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Biomedical subjects

R M Chanock

Publications and source records attributed to R M Chanock.

At least 55 records · Page 3Linked to original sources
PubMed1990

Evaluation in chimpanzees of vaccinia virus recombinants that express the surface glycoproteins of human respiratory syncytial virus.

The immunogenicity and protective efficacy of recombinant vaccinia viruses that express the two major protective antigens of human respiratory syncytial virus (RSV), the F and G glycoproteins, were evaluated in chimpanzees. In previous studies in rodents and monkeys the F and G proteins expressed by the same recombinants were highly immunogenic and induced high levels of resistance to RSV replication following subsequent challenge. In contrast, in chimpanzees, a single intradermal immunization induced only moderate levels of F and G-specific serum antibodies as measured by an enzyme-linked immunosorbent assay, and these antibodies did not efficiently neutralize RSV infectivity in vitro. This poor antibody response in chimpanzees to the F and G glycoproteins occurred despite efficient replication of the vaccinia virus vector as evidenced by lesion size and serum antibody response to vaccinia virus. Upon intranasal RSV challenge, it was observed that prior immunization with the F and G recombinants effected only a marginal reduction in the magnitude and duration of RSV shedding from the nose and trachea and did not reduce illness. However, the RSV challenge induced a strong secondary antibody response, resulting in very high titres (greater than 8000 reciprocal mean titre) of serum neutralizing antibodies. The poor protective efficacy observed here is discussed with regard to the permissiveness of the chimpanzee to RSV replication, the general requirements for effective immunization against RSV, and the limitations of experimental animals for evaluating candidate RSV vaccines.

Animals↗
PubMed1990

Role of early genes in pathogenesis of adenovirus pneumonia.

Intranasal inoculation of type 5 adenovirus into the cotton rat Sigmodon hispidus produces a pneumonia pathologically similar to that in humans, and it, therefore, provides an excellent animal model to investigate the pathogenesis of this disease. The goal of this study was to test the hypothesis that accumulation of viral structural proteins is responsible for a major portion of the cell-damage-producing disease. Since viral DNA replication is essential for synthesis of the viral structural proteins, which are products of late genes, the hypothesis was tested using mutants defective in genes required for DNA synthesis. Most experiments were done with the conditionally lethal temperature-sensitive (ts) mutant H5ts125, which contains a mutation in the early region 2A (E2A) gene encoding the DNA-binding protein. The data show that infection with 1 x 10(9.0) plaque-forming units of H5ts125 induced a pneumonia that was as extensive and qualitatively the same as that after wild-type adenovirus type 5 infection, although H5ts125 did not replicate to produce infectious virus. When cotton rats were infected with 1 x 10(8.0) plaque-forming units of wild-type adenovirus type 5 or H5ts125, the pneumonias that followed were pathologically similar; in the latter phases, however, wild-type virus produced slightly more extensive pneumonia than did H5ts125, probably because its replication permitted infection of more susceptible cells.

Adenoviridae Infections↗
PubMed1990

Antigenic relationships among human rotaviruses as determined by outer capsid protein VP4.

