Epidemic of complicated mumps in previously vaccinated young adults in the South-West of France.

OBJECTIVE: We report the features and diagnosis of complicated mumps in previously vaccinated young adults. PATIENTS AND METHODS: We retrospectively studied 7 cases of complicated mumps managed during 1 year at the Bordeaux University Hospital. The diagnosis was suggested by the clinical presentation and confirmed using specific RT-PCR. RESULTS: Five cases of meningitis, 1 of orchitis, and 1 of unilateral hearing impairment were identified. Each of the 7 patients had been previously vaccinated with MMR, 4 had received 2 doses of this vaccine. Blood tests revealed high rates of IgG antibodies, usually considered as sufficient for immunological protection, and every patient had at least 1 positive RT-PCR test for mumps. CONCLUSION: Outbreaks of complicated mumps may still occur despite a broad coverage of MMR vaccination. The clinical presentation suggested mumps but the final diagnosis could only be confirmed by genomic detection of the virus. Unusual viral strains with increased neurovirulence, insufficient population coverage associated with immunity decrease over time may explain outbreaks of complicated mumps. A full vaccine scheme of contact people or a third injection of vaccine for previously vaccinated people who are at risk of developing mumps are required to prevent further spreading of the disease during the outbreak.

Increased levels of cytokines in cerebrospinal fluid of children with aseptic meningitis caused by mumps virus and echovirus 30.

We measured levels of pro-inflammatory cytokines in the cerebrospinal fluid (CSF) of patients with mumps meningitis, enteroviral echovirus 30 meningitis and children without central nervous system infection to investigate whether these molecules were involved in the pathogenesis of viral meningitis. The CSF was obtained from 62 children suspected with meningitis. These patients were classified to the mumps meningitis (n = 19), echovirus 30 meningitis (n = 22) and non-meningitis (n = 21) groups. The concentrations of interleukin-1 (IL-1), interleukin-1 soluble receptor type 2 (IL-1R2), interleukin-8 (IL-8), human interferon gamma (IFN-γ) and human tumour necrosis factor alpha (TNF-α) were determined by immunoassay. A significant increase was noted in the levels of IL-8, TNF-α and IL-1R2 in the CSF of both meningitis groups as compared to controls. The concentrations of IFN-γ and IL-1 differed significantly only between the mumps group and control. The levels of IL-1, IFN-γ and TNF-α were significantly higher in mumps meningitis when compared to the echovirus 30 group. Of all cytokines examined, only IFN-γ correlated with pleocytosis (r = 0.58) in the mumps meningitis group. The increased CSF cytokine levels are markers of meningeal inflammation, and each virus may cause a specific profile of the cytokine pattern.

Enhanced expression of cytokines/chemokines in cerebrospinal fluids in mumps meningitis in children.

BACKGROUND: The mumps virus is frequently the causative agent in aseptic meningitis and mumps has still prevailed in Japan. We compared data obtained from patients with mumps meningitis and patients with aseptic meningitis caused by other viruses in order to identify mumps meningitis-specific cytokine/chemokine alterations in cerebrospinal fluid (CSF). METHODS: We elucidated the cytokine/chemokine network based on the cytokine/chemokine profiles in CSF from children with mumps meningitis and meningitis due to other viral infections using multiplex cytokine measurement. Seventeen cytokines/chemokines, namely interleukin (IL)-1ß, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p70), IL-13, IL-17, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, granulocyte colony-stimulating factor (G-CSF), granulocyte monocyte colony-stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1ß (MIP-1ß), were measured simultaneously in CSF supernatants from eight children with mumps meningitis, 11 children with other types of viral meningitis and eight children with fever without neurological complications such as convulsion. RESULTS: We found that IL-8, IL-10, IL-12, IL-13 and IFN-γ showed a statistically significant increase in CSF from mumps meningitis when compared to other types of viral meningitis and fever without neurological complications. CONCLUSION: Mumps meningitis may induce a distinct immunological response when compared with other types of viral meningitis.

