A vectored measles virus induces hepatitis B surface antigen antibodies while protecting macaques against measles virus challenge

Hepatitis B virus (HBV) acute and chronic infections remain a major worldwide health problem. Towards developing an anti-HBV vaccine with single-dose scheme potential, we engineered infectious measles virus (MV) genomic cDNAs with a vaccine strain background and expression vector properties. Hepatitis B surface antigen (HBsAg) expression cassettes were inserted into this cDNA and three MVs expressing HBsAg at different levels generated. All vectored MVs, which secrete HBsAg as subviral particles, elicited humoral responses in MV-susceptible genetically modified mice. However, small differences in HBsAg expression elicited vastly different HBsAg antibody levels. The two vectors inducing the highest HBsAg antibody levels were inoculated into rhesus monkeys (Macaca mulatta). After challenge with a pathogenic MV strain (Davis87), control naive monkeys showed a classic measles rash and high viral loads. In contrast, all monkeys immunized with vaccine or a control nonvectored recombinant vaccine or HBsAg-expressing vectored MV remained healthy, with low or undetectable viral loads. After a single vaccine dose, only the vector expressing HBsAg at the highest levels elicited protective levels of HBsAg antibodies in two of four animals. These observations reveal an expression threshold for efficient induction of HBsAg humoral immune responses. This threshold is lower in mice than in macaques. Implications for the development of divalent vaccines based on live attenuated viruses are discussed.     

Mice immunized with measles virus develop antibodies to a cell surface receptor for binding virus.

Mice were immunized with measles virus to determine whether an auto-anti-idiotypic antireceptor response could be generated as a probe for measles virus receptors. Mice initially responded to viral antigens (days 11 to 18) and subsequently developed antibodies to a putative measles virus receptor (peak at day 30 to 35) by three criteria: the sera (1) agglutinated erythrocytes which virus agglutinates, (2) reacted with Vero cells, and (3) inhibited virus attachment to Vero cells. Additionally, select sera inhibited virus infection of Vero cells. The cell-reactive activity was identified as immunoglobulin G antibody and was neutralized by sera reacting with virus (idiotype). The application of this anti-idiotypic antibody to identify measles virus-binding sites on Vero cells was revealed by the ability of sera to immunoprecipitate 20- and 30.5-kilodalton proteins from metabolically labeled ([35S]methionine) Vero cells.     

Development of a guinea pig colony free of complement-fixing antibodies to parainfluenza virus.

Complement-fixing antibodies to parainfluenza 3 virus were found in Hartley strain [Cds: (HA)] guinea pigs from the authors' production colony. The prevalence and distribution of these antibodies were determined by testing guinea pigs of five age categories: 4 weeks, 8 weeks, 12 weeks, 6 months to 1 year, and over 1 year of age. Forty-seven percent (28 of 60) were positive to parainfluenza 3 antigen. Positive reactors were found in all age groups except those 8 weeks old. The 12-week-old group had the highest titers; the group over 1 year of age had the highest percentage of positives (92%). When 8-week-old guinea pigs were isolated, 55% were positive at some time between 8 and 34 weeks of age. The titers characteristically rose rapidly and then dropped slowly to low or undetectable levels. Four pairs of breeders over 6 months of age (most of whom were positive for parainfluenza 3 virus antibodies and, therefore, presumed to be immune to the virus) were isolated and allowed to breed. Their offspring were found to be free of complement-fixing antibodies to parainfluenza 3 virus.     

Three monoclonal antibodies against measles virus F protein cross-react with cellular stress proteins.

In a group of 11 monoclonal antibodies specifically reacting with the measles virus fusion protein, three antibodies also immunoprecipitated other proteins, in particular a 79,000-molecular-weight protein from virus-infected cells. The cross-reacting 79,000-molecular-weight protein was shown to be a virus-induced host stress protein. This protein could be induced by (i) different paramyxoviruses, (ii) heat shock of uninfected HeLa cells, and (iii) 2-deoxyglucose, tunicamycin, or L-canavanine treatment of different mammalian cell lines. Immunofluorescence of stressed HeLa cells localized the cross-reacting host protein(s) mainly in the cytoplasm. The significance of these results in relation to autoimmunity is discussed.     

A novel role for non-neutralizing antibodies against nucleoprotein in facilitating resistance to influenza virus

Current influenza vaccines elicit Abs to the hemagglutinin and neuraminidase envelope proteins. Due to antigenic drift, these vaccines must be reformulated annually to include the envelope proteins predicted to dominate in the following season. By contrast, vaccination with the conserved nucleoprotein (NP) elicits immunity against multiple serotypes (heterosubtypic immunity). NP vaccination is generally thought to convey protection primarily via CD8 effector mechanisms. However, significant titers of anti-NP Abs are also induced, yet the involvement of Abs in protection has largely been disregarded. To investigate how Ab responses might contribute to heterosubtypic immunity, we vaccinated C57BL/6 mice with soluble rNP. This approach induced high titers of NP-specific serum Ab, but only poorly detectable NP-specific T cell responses. Nevertheless, rNP immunization significantly reduced morbidity and viral titers after influenza challenge. Importantly, Ab-deficient mice were not protected by this vaccination strategy. Furthermore, rNP-immune serum could transfer protection to naive hosts in an Ab-dependent manner. Therefore, Ab to conserved, internal viral proteins, such as NP, provides an unexpected, yet important mechanism of protection against influenza. These results suggest that vaccines designed to elicit optimal heterosubtypic immunity to influenza should promote both Ab and T cell responses to conserved internal proteins.     

The prevalence of islet-cell antibodies and complement-fixing islet-cell antibodies in Japanese diabetics

The presence of complement-fixing islet-cell antibodies (CF-ICA) and islet-cell antibodies (ICA) was examined in 355 patients with different types of diabetes mellitus in the Japanese population by an indirect immunofluorescence test (IFT). The overall prevalence of ICA, which were stained as a homogenous cytoplasmic fluorescence in islet cells, was 7 per cent (5/67) in insulin-dependent (Type I) diabetics, 4 per cent (6/137) in noninsulin-dependent (Type II) diabetics treated with insulin and 2 per cent (1/58) in Type II diabetics treated with oral hypoglycemic agents. None of 84 Type II diabetics receiving diet alone and 9 diabetics associated with chronic pancreatitis had ICA. CF-ICA, which were stained as a "ring-shaped" fluorescence in a part of the cytoplasma, were demonstrated in 5 out of 12 cases (42%) whose sera possessed ICA. The lower prevalence and remarkably shorter persistence of ICA and CF-ICA in Japanese diabetics than those observed in Caucasian diabetics may be explained by the heterogenous immunological response in different races or possible heterogeneity of Type I diabetics.     

Relative contributions of measles virus hemagglutinin- and fusion protein-specific serum antibodies to virus neutralization

The relative contribution of measles virus hemagglutinin (H)- or fusion protein (F)-specific antibodies to virus neutralization (VN) has not been demonstrated. We have depleted these specific antibodies from sera collected from young adults, who had been vaccinated during childhood, by prolonged incubation with viable transfected human melanoma cells expressing H or F. Simultaneous depletion of antibodies of both specificities completely abrogated VN activity. Depletion of F-specific antibodies only had a minimal effect, whereas removal of H-specific antibodies resulted in almost complete reduction of VN activity. These results demonstrate that measles virus neutralizing antibodies are mainly directed to the H protein.