cDNA clones representing the VP4 gene of symptomatic human rotavirus strain KU (VP7 serotype 1) or DS-1 (VP7 serotype 2) or asymptomatic human rotavirus strain 1076 (VP7 serotype 2) were constructed and inserted into a baculovirus expression vector under the control of the polyhedrin promoter. The resulting recombinants expressed the appropriate authentic VP4 rotavirus outer capsid protein. Guinea pigs immunized with these VP4 proteins developed antibodies that neutralized infectivity of the rotavirus from which the immunizing VP4 was derived. These antisera were then used in neutralization tests to define the extent and distribution of VP4 antigenic polymorphism among human rotaviruses. Three distinct serotypes and one subtype of the VP4 outer capsid protein were identified among 17 human rotavirus strains that had previously been assigned to five distinct VP7 serotypes. For the most part, VP4 serotype segregated independently of VP7 serotype. Ten strains of human rotavirus that were associated with symptomatic infection and that exhibited VP7 serotype 1, 3, 4, or 9 specificity, each possessed a VP4 of the same serotype and subtype, designated VP4 serotype 1A. Both symptomatic human rotavirus strains with VP7 serotype 2 specificity were related by neutralization to the VP4 serotype 1A strains and were classified as a subtype of VP4 serotype 1--i.e., serotype 1B--since viruses of serotype 1A appeared to be prime strains. Four human rotavirus strains that were recovered from healthy infants in newborn nurseries in which virus transmission persisted over a long interval, belonged to VP7 serotype 1, 2, 3, or 4, but each strain possessed the same VP4 antigenic specificity that was designated VP4 serotype 2. Finally, a single strain of symptomatic human rotavirus of VP7 serotype 1 specificity possessed a unique VP4 that was provisionally classified as VP4 serotype 3 but this remains to be confirmed because neutralization tests were performed in only one direction. Among the 10 rotavirus strains whose VP4 gene was previously sequenced, there was complete concordance between assignment of VP4 serotype by neutralization and classification according to VP4 amino acid homology. Thus, rotaviruses that exhibited a VP4 amino acid homology of 89% or greater belonged to the same VP4 serotype and subtype as determined by neutralization. Finally, evidence was obtained that the serotype-specific domain is located on the VP8 subunit of VP4.

Animals↗
PubMed1990

Antibody responses of humans and nonhuman primates to individual antigenic sites of the hemagglutinin-neuraminidase and fusion glycoproteins after primary infection or reinfection with parainfluenza type 3 virus.

An unusual feature of human parainfluenza virus type 3 (PIV3) is ita ability to cause reinfection with high efficiency. The antibody responses of 45 humans and 9 rhesus monkeys to primary infection or subsequent reinfection with PIV3 were examined to identify deficiencies in host immunologic responses that might contribute to the ability of the virus to cause reinfection with high frequency. Antibody responses in serum were tested by using neutralization and hemagglutination inhibition (HI) assays and a monoclonal antibody blocking immunoassay able to detect antibodies to epitopes within six antigenic sites on the PIV3 hemagglutinin-neuraminidase (HN) glycoprotein and eight antigenic sites on the fusion (F) protein. Primary infection of seronegative infants or children with PIV3 stimulated strong and rather uniform HI and neutralizing antibody responses. More than 90% of the individuals developed antibodies to four of the six HN antigenic sites (including three of the four neutralization sites), but the responses to F antigenic sites were of lesser magnitude and varied considerably from person to person. Young infants who possessed maternally derived antibodies in their sera developed lower levels and less frequent HI, neutralizing, and antigenic site-specific responses to the HN and F glycoproteins than did seronegative infants and children. In contrast, children reinfected with PIV3 developed even higher HI and neutralizing antibody responses than those observed during primary infection. Reinfection broadened the HN and F antigenic site-specific responses, but the latter remained relatively restricted. Adults possessed lower levels of HI, neutralizing, and antigenic site-specific antibodies in their sera than did children who had been reinfected, suggesting that these antibodies decay with time. Rhesus monkeys developed more vigorous primary and secondary antibody responses than did humans, but even in these highly responsive animals, response to the F glycoprotein was relatively restricted following primary infection. Bovine PIV3 induced a broader response to human PIV3 in monkeys than was anticipated on the basis of their known relatedness as defined by using monoclonal antibodies to human PIV3. These observations suggest that the restricted antibody responses to multiple antigenic sites on the F glycoprotein in young seronegative infants and children and the decreased responses to both the F and HN glycoproteins in young infants and children with maternally derived antibodies may play a role in the susceptibility of human infants and young children to reinfection with PIV3.

Adult↗
PubMed1989

VP4 protein of porcine rotavirus strain OSU expressed by a baculovirus recombinant induces neutralizing antibodies.

