Biomedical subjects

W J Bellini

Publications and source records attributed to W J Bellini.

At least 37 records · Page 2Linked to original sources

PubMed1999

Timing of development of measles-specific immunoglobulin M and G after primary measles vaccination.

A standard method for diagnosing measles is to detect measles-specific immunoglobulin M (IgM) in the serum of infected persons. Interpreting a positive IgM result from a person with suspected measles can be difficult if the person has recently received a measles vaccine. We have previously demonstrated that measles-specific IgM may persist for at least 8 weeks after primary vaccination, but it is unknown how quickly IgM appears. This study determined the timing of the rise of measles-specific IgM and IgG after primary measles vaccination with Schwartz vaccine. Two hundred eighty 9-month-old children from Ethiopia presenting for routine measles vaccination were enrolled. Sera were collected before and either 1, 2, 3, or 4 weeks after vaccination and tested for measles-specific antibodies by an IgM capture enzyme immunoassay (EIA) and by an indirect IgG EIA. A total of 209 of the 224 children who returned for the second visit had prevaccination sera that were both IgM and IgG negative. The postvaccination IgM positivity rates for these 209 children were 2% at 1 week, 61% at 2 weeks, 79% at 3 weeks, and 60% at 4 weeks. The postvaccination IgG positivity rates were 0% at 1 week, 14% at 2 weeks, 81% at 3 weeks, and 85% at 4 weeks. We conclude that an IgM-positive result obtained by this antibody capture EIA is difficult to interpret if serum is collected between 8 days and 8 weeks after vaccination; in this situation, the diagnosis of measles should be based on an epidemiologic linkage to a confirmed case or on the detection of wild-type measles virus.

Antibodies, Viral↗

PubMed1999

Spontaneous mutation rate of measles virus: direct estimation based on mutations conferring monoclonal antibody resistance.

High mutation rates typical of RNA viruses often generate a unique viral population structure consisting of a large number of genetic microvariants. In the case of viral pathogens, this can result in rapid evolution of antiviral resistance or vaccine-escape mutants. We determined a direct estimate of the mutation rate of measles virus, the next likely target for global elimination following poliovirus. In a laboratory tissue culture system, we used the fluctuation test method of estimating mutation rate, which involves screening a large number of independent populations initiated by a small number of viruses each for the presence or absence of a particular single point mutation. The mutation we focused on, which can be screened for phenotypically, confers resistance to a monoclonal antibody (MAb 80-III-B2). The entire H gene of a subset of mutants was sequenced to verify that the resistance phenotype was associated with single point mutations. The epitope conferring MAb resistance was further characterized by Western blot analysis. Based on this approach, measles virus was estimated to have a mutation rate of 9 x 10(-5) per base per replication and a genomic mutation rate of 1.43 per replication. The mutation rates we estimated for measles virus are comparable to recent in vitro estimates for both poliovirus and vesicular stomatitis virus. In the field, however, measles virus shows marked genetic stability. We briefly discuss the evolutionary implications of these results.

Animals↗

PubMed1999

Management of a measles outbreak among Old World nonhuman primates.

BACKGROUND AND PURPOSE: A measles outbreak in a facility housing Old World nonhuman primates developed over a 2-month period in 1996, providing an opportunity to study the epidemiology of this highly infectious disease in an animal-handling setting. METHODS: Serum and urine specimens were collected from monkeys housed in the room where the initial measles cases were identified, other monkeys with suspicious measles-like signs, and employees working in the affected areas. Serum specimens were tested for measles virus-specific IgG and IgM antibodies, and urine specimens were tested for measles virus by virus isolation or reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: A total of 94 monkeys in two separate facilities had evidence of an acute measles infection. The outbreak was caused by a wild-type virus that had been associated with recent human cases of acute measles in the United States; however, an investigation was unable to identify the original source of the outbreak. Quarantine and massive vaccination helped to control further spread of infection. CONCLUSIONS: Results emphasize the value of having a measles control plan in place that includes a preventive measles vaccination program involving human and nonhuman primates to decrease the likelihood of a facility outbreak.

Animals↗

PubMed1998

Nonclassic measles infections in an immune population exposed to measles during a college bus trip.

