Receptor Gene Family Research Articles

Association analysis of polymorphisms in EGFR, HER2, ESR1 and THRA genes with coronary artery diseases

BackgroundResearch in the genetic basis of coronary artery diseases (CAD) has identified some genes and pathways associated with diseases that would not be considered to underlie conventional risk factors. Among these genes there are the EGFR (epidermal growth factor receptor) receptor family genes and the regulation factor genes (such as thyroid hormone receptor α (THRA) and estrogen receptor α (ESR1)). AimIn this study we investigated the relation between 4 polymorphisms within EGFR, HER2 (human epidermal growth factor receptor 2), ESR1 and THRA genes and CAD. Subjects and methodsThe association analysis was performed with 151 healthy individuals and 151 CAD patients documented by angiography. ResultsNo significant difference was found in the allelic and genotypic frequency distribution of the four variants studied between the control and patient groups. We have also investigated the relationship of these polymorphic sites with clinical and biochemical parameters such as smoking habit, diabetes mellitus, hypertension, dyslipidemia, CAD severity, glucose, triglyceride, total cholesterol and urea levels. The EGFR and THRA variants were associated with glycemia and triglyceride levels, respectively. Also a significant correlation was found between the ESR1 polymorphism and the levels of urea and triglyceride. ConclusionOur results suggest the absence of any significant association between the four polymorphisms analyzed and CAD risk as well as disease severity.

P036 Whole gene sequencing of KIR-3DL1 with PacBio RSII and the distribution of alleleic variants in different ethic groups

Aim The killer-cell immunoglobulin-like receptor (KIR) gene family are involved in immune modulation during viral infection, autoimmune disease and in allogeneic stem cell transplantation. Most KIR gene diversity studies and their impact on the transplant outcome is performed by gene absence/presence assays. However it is well known that KIR gene allelic variations have biological significance. Allele level typing of KIR genes has been very challenging until recently due to the homologous nature of those genes and very long intronic sequences. PacBio RSII platform’s Single Molecule, Real-Time (SMRT®) sequencing, generates average long reads of 10 to 15 kb and allows us to obtain in-phase long sequence reads. We have developed a PCR assay for PacBio RSII platform in our lab for 3DL1 whole gene sequencing. This approach allows us to obtain allele level typing for 3DL1 genes and could serve as a model to type other KIR genes at allelic level. Methods DNA samples were obtained from 41 cell lines of Caucasian ethnicity and 97 volunteers with Korean and South Asian origin. Gene absence/presence assays were done on Illumina MiSeq platform using in-house protocol.We performed whole gene 3DL1 sequencing on the genomic DNA of samples using two gene-specific primers to get two amplicons (5.5 kb and 5.3 kb, respectively) that will span all the exons and most of the Intron 5. These amplicons were sequenced on the PacBio RSII platform. We analyzed the data based on current KIR DB v 2.6.0 using our in house program. Results A total of 123 out of 138 samples had 3DL1 gene by gene absence/presence assay. Based on the exon sequences alone, 123 samples had 47 of the 110 known alleles. However when we included intron sequences to our analysis only 23 sequences out of 47 unique alleles had a match to the database for intronic sequences. The remainder 24 sequences had novel variations in the introns. The majority were in intron 6. 3DL1 ∗ 002 that has stronger inhibitory receptor was found in 13 samples and mostly in CAU/EURO samples in this study. Some ethnic clustering of certain alleles (ie: 3DL1 ∗ 01501/2 group) were observed in Korean samples. Conclusion Whole gene sequencing of KIR genes is now feasible. Allelic typing of KIR genes will allow us to better dissect their modulatory role during immune response and their distribution among diverse populations. S. Yang: Stock Shareholder; Company/Organization; PacBio.

Population differences in olfaction accompany host shift in Drosophila mojavensis.

Evolutionary shifts in plant-herbivore interactions provide a model for understanding the link among the evolution of behaviour, ecological specialization and incipient speciation. Drosophila mojavensis uses different host cacti across its range, and volatile chemicals emitted by the host are the primary cue for host plant identification. In this study, we show that changes in host plant use between distinct D. mojavensis populations are accompanied by changes in the olfactory system. Specifically, we observe differences in olfactory receptor neuron specificity and sensitivity, as well as changes in sensillar subtype abundance, between populations. Additionally, RNA-seq analyses reveal differential gene expression between populations for members of the odorant receptor gene family. Hence, alterations in host preference are associated with changes in development, regulation and function at the olfactory periphery.

