Pongsak Utaisincharoen

Publications (12)42.5 Total impact

• Article: Involvement of GRIM-19 in apoptosis induced in H5N1 virus-infected human macrophages.

  Peeraya Ekchariyawat, Arunee Thitithanyanont, Stitaya Sirisinha, Pongsak Utaisincharoen
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  ABSTRACT: The fatal H5N1 infection has a high mortality rate among infected patients. The pathogenesis of H5N1 viral infection is associated with the ability of the virus to induce apoptotic cell death. However, the molecular mechanism of apoptosis induced by H5N1 remains unclear. In the present study we demonstrate that H5N1 virus is able to up-regulate the expression of gene associated with retinoid and interferon induced mortality-19 (GRIM-19) in human monocyte-derived macrophages (hMDMs). GRIM-19 has been identified as a novel gene with apoptotic effects in virus-infected cells. The percentage of apoptotic cells is significantly decreased in H5N1-infected GRIM-19 depleted hMDMs, which is also associated with a decrease of BH3-interacting domain death agonist cleavage and apoptosis-inducing factor (AIF) release to the cytosol. These results suggested the involvement of GRIM-19 in apoptosis induced by H5N1 virus. Furthermore, neutralizing-IFN-β Ab is able to suppress GRIM-19 expression in H5N1-infected cells resulting in a decrease in apoptotic cell number, indicating that IFN-β secreted by H5N1-infected hMDMs regulates GRIM-19 expression leading to apoptosis. Altogether, the results presented here provide additional insight on the regulatory mechanism of H5N1 viral-induced apoptotic cell death in hMDMs.
  Innate Immunity 03/2013; · 4.00 Impact Factor
• Article: Involvement of Signal Regulatory Protein α, a Negative Regulator of Toll-Like Receptor Signaling, in Impairing MyD88-Independent Pathway and Intracellular Killing of B. pseudomallei-Infected Mouse Macrophages.

  Pankaj Baral, Pongsak Utaisincharoen
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  ABSTRACT: The facultative intracellular Gram-negative bacterium Burkholderia pseudomallei is the causative agent of melioidosis and is known for its ability to evade Toll-like receptor (TLR)-mediated innate immune response. Previously it has been demonstrated that this bacterium was able to suppress MyD88-independent pathway and can survive macrophage intracellular killing. However the underlying mechanisms responsible for the suppression of this pathway are not fully understood. In the present report, we showed that both living and heat-killed B. pseudomallei restricts the TLR signaling response, particularly macrophage inducible nitric oxide synthase (iNOS) expression by preventing downregulation of constitutively expressed signal regulatory protein α (SIRPα) molecule, a known negative regulator of TLR signaling. In contrast, an LPS mutant of B. pseudomallei, a less virulent strain, was able to downregulate SIRPα expression in mouse macrophages. However, depletion of constitutively expressed SIRPα was able to induce the gene expression downstream of TLR signaling pathways, particularly MyD88-independent pathway such as inos, leading to enhanced macrophage intracellular killing of B. pseudomallei. Induction of gene expression was consistent with the enhanced degradation pattern of IκBα with SIRPα depletion. Additionally, the downregulation of SIRPα expression with upregulation of iNOS was observed when the macrophages were pretreated with interferon (IFN)-γ prior to the infection, suggesting the enhanced intracellular killing of bacteria by IFN-γ is associated with the decreased SIRPα expression. Altogether our findings demonstrate that B. pseudomallei evades macrophage intracellular killing by preventing the downregulation of SIRPα that results the inhibition of gene expression downstream of MyD88-independent pathway.
  Infection and immunity 09/2012; · 4.21 Impact Factor
• Article: Apoptosis induced by avian H5N1 virus in human monocyte-derived macrophages involves TRAIL-inducing caspase-10 activation.

