MIP-1α level in nasopharyngeal aspirates at the first wheezing episode predicts recurrent wheezing.

BACKGROUND: Respiratory virus-induced wheezing, such as that induced by respiratory syncytial virus (RSV) and human rhinovirus, is an important risk factor for recurrent wheezing and childhood asthma. However, no biomarkers for predicting recurrent wheezing have been identified. OBJECTIVE: We searched for predictors of recurrent wheezing using nasopharyngeal aspirates obtained from patients during the first wheezing episode who were hospitalized with an acute lower respiratory tract illness. METHODS: We enrolled 82 infants during the first wheezing episode (median age, 5.0 months) who were hospitalized for acute lower respiratory tract illness between August 2009 and June 2012 and followed these patients for 2.5 years. Nasopharyngeal aspirates and blood samples were obtained on the first day of hospitalization. Viral genomes were identified by using RT-PCR and sequencing. Levels of 33 cytokines, tryptase, IgE, anti-RSV IgE, and anti-RSV IgG were measured by using ELISAs or the Bio-Plex multiplex assay. Predictors of recurrent wheezing were examined by using a stepwise logistic regression model with backward elimination. RESULTS: Sixty percent of the patients experienced recurrent wheezing episodes. One or more viruses were detected in the nasopharynxes of 93% of the patients during the first wheezing episode. IFN-γ, IL-2, IL-9, MIP-1α, and MIP-1ß levels were significantly higher among patients with recurrent wheezing than among those without recurrent wheezing (P < .05 or .01). The stepwise model demonstrated that the MIP-1α level (odds ratio, 7.72; 95% CI, 1.50-39.77; P = .015) was the strongest independent predictor of the occurrence of recurrent wheezing. CONCLUSION: An increased MIP-1α level in nasopharyngeal aspirates from patients with acute respiratory symptoms during the first wheezing episode caused by viral infections might predict recurrent wheezing.

Triplex real-time polymerase chain reaction assay for detection and quantification of norovirus (GI and GII) and sapovirus.

To improve detection of norovirus (NoVGI, NoVGII) and sapovirus (SaV), a simultaneous quantitative RT-PCR method was established. This triplex real-time PCR method was evaluated using a combination of optimized specific primers and probes. The performance of the developed PCR assay was equivalent to that of monoplex real-time PCR across a broad dynamic range of 10(2) -10(7) copies/assay using plasmid DNA standards. The limit of detection was 10(2) copies/assay. The quantitative value was comparable with that of monoplex real-time PCR of stool samples. Our triplex real-time PCR is useful for detection of NoV and SaV infections.

Evidence for reactivation of human herpesvirus 6 in generalized lymphadenopathy in a patient with drug-induced hypersensitivity syndrome.

The present case provides direct evidence of human herpesvirus 6 reactivation in resected lymph node tissue in a patient with drug-induced hypersensitivity syndrome. This case clearly demonstrates that appropriate pathological evaluation of lymphadenopathy for drug-induced hypersensitivity syndrome, which mimics malignant lymphoma in clinical, radiological, and pathological findings, is required.

Elevated macrophage inflammatory protein 1α and interleukin-17 production in an experimental asthma model infected with respiratory syncytial virus.

BACKGROUND: Respiratory syncytial virus (RSV) infection is associated with both the development and exacerbation of bronchial asthma. We examined eosinophil infiltration and the cytokine profiles of both airway and peripheral blood in antigen-sensitized mice infected with RSV to investigate the pathogenesis of exacerbations of asthma due to RSV infection. METHODS: Ovalbumin (OVA)-sensitized mice were challenged by OVA inhalation 3 times and then infected with RSV [10(5) TCID50 (50% of tissue culture infectious dose)/25 g body weight] or mock infection immediately after the last challenge. Animals from each group, namely, the control (PBS instead of OVA inhalation plus mock infection), RSV (PBS plus RSV), OVA (OVA plus mock) and OVA/RSV (OVA plus RSV) were analyzed. Analysis included evaluation of airway responsiveness to methacholine, pathological findings in the airway by hematoxylin and eosin (HE) and Luna staining, bronchoalveolar fluid (BALF) and peripheral leukocytes counts, and concentrations of multiple cytokines/chemokines in both BALF and serum. RESULTS: Airway responsiveness was significantly enhanced in the OVA and OVA/RSV groups compared with the control group. Levels of tissue and BALF eosinophils were higher in the OVA and OVA/RSV groups than in the RSV or control group. Significantly higher levels of macrophage inflammatory protein (MIP)-1α in BALF were observed in the OVA/RSV group compared with the 3 other groups. Production of serum IL-17 was also significantly elevated in the OVA/RSV group compared with the control or OVA group. CONCLUSIONS: These findings suggest that MIP-1α and IL-17 may play important roles in acute exacerbation of asthma induced by RSV in an animal model.