Variations in TAS1R taste receptor gene family modify food intake and gastric cancer risk in a Korean population.

Human taste receptor type 1 (TAS1R, T1R) gene family mediates the perception of umami and sweet tastes and regulates metabolism. This study investigated whether variants in the TAS1R are associated with food intake and susceptibility to gastric cancer (GC). A total of 1131 Koreans including 377 GC cases were analyzed to determine the dietary intake and genotype for 25 variants in TAS1R genes. Among the genotyped polymorphisms, six loci with a minor allele frequency >0.05 were tested for an association with dietary intake and GC risk. Findings suggest that TAS1R polymorphisms were associated with a variety of food intake that is beyond the known T1R-taste association. Furthermore, differential consumption for cigarettes and citrus fruits by TAS1R genetic variants may be linked to GC risk in males and females, respectively. Additionally, in males, TAS1R1 rs34160967 heterozygote showed an increased risk of GC (odds ratio: 1.52, 95% confidence interval: 1.10-2.09), independent of dietary intake. However, this GC-risk effect of TAS1R variants was not evident in females. Genetic variants in TAS1R were associated with dietary consumption, which may be associated with GC risk. TAS1R1 rs34160967 may also modify the risk for GC independent of diet in Korean males.

G Protein-Coupled Receptors in Macrophages.

As the largest receptor gene family in the human genome, with >800 members, the signal-transducing G protein-coupled receptors (GPCRs) play critical roles in nearly all conceivable physiological processes, ranging from the sensing of photons and odorants to metabolic homeostasis and migration of leukocytes. Unfortunately, an exhaustive review of the several hundred GPCRs expressed by myeloid cells/macrophages (P.J. Groot-Kormelink, L .Fawcett, P.D. Wright, M. Gosling, and T.C. Kent, BMC Immunol 12:57, 2012, doi:10.1186/1471-2172-13-57) is beyond the scope of this chapter; however, we will endeavor to cover the GPCRs that contribute to the major facets of macrophage biology, i.e., those whose expression is restricted to macrophages and the GPCRs involved in macrophage differentiation/polarization, microbial elimination, inflammation and resolution, and macrophage-mediated pathology. The chemokine receptors, a major group of myeloid GPCRs, will not be extensively covered as they are comprehensively reviewed elsewhere.

Pangolin genomes and the evolution of mammalian scales and immunity.

Pangolins, unique mammals with scales over most of their body, no teeth, poor vision, and an acute olfactory system, comprise the only placental order (Pholidota) without a whole-genome map. To investigate pangolin biology and evolution, we developed genome assemblies of the Malayan (Manis javanica) and Chinese (M. pentadactyla) pangolins. Strikingly, we found that interferon epsilon (IFNE), exclusively expressed in epithelial cells and important in skin and mucosal immunity, is pseudogenized in all African and Asian pangolin species that we examined, perhaps impacting resistance to infection. We propose that scale development was an innovation that provided protection against injuries or stress and reduced pangolin vulnerability to infection. Further evidence of specialized adaptations was evident from positively selected genes involving immunity-related pathways, inflammation, energy storage and metabolism, muscular and nervous systems, and scale/hair development. Olfactory receptor gene families are significantly expanded in pangolins, reflecting their well-developed olfaction system. This study provides insights into mammalian adaptation and functional diversification, new research tools and questions, and perhaps a new natural IFNE-deficient animal model for studying mammalian immunity.

A targeted sequencing study of glutamatergic candidate genes in suicide attempters with bipolar disorder

Suicidal behavior has been shown to have a heritable component that is partly driven by psychiatric disorders [Brent and Mann, 2005]. However, there is also an independent factor contributing to the heritability of suicidal behavior. We previously conducted a genome-wide association study (GWAS) of bipolar suicide attempters and bipolar non-attempters to assess this independent factor [Willour et al., 2012]. This GWAS implicated glutamatergic neurotransmission in attempted suicide. In the current study, we have conducted a targeted next-generation sequencing study of the glutamatergic N-methyl-D-aspartate (NMDA) receptor, neurexin, and neuroligin gene families in 476 bipolar suicide attempters and 473 bipolar non-attempters. The goal of this study was to gather sequence information from coding and regulatory regions of these glutamatergic genes to identify variants associated with attempted suicide. We identified 186 coding variants and 4,298 regulatory variants predicted to be functional in these genes. No individual variants were overrepresented in cases or controls to a degree that was statistically significant after correction for multiple testing. Additionally, none of the gene-level results were statistically significant following correction. While this study provides no direct support for a role of the examined glutamatergic candidate genes, further sequencing in expanded gene sets and datasets will be required to ultimately determine whether genetic variation in glutamatergic signaling influences suicidal behavior. © 2016 Wiley Periodicals, Inc.