  Peeraya Ekchariyawat, Arunee Thitithanyanont, Stitaya Sirisinha, Pongsak Utaisincharoen
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  ABSTRACT: Avian influenza virus H5N1 is a potentially fatal disease not only in birds, but also in humans. The virus is able to induce apoptosis in many cell types including macrophages and dendritic cells. In the present study, we demonstrated that TNF-related apoptosis-inducing ligand (TRAIL) is involved in apoptosis-associated mechanisms of apoptosis downstream of the TRAIL receptor in H5N1 virus-infected human monocyte-derived macrophages (MDMs). Activation of caspase-10 was also observed in avian virus H5N1-infected MDMs. In the presence of caspase-10 inhibitor, Z-AEVD-FMK, the activation of Bid and a release of apoptotic-inducing factor (AIF) from mitochondria were markedly reduced, resulting in a significant decrease of apoptotic cells which suggested the involvement of caspase-10 activation in mitochondria leakage. Furthermore, neutralizing Ab against TRAIL significantly reduced caspase-10 activities, which paralleled with a decrease in the number of apoptotic cells. Together, this study demonstrated that apoptosis in avian virus H5N1-infected MDMs was induced by TRAIL-activated caspase-10, resulting in the activation of Bid and the release of AIF from mitochondria.
  Innate Immunity 09/2011; 18(3):390-7. · 4.00 Impact Factor
• Article: Nucleotide-binding oligomerization domain-containing protein 2 regulates suppressor of cytokine signaling 3 expression in Burkholderia pseudomallei-infected mouse macrophage cell line RAW 264.7.

  Matsayapan Pudla, Anchasa Kananurak, Kornkaew Limposuwan, Stitaya Sirisinha, Pongsak Utaisincharoen
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  ABSTRACT: Burkholderia pseudomallei, a causative agent of melioidosis, is a facultative intracellular Gram-negative bacterium that can survive and multiply inside the macrophages. Toll-like receptors are one class of pattern recognition receptors (PRRs) that have been documented to play significant role in B. pseudomallei infection. In the present study, we investigated a potential role of nucleotide-binding oligomerization domain-containing protein 1 and 2 (NOD1 and NOD2), cytoplasmic pattern recognition receptors, in B. pseudomallei-infected mouse macrophage cell line RAW 264.7. Both live and heat-killed B. pseudomallei were able to up-regulate NOD1 and NOD2 expression in a time-dependent manner. Marked reduction of a negative regulator, suppressor of cytokine signaling 3 (SOCS3), expression was observed only in B. pseudomallei-infected NOD2-depleted macrophages and not in NOD1-depleted macrophages. The decrease in SOCS3 expression also led to an increase in IFN-γ responsiveness as judged by an enhanced STAT-1 phosphorylation on tyrosine 701 in the B. pseudomallei-infected macrophages. Together, these results suggested that, in addition to using other PRRs to evade macrophage defense, B. pseudomallei may also use NOD2 to regulate a negative regulator like SOCS3.
  Innate Immunity 11/2010; 17(6):532-40. · 4.00 Impact Factor
• Article: Antiviral immune responses in H5N1-infected human lung tissue and possible mechanisms underlying the hyperproduction of interferon-inducible protein IP-10.

