Yasuyo Kashiwagi

Single genotype of measles virus is dominant whereas several genotypes of mumps virus are co-circulating.

We have reported that in Japan measles virus strains have been classified into three distinct different genotypes (C1, D3 and D5) under the new international genotype classification since 1984. Similarly, mumps virus strains have been divided into two genotypes with three subtypes (B1, B2, B3, and D) under the proposed international classification since 1976. To differentiate these genotypes we developed a restriction fragment length polymorphism assay in the hemagglutinin (H) region for measles virus and in the hemagglutinin‐neuraminidase (HN) region for mumps virus to facilitate the expanded molecular epidemiology. In the Sapporo 1995/1996 measles outbreak, all 26 strains were classified as D5. Among 32 samples from patients with measles from 1994 to 1997 in Tokyo, 28 were identified as D5 and four were D3; these D3 strains were ascertained as a same hospital acquired infection. Among 45 strains obtained in the Tokyo 1999 outbreak, 38 were D3 and the remaining seven were D5. The dominant genotype of measles in Tokyo has replaced from D5 to D3 similar to the Chicago1/89 strain. We obtained 220 samples from patients with mumps from 1993 to 1997 and they were classified into one strain of B1, 14 strains of B2, 151 strains of B3, and 54 strains of D. Therefore, we suggest that two or three subtypes of mumps virus are co‐circulating with a different geographic pattern in genotype distribution, whereas a single measles virus genotype is dominantly observed, showing different epidemiological patterns. J. Med. Virol. 62:278–285, 2000.

Cytokine and chemokine response in children with the 2009 pandemic influenza A (H1N1) virus infection

We report the systemic cytokine and chemokine response in children with the 2009 pandemic influenza A (H1N1) virus infection. In patients with pneumonia, the serum levels of IFN-γ and IL-5 were significantly higher than those in patients without pneumonia. This tendency was also present for IL-6, IL-8, IL-10, IL-13, and MCP-1 in patients with pneumonia. Among patients with pneumonia, the levels of MCP-1 were significantly higher in the group of patients with pneumonia with severe respiratory failure than patients with mild pneumonia.

Combined Treatment of Steroids and Cyclosporine in Kimura Disease

Kimura disease is a rare but distinctive chronic eosinophilic inflammatory disorder that is characterized by tumor-like lesions in the soft tissue and lymph nodes of the head and neck or parotid gland. Recently, many immunopathogenetic features of underlying T lymphocytes and related cytokines have been noted in Kimura disease. However, few previous studies have investigated the serial levels of cytokines in children. In this report we describe an 11-year-old Japanese boy with relapsing Kimura disease. Before the diagnosis of Kimura disease, the patient had a swelling on his left neck. Steroids were effective, but the tumor relapsed within a few months as the steroids were tapered. He was treated with steroids and cyclosporine. This treatment was done by measuring serial levels of serum soluble interleukin-2 receptor, interleukin-4, interleukin-5, and eosinophil cationic protein. These results suggest the activation of T-helper cells and T-helper 2 cytokines, that after activated B cells and eosinophilic infiltration play an important role in Kimura disease, and that cyclosporine suppresses the activity of this disease.

Inflammatory cytokines as predictors of resistance to intravenous immunoglobulin therapy in Kawasaki disease patients.

Kawasaki disease (KD) is an acute systemic vasculitis. Activation of the immune system is a central feature of KD. Some KD patients are resistant to initial high‐dose intravenous immunoglobulin (IVIG) treatment. The study aimed to determine the predictors of IVIG resistance.

Sequence analysis of F, SH, and HN genes among mumps virus strains in Japan

SummaryThe fusion (F), small hydrophobic (SH), and hemagglutinin-neuram- inidase (HN) regions of 15 mumps virus (MuV) strains were sequenced to demon- strate the genetic variability since 1976 in Japan. The MuV strains were classified into 2 major genotypes, A and B, and genotype A was subdivided into three subtypes (Aff1, Aff2, and Aff3). Aff1 and Aff2 strains were mainly isolated in 1977 and 1980. Aff3 strains were isolated in 1985 and 1989 and genotype B strains in 1993 and 1994. Genotypes Aff1, Aff2, and Aff3 were closely related indigenous lineages in Japan but genotype B was in the similar cluster in Europe and North America.

Detection of mumps virus genome directly from clinical samples and a simple method for genetic differentiation of the Hoshino vaccine strain from wild strains of mumps virus.

A simple and sensitive method was developed for the differentiation of the Hoshino vaccine strain from wild strains with a restriction fragment length polymorphism (RFLP) analysis in the part of hemagglutinin‐neuraminidase (HN) gene. The virus genome was amplified by using a reverse transcriptase‐polymerase chain reaction (RT‐PCR) directly from clinical samples. The PCR product of the Hoshino vaccine strain was cleaved into 2 fragments after digestion with Sca I and Afl II. All wild strains showed 2 RFLP profiles, A and B, different from that of vaccine strain. Wild A strains were cut into 2 fragments after digestion with Sca I only, while wild B strains were cleaved neither with Sca I nor Afl II. This molecular approach provides an effective method for differentiation of the Hoshino vaccine strain from wild strains of mumps virus in patients after vaccination. J. Med. Virol. 52:195–199, 1997.