SorCS2 is required for BDNF-dependent plasticity in the hippocampus.

SorCS2 is a member of the Vps10p-domain receptor gene family receptors with critical roles in the control of neuronal viability and function. Several genetic studies have suggested SORCS2 to confer risk of bipolar disorder, schizophrenia and attention deficit-hyperactivity disorder. Here we report that hippocampal N-methyl-d-aspartate receptor-dependent synaptic plasticity is eliminated in SorCS2-deficient mice. This defect was traced to the ability of SorCS2 to form complexes with the neurotrophin receptor p75NTR, required for pro-brain-derived neurotrophic factor (BDNF) to induce long-term depression, and with the BDNF receptor tyrosine kinase TrkB to elicit long-term potentiation. Although the interaction with p75NTR was static, SorCS2 bound to TrkB in an activity-dependent manner to facilitate its translocation to postsynaptic densities for synaptic tagging and maintenance of synaptic potentiation. Neurons lacking SorCS2 failed to respond to BDNF by TrkB autophosphorylation, and activation of downstream signaling cascades, impacting neurite outgrowth and spine formation. Accordingly, Sorcs2-/- mice displayed impaired formation of long-term memory, increased risk taking and stimulus seeking behavior, enhanced susceptibility to stress and impaired prepulse inhibition. Our results identify SorCS2 as an indispensable coreceptor for p75NTR and TrkB in hippocampal neurons and suggest SORCS2 as the link between proBDNF/BDNF signaling and mental disorders.

Progesterone inhibits proliferation and modulates expression of proliferation—Related genes in classical progesterone receptor-negative human BxPC3 pancreatic adenocarcinoma cells

Recent studies suggest that progesterone may possess anti-tumorigenic properties. However, a growth-modulatory role of progestins in human cancer cells remains obscure. With the discovery of a new class of membrane progesterone receptors (mPRs) belonging to the progestin and adipoQ receptor gene family, it becomes important to study the effect of this hormone on proliferation of tumor cells that do not express classical nuclear progesterone receptors (nPRs). To identify a cell line expressing high levels of mPRs and lacking nPRs, we examined mRNA levels of nPRs and three forms of mPRs in sixteen human tumor cell lines of different origin. High expression of mPR mRNA has been found in pancreatic adenocarcinoma BxPC3 cells, while nPR mRNA has not been detected in these cells. Western blot analysis confirmed these findings at the protein level. We revealed specific binding of labeled progesterone in these cells with affinity constant similar to that of human mPR expressed in yeast cells. Progesterone at high concentration of 20 μM significantly reduced the mRNA levels of proliferation markers Ki67 and PCNA, as well as of cyclin D1, and increased the mRNA levels of cyclin dependent kinase inhibitors p21 and p27. Progesterone (1 μM and 20 μM) significantly inhibited proliferative activity of BxPC3 cells. These results point to anti-proliferative effects of the progesterone high concentrations on BxPC3 cells and suggest that activation of mPRs may mediate this action. Our data are a starting point for further investigations regarding the application of progesterone in pancreatic cancer.

Molecular characterization and expression profiles of nicotinic acetylcholine receptors in the rice striped stem borer, Chilo suppressalis (Lepidoptera: Crambidae).

Nicotinic acetylcholine receptors (nAChRs) are members of the cys-loop ligand-gated ion channel (cysLGIC) superfamily, mediating fast synaptic cholinergic transmission in the central nervous system in insects. Insect nAChRs are the molecular targets of economically important insecticides, such as neonicotinoids and spinosad. Identification and characterization of the nAChR gene family in the rice striped stem borer, Chilo suppressalis, could provide beneficial information about this important receptor gene family and contribute to the investigation of the molecular modes of insecticide action and resistance for current and future chemical control strategies. We searched our C. suppressalis transcriptome database using Bombyx mori nAChR sequences in local BLAST searches and obtained the putative nAChR subunit complementary DNAs (cDNAs) via reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends methods. Similar to B. mori, C. suppressalis possesses 12 nAChR subunits, including nine α-type and three β-type subunits. Quantitative RT-PCR analysis revealed the expression profiles of the nAChR subunits in various tissues, including the brain, subesophageal ganglion, thoracic ganglion, abdominal ganglion, hemocytes, fat body, foregut, midgut, hindgut and Malpighian tubules. Developmental expression analyses showed clear differential expression of nAChR subunits throughout the C. suppressalis life cycle. The identification of nAChR subunits in this study will provide a foundation for investigating the diverse roles played by nAChRs in C. suppressalis and for exploring specific target sites for chemicals that control agricultural pests while sparing beneficial species.