Cytokine profiles, signalling pathways and effects of fluticasone propionate in respiratory syncytial virus-infected human foetal lung fibroblasts.

To examine cytokine production in response to RSV infection, we assessed the levels of 29 cytokines released from RSV-infected human foetal lung fibroblasts. We also examined the relationships between the effects of fluticasone propionate and various signalling pathways in the cells. Twenty-four hours after infection (1MOI), RSV-infected cells released cytokines, for example proinflammatory cytokines (IL-1ß, IL-6 and TNF-α), anti-inflammatory (IL-1ra), Th1 (IFN-γ, IFN-λ1a, IL-2 and IL-12), Th2 (IL-4, IL-5, IL-10 and IL-13), granulopoiesis-inducing (G-CSF and GM-CSF), eosinophil recruitment-inducing (eotaxin and RANTES) and neutrophil recruitment-inducing cytokines (IL-8, IP-10, MCP-1 and MIP-1α). Aberrant release of most was significantly suppressed by fluticasone propionate. Twelve hours after RSV infection, increased phosphorylation of Akt, p38 MAPK, ERK1/2 and IκB-α was noted. Fluticasone propionate suppressed the phosphorylation of Akt, p38 MAPK, and ERK1/2, but not IκB-α, in virus-infected cells. TLR-4 expression was unchanged in control and RSV-infected cells, and TLR-3 and RIG-I expression was not detected. The results indicate that RSV infection induces aberrant production and release of certain cytokines through these signalling pathways in human lung fibroblasts. Overproduction and imbalance of these cytokines may be associated with the pathophysiology of RSV-induced excessive and allergic inflammation.

Temporal alteration of serum G-CSF and VEGF levels in perinatal asphyxia treated with head cooling.

OBJECTIVE: Granulocyte-colony stimulating factor (G-CSF) and vascular endothelial growth factor (VEGF) are thought to be associated with the pathophysiology of perinatal asphyxia. To clarify any such association, we analyzed the serum levels in neonates with perinatal asphyxia treated with head cooling. STUDY DESIGN: Temporal alterations of serum G-CSF and VEGF levels were measured within 24h of birth in five neonatal cases of severe asphyxia treated with head cooling, five neonatal cases without head cooling, and four healthy neonatal cases. RESULTS: G-CSF in sera markedly increased and sustained in severely asphyxiated neonates treated with head cooling, while VEGF decreased and remained low. CONCLUSION: G-CSF and VEGF levels in sera might be associated with an early phase of brain protection after birth in severe asphyxia treated with head cooling.

Different cytokine profile and eosinophil activation are involved in rhinovirus- and RS virus-induced acute exacerbation of childhood wheezing.

Because little information is available on eosinophil activation and cytokine response in virus-induced wheezing, we attempted to detect respiratory viruses and measure eosinophil cationic protein (ECP), and 27 types of cytokines/chemokines in both serum and nasal secretions from children with wheezing. This study was an observational, case-control investigation of 267 subjects, who were visited and/or hospitalized with acute respiratory symptoms (with wheezing: men, 115; women, 59; mean/median age, 3.6/3.0 years) or who were visited for regular physical examination and treatment (non-symptomatic wheezing: men, 48; women, 31; mean/median, 5.0/4.7 years), and 14 control subjects (controls: men, 9; women, 5; mean/median, 3.6/3.7 years). We detected viruses in nasal secretions from 174 patients with acute exacerbations of wheezing using antigen detection kits or reverse transcription-polymerase chain reaction, followed by direct DNA sequencing analysis. We measured peripheral eosinophil counts, and serum concentrations of ECP and 27 cytokines/chemokines using a multiplex bead-based assay in patients with wheezing or non-symptomatic wheezing. We also examined nasal ECP and 27 cytokines/chemokines in patients with wheezing. Of 174 samples from wheezing exacerbations, rhinovirus was detected in 59; respiratory syncytial (RS) virus in 44; enterovirus in 17; other viruses in 19; and no viruses in 35. Serum concentrations of ECP, IL-5, IL-6, IL-1ra, and IP-10 were significantly elevated in rhinovirus-induced wheezing compared with non-symptomatic wheezing. Similarly, serum ECP, IL-5, and IP-10 were significantly higher in rhinovirus-induced wheezing than in controls. On the other hand, IL-1ra and IP-10, but not ECP and IL-5 were significantly higher in RS virus-induced wheezing than in controls. Furthermore, only IL-5 was significantly elevated in the rhinovirus group compared with the RS virus group in both serum and nasal secretions. Different cytokine profile and eosinophil activation might be involved in rhinovirus- and RS virus-induced acute exacerbation of childhood wheezing.