Elevated levels of MMP-9 and TIMP-1 in the cerebrospinal fluid of children with echovirus type 30 and mumps meningitis.

Matrix metalloproteinases are involved in leucocyte invasion into the central nervous system (CNS) during meningitis. The aim of the study was to determine whether there are differences in the expression patterns of matrix metalloproteinase-9 (MMP-9) and the tissue inhibitor of metalloproteinase-1 (TIMP-1) in the cerebrospinal fluid (CSF) of patients with meningitis caused by one of two known distinct viral agents. Concentrations were measured by using an enzyme-linked immunosorbent assay (ELISA) in 16 children with mumps meningitis, in 25 children with echovirus type 30 meningitis and in a control group of 23 children without any CNS infection. Increased levels of MMP-9 were found in children with mumps (median 0.48 ng/ml; P < 0.001) and enteroviral meningitis (median 2.76 ng/ml; P < 0.001) compared with that in controls (median: 0.01 ng/ml). Concentrations of TIMP-1 greatly exceeded concentrations of MMP-9 and were elevated in children with mumps (median: 56 ng/ml) and echovirus type 30 meningitis (median: 55 ng/ml) compared to controls (median: 17 ng/ml). No significant differences in MMP-9 or TIMP-1 levels were detected between the two meningitis groups. The MMP-9/TIMP-1 ratio was greater in children with echovirus type 30 than in those with mumps meningitis. There was no correlation between MMP-9 levels and total CSF cell count. MMP-9 correlated with CSF absolute neutrophil count in children with echovirus type 30 meningitis (r = 0.431; P < 0.05). The concentration of MMP-9 is higher in children with viral meningitis, possibly because of infiltrating polymorphonuclear cells present in the initial phase of the disease.

Increase in adhesion molecules in cerebrospinal fluid of children with mumps and mumps meningitis.

Adhesion molecules play a key role in leucocyte migration into the central nervous system (CNS). Concentrations of endothelial-derived soluble intercellular adhesion molecule-1 (sICAM-1) and leucocyte-originated soluble L-selectin (sL-selectin) in cerebrospinal fluid (CSF) of children with mumps meningitis (mononuclear pleocytosis, n = 33) and mumps (absence of pleocytosis, n = 9) were compared with values from age-matched control group (n = 19). In 14 patients from the meningitis group, adhesion molecule levels together with albumin concentration were estimated in paired CSF/serum samples to calculate concentration quotients and determine molecule intrathecal release. Both sICAM-1 (median 3.44 versus 0.86 ng/ml; P < 0.0001) and sL-selectin (median 29.91 versus 8.52 ng/ml; P < 0.0001) concentrations in CSF were increased in mumps meningitis patients compared with controls. Increased levels of the selected adhesion molecules were also observed in mumps patients without CNS involvement when compared with controls (median sICAM-1: 1.14 versus 0.86 ng/ml, sL-selectin: 13.54 versus 8.52 ng/ml; P < 0.01). Additionally, the concentration of adhesion molecules was found to correlate with CSF leucocyte count. Considerable correlation of sICAM-1 and sL-selectin quotients and corresponding albumin quotients suggests that a majority of the soluble adhesion molecules originated from the bloodstream. Analysis of adhesion molecule levels demonstrated indirect evidence of brain-derived fractions. Our results suggest the involvement of adhesion molecules during the early phase of mumps meningitis.

Real-time PCR for mumps diagnosis on clinical specimens--comparison with results of conventional methods of virus detection and nested PCR.

