Disease burden of congenital cytomegalovirus infection at school entry age: study design, participation rate and birth prevalence.

Congenital cytomegalovirus infection (cCMV) may lead to symptoms at birth and long-term consequences. We present a nationwide, retrospective cohort study on the outcome of cCMV up to age 6 years. For this study we identified cCMV, using polymerase chain reaction, by analysing dried blood spots, which are taken shortly after birth for neonatal screening. The group of children with cCMV were compared to a group of children who were cCMV negative at birth. Data were collected about their health and development up to age 6 years. Parents of 73 693 children were invited to participate, and 32 486 (44·1%) gave informed consent for testing of their child's dried blood spot for CMV. Of the 31 484 dried blood spots tested, 156 (0·5%) were positive for cCMV. Of these, four (2·6%) children had been diagnosed with cCMV prior to this study. This unique retrospective nationwide study permits the estimation of long-term sequelae of cCMV up to the age of 6 years. The birth prevalence of cCMV in this study was 0·5%, which is in line with prior estimates. Most (97·4%) children with cCMV had not been diagnosed earlier, indicating under-diagnosis of cCMV.

High risk of a large measles outbreak despite 30 years of measles vaccination in The Netherlands.

Our aim was to assess progress towards measles elimination from The Netherlands by studying humoral measles immunity in the Dutch population. A population-based seroepidemiological study was conducted in 2006-2007 (N = 7900). Serum samples were analysed by a bead-based multiplex immunoassay. IgG levels ⩾0·2 IU/ml were considered protective. The overall seroprevalence in the Dutch population was 96%. However, 51% of socio-geographically clustered orthodox Protestant individuals aged <10 years were susceptible. Infants might be susceptible to measles between ages 4 months and 14 months, the age at which maternal antibodies have disappeared and the first measles, mumps, rubella (MMR) vaccination is administered, respectively. Waning of antibody concentrations was slower after the second MMR vaccination than after the first. The Netherlands is at an imminent risk of a measles outbreak in the orthodox Protestant minority. To prevent subsequent transmission to the general population, efforts to protect susceptible age groups are needed.

Characterization of the human CD8⁺ T cell response following infection with 2009 pandemic influenza H1N1 virus.

The 2009 H1N1 influenza pandemic provided an opportunity to study human virus-specific T cell responses after infection with a novel influenza virus against which limited humoral immunity existed in the population. Here we describe the magnitude, kinetics, and nature of the virus-specific T cell response using intracellular gamma interferon (IFN-γ) staining and fluorochrome-labeled major histocompatibility complex (MHC) class I-peptide complexes. We demonstrate that influenza virus-infected patients develop recall T cell responses that peak within 1 week postinfection and that contract rapidly. In particular, effector cell frequencies declined rapidly postinfection in favor of relatively larger proportions of central memory cells.

Two cases of mild IgM-negative measles in previously vaccinated adults, the Netherlands, April and July 2011.

We describe two cases of mild, modified measles in fully vaccinated adults in the Netherlands. The mildness of disease, the lack of an IgM antibody response, the relatively low amounts of virus detected and the fact that no additional cases were reported, suggests that these vaccinated patients were less contagious than unvaccinated patients.

Mumps epidemic in orthodox religious low-vaccination communities in the Netherlands and Canada, 2007 to 2009.

We assessed the epidemiological characteristics of a mumps virus epidemic (genotype D) that occurred in the Netherlands between August 2007 and May 2009 and its association with a subsequent mumps outbreak in Canada. In the Netherlands, five data sources were used: notifications (only mandatory since the end of 2008) (56 cases), laboratory confirmation data (177 cases), a sentinel general practitioner (GP) database (275 cases), hospitalisation data (29 cases) and weekly virological reports (96 cases). The median age of cases in the notification, laboratory and GP databases ranged from 13 to 15 years. The proportion of cases that were unvaccinated ranged from 65% to 85% in the notification, laboratory and GP databases. Having orthodox Protestant beliefs was the main reason for not being vaccinated. In Canada, a mumps virus strain indistinguishable from the Dutch epidemic strain was detected between February and October 2008 in an orthodox Protestant community with historical and family links to the affected community in the Netherlands, suggesting that spread to Canada had occurred. Prevention and control of vaccine-preventable diseases among population subgroups with low vaccination coverage remains a priority.

Measles virus transmembrane fusion protein synthesized de novo or presented in immunostimulating complexes is endogenously processed for HLA class I- and class II-restricted cytotoxic T cell recognition.

The routes used by antigen-presenting cells (APC) to convert the transmembrane fusion glycoprotein (F) of measles virus (MV) to HLA class I and class II presentable peptides have been examined, using cloned cytotoxic T lymphocytes in functional assays. Presentation by Epstein-Barr virus-transformed B lymphoblastoid cell lines was achieved using live virus, ultraviolet light-inactivated virus, and purified MV-F delivered either as such or incorporated in immunostimulating complexes (MV-F-ISCOM). Only live virus and MV-F-ISCOM allow presentation by class I molecules, while all antigen preparations permit class II-restricted presentation. We observe presentation of MV-F from live virus and as MV-F-ISCOM by class II molecules in a fashion that is not perturbed by chloroquine. Our studies visualize novel presentation pathways of type I transmembrane proteins.