This study investigated the frequency of mild or asymptomatic measles infections among 44 persons exposed to a student with measles during a 3-day bus trip using two buses. Questionnaires and serum samples were obtained 26-37 days after the trip. All participants had detectable measles-neutralizing antibodies, and none developed classic measles symptoms. Ten persons (23%) were IgM positive for measles, indicating recent infection. Among previously vaccinated IgM-negative persons, those who rode on bus A with the index case-patient had significantly higher microneutralization titers than those on bus B (P= .001), suggesting that some persons on bus A were infected but were IgM negative at the time of the study. Mild or asymptomatic measles infections are probably very common among measles-immune persons exposed to measles cases and may be the most common manifestation of measles during outbreaks in highly immune populations.

Adolescent↗

PubMed1998

Genetic and antigenic characterisation of the haemagglutinin protein of measles virus strains recently circulating in the UK.

The complete nucleotide sequence of the H protein gene of seven measles virus (MV) strains, representing three MV genotypes circulating in the UK in recent years, was determined. Compared to the MV vaccine strain Moraten (Mor-v), the divergence of the coded H gene (aal-600) of the seven UK strains was between 1.8% and 2.8%. Representative isolates from each of the genotypes were tested by radio-immunoprecipitation using a panel of H protein-specific MAbs. Different patterns of MAb reactivity were shown between the three genotypes and between the wild-type strains and the vaccine strain. Plaque reduction neutralising antibody titres against strains UK350/94 (genotype I) and UK226/94 (genotype III) were measured in sera from 11 vaccinees. Vaccine derived antibody neutralised both strains and the GMTs were not significantly lower against the wild-type strains than against strain Mor-v.

Amino Acid Substitution↗

PubMed1998

New genetic group of measles virus isolated in the People's Republic of China.

Genetic and antigenic characterization of 14 wild-type measles viruses isolated from four provinces in the People's Republic of China during 1993 and 1994 was conducted. Sequence analyses of the hemagglutinin (H) and nucleoprotein (N) genes indicated that 13 of the 14 Chinese viruses comprised a previously undescribed genetic group. Viruses from this unique group were the most genetically diverse measles viruses described, so far. The Chinese viruses differed from other wild-type viruses by as much as 6.9% in the H gene and 7.0% in the N gene at the nucleotide level. One of the 14 viruses was a member of the same genetic group that contains the Edmonston strain. Antigenic analysis using monoclonal antibodies to the H protein did not detect significant differences in binding patterns between the Chinese viruses and other wild-type measles viruses. In addition, representative viruses from the unique Chinese group were neutralized by both human post-vaccination antiserum and mouse antiserum against the H protein of the Edmonston vaccine virus. Viruses closely related to these Chinese viruses were also associated with importations of measles into the United States during 1997 from Vietnam and Hong Kong suggesting that viruses from this new genetic group continue to circulate in China and possibly other parts of Asia.

Antigens, Viral↗

PubMed1998

Genetic analysis of measles viruses isolated in the United States, 1995-1996.

Genetic analysis was conducted on 28 wild type measles viruses isolated from outbreaks or cases in the United States during 1995-1996. These viruses were members of at least 6 distinct genetic groups. However, none of these viruses was related to the group 2 viruses that were associated with the resurgence of measles in the United States between 1989 and 1992 except for a single importation from the Philippines. The sequence data support and extend previous findings showing that transmission of group 2 viruses within the United States was interrupted after 1993. The data also suggest that all measles cases that occurred in the United States in 1995-1996 were the result of importation of virus, even in instances when the source was unknown. Molecular epidemiologic studies can provide a means to measure the success of measles control programs by helping to identify the transmission pathways of the virus.

Disease Outbreaks↗

PubMed1998

Population biology, evolution, and immunology of vaccination and vaccination programs.

The purpose of prophylactic vaccination is to reduce morbidity and mortality in a population. Many questions related to the design of vaccines and vaccination programs require a population standpoint for their sharp formulation and laboratory and field studies to understand their immunologic background. Practical suggestions of the workshop included increased studies of age-specific immunity, better immunoepidemiologic surveillance, better design of efficacy studies, and more systematic sampling of parasite strains to study the evolutionary pressure exerted by vaccines. Theoretical immunology has much to contribute. One of the realizations of the workshop was the value of a strong interdisciplinary approach in vaccine development, utilizing relevant contributions from immunology, population biology, mathematical modeling, epidemiology, molecular biology, and virology.