BACKGROUND: Since November 2003, the UK has seen a dramatic rise in the number of mumps cases, resulting in increasing demands on virology laboratories to confirm mumps infection in a timely and efficient manner. Traditional mumps virus detection methods are often insensitive, lengthy, and cumbersome. Some laboratories in the UK now use molecular methods that are based on nested polymerase chain reaction (PCR). Early serological diagnosis often relies on detection of anti-mumps IgM, which may be absent in the first 10 days of illness. OBJECTIVES: We compared a one-step real-time RT-PCR with an established nested PCR (SH-PCR) and virus detection by culture and antigen detection, and assessed the clinical usefulness of mumps real-time PCR for diagnosis from CSF. STUDY DESIGN: In total, 280 clinical samples were investigated by real-time PCR, nested PCR and a combination of traditional virus detection methods (antigen detection on oral samples, cell culture on all samples). Furthermore, 88 CSF samples submitted for diagnosis of possible viral meningitis were analysed by real-time PCR. RESULTS: The real-time PCR detected the highest number of positive oral samples (119/180) compared to SH-PCR (92/180) and combined virus culture and antigen detection procedures (90/180). Sensitivity of mumps virus detection in urine was poor for all three methods: 34.0% (traditional detection), 29.8% (real-time PCR) and 2.1% (SH-PCR), respectively. Real-time PCR on 88 CSF samples identified five patients with mumps meningitis, significantly increasing viral diagnosis in this cohort. CONCLUSION: Real-time PCR on oral samples is the investigation of choice for mumps infection. Mumps virus detection in urine by any of the PCRs used was clearly less successful. Real-time PCR on CSF samples seems a promising adjunct for diagnosis of mumps meningitis, especially in an age group with high incidence of mumps.

Analysis of cytokine levels in cerebrospinal fluid in mumps meningitis: comparison with echovirus type 30 meningitis.

BACKGROUND: It is unclear whether or not the CSF cytokine profiles in viral meningitis differ with the kind of causative virus. METHODS: We measured the concentrations of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin-2 (IL-2), IL-4, IL-6, and IL-10 in CSF during the acute stage in 15 children with mumps meningitis (MM), and 34 with echovirus type 30 meningitis (EM). RESULTS: The CSF IFN-gamma, IL-2, IL-6, and IL-10 levels were elevated in MM, and the CSF IFN-gamma, IL-2, and IL-6 levels were elevated in EM. The CSF IFN-gamma, IL-2, and IL-10 levels in MM were significantly higher than those in EM (p<0.0001, p<0.0001, and p<0.0001, respectively). The CSF IL-6 levels in EM were significantly higher than those in MM (p=0.0255). The CSF TNF-alpha and IL-4 levels were not elevated in MM or EM. In MM, the IL-6 level was correlated with the IL-2 and IL-10 levels in CSF (p=0.0347 and p=0.0120, respectively). In EM, the IFN-gamma level was correlated with the IL-10 level in CSF (p=0.0002). CONCLUSION: CSF cytokine profiles in MM were different from those in EM. Therefore, it is likely that the pathogenesis of MM is different from that of EM.

Frequency of lymphocytic meningitis associated with mumps before and after a mass campaign for mumps vaccination in children from Salvador, Northeast Brazil.

OBJECTIVE: To describe the frequency of lymphocytic meningitis(LM) concomitant with mumps, before and after the mumps mass immunization campaign in 1997. METHOD: Demographic, clinical and cerebrospinal fluid(CSF) information was collected from the chart of all patients aged from 2 to 59 months, whose CSF exam was performed at the CSF Lab/FJS, between 1989 and 2001. LM was defined as pleocytosis composed by lymphomononuclear cells and negative exams for bacterial or mycologic infection. RESULTS: Of 1,519 patients, 894(58.9%) had normal exams. LM was present in 301(19.8%) patients, out of which 22(7.3%) had concomitant mumps. The frequency of LM ranged from 15.8% in 1989 to 19.7% in 2001 and of LM with concomitant Mumps ranged from 10.5% in 1989 to 4.7% in 1995, when the last cases were registered. CONCLUSION: It is probable that the mumps vaccine campaign has influenced the absence of LM with concomitant Mumps, from 1996 to 2001.

Specific humoral immunity in mumps meningitis in children.

