binding sites at -380 bp and -160 bp from the transcriptional start site of the TLR-5 gene. Sp-1/GC is primarily responsible for the constitutive TLR-5 expression, and may also contribute to NF-kB at -160 bp to induce TLR-5 after atRA stimulation in THP-1 cells. The role of NF-kB in TLR-5 expression was further confirmed by inhibitor pyrrolidine dithiocarbamate (PDTC) experiments, which greatly reduced the TLR-5 transcription by 70-80%. Conclusion: atRA induces the expression of the human TLR-5 gene and NF-kB is a critical transcription factor for the atRA-induced expression of TLR-5. Accordingly, it is conceivable that retinoids are required for adequate innate and adaptive immune responses to agents of infectious diseases. atRA and various synthetic retinoids have been used therapeutically in human diseases, such as leukemia and other cancers due to the antiproliferative and apoptosis inducing effects of retinoids. Therefore, understanding the molecular regulatory mechanism of TLR-5 may assist in the design of alternative strategies for the treatment of infectious diseases, leukemia and cancers.

• BMB Reports
• /
• v.53 no.9
• /
• pp.478-483
• /
• 2020

Kudoa septempunctata is a myxozoan parasite that causes food poisoning in individuals consuming olive flounder. The present study aimed to investigate the currently insufficiently elucidated early molecular mechanisms of inflammatory responses in the intestine owing to parasite ingestion. After Kudoa spores were isolated from olive flounder, HT29 cells were exposed to spores identified to be alive using SYTO-9 and propidium iodide staining or to antigens of Kudoa spores (KsAg). IL-1β, IL-8, TNF-α and NFKB1 expression and NF-κB activation were assessed using real-time PCR, cytokine array and western blotting. The immunofluorescence of FITC-conjugated lectins, results of ligand binding assays using Mincle-Fc and IgG-Fc, CLEC4E expressions in response to KsAg stimulation, and Mincle-dependent NF-κB activation were assessed to clarify the early immune-triggering mechanism. Inflammatory cytokines (IL-1β, GM-CSF and TNF-α), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-κB activation (pNF-κB/NF-κB) in HT29 cells increased following stimulation by KsAg. The immunofluorescence results of spores and lectins (concanavalin A and wheat germ agglutinin) suggested the importance of Mincle in molecular recognition between Kudoa spores and intestinal cells. Practically, data for Mincle-Fc and KsAg binding affinity, CLEC4E mRNA expression, Mincle immunofluorescence staining and hMincle-dependent NF-κB activation demonstrated the involvement of Mincle in the early immune-triggering mechanism. The present study newly elucidated that the molecular recognition and immune-triggering mechanism of K. septempunctata are associated with Mincle on human intestinal epithelial cells.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.42 no.6
• /
• pp.614-620
• /
• 2009

We report non-specific immune responses and its disease resistance against Edwardsiella tarda by alga mixture (HE; Hizikia:Ecklonia) in olive flounder for the first time. Five isonitrogenous (44% crude protein) and isocaloric (17.1 MJ $kg^{-1}$) diets were formulated to have 0%, 2%, 4%, 6% and 8% of the alga mixture. One of five experimental diets was fed triplicate groups of fish (30 fish/group) to apparent satiation in a flow through system. After a two week feeding, blood was sampled at 3, 6, 12, 24 h after the last feeding for a kinetic measurement of nitroblue tetrazolium (NBT) activity and healthy fish with similar sizes in each tank were selected and injected with 1 mL of E. tarda suspension ($1.0\times10^7$ CFU/mL) to evaluate the disease resistance of the fish. Dietary supplementation of alga mixtures resulted in significantly higher non-specific immune responses compared with the fish fed the control diet. The cumulative mortality was significantly lower in the fish groups fed alga mixture containing diets than control group in the challenge test with E. tarda. Therefore, the results in this study indicate that dietary supplementation of Hizikia and Ecklonia mixtures enhance the non-specific immune responses and a disease resistance of olive flounder.

• Journal of fish pathology
• /
• v.27 no.1
• /
• pp.1-9
• /
• 2014

The olive flounder, Paralichthys olivaceus is susceptible to viral hemorrhagic septicaemia virus (VHSV) at $15^{\circ}C$ but no mortality at $20^{\circ}C$ even though the virus can grow well in vitro at $20^{\circ}C$. Thus, we designed an experiment to know immune response of olive flounder against VHSV when the host reared at $15^{\circ}C$ or $20^{\circ}C$. cDNA microarray analysis was performed to compare the gene expression patterns of the kidney cells between the host reared at $15^{\circ}C$ or $20^{\circ}C$. The expression of MHC class I, IL-8, myeloperoxidae and endonuclease G-like having function for the antigen presentation and chemokine-factor were up-regulted both the $15^{\circ}C$ and $20^{\circ}C$ during VHSV infection. MHC class II gene existing on antigen-presenting cells and B cell lymphocytes, immunoglobulin (Ig) genes and phagocytosis related genes were down-regulated at $15^{\circ}C$ but highly expressed at $20^{\circ}C$. It can be thought that innate immune related antigen presentation by MHC class I and phagocytosis reaction against VHSV are efficiently occur both the temperature but macrophage or B cell related antigen presentation via MHC class II fails to induce downstream immune reactions (adaptive immunity) to make antibody, and it can be one of the reason that causes high mortality only at $15^{\circ}C$.

• Korean Journal of Food Science and Technology
• /
• v.47 no.4
• /
• pp.511-516
• /
• 2015

In the present study, we investigated changes in the nutritional components of pre- and post-germinated barley seeds and also investigated their immune-enhancing effects such as production of nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)-${\alpha}$ on the RAW 264.7 macrophage cell line. Protein and total sugar contents increased slightly with increase in germination time. The major neutral sugars of germinated balrey seeds were arabinose, glucose, and xylose. Glucose content decreased during germination, whereas arabinose and xylose contents increased during germination. Amino acid contents of barley germinated for 24 and 48 hours increased 1.03-fold and 1.24-fold, respectively, compared to that of pre-germinated barley. Moreover, RAW 264.7 macrophages stimulated with barley germinated for 24 and 48 hours showed higher production of NO, IL-6, and TNF-${\alpha}$ compared to that observed in pre-germinated barley. The results of the present study indicate that germinated barley may have immune-enhancing effects derived from its ability to activate RAW 264.7 macrophages, which play a major role in innate immunity.