Animals↗

PubMed1998

Decline of measles-specific immunoglobulin M antibodies after primary measles, mumps, and rubella vaccination.

Detection of measles-specific immunoglobulin M (IgM) has become the standard diagnostic method for laboratory confirmation of measles. In outbreaks, the interpretation of an IgM-positive result can be complicated when persons with suspected measles receive a dose of measles vaccine as part of outbreak control measures. This investigation evaluated the decay of measles-specific IgM antibodies 1 to 4 months after primary vaccination with measles, mumps, and rubella vaccine (MMRII). Serum samples were obtained from 536 infants vaccinated when they were 15 months old as part of a study to assess primary and secondary measles vaccine failure. Sixty serum specimens per week were selected from specimens collected between 4 and 9 weeks after MMRII vaccination; all 176 available serum specimens collected between 10 and > or = 16 weeks were included. Specimens were tested for the presence of measles-specific IgM by an antibody-capture enzyme immunoassay. The proportion of IgM-positive specimens dropped from 73% at 4 weeks after vaccination to 52% at 5 weeks after vaccination and then declined to 7% by 8 weeks after vaccination. Less than 10% of children remained IgM positive between 9 and 11 weeks. An IgM-negative result helps rule out the diagnosis of measles in a person with suspected infection and a history of recent vaccination. The interpretation of a positive IgM result from a person with a clinically suspected case of measles and a recent history of measles vaccination (especially within 8 weeks) is problematic, and the diagnosis of measles should be based on epidemiologic linkage to a confirmed case or on detection of wild-type measles virus.

Antibodies, Viral↗

PubMed1998

Strain-specific differences in LFA-1 induction on measles virus-infected monocytes and adhesion and viral transmission to endothelial cells.

Measles virus (MV) infection of monocytes induces leukocyte function-associated antigen-1 (LFA-1), an integrin that mediates intercellular adhesion to the endothelium. Thus, an increase in LFA-1 expression could lead to enhanced monocyte adherence and virus dissemination to endothelial cells (ECs) and potentially be an important means of distinction between MV strains. We identified both vaccine and wild-type strains that induced LFA-1 and others that failed to induce. Although adhesion of MV-infected monocytes and viral transmission to ECs was demonstrated, strain-specific differences were not correlated with LFA-1 induction. MV infection of ECs was dramatically reduced in the absence of cell contact, suggesting virus dissemination by cell-cell transmission.

Cell Adhesion↗

PubMed1998

A single amino acid change in the hemagglutinin protein of measles virus determines its ability to bind CD46 and reveals another receptor on marmoset B cells.

This paper provides evidence for a measles virus receptor other than CD46 on transformed marmoset and human B cells. We first showed that most tissues of marmosets are missing the SCR1 domain of CD46, which is essential for the binding of Edmonston measles virus, a laboratory strain that has been propagated in Vero monkey kidney cells. In spite of this deletion, the common marmoset was shown to be susceptible to infections by wild-type isolates of measles virus, although they did not support Edmonston measles virus production. As one would expect from these results, measles virus could not be propagated in owl monkey or marmoset kidney cell lines, but surprisingly, both a wild-type isolate (Montefiore 89) and the Edmonston laboratory strain of measles virus grew efficiently in B95-8 marmoset B cells. In addition, antibodies directed against CD46 had no effect on wild-type infections of marmoset B cells and only partially inhibited the replication of the Edmonston laboratory strain in the same cells. A direct binding assay with insect cells expressing the hemagglutinin (H) proteins of either the Edmonston or Montefiore 89 measles virus strains was used to probe the receptors on these B cells. Insect cells expressing Edmonston H but not the wild-type H bound to rodent cells with CD46 on their surface. On the other hand, both the Montefiore 89 H and Edmonston H proteins adhered to marmoset and human B cells. Most wild-type H proteins have asparagine residues at position 481 and can be converted to a CD46-binding phenotype by replacement of the residue with tyrosine. Similarly, the Edmonston H protein did not bind CD46 when its Tyr481 was converted to asparagine. However, this mutation did not affect the ability of Edmonston H to bind marmoset and human B cells. The preceding results provide evidence, through the use of a direct binding assay, that a second receptor for measles virus is present on primate B cells.