BACKGROUND: The pathogenesis of mumps meningitis remains unclear. The aim of this study was to assess the relation between IgM and IgG levels in blood and their relationship with the picture of CSF. MATERIAL AND METHODS: We tested 40 children of both sexes aged 2-14 years. CSF was examined at admission (I) and after 12-14 days (II). Antibodies were measured four times: at admission (I), and after 2 (II), 4 (III), and 8 weeks (IV) by EIA (Behring). The results were expressed as delta absorbency. RESULTS: The mean IgM level was respectively 0.403; 0.424; 0.317; 0.220. Significant differences were demonstrated between tests I and III and between III and IV. The mean IgG level was respectively 0.923; 1.322; 1.381 and 1.257. Significant differences were noted between tests I and II and between III and IV. We observed significant positive correlations between the IgM levels in tests I and II, II and III, and III and IV. The IgG concentrations significantly correlated in tests I and II, II and III, and II and IV. Significant negative correlations were noted between the IgG levels at test I and the IgM levels at test II. A negative correlation also appeared between the IgM level and CFS pleocytosis at admission, and between the IgM concentration and CSF pleocytosis after two weeks. A positive correlation was found between the IgG level at test I and the CSF glucose level in test II. In this study a high IgG level probably resulted in a lower IgM level at the next test. The IgM concentration at week 8 weeks was about 50% lower than its highest concentration (week 2), while the IgG level decreased simultaneously. The IgM concentration at hospital admission and two weeks later influenced CSF pleocytosis during these periods (higher IgM level - lower pleocytosis). A high IgG level at admission resulted in a lower CSF glucose concentration at the second examination.

Characterization of mumps virus isolated in saitama prefecture, Japan, by sequence analysis of the SH gene.

Mumps virus (MuV) strains isolated from cerebrospinal fluid and throat swabs from patients in Saitama Prefecture and Tokyo, Japan, from 1997 to 2000 were examined by analyzing the SH gene nucleotide sequence (316-nt). Eighteen of the 20 strains studied were divided into three genotypes, recognized as B, G, and H in previous reports. Two genotypes (G and H) are believed to be new in Japan. Two of the 20 strains belonged to none of the previously reported genotypes (A-I), but were closely related to two known strains, MP94-H and Loug1/UK97. We propose that the two strains identified in this study together with the previously reported strains, MP94-H and Loug1/UK97, form a new genotype, designated J, based on the divergence of the SH gene nucleotide sequences between these four strains and other strains reported (genotypes A-I). Our results also suggest that more than two genotypes circulated in Saitama Prefecture from 1997 to 1999, but only one, genotype G, was in evidence in 2000. Genotype B was earlier reported as the predominant strain in Japan, but it became undetectable by the year 2000. These results provide important epidemiological data on mumps in Japan.

Nested PCR for rapid detection of mumps virus in cerebrospinal fluid from patients with neurological diseases.

In this study, we have developed a reverse transcription (RT)-nested polymerase chain reaction (n-PCR) for the detection of mumps virus RNA in cerebrospinal fluid (CSF) from patients with neurological infections. A specific 112-bp fragment was amplified by this method with primers from the nucleoprotein of the mumps virus genome. The mumps virus RT-n-PCR was capable of detecting 0.001 PFU/ml and 0.005 50% tissue culture infective dose/ml. This method was found to be specific, since no PCR product was detected in each of the CSF samples from patients with proven non-mumps virus-related meningitis or encephalitis. Mumps virus RNA was detected in all 18 CSF samples confirmed by culture to be infected with mumps virus. Positive PCR results were obtained for the CSF of 26 of 28 patients that were positive for signs of mumps virus infection (i.e., cultivable virus from urine or oropharyngeal samples or positivity for anti-mumps virus immunoglobulin M) but without cultivable virus in their CSF. Overall, mumps virus RNA was detected in CSF of 96% of the patients with a clinical diagnosis of viral central nervous system (CNS) disease and confirmed mumps virus infection, while mumps virus was isolated in CSF of only 39% of the patients. Furthermore, in a retrospective study, we were able to detect mumps virus RNA in 25 of 55 (46%) CSF samples from patients with a clinical diagnosis of viral CNS disease and negative laboratory evidence of viral infection including mumps virus infection. The 25 patients represent 12% of the 236 patients who had a clinical diagnosis of viral CNS infections and whose CSF was examined at our laboratory for a 2-year period. The findings confirm the importance of mumps virus as a causative agent of CNS infections in countries with low vaccine coverage rates. In summary, our study demonstrates the usefulness of the mumps virus RT-n-PCR for the diagnosis of mumps virus CNS disease and suggests that this assay may soon become the "gold standard" test for the diagnosis of mumps virus CNS infection.