Dual targeting of EGFR and ERBB2 pathways produces a synergistic effect on cancer cell proliferation and migration in vitro

Members of the epidermal growth factor receptor (EGFR/ERBB) gene family are frequently dysregulated in a range of human cancers, and therapeutics targeting these proteins are in clinical use. We hypothesized that similar pathways are involved in feline and canine tumours and that the same drugs may be of clinical use in veterinary patients. We investigated EGFR and ERBB2 targeting using a panel of feline and canine cell lines. EGFR and ERBB2 were targeted with siRNAs or tyrosine kinase inhibitors (TKIs) and their effect on cellular proliferation, colony formation and migration was investigated in vitro. Here we report that EGFR and ERBB2 combined siRNA targeting produced synergistic effects in feline and canine cell lines similar to that reported in human cell lines. We conclude that dual EGFR and ERBB2 targeting using TKIs should be further evaluated as a potential new therapeutic strategy in feline head and neck and mammary tumours and canine mammary tumours.

Effects of killer cell immunoglobulin like receptor gene on common complications and recurrence after allogeneic hematopoietic stem cell transplantation

Hematopoietic stem cell transplantation(HSCT) is an effective treatment for some hematologic diseases, immunodeficiency diseases and hereditary metabolic diseases. Accompanied by increased transplantations, the complications of HSCT aroused people's attention, including virus infection, graft versus host disease(GVHD) and recurrence are the main reasons for the failure of transplantation. Natural killer(NK) cells are important part of human nonspecific cellular immunity, as the first line of defense against foreign pathogens. NK cells are the first present lymphatic cells in peripheral blood after hematopoietic reconstitution.The NK cells' killer cell immunoglobulin receptor(KIR) gene family encode KIR, some KIR proteins are involved in human leakocyte omtigen(HLA) class Ⅰ ligand binding.In recent years, more and more studies have shown that donor KIR genes or KIR haplotype are correlated with virus infection, GVHD and recurrence after transplantation. Key words: Receptors, natural killer cell; Hematopoietic stem cell transplantation; Graft host disease; Recurrence

The role of the low-density lipoprotein receptor-related protein 1 (LRP-1) in regulating blood-brain barrier integrity.

The blood-brain barrier (BBB) is a protective structure that helps maintaining the homeostasis in cerebral microenvironment by limiting the passage of molecules into the brain. BBB is formed by closely conjugated endothelial cells, with astrocytic endfeet surrounded and extracellular matrix (ECM) consolidated. Numerous neurological diseases can cause disturbance of BBB, leading to brain edema and neurological dysfunctions. The low-density lipoprotein (LDL) receptor-related protein 1 (LRP-1), a member of the LDL receptor gene family, is involved in a lot of important processes in the brain under both physiological and pathological conditions. As a membrane receptor, LRP-1 interacts with a variety of ligands and mediates the internalization of several important substances. LRP-1 is found responsible for inducing the opening of BBB following ischemic attack. It has also been reported that LRP-1 regulates several tight junction proteins and mediates the clearance of major ECM-degrading proteinases. In this review, we briefly discussed the role of LRP-1 in regulating BBB integrity by modulating tight junction proteins, endothelial cells and the remodeling of ECM.

Molecular Characterization and Sex Distribution of Chemosensory Receptor Gene Family Based on Transcriptome Analysis of Scaeva pyrastri.