Amino Acid Sequence↗

PubMed1997

Use of synthetic peptides to identify measles nucleoprotein T-cell epitopes in vaccinated and naturally infected humans.

Recombinant measles nucleoprotein (N) and synthetic peptides spanning the length of the N-protein-coding region were used with a proliferation assay to identify human T-cell epitopes in vaccinated and naturally infected adults. A number of epitopes were mapped to specific regions of the measles virus N. The proliferative response of at least two donors was mediated by CD4(+) T cells in association with HLA DR antigens. Over 70% of all donors tested responded to peptides representing amino acids 271-290, 367-386, 400-420, and 483-502, suggesting that these peptides may be broadly recognized within an HLA diverse population. The most frequently recognized T-cell epitopes in both naturally infected and vaccinated donors were located in the genetically heterogeneous carboxy-terminal half of the N. Analysis of patterns of peptide reactivity among vaccinated and naturally infected subjects identified several regions of potential difference between these two groups. Peptides 221-240 and 237-256 were recognized among 100% of naturally infected donors but among only 37.5% of vaccinated donors and therefore may be of further interest in studies to investigate induction of lifelong versus transient immunity to measles. Use of chimeric molecules containing multiple well-characterized T- and B-cell epitopes or genetic alteration of attenuated vaccine virus to enhance critical T-cell responses may eventually lead to the development of a vaccine candidate that can more closely model the patterns of immune response elicited by wild-type virus.

Adult↗

PubMed1997

Experimental measles. I. Pathogenesis in the normal and the immunized host.

An animal model to study measles pathogenesis and the correlates of protective immunity was established using rhesus monkeys. A measles isolate, obtained during an epidemic of measles in the primate colony at the University of California, Davis, was passaged through rhesus monkeys and amplified in rhesus mononuclear cells to create a pathogenic virus stock. Sequence analysis of the nucleoprotein and hemagglutinin genes of this isolate revealed strong homology with the Chicago 89 strain of measles virus. Conjunctival/intranasal inoculation of juvenile rhesus monkeys with this virus resulted in skin rash, pneumonia, and systemic infection with dissemination to other mucosal sites and to the lymphoid tissues. Inflammation and necrosis occurred in the lungs and lymphoid tissues and many cell types were infected with measles virus on Day 7 postinoculation (p.i.). The most commonly infected cell type was the B lymphocyte in lymphoid follicles. Measles antigen was found in follicular dendritic cells on Day 14 p.i. In contrast to naive monkeys infected with measles virus, animals vaccinated with the attenuated Moraten strain did not develop clinical or pathologic signs of measles after challenge. However, moderate to marked hyperplasia occurred in the lymph nodes and spleen of a vaccinated animal on Day 7 after pathogenic virus challenge, suggesting that an effective measles vaccine limits but does not prevent infection with wild-type measles virus.

Animals↗

PubMed1997

Diagnosis of measles with an IgM capture EIA: the optimal timing of specimen collection after rash onset.

The optimal timing for collection of a single serum specimen to diagnose measles by using a monoclonal antibody-capture EIA was evaluated. Results of testing paired serum samples from 166 measles cases with at least 1 IgM-positive specimen were analyzed. Among persons whose second samples were IgM-positive, the seropositivity rate for first samples was 77% when collected within 72 h and 100% when collected 4-11 days after rash onset. Among unvaccinated persons whose first samples were IgM-positive, the rate for IgM positivity of second specimens declined from 100% at 4 days to 94% at 4 weeks after rash onset, then declined further to 63% at 5 weeks. Some previously vaccinated persons became IgM-negative during the third week after rash onset. In general, a single serum specimen collected between 72 h and 4 weeks after rash onset can be used to diagnose most cases of measles with an IgM capture EIA.

Adolescent↗

PubMed1997

The diversity of measles virus in the United Kingdom, 1992-1995.