A molecular epidemiological study of respiratory viruses detected in Japanese children with acute wheezing illness.

BACKGROUND: Recent studies strongly suggest that some respiratory viruses are associated with the induction of acute wheezing and/or exacerbation of bronchial asthma. However, molecular epidemiology of these viruses is not exactly known. METHODS: Using PCR technology, we attempted to detect various respiratory viruses from 115 Japanese children. Furthermore, the detected viruses were subjected to homology, pairwise distance, and phylogenetic analysis. RESULTS: Viruses were detected from 99 (86.1%) patients. Respiratory syncytial virus (RSV) alone and human rhinovirus (HRV) alone were detected in 47 (40.9%) and 36 (31.3%) patients, respectively. Both RSV and HRV were detected in 14 (12.2%) patients. Human metapneumovirus (HMPV) alone and human parainfluenza virus (HPIV) alone were detected in 1 (0.9%) patient each, respectively. Homology and phylogenetic analyses showed that the RSV and HRV strains were classified into genetically diverse species or subgroups. In addition, RSV was the dominant virus detected in patients with no history of wheezing, whereas HRV was dominant in patients with a history of wheezing. CONCLUSIONS: The results suggested that these genetically diverse respiratory viruses, especially RSV and HRV, might be associated with wheezing in Japanese children.

Release of cytokines/chemokines and cell death in UVB-irradiated human keratinocytes, HaCaT.

Ultraviolet (UV) B can lead to inflammatory responses such as sunburn, which involves the production of various inflammatory cytokines and chemokines, and the induction of cell death. Keratinocytes in the skin has one of the highest risks of exposure to UV. However, the detailed mechanisms underlying UVB irradiation-induced inflammation and cell death are not well known. Thus, we investigated the effect of UVB irradiation on the production of various cytokines/chemokines and the induction of cell death in UVB-irradiated human keratinocytes (HaCaT cells). We evaluated 11 cytokines/chemokines in cell culture supernatants from HaCaT cells exposed to 0-400 mJ/cm(2) UVB irradiation. UVB at a dose 400 mJ/cm(2) induced the release of various cytokines; interleukin (IL)-1beta, IL-6, IL-8, interferon (IFN)-gamma, granulocyte-colony stimulating factor (G-CSF), macrophage inflammatory protein (MIP)-1beta, and tumor necrosis factor (TNF)-alpha. These results suggest that UVB irradiation-induced the release of several cytokines/chemokines and led to cell death in human keratinocytes. UV exposure may be associated with multiple physiological events in the human skin.

Sequence and phylogenetic analysis of the nucleoprotein (N) gene in measles viruses prevalent in Gunma, Japan, in 2007.

In 2007, relatively large outbreaks of measles occurred in the Kanto region of Japan, including Gunma Prefecture. We performed sequence and phylogenetic analysis of the nucleoprotein gene (N gene) of measles viruses from 3 measles patients in this area in May 2007. The N gene sequences of the present strains were identical to each other, and phylogenetic analysis showed these viruses were classified into genotype D5. The results suggest that highly homologous measles viruses may be associated with outbreaks of measles in Gunma, Japan.

Tuberculosis-like peritonitis due to an atypical Mycobacterium infection in a Japanese woman.

It is not known whether atypical Mycobacterium (AM) causes peritonitis in humans. We described a case of tuberculosis-like peritonitis caused by an AM. Genetic analysis of the biopsy specimens suggested an AM infection. Thus, we concluded that peritonitis in humans can be caused by some AM species as well as by Mycobacterium tuberculosis complex.

Detection and sequence analysis of DNA polymerase and major envelope protein genes in koi herpesviruses derived from Cyprinus carpio in Gunma prefecture, Japan.