  Arunee Thitithanyanont, Anneke Engering, Monkol Uiprasertkul, Peeraya Ekchariyawat, Suwimon Wiboon-Ut, Romchat Kraivong, Amporn Limsalakpetch, Utaiwan Kum-Arb, Kosol Yongvanitchit, Noppadol Sa-Ard-Iam, Pimprapa Rukyen, Rangsini Mahanonda, Kamon Kawkitinarong, Prasert Auewarakul, Pongsak Utaisincharoen, Stitaya Sirisinha, Carl J Mason, Mark M Fukuda, Sathit Pichyangkul
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  ABSTRACT: Information on the immune response against H5N1 within the lung is lacking. Here we describe the sustained antiviral immune responses, as indicated by the expression of MxA protein and IFN-alpha mRNA, in autopsy lung tissue from an H5N1-infected patient. H5N1 infection of primary bronchial/tracheal epithelial cells and lung microvascular endothelial cells induced IP-10, and also up-regulated the retinoic acid-inducible gene-I (RIG-I). Down-regulation of RIG-I gene expression decreased IP-10 response. Co-culturing of H5N1-infected pulmonary cells with TNF-alpha led to synergistically enhanced production of IP-10. In the absence of viral infection, TNF-alpha and IFN-alpha also synergistically enhanced IP-10 response. Methylprednisolone showed only a partial inhibitory effect on this chemokine response. Our findings strongly suggest that both the H5N1 virus and the locally produced antiviral cytokines; IFN-alpha and TNF-alpha may have an important role in inducing IP-10 hyperresponse, leading to inflammatory damage in infected lung.
  Biochemical and Biophysical Research Communications 08/2010; 398(4):752-8. · 2.48 Impact Factor
• Source

  Available from: Anan Jongkaewwattana

  Article: Cross-reactive Antibodies against avian influenza virus A (H5N1).

  Sathit Pichyangkul, Anan Jongkaewwattana, Arunee Thitithanyanont, Peeraya Ekchariyawat, Suwimon Wiboon-ut, Amporn Limsalakpetch, Kosol Yongvanitchit, Utaiwan Kum-Arb, Rangsini Mahanonda, Pongsak Utaisincharoen, Stitaya Sirisinha, Carl J Mason, Mark M Fukuda
  Emerging Infectious Diseases 09/2009; 15(9):1537-9. · 6.79 Impact Factor
• Source

  Available from: Suwimol Taweechaisupapong

  Article: Comparative in vivo and in vitro analyses of putative virulence factors of Burkholderia pseudomallei using lipopolysaccharide, capsule and flagellin mutants.

  Chanthiwa Wikraiphat, Jaruek Charoensap, Pongsak Utaisincharoen, Surasakdi Wongratanacheewin, Suwimol Taweechaisupapong, Donald E Woods, Jan G M Bolscher, Stitaya Sirisinha
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  ABSTRACT: Burkholderia pseudomallei is a gram-negative bacillus that is the causative agent of melioidosis. We evaluated host-pathogen interaction at different levels using three separate B. pseudomallei mutants generated by insertional inactivation. One of these mutants is defective in the production of the polysaccharide side chains associated with lipopolysaccharide; one does not produce the capsular polysaccharide with the structure -3)-2-O-acetyl-6-deoxy-beta-d-manno-heptopyranose-(1-; and the third mutant does not produce flagellin. We compared the in vivo virulence in BALB/c mice, the in vitro fate of intracellular survival inside human polymorphonuclear cells (PMNs) and macrophages (Mphis) and the susceptibility to killing by 30% normal human serum, reactive nitrogen and oxygen intermediates and antimicrobial peptides with that of their wild-type counterpart. The lipopolysaccharide and capsule mutants demonstrated a marked reduction in virulence for BALB/c mice, but the flagellin mutant was only slightly less virulent than the parent strain. The results from the BALB/c mice experiments correlated with survival in Mphis. The lipopolysaccharide and capsule mutants were also more susceptible to killing by antimicrobial agents. All bacteria were equally susceptible to killing by PMNs. Altogether, the data suggest that lipopolysaccharide and capsule and, to a much lesser extent, flagella, are most likely associated with the virulence of this bacterium and highlight the importance of intracellular killing by PMNs and Mphis in disease pathogenesis.
  FEMS Immunology & Medical Microbiology 06/2009; 56(3):253-9. · 2.44 Impact Factor
• Source

  Available from: PubMed Central

  Article: Differential intracellular fate of Burkholderia pseudomallei 844 and Burkholderia thailandensis UE5 in human monocyte-derived dendritic cells and macrophages.