Chemosensory receptors play key roles in insect behavior. Thus, genes encoding these receptors have great potential for use in integrated pest management. The hover fly Scaeva pyrastri (L.) is an important pollinating insect and a natural enemy of aphids, mainly distributed in the Palearctic and Nearctic regions. However, a systematic identification of their chemosensory receptor genes in the antennae has not been reported. In the present study, we assembled the antennal transcriptome of S. pyrastri by using Illumina sequencing technology. Analysis of the transcriptome data identified 60 candidate chemosensory genes, including 38 for odorant receptors (ORs), 16 for ionotropic receptors (IRs), and 6 for gustatory receptors (GRs). The numbers are similar to those of other Diptera species, suggesting that we were able to successfully identify S. pyrastri chemosensory genes. We analyzed the expression patterns of all genes by using reverse transcriptase PCR (RT-PCR), and found that some genes exhibited sex-biased or sex-specific expression. These candidate chemosensory genes and their tissue expression profiles provide information for further studies aimed at fully understanding the molecular basis behind chemoreception-related behaviors in S. pyrastri.

25 - A Rare Case of Russell-Silver and 1p36 Deletion Syndromes

receptor gene family, which have been implicated in the etiology of autism and susceptibility to alcohol dependence, and may in tern play a role in the family’s problems with drugs and alcohol. The maternal great grandmother (I.1) did not have the duplications of 16q observed on array in the other family members. However, she was found to have a 461 kb loss on 16q21. This appears to be the same region which separates two of the duplicated regions observed in other family members. FISH analysis has determined the chromosome 16 duplicated regions have been inserted into the short arm of chromosome 4. Further molecular characterization of the duplication, deletions, and insertion are needed for a better understanding of the genomic architecture and mechanism of this complex rearrangement of chromosome 16. The complex rearrangement and copy number aberrations observed in the majority of this family appear to have begun as a unique deletion on chromosome 4p and a unique deletion of chromosome 16q21 in generation I as revealed by microarray analysis. Since at least two of her children received the insertion duplication, she most likely has a balanced version of the insertion. The complex duplication is speculated to have emerged by either nonhomologous end joining recombination (NHEJ) such as in a chromothripsis type event or non-allelic homologous recombination (NAHR) due to microhomology. NAHR mediated by regions of sequence homology at the breakpoints of the 4p deletion and the proximal and distal breakpoints of chromosome 16 segments might best explain the insertion of the 16 into chromosome 4. Further molecular characterization of the breakpoints involved will help elucidate the mechanism which produced this complex aberration.

Crypt cells are involved in kin recognition in larval zebrafish.

Zebrafish larvae imprint on visual and olfactory kin cues at day 5 and 6 postfertilization, respectively, resulting in kin recognition later in life. Exposure to non-kin cues prevents imprinting and kin recognition. Imprinting depends on MHC class II related signals and only larvae sharing MHC class II alleles can imprint on each other. Here, we analyzed which type of olfactory sensory neuron (OSN) detects kin odor. The single teleost olfactory epithelium harbors ciliated OSNs carrying OR and TAAR gene family receptors (mammals: main olfactory epithelium) and microvillous OSNs with V1R and V2R gene family receptors (mammals: vomeronasal organ). Additionally, teleosts exhibit crypt cells which possess microvilli and cilia. We used the activity marker pERK (phosphorylated extracellular signal regulated kinase) after stimulating 9 day old zebrafish larvae with either non-kin conspecific or food odor. While food odor activated both ciliated and microvillous OSNs, only the latter were activated by conspecific odor, crypt cells showed no activation to both stimuli. Then, we tested imprinted and non-imprinted larvae (full siblings) for kin odor detection. We provide the first direct evidence that crypt cells, and likely a subpopulation of microvillous OSNs, but not ciliated OSNs, play a role in detecting a kin odor related signal.

KIR2DL2/2DL3-E(35) alleles are functionally stronger than -Q(35) alleles.

KIR2DL2 and KIR2DL3 segregate as alleles of a single locus in the centromeric motif of the killer cell immunoglobulin-like receptor (KIR) gene family. Although KIR2DL2/L3 polymorphism is known to be associated with many human diseases and is an important factor for donor selection in allogeneic hematopoietic stem cell transplantation, the molecular determinant of functional diversity among various alleles is unclear. In this study we found that KIR2DL2/L3 with glutamic acid at position 35 (E35) are functionally stronger than those with glutamine at the same position (Q35). Cytotoxicity assay showed that NK cells from HLA-C1 positive donors with KIR2DL2/L3-E35 could kill more target cells lacking their ligands than NK cells with the weaker -Q35 alleles, indicating better licensing of KIR2DL2/L3+ NK cells with the stronger alleles. Molecular modeling analysis reveals that the glutamic acid, which is negatively charged, interacts with positively charged histidine located at position 55, thereby stabilizing KIR2DL2/L3 dimer and reducing entropy loss when KIR2DL2/3 binds to HLA-C ligand. The results of this study will be important for future studies of KIR2DL2/L3-associated diseases as well as for donor selection in allogeneic stem cell transplantation.