High concentration of serum nitrite/nitrate obtained from patients with influenza‐associated encephalopathy

According to a marked decrease in the vaccination rate of influenza virus in Japan during the last 10 years, it has become evident that a number of patients with a new type of influenza-associated encephalopathy is increasing. 1 The onset of the seizures and unconsciousness is abrupt, usually within 12–24 h after the first symptoms, that is severe fever, and findings of severe brain edema by brain computed tomography (CT) scanning which is characteristic but not the specified localization of necrosis and bleeding. Virus isolation from serum and cerebrospinal fluid is not always successful and the viremic condition is not always demonstrated. 2,3 High levels of cytokines in the serum or central nervous system of influenza patients were reported from several institutes. 4,5 In such cytokines, tumor necrosis factorα (TNFα ) is thought to be one of the cause of neural damage. Additionally, one of the biological gases, nitric oxide (NO), elicits the immune response related to the cytokine network under inflammatory condition. However, these pathophysiological correlations are actually still unknown. In this study, we measured the nitrite/nitrate (NOx) levels in serum of influenza patients in order to evaluate the correlation between the NO production and the process of influenza-associated encephalopathy assessed by the sensitive high-performance liquid chromatographyultra violet (HPLC-UV) system. Study subjects

PRIMARY BIOMARKERS IN CEREBRAL SPINAL FLUID OBTAINED FROM PATIENTS WITH INFLUENZA-ASSOCIATED ENCEPHALOPATHY ANALYZED BY METABOLOMICS

In order to search for the specific biomarkers of patients with influenza-associated encephalopathy this article analyzed all metabolites in cerebrospinal fluid (CSF) by using metabolome analysis. In all metabolites, the peaks of two molecular weights, 246.0092 and 204.0611, were significantly higher than those in other diseases including influenza without convulsion (p < .05). The peak of a molecular weight 228.0247 in all of the patients except one was less than that in other patients. These results indicate that the new metabolites detected in CSF would be primary markers for the diagnosis of influenza-associated encephalopathy.

Inflammatory responses following intramuscular and subcutaneous immunization with aluminum-adjuvanted or non-adjuvanted vaccines.

Aluminum-adjuvanted vaccines are administered through an intramuscular injection (IM) in the US and EU, however, a subcutaneous injection (SC) has been recommended in Japan because of serious muscle contracture previously reported following multiple IMs of antibiotics. Newly introduced adjuvanted vaccines, such as the human papillomavirus (HPV) vaccines, have been recommended through IM. In the present study, currently available vaccines were evaluated through IM in mice. Aluminum-adjuvanted vaccines induced inflammatory nodules at the injection site, which expanded into the intra-muscular space without any muscle degeneration or necrosis, whereas non-adjuvanted vaccines did not. These nodules consisted of polymorph nuclear neutrophils with some eosinophils within the initial 48h, then monocytes/macrophages 1 month later. Inflammatory nodules were observed 6 months after IM, had decreased in size, and were absorbed 12 months after IM, which was earlier than that after SC. Cytokine production was examined in the injected muscular tissues and AS04 adjuvanted HPV induced higher IL-1β, IL-6, KC, MIP-1, and G-CSF levels in muscle tissues than any other vaccine, but similar serum cytokine profiles were observed to those induced by the other vaccines. Currently available vaccines did not induce muscular degeneration or fibrotic scar as observed with muscle contracture caused by multiple IMs of antibiotics in the past.

Expressions of Brain-Derived Neurotrophic Factor (BDNF) in Cerebrospinal Fluid and Plasma of Children With Meningitis and Encephalitis/Encephalopathy

ABSTRACT Many reports in the field of childhood brain disorders have documented that brain-derived neurotrophic factor (BDNF) affects central nervous system (CNS) functions. In this clinical study, BDNF levels were evaluated in association with pediatric CNS infections. BDNF levels in the serum and cerebrospinal fluid (CSF) of 42 patients admitted during 5-year period, due to CNS infections, were measured by enzyme-linked immunosorbent assays (ELISAs). Control samples were collected from 108 patients with non-CNS infections (urinary tract infection, acute upper respiratory infection, acute gastroenteritis, etc.). Mean values of BDNF levels, at various ages, were determined and compared. BDNF levels were below the sensitivity of the ELISA in most CSF samples from the control group, but were significantly elevated in the patients with bacterial meningitis. The serum BDNF levels were elevated in all subgroups of patients with CNS infections, and the elevation was particularly notable in those with bacterial meningitis. BDNF expression in the CSF was correlated with CSF interleukin (IL)-6 levels as well as with blood platelet counts and neurological prognoses in those with bacterial meningitis. No correlation was found between BDNF levels and serum leukocyte numbers or C-reactive protein (CRP) levels. BDNF levels were found to be elevated in the serum and CSF of pediatric patients with CNS infections, particularly those with bacterial meningitis. Monitoring the changes in serum and CSF levels of BDNF may facilitate the diagnosis of acute meningitis and acute encephalopathy and allow the differential diagnosis of specific CNS infections.