  Jaruek Charoensap, Pongsak Utaisincharoen, Anneke Engering, Stitaya Sirisinha
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  ABSTRACT: Burkholderia pseudomallei (Bp) is a category B biothreat organism that causes a potentially fatal disease in humans and animals, namely melioidosis. Burkholderia thailandensis (Bt) is another naturally occurring species that is very closely related to Bp. However, despite this closely related genotype, Bt is considered avirulent as it does not cause the disease. In the present study, we compared the growth kinetics of B. pseudomallei strain 844 (Bp-844) in human monocyte-derived dendritic cells (MoDCs) and macrophages (Mphis), as well as its ability to stimulate host cell responses with those of B. thailandensis strain UE5 (Bt-UE5). Primary human MoDCs and Mphis were infected with Bp-844 and its intracellular growth kinetics and ability to induce host cell responses were evaluated. The results were compared with those obtained using the Bt-UE5. In human MoDCs, both bacteria were similar in respect to their ability to survive and replicate intracellularly, induce upregulation of costimulatory molecules and cytokines and bias T helper cell differentiation toward a Th1 phenotype. By contrast, the two bacteria exhibited different growth kinetics in human Mphis, where the intracellular growth of Bt-UE5, but not Bp-844, was significantly suppressed. Moreover, the ability of Mphis to kill Bp-844 was markedly enhanced following stimulation with IFN-gamma. The data presented showed that while both strains were similar in their ability to survive and replicate in human MoDCs, only Bp-844 could readily replicate in human Mphis. Both bacteria induced similar host cellular responses, particularly with regard to their ability to bias T cell differentiation toward a Th1 phenotype.
  BMC Immunology 05/2009; 10:20. · 2.53 Impact Factor
• Article: A role of Burkholderia pseudomallei flagella as a virulent factor.

  Tippamas Chuaygud, Sumalee Tungpradabkul, Stitaya Sirisinha, Kim Lee Chua, Pongsak Utaisincharoen
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  ABSTRACT: Burkholderia pseudomallei is an agent of melioidosis and is closely related to avirulent B. thailandensis. Burkholderia thailandensis has a 15-bp deletion within the variable region of the flagellin gene fliC compared with B. pseudomallei. The difference in the fliC gene might be related to virulence. In the present study, the invasion, internalization and intracellular replication of both phagocytic (mouse macrophage cell line RAW264.7) and non-phagocytic cells (human lung epithelial cell line A549) of B. pseudomallei fliC knockout mutant (MM35) complemented with its own fliC (Cp) or with B. thailandensis fliC (Ct) was compared with those of the wild-type strains of B. pseudomallei (1026b) and B. thailandensis (E257). In phagocytic cells, there was no significant difference in bacterial uptake between Cp and Ct, but MM35 was internalized significantly less compared with 1026b, Cp, Ct and E257. The results suggest that flagella are involved in macrophage invasion. In non-phagocytic cells, Cp and Ct showed similar invasive capacities while 1026b, Cp and Ct showed significantly higher invasiveness than MM35, suggesting that flagella facilitate the non-phagocytic cell invasion. However, the invasive capacity of MM35 was significantly higher than that of E257, suggesting that in addition to the flagella, B. pseudomallei may need other factor(s) to facilitate invasion in non-phagocytic cells.
  Transactions of the Royal Society of Tropical Medicine and Hygiene 01/2009; 102 Suppl 1:S140-4. · 2.16 Impact Factor
• Article: Activation of human monocyte-derived dendritic cells by Burkholderia pseudomallei does not require binding to the C-type lectin DC-SIGN.