Characterization of five conserved genotypes of the mumps virus small hydrophobic (SH) protein gene.

Twenty-one different mumps virus isolates from Sweden and Japan collected over 25 years were compared by nucleotide sequence analysis of the small hydrophobic (SH) protein gene, and the deduced 57 amino acid sequences of the coding part of the gene were aligned with published sequences of viral isolates from the USA, the UK, Sweden and Japan. Five genotypes were found which, in accordance with previously used nomenclature, were named A to E. Genotypes A, C, D and E were found in Europe and genotype B was found in Japan. Amino acid signature sequence motifs specific for each genotype were identified. A triplet of three amino acids at positions 28-30 was the most characteristic. Different genotypes can circulate simultaneously in a given geographical location. In Stockholm, genotypes A and D or C and D were found over different time periods. In contrast, only genotype B was found in Japan.

[Beta 2 microglobulin in children with neuroinfections]. / Beta2 mikroglobulina w neuroinfekcjach u dzieci.

CSF and plasma beta2 microglobulin (B2M) concentrations were determined by an enzyme linked fluorescent assay (ELFA) (Vidas-bioMerieux) in children with bacterial (B2M), viral (mumps and enteroviral) meningitis and in the controls. CSF B2M concentrations in children with B2M at admission, at 24-48 hrs of treatment and at recovery (day 10), in children with viral meningitis at admission and at recovery were significantly higher in comparison with the control group of children with non-pleiocytic CSF. The levels of CSF B2M at 24-48 hrs of treatment of B2M cases were significantly higher than those at the beginning of both mumps and enteroviral meningitis cases which may be helpful in differential diagnosis of meningitis, especially in cases of retarded diagnosis or partially treated B2M. Plasma levels of B2M during bacterial and mumps meningitis did not differ from those in healthy children but in children with enteroviral meningitis were significantly higher. There was a positive correlation between CSF B2M at the beginning of B2M and some laboratory findings of inflammatory response (CRP, ERS). The CSF B2M levels were significantly higher than its plasma levels in patients with B2M at 24-48 hours (second stage) of disease, mumps meningitis on admission and recovery which may suggest intrathecal production of B2M during central nervous system infection.

Gamma delta T lymphocytes in mumps meningitis patients.

Flowcytometric analysis on T-cell subpopulations in the blood and cerebrospinal fluid (CSF) of 10 children with mumps meningitis (MM) revealed that the proportion of lymphocytes which bore the gamma delta T-cell receptor for antigen was significantly higher in CSF than in autologous and heterologous blood samples. gamma delta T-cell values in CSF of MM patients were also significantly higher than those calculated in CSF specimens from nine children with non-inflammatory neurological disorders. Whether and how the expanded CSF gamma delta T-lymphocyte subset, either CD8+ or double-negative (CD4-/CD8-), contributes to the eradication of mumps virus infection from the central nervous system represents a central focus of our ongoing research.

[Cytologic characteristics of cerebrospinal fluid in patients with serous meningitis caused by enteroviruses, mumps virus or Koch's bacillus]. / Citoloske karakteristike likvora kod bolesnika sa seroznim meningitisom uzrokovanim enterovirusim, mumps virusom i bacilom Koch.