Mumps: not an innocent bystander in solid organ transplantation.

Recently two major outbreaks of mumps have occurred: in the UK more than 56,000 cases were notified between 2004 and 2005, and in the United States, 6,584 cases were reported in 2006. Most patients were young healthy adults, in whom mumps normally has a benign course. Little is known about mumps in the immunocompromised patient. Here, we report a case of a 56-year renal transplant recipient who developed acute irreversible transplant failure due to interstitial nephritis caused by mumps. RNA of the mumps virus was detected in the urine as well as in a renal biopsy. In view of the ongoing presence of the mumps virus in the population, one should be aware of the possible occurrence of this infection in immunocompromised patients.

[An outbreak of measles at an emergency room]. / Een uitbraak van mazelen op de Spoedeisende Hulp.

A small outbreak of measles occurred after a 33-year-old female aircrew (cabin) member presented at an emergency room with fever. Three members of the hospital staff were infected: a 42-year-old man, a 33-year-old woman, and a 26-year-old woman. The first 2 patients had not been immunised, and the third had received 2 immunisations according to the Dutch National Immunisation Programme. Vaccination of the 2 sero-negative patients within 48 h after exposure with the measles-mumps-rubella vaccine (MMR) did not prevent the development of measles. Vaccination was deemed unnecessary in the third patient. No tertiary cases occurred. The same measles virus (genotype D5) was detected by PCR and sequencing in all 4 patients. Measles remains a risk for hospital staff members who have not acquired natural immunity. The current policy of immunising patients within 72 h after exposure to measles may not be sufficient. It also appears that immunisation through the Dutch National Immunisation Programme does not always protect against nosocomial infection. Providing MMR vaccination or boosters to hospital staff in certain departments might be beneficial.

[Two patients with mumps]. / Twee patiënten met de bof.

Two patients, men aged 17 and 19 years respectively, were admitted with parotitis epidemica and orchitis caused by mumps. The second patient also had meningitis. PCR analysis revealed that, in both cases, the causative agentwas a mumps virus that was genetically related to a wild-type virus responsible for an outbreak in Singapore. This viral strain was also responsible for a mumps outbreak at Hotel School The Hague in September 2004. Both patients were not fully vaccinated. Both patients were from regions in which clustering of patients with clinical signs of mumps has been seen. Interestingly, a number of patients with confirmed mumps had been fully vaccinated. Possible explanations for the increase in mumps cases include low vaccination and immunity levels, primary and secondary vaccine failure and the emergence of genetically disparate mumps viruses.

[Rubella epidemic in the Netherlands, 2004/'05: awareness of congenital rubella syndrome required]. / Rubella-epidemie in Nederland in 2004/'05: alertheid op congenitaal rubellasyndroom vereist.

Rubella is a public health problem due to the teratogenic effects associated with primary rubella infection during pregnancy (congenital rubella syndrome). Following universal rubella vaccination of infants in the Netherlands, the incidence of rubella has declined dramatically. However, since September 2004, an outbreak has occurred among unvaccinated individuals, most of whom declined vaccination based on religious beliefs. In the period 1 September 2004-22 March 2005, 166 cases of rubella were reported, including 12 pregnant women. Monitoring for signs that the epidemic has spread to other populations in the Netherlands is important because this might indicate the need for additional interventions. Awareness among health-care workers of the possible occurrence of congenital rubella syndrome should be raised. The clinical manifestations of congenital rubella syndrome are diverse, can be transient or permanent, and may not present until adolescence or adulthood. All cases of laboratory-confirmed rubella infection and congenital rubella syndrome should be reported to municipal health authorities. There is a possibility that this outbreak will spread abroad. The WHO aims to reduce the incidence of congenital rubella syndrome to < 1/100,000 live births. Health-care workers in the Netherlands should be extra alert to detect and notify rubella in a timely manner.

Inhibition of measles virus-induced cell-cell fusion with a monoclonal antibody directed against the haemagglutinin.

A neutralizing monoclonal antibody (C26-15) against the haemagglutinin (H protein) of measles virus was generated which caused cell-cell fusion inhibition in cultures of measles virus-infected cells. It was shown that this phenomenon coincided with a down-regulation of the expression of both the H protein and the fusion (F) protein. We also showed cell-cell fusion inhibition with a polyclonal rabbit serum directed against Tween-ether inactivated measles virus, which did not contain biologically active antibodies against the F protein. Cell-cell fusion inhibition caused by anti-H antibodies is distinct from cell-cell fusion inhibition induced by a direct interaction of anti-F antibody with the F protein in the membrane of infected cells. Since both mechanisms may also be involved in the in vivo situation, the exclusive role for the generation of anti-F antibody to prevent virus spread by cell-cell fusion in vivo is questioned. It is speculated that the observed down-regulation of both glycoproteins may lead to a less efficient killing of infected cells by cytotoxic T-lymphocytes, which may constitute an alternative explanation for the insufficient protection after vaccination with an inactivated measles vaccine.