The chemosensory receptors of codling moth Cydia pomonella-expression in larvae and adults.

Olfaction and gustation play critical roles in the life history of insects, mediating vital behaviors such as food, mate and host seeking. Chemosensory receptor proteins, including odorant receptors (ORs), gustatory receptors (GRs) and ionotropic receptors (IRs) function to interface the insect with its chemical environment. Codling moth, Cydia pomonella, is a worldwide pest of apple, pear and walnut, and behavior-modifying semiochemicals are used for environmentally safe control. We produced an Illumina-based transcriptome from antennae of males and females as well as neonate head tissue, affording a qualitative and quantitative analysis of the codling moth chemosensory receptor repertoire. We identified 58 ORs, 20 GRs and 21 IRs, and provide a revised nomenclature that is consistent with homologous sequences in related species. Importantly, we have identified several OR transcripts displaying sex-biased expression in adults, as well as larval-enriched transcripts. Our analyses have expanded annotations of the chemosensory receptor gene families, and provide first-time transcript abundance estimates for codling moth. The results presented here provide a strong foundation for future work on codling moth behavioral physiology and ecology at the molecular level, and may lead to the development of more precise biorational control strategies.

Biochemical and Functional Insights into the Integrated Regulation of Innate Immune Cell Responses by Teleost Leukocyte Immune-Type Receptors.

Across vertebrates, innate immunity consists of a complex assortment of highly specialized cells capable of unleashing potent effector responses designed to destroy or mitigate foreign pathogens. The execution of various innate cellular behaviors such as phagocytosis, degranulation, or cell-mediated cytotoxicity are functionally indistinguishable when being performed by immune cells isolated from humans or teleost fishes; vertebrates that diverged from one another more than 450 million years ago. This suggests that vital components of the vertebrate innate defense machinery are conserved and investigating such processes in a range of model systems provides an important opportunity to identify fundamental features of vertebrate immunity. One characteristic that is highly conserved across vertebrate systems is that cellular immune responses are dependent on specialized immunoregulatory receptors that sense environmental stimuli and initiate intracellular cascades that can elicit appropriate effector responses. A wide variety of immunoregulatory receptor families have been extensively studied in mammals, and many have been identified as cell- and function-specific regulators of a range of innate responses. Although much less is known in fish, the growing database of genomic information has recently allowed for the identification of several immunoregulatory receptor gene families in teleosts. Many of these putative immunoregulatory receptors have yet to be assigned any specific role(s), and much of what is known has been based solely on structural and/or phylogenetic relationships with mammalian receptor families. As an attempt to address some of these shortcomings, this review will focus on our growing understanding of the functional roles played by specific members of the channel catfish (Ictalurus punctatus) leukocyte immune-type receptors (IpLITRs), which appear to be important regulators of several innate cellular responses via classical as well as unique biochemical signaling networks.

Immune responses to alum in the Japanese pufferfish (Takifugu rubripes)

To elucidate the basis of innate immune responses against insoluble compounds (such as silica and alum) through the inflammasome activation pathway in fish, biological assays and expression analysis of immune-related genes were conducted in the Japanese pufferfish (Takifugu rubripes) head kidney (HK) cells. At first, the Japanese pufferfish was stimulated by intraperitoneal injection of alum (4 mg/fish). After 0, 6, 12, 24 and 48 h post-injection, HK cells were prepared from the injected fish. To understand functionality of activated inflammatory response, phagocytic activity and superoxide anion production (NBT assay) in the HK cells were assessed. As results, the phagocytic and NBT reduction activities were significantly increased in alum-stimulation. To further clarify the immune response through transcriptomic analysis, expression of 19 cytokine genes was checked by the multiplex reverse transcription (RT)-PCR method. Results showed that expression of IL-6 and type I-IFN genes was significantly elevated in the alum-stimulated fish. Furthermore, expression of NLR-C9 and NLR-C12 genes [i.e., NOD-like receptor (NLR) family genes as sensor molecules of inflammasome], ASC and caspase-1 genes (inflammasome associated molecules) were examined by qPCR. Results indicated an increased expression of four different inflammasome associated genes in the HK cells of fish injected with alum. The results suggested activation of inflammatory response to the alum with a likely involvement of inflammasome components in the Japanese pufferfish.