The qualitative and quantitative analysis of the cytogram of cerebrospinal fluid (CSF) was performed in the acute phase of enteroviral meningitis (ENTERO MGT. 31 patients), mumps meningitis (MUMPS MGT, 25 patients) and tubercular meningoencephalitis (TBC ME, 15 patients), to observe the basic characteristics and answer the question whether the cytological response of the central nervous system (CNS) is ethyologically determined. The CSF sediment was obtained in Saykov chamber and was dyed according to Pappenheim. In the group of ENTERO MGT the significant representation of neutrophils with the presence of basophile was observed. The main characteristic of MUMPS MGT was mononuclear response with various morphological forms of lymphocyte cells and neglectable presence of neutrophils. TBC ME was characterized by the neutrophilia in the first 12 days of illness; till the end of the fourth week there was an equal representation of neutrophils and mononuclear cells, followed by a significant lymphocytosis. The cells of the monocytemacrophage system and the plasma-cells were constantly present. It could be concluded that differences in cytological response of CNS between ENTERO and MUMPS group were mainly quantitative and dynamic, probably caused by the various pathogenesic mechanisms of inflammation. The cytological picture of CSF during TBC ME in relation to other two groups had certain qualitative, quantitative and dynamic characteristics which could indicate the diagnosis of the specific meningoencephalitis, making this analysis very useful in the early diagnostics of this illness.

T-cell receptor V alpha region usage in the cerebrospinal fluid of patients with mumps meningitis.

To investigate clonality of local T cells in viral infections of the central nervous system, the T-cell receptor (TCR) repertoire was evaluated in T cells from the cerebrospinal fluid (CSF) of nine patients with mumps meningitis, using the polymerase chain reaction (PCR) assay. The usage of the variable region of TCR alpha chain gene (V alpha gene) in the CSF was widespread, and an average of 13 out of the 18 V alpha families were expressed. Quantitative PCR analyses showed that the V alpha gene expression was biased toward three or less V alpha families in the CSF of each patient. When compared with peripheral blood T cells, the average percentages of V alpha 11 and V alpha 12 gene expression were significantly higher in the CSF than in the peripheral blood. These results suggested that mumps-specific T lymphocytes with a restricted TCR repertoire are selectively recruited to the central nervous system in mumps meningitis, although polyclonal, probably nonspecific, T-cell populations are present in the CSF.

Laboratory surveillance of viral meningitis by examination of cerebrospinal fluid in Cape Town, 1981-9.

Nine years accumulated laboratory data derived from the culture of the cerebrospinal fluid of 11,360 aseptic meningitis cases were retrospectively reviewed to establish the epidemiology of viral meningitis in Cape Town. Virus was isolated from 3406 of the cases (91% enteroviruses and 9% mumps). Five major summer viral meningitis episodes were documented: two of echovirus 4 (706 and 445 cases), echovirus 9 (223), coxsackie A9 (104) and one of unidentified enterovirus (324 cases--probably echo 9). Although coxsackie B was endemic, clusters of one or other type were dominant at any one time. Mumps was endemic. Sixty-two percent of all viral cases were < 5 years old. The median ages of 4 and 5 years in echoviruses 9 and 4 (the epidemic strains) contrasted with that of 1 year in coxsackie B (with many cases < 3 months old). Mumps peaked at 3-4 years of age. Males dominated overall, particularly in mumps.

Nucleotide sequence analysis of Urabe mumps vaccine strain that caused meningitis in vaccine recipients.

Mumps virus was isolated from the cerebrospinal fluid of eight patients who acquired meningitis within 4 weeks of immunization with live Trivirix vaccine that contains mumps (Urabe Am 9), measles (Schwarz), and rubella (RA 27/3) viruses. Part of the haemagglutinin-neuraminidase (HN) gene from three postvaccination isolates of mumps virus, three wild strains and the Urabe and Jeryl Lynn vaccine strains was cloned following polymerase chain reaction (PCR) amplification, for the purpose of sequence analysis. A 200-nucleotide portion of the cloned HN genes was sequenced and compared to published sequences of two other strains (RW and SBL-1). The postvaccination mumps strains were identical in sequence to Urabe and were distinguishable from the wild and the Jeryl Lynn vaccine strains. Twenty-two out of 200 positions were seen to vary among the group of viruses. It was concluded that the Urabe vaccine strain was the cause of postvaccination meningitis. Therefore, with effect from 1990, Trivirix measles, mumps and rubella vaccine is no longer licensed for sale in Canada.