Koi herpesvirus (KHV), which is believed to be an emerging virus, causes fatal diseases in carps. Since the 1990s, the presence of KHV has been confirmed in several countries. In Japan, from 2003 to 2004, large outbreaks of KHV infection in farmed carps resulted in the death of numerous fishes. From April to May 2004, we collected 43 dead or dying carps exhibiting typical symptoms of KHV infection in Gunma prefecture. To conduct a molecular epidemiologic study of KHV in our prefecture, we amplified DNA polymerase and the major envelope protein genes of KHV derived from carp gills using newly designed primers. We also performed sequence analysis of both genes of KHV. Sensitivity of our PCR method for amplification of DNA polymerase and the major envelope protein genes of KHV was 3 x 10(2) (100 fg) and 3 x 10(3) (1000 fg) copies of KHV genome, respectively. We detected both DNA polymerase and major envelope protein genes in 37 of 43 carps (86%). No mutation was found in both the genes sequenced from 11 strains, which included two foreign strains and one domestic strain. The results suggested that KHV strains derived from carps in our prefecture were closely related genetically to the other KHV strains.

Alteration in serum levels of inflammatory cytokines during parainfluenza virus type 1 infection in patients with severe multiple disabilities.

An epidemic of parainfluenza virus type 1 (PIV1) infection occurred in a hospital ward housing patients with severe motor and intellectual disabilities. Twenty-three infected patients exhibited persistent high fever for 4-16 days and decreased lymphocyte counts. One-half of the symptomatic patients had increased blood monocyte counts and the other half progressed to bronchitis or pneumonia. We also compared levels of 27 cytokines in the sera of 21 patients during the acute and normal phases of infection. Cytokine levels were measured with a bead immunoassay performed using the Luminex Multiplex System. Serum levels of interleukin (IL)-1Ra, C-C-motif chemokine (CCL) 2, and C-X-C-motif chemokine (CXCL) 10 significantly increased during the acute phase. In contrast, the serum level of CXCL8 decreased slightly. These results suggest the involvement of monocytes/macrophages and respiratory epithelial cells in the initial stage of PIV1 infection. A previous report using nasal wash samples also found a significant increase in levels of CXCL10 during the acute phase. Hence, CXCL10 may be a useful marker of a cytokine storm produced upon viral infection. However, alterations in levels of IL-1Ra, CCL2, and other cytokines differed between the 2 studies, suggesting that the cytokine profile produced systemically at viral infection is different from that produced at mucosal sites. Further analysis is required to clarify the mechanisms underlying cytokine production during PIV1 infections.

Cytokine production and signaling pathways in respiratory virus infection.

It has been confirmed that respiratory virus infections can induce abberant cytokine production in the host. These cytokines may be associated with both elimination of the virus and complications in the host, such as virus-induced asthma. Representative host defense mechanisms against pathogens, including bacteria and viruses, are mediated by the innate immune system. Cells of the innate immune system express essential molecules, namely pattern recognition receptors (PRRs), such as Toll-like receptors, nucleotide-binding oligomerization domain-like receptors, and retinoic acid-inducible gene-I-like receptors. These PRRs can recognize components of pathogens such as bacterial lipopolysaccharide, viral antigens, and their genomes (DNA and RNA). Furthermore, PRRs activate various signaling pathways resulting in cytokine production against pathogen infection. However, the exact mechanisms remain unknown. In this review, we mainly focus on the representative mechanisms of cytokine production through PRRs and signaling pathways due to virus infections, including respiratory virus infections. In addition, we describe the relationships between respiratory infections and virus-induced asthma.

Serum cytokine profiles in patients with prostate carcinoma.

It has been suggested that various cytokines are associated with the pathophysiology of prostate carcinoma (Pca). We profiled ten cytokines (IL-1ß, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, IFN-γ and TNF-α) in the serum levels of 11 patients with organ-confined Pca, 15 with advanced Pca without cachexia, 8 with advanced Pca with cachexia (cachexia group) and 5 healthy males as controls. Cytokines were measured using a highly sensitive fluorescence microsphere system. Compared to the control group, serum levels of all cytokines were significantly higher in the cachexia group, and six cytokines (IL-1ß, IL-2, IL-8, IL-12, TNF-α and IFN-γ) were significantly higher in the group with advanced Pca without cachexia. In the group with organ-confined Pca, only IL-1ß and IL-12 levels were significantly higher compared to the control group. In the cachexia group, levels of all cytokines apart from TNF-α were significantly higher compared to the group with organ-confined Pca, and levels of four cytokines (IL-2, IL-4, IL-8 and IL-10) were significantly higher compared to the group with advanced Pca without cachexia. These results indicate that i) an aberrance imbalance of cytokine production was associated with the pathophysiology of Pca and cachexia, ii) cytokine profiles in Pca patients were distinct by disease stage, and iii) IL-1ß and IL-12 may be applicable as early diagnostic indicators.