Three distinct genotypes were identified amongst 50 measles virus (MV) strains characterized in the UK between 1992 and 1995 by direct sequencing of the RT-PCR products amplified from clinical specimens. All three genotypes were related to viruses previously reported in the United States or in continental Europe. Phylogenetic analyses of 255 and 152 nucleotide sequences from the N and M genes, respectively, generated very similar lineages. The degree of divergence between genotypes was 3.5-16% and 2.6-7.9% in the N and the M genes, respectively. MV genotypes which circulated during the 1970s and 1980s in the UK were not detected in this period. Comparison of the C-terminal regions of the N gene sequences of UK strains isolated or detected between 1974 and 1995 suggests that there have been multiple genotypes of MV circulating in the UK over the past 20 years.

Base Sequence↗

PubMed1996

Generation of defective interfering particles by two vaccine strains of measles virus.

A systematic study was made to measure the generation of defective interfering particles upon up to 13 serial passages of two measles vaccine strains, Edmonston and Edmonston-Zagreb, through either simian (Vero) or human (WI-38) cell lines. Results for the Vero cell passage were nearly identical for both viruses. Infectivity titers dropped by nearly 8 logs to undetectable levels at passage 4 and cycled between maximum and minimum levels every 4 passages. Samples with the lowest infectivity titers produced the greatest reduction in titer of standard virus and contained an approximately 900-nucleotide subgenomic RNA for the Edmonston strain and two subgenomic RNAs of 4300 and 3000 nucleotides for the Edmonston-Zagreb vaccine strain. A defective interfering RNA-specific reverse transcription-polymerase chain reaction (RT-PCR) detected subgenomic RNAs at all passage levels. In contrast, samples obtained after passage of these viruses in WI-38 did not reduce the yield of standard virus and did not contain subgenomic RNAs in both Northern blot and RT-PCR assays. These results clearly show that cell type rather than virus strain affects defective interfering particle generation for measles virus.

Animals↗

PubMed1996

Antibody response to measles-mumps-rubella vaccine of children with mild illness at the time of vaccination.

OBJECTIVE: To examine the response to measles-mumps-rubella (MMR) vaccine among children with and without mild illness. DESIGN: Prospective cohort. PARTICIPANTS: A total of 386 children aged 15 to 23 months. MAIN OUTCOME MEASURES: Seroconversion rates to measles, mumps, and rubella in ill and well children. SETTING: Six public health immunization clinics in two counties in the greater metropolitan Atlanta, Ga, area from February 1992 to April 1993. RESULTS: Acute upper respiratory tract infection, otitis media, and diarrhea were observed in 128 (33%), 41 (11%), and 13 (3%) of the children (groups are not mutually exclusive); 157 children had one of these mild illnesses and 229 were well. Overall seroconversion rates were 98% for measles, 83% for mumps, and 98% for rubella antigens. Measles seroconversion rates for ill children compared with well children, respectively, were as follows: upper respiratory tract infection, 99% vs 97%; mild fever, 98% vs 97%; otitis media, 98% vs 98%; diarrhea, 100% vs 98%; and any mild illness, 99% vs 97%. Estimates of the magnitude of antibody response to measles, mumps, and rubella antigens were the same for children with and without mild illness. There was no association of mild illness with increased rates and severity of adverse events reported in the 2 weeks after vaccination. CONCLUSIONS: Vaccinating children who present with mild illnesses with MMR vaccine is a safe and efficacious practice. These results support recommendations of the Advisory Committee on Immunization Practices and the American Academy of Pediatrics.

Antibodies, Viral↗

PubMed1996

Molecular epidemiology of measles virus: identification of pathways of transmission and implications for measles elimination.

The nucleotide sequences of either the hemagglutinin or nucleoprotein genes from wild type measles viruses isolated in the United States between 1989 and 1992 differed by < 0.5%. This suggests that the majority of viruses associated with resurgence of measles in the United States belonged to a single indigenous genotype. In contrast, wild type viruses isolated from sporadic outbreaks of measles in the United States during 1994 were genetically heterogeneous. These viruses were more closely related to wild type viruses previously circulating in Europe, Africa, or Japan and were epidemiologically linked to importations or no known source. In addition to demonstrating the utility of genetic analysis in understanding the epidemiology of measles, these data suggest that the transmission of the indigenous virus was interrupted after the 1989-1992 epidemic. Measures to further reduce the incidence of measles in the United States should include efforts to control importation and subsequent spread of measles.

Base Sequence↗