  Jaruek Charoensap, Anneke Engering, Pongsak Utaisincharoen, Yvette van Kooyk, Stitaya Sirisinha
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  ABSTRACT: Dendritic cells (DCs) are essential in regulating adaptive immunity. DC-SIGN (DC-specific ICAM-grabbing nonintegrin) is a C-type lectin receptor that is expressed mainly by DCs. Accumulating evidence supports that certain pathogens target DC-SIGN to escape host immunity. To investigate a possible role of DC-SIGN in Burkholderia pseudomallei infection, we initially screened its DC-SIGN binding activity by an ELISA method utilizing a DC-SIGN-Fc chimeric protein and found that all of the B. pseudomallei strains tested failed to bind DC-SIGN. However, one strain, the LPS mutant SRM117, which lacks the type II O-polysaccharide expression, actually bound DC-SIGN, in contrast to its wild-type counterpart 1026b (P<0.001). We also found that, although the LPS mutant could readily activate monocyte-derived human DCs, it induced lower levels of IL-12p70 and IL-10 production than its wild-type counterpart (P<0.01). By contrast, the wild-type and the LPS mutants were indistinguishable from one another in terms of T(H)1/T(H)2 differentiation. Altogether, these data suggest that, unlike other certain host pathogen interactions, activation of DCs by B. pseudomallei is not dependent on DC-SIGN. We also found evidence that the LPS mutant that binds DC-SIGN has a suppressive effect on DC cytokine production.
  Transactions of the Royal Society of Tropical Medicine and Hygiene 12/2008; 102 Suppl 1:S76-81. · 2.16 Impact Factor
• Article: High susceptibility of human dendritic cells to avian influenza H5N1 virus infection and protection by IFN-alpha and TLR ligands.

  Arunee Thitithanyanont, Anneke Engering, Peeraya Ekchariyawat, Suwimon Wiboon-ut, Amporn Limsalakpetch, Kosol Yongvanitchit, Utaiwan Kum-Arb, Watcharoot Kanchongkittiphon, Pongsak Utaisincharoen, Stitaya Sirisinha, Pilaipan Puthavathana, Mark M Fukuda, Sathit Pichyangkul
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  ABSTRACT: There is worldwide concern that the avian influenza H5N1 virus, with a mortality rate of >50%, might cause the next influenza pandemic. Unlike most other influenza infections, H5N1 infection causes a systemic disease. The underlying mechanisms for this effect are still unclear. In this study, we investigate the interplay between avian influenza H5N1 and human dendritic cells (DC). We showed that H5N1 virus can infect and replicate in monocyte-derived and blood myeloid DC, leading to cell death. These results suggest that H5N1 escapes viral-specific immunity, and could disseminate via DC. In contrast, blood pDC were resistant to infection and produced high amounts of IFN-alpha. Addition of this cytokine to monocyte-derived DC or pretreatment with TLR ligands protected against infection and the cytopathic effects of H5N1 virus.
  The Journal of Immunology 11/2007; 179(8):5220-7. · 5.79 Impact Factor
• Article: Virulent Burkholderia pseudomallei is more efficient than avirulent Burkholderia thailandensis in invasion of and adherence to cultured human epithelial cells.

  Wannapa Kespichayawattana, Pakamas Intachote, Pongsak Utaisincharoen, Stitaya Sirisinha
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  ABSTRACT: Burkholderia pseudomallei, a causative agent of melioidosis, is a facultative intracellular gram-negative bacillus that is closely related to its avirulent counterpart, Burkholderia thailandensis. However, pathogenic mechanisms and virulence factors of B. pseudomallei remain elusive. In the present study, we compared the invasiveness, adherence, and replication of B. pseudomallei and B. thailandensis in human respiratory epithelial cells A549. Invasion was determined after 4 h of coculturing using antibiotic protection assay. Adherence was demonstrated by coculturing the cells with fluorescein-labeled bacteria for 1 h and the number of positive cells was analyzed by flow cytometry. The results obtained with this in vitro study demonstrated that compared with its avirulent counterpart, B. pseudomallei is significantly more efficient (P<0.01) in invasion, adherence and inducing cellular damage, as represented by plaque formation.
  Microbial Pathogenesis 06/2004; 36(5):287-92. · 1.94 Impact Factor