Peripheral lymphocyte subset and serum cytokine profiles of patients with West syndrome.

OBJECTIVE: To clarify the immune pathophysiology of West syndrome (WS). STUDY DESIGN: We measured peripheral blood lymphocyte subset and serum cytokine profiles in 76 WS patients and 26 age-matched controls. Adrenocorticotropic hormone (ACTH) is one of the most effective therapy for WS and presumably immune-modulating; therefore, we compared the measured parameters between before ACTH (pre-ACTH) WS patients and controls, between cryptogenic and symptomatic WS patients before ACTH (pre-ACTH), and between before (pre-ACTH) and after (post-ACTH) ACTH WS patients. The post-ACTH group included those who received the last ACTH dose within 1 month of sampling. RESULTS: CD3+ CD25+, CD19+, and CD19+ CD95+ cells were found to be significantly lower in the pre-ACTH group than in the controls. Interleukin (IL)-1 receptor antagonist (RA), 5, 6, and 15; eotaxin; basic fibroblast growth factor (bFGF); and interferon gamma-inducible protein (IP)-10 levels were higher in pre-ACTH group than in the controls. No significant differences were found between the pre-ACTH cryptogenic and symptomatic groups. CD4+ cells, CD3+ cells, CD4+/8+ ratio, IL-1 beta, IL-12, and macrophage inflammatory protein (MIP)-1 beta were significantly higher in pre-ACTH group than in the post-ACTH group. CONCLUSIONS: Our study revealed immunological alterations in WS patients, and these responses were modified by ACTH therapy. Further study is needed to elucidate whether or how the immune system alteration is involved in the pathophysiology of WS.

Relationships between Cytokine Profiles and Signaling Pathways (IκB Kinase and p38 MAPK) in Parainfluenza Virus-Infected Lung Fibroblasts.

Respiratory viruses such as parainfluenza virus (PIV) in individuals with certain genetic predispositions in early life are associated with the induction of wheezing, which can progress to the development of asthma. It has been suggested that aberrant production of various cytokines due to viral infection are associated with virus-induced asthma. However, the mechanisms of how respiratory viruses induce and exacerbate asthma have yet to be clarified. To examine cytokine responses to PIV infection, we assessed 27 cytokine levels released from PIV-infected human fetal lung fibroblasts. In addition, we examined relationships between these cytokine responses and signaling pathways (IκB kinase and p38 MAPK) in PIV-infected cells. At 24 h after infection, PIV-infected cells significantly released a number of cytokines, namely, proinflammatory cytokines [interleukins (IL)-1ß, IL-6, and tumor necrosis factor-α], anti-inflammatory cytokine (IL-1ra), Th1 cytokines (interferon-γ, and IL-2), Th2 cytokines (IL-4, IL-5, and IL-10), granulopoiesis-inducing cytokines (granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor), neutrophil recruitment-inducing cytokines (IL-8 and interferon-inducible protein-10), and eosinophil recruitment-inducing cytokines (eotaxin and regulated on activation normal T-cell expressed and secreted). PIV infection enhanced phosphorylation of both IκB and p38 MAPK, but not Akt, in the cells. Signaling pathway inhibitors, BMS-345541 (a specific IκB kinase inhibitor) and SB203580 (a specific p38 MAPK inhibitor), significantly suppressed release of these cytokines from PIV-infected cells. The results indicate that PIV infection induces aberrant production and release of various cytokines through IκB kinase and p38 MAPK pathways in human lung fibroblasts. Overproduction and imbalance of these cytokines may be partially associated with the pathophysiology of virus-induced asthma.

Differential diagnosis of primary versus metastatic pulmonary adenocarcinomas using gene mutation analyses: a case report.

A 61-year-old Japanese woman underwent a partial mastectomy for cancer of the right breast (pT1cN0M0, stage I). Eight months later, chest computed tomography revealed two small nodules in the left lower lobe (Segmentum basale laterale and Segmentum basale posterius; S9 and S10). She thereafter underwent partial pulmonary resections for both diagnostic and treatment purposes. The nodule of S10 was pathologically diagnosed to be primary lung cancer. The nodule of S9 was pathologically diagnosed to be poorly differentiated adenocarcinoma. The same pattern of the distribution of the p53 mutation was observed in the DNA samples of the S9 nodule and the treated breast cancer. We therefore finally diagnosed the S9 nodule to be metastatic pulmonary carcinoma. A mutation analysis of the p53 gene is thus considered to be a good modality for differentiating metastases from primary carcinomas of the lung.