Measles virus fusion protein- and hemagglutinin-transfected cell lines are a sensitive tool for the detection of specific antibodies by a FACS-measured immunofluorescence assay.

A FACS-measured immunofluorescence assay was developed for the detection of antibodies directed against the hemagglutinin (H) and fusion (F) glycoproteins of measles virus (MV). Human melanoma cell lines transfected with either the MV H or F genes, which showed a high surface expression of the respective proteins in their native conformation, were used as target cells. The cells were incubated with diluted plasma samples, and stained subsequently with FITC-conjugated secondary antibodies. The FACS-measured fluorescence signals correlated directly with the amount of specific immunoglobulins over a wide concentration range. The use of different conjugates enabled the separate detection of MV-specific IgG, IgM, IgA and IgG subclasses, with relatively low backgrounds. Hemagglutinin-specific IgG, IgM and IgA fluorescence signals were shown to correlate well with MV-specific IgG ELISA titers and MV-specific IgM or IgA capture ELISA OD450-values, respectively. The polyclonal conjugates with specificity for human immunoglobulins offered sufficient cross-reactivity to detect MV-specific IgG, IgM and IgA in plasma samples of cynomolgus macaques, making this technique a useful tool for studying serological responses in vaccination and challenge experiments in non-human primate models.

[A measles epidemic in an adequately vaccinated middle school population]. / Een mazelenepidemie in een goed gevaccineerde middelbare-schoolpopulatie.

OBJECTIVE: To assess the extent of a measles epidemic in a secondary school. DESIGN: Retrospective and questionnaire investigation. SETTING: Secondary school, Bilthoven. METHOD: Questionnaire followed by laboratory testing for measles and other infectious diseases with exanthema. RESULTS: The response rate was 99% (935/949 pupils, aged 12-21 years, vaccination rate 92%). Seventy-seven students underwent laboratory investigations. Measles virus was isolated in 2 suspected patients. Thirty-three of 37 patients with clinical or laboratory criteria of measles had been vaccinated. Complications of measles were not detected. Infection was also detected in patients with relatively few or atypical symptoms. The protective efficacy of measles vaccine could be determined because the attack rate of the school population was less than 5%. CONCLUSION: Primary failure of the measles vaccine might be the cause of the minor epidemic but the results do not cast doubt on the efficacy of the current measles vaccination programme.

[Measles epidemic in the Netherlands, 1999-2000]. / Mazelenepidemie in Nederland, 1999-2000.

OBJECTIVE: Description of measles epidemic in the Netherlands, 1999-2000. DESIGN: Observational descriptive study. METHODS: Intensified surveillance of measles cases by means of a case register established at the 'Landelijke Coördinatiestructuur Infectieziektebestrijding (LCI)' [National Co-ordination Centre for Communicable Disease Outbreak Management]. RESULTS: There were 3,292 reported measles patients, most of whom came from areas with low vaccine coverage. Of these patients, 94% had not been vaccinated; in 85% of cases this was for religious reasons. Of the 158 (5%) vaccinated patients, 157 had not (yet) received a second dose of vaccine. The incidence of measles increased with decreasing vaccine coverage in a municipality, both for unvaccinated and vaccinated persons. Three of the reported patients died. The percentage of patients with one or more complications was 22% in the group < 15 months of age, 19% in the group 15 months-4 years, 16% in the group 5-9 years, 11% in the group 10-19 years, and 15% in the group > 19 years of age. CONCLUSION: Considering the complications observed, the epidemic described involved a serious disease. Vaccination was accompanied by effective protection against measles infection and its complications. Herd immunity outside the unvaccinated groups was sufficient to prevent an epidemic there. However, incidental spread to vaccinated children did occur; the greatest risk factor for acquiring measles for vaccinated children is a stay in an area with low vaccine coverage.

Long-term presence of memory B-cells specific for different vaccine components.

Vaccination against infectious diseases ideally should provide lifelong immunity, but in many cases waning of antibody titers has been observed over time. In this study we describe the identification of antigen-specific memory B-cells in peripheral blood of persons born between 1940 and 2004 in The Netherlands. Polyclonal stimulation of either PBMCs or purified B-cells induced proliferation and differentiation of B-cells of the memory phenotype (CD19(+)/CD27(+)) into antibody secreting cells (ASC). Memory B-cells against components of bacterial vaccines (Bordetella pertussis and tetanus) as well as viral vaccines (measles and influenza) were thus identified, even in persons with low serum antibody titers. Enrichment of B-cells increased the sensitivity of memory B-cell detection when compared to PBMCs. Low, but significant correlations between numbers of antigen-specific memory B-cells and the corresponding circulating antibody titers were found for the pertussis proteins and measles virus, but not for tetanus. The identification of the numbers and specificities of peripheral memory B-cells and their relationship with circulating antibodies may be very useful to determine the long-term efficacy of vaccination.