The complete VP4 gene of porcine rotavirus strain OSU has been inserted into a baculovirus expression vector under the control of the polyhedrin promoter. The VP4 outer capsid protein, which is a major neutralization antigen in rotavirus, was expressed in high yield in Spodoptera frugiperda cells. Reactivity with polyclonal and monoclonal antibodies suggested that neutralizing epitopes were functionally unaltered on the expressed VP4. The VP4 produced in this system also induced antibodies in guinea pigs which inhibited hemagglutination of OSU and neutralized its infectivity to high titer. The available evidence suggests that the VP4 expressed in insect cells maintained its antigenic configuration and may prove useful in elucidation of (1) the extent of VP4 polymorphism among human and animal rotaviruses and (2) the distribution of VP4 among these viruses.

Animals↗
PubMed1989

Rotavirus VP7 neutralization epitopes of serotype 3 strains.

Sequence analysis of the gene encoding the major neutralization glycoprotein (VP7) was performed on 27 human and animal rotavirus strains of serotype 3 in order to examine genetic variation within strains of identical serotype. Comparisons of the deduced amino acid sequences of the VP7s showed overall sequence identities of 85% or higher. A higher degree of overall VP7 sequence similarity was observed among strains from the same animal species when compared to strains from different animal species, suggesting that there are species-specific sequences in the VP7 protein. Alignment of the amino acid sequences demonstrated that amino acid sequence divergence among serotype 3 strains from different species was located primarily in previously established VP7 serotype-specific regions where genetic variation was identified among strains of different serotype. These regions were highly conserved among serotype 3 strains derived from the same species. The varying reactivities of three anti-VP7 monoclonal antibodies with the 27 strains was consistent with the occurrence of antigenic variation among serotype 3 strains. Moreover the reactivity of monoclonal antibodies correlated with the amino acid sequence found in two serotype-specific regions (VR5 and VR8). A computer-derived predicted phylogenetic tree suggests that rotavirus strains from different animal species belonging to serotype 3 are more closely related to each other than to rotavirus strains of different serotypes.

Amino Acid Sequence↗
PubMed1989

Immunization of cotton rats with the fusion (F) and large (G) glycoproteins of respiratory syncytial virus (RSV) protects against RSV challenge without potentiating RSV disease.

A formalin-inactivated respiratory syncytial virus (RSV) vaccine tested 22 years ago failed to protect infant vaccinees against RSV infection or disease. Instead, lower respiratory tract disease was enhanced during subsequent infection by RSV. Enhancement of pulmonary pathology is also observed when cotton rats are immunized with formalin-inactivated RSV and subsequently infected with this virus. A major question that must be addressed for each new paramyxovirus vaccine is whether the immunogen possesses the capacity to potentiate disease. In the present study, we evaluated a newly developed purified F and G glycoprotein vaccine over a wide dosage range for immunogenicity, efficacy and capacity to potentiate pulmonary pathology in cotton rats. In addition, a formalin-inactivated RSV vaccine, which served as a positive control for enhancement of pulmonary pathology, was evaluated simultaneously. The results of these comparisons indicate that the purified F and G glycoprotein vaccine was highly immunogenic and was efficacious even in animals that developed low levels of serum-neutralizing antibodies. Furthermore, the F and G vaccine did not induce potentiation of pulmonary pathology. In contrast, formalin-inactivated RSV potentiated RSV pulmonary histopathology, but there was a sparing of potentiation at high and low doses. Both the formalin-inactivated RSV and purified F and G preparations induced a high level of serum antibodies capable of binding to purified F and G glycoproteins but both sets of antibodies had significantly reduced neutralizing activity. These results are encouraging because they suggest that purified paramyxovirus glycoproteins might be used safely as a vaccine.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Role of early region 3 (E3) in pathogenesis of adenovirus disease.

The cotton rat Sigmodon hispidus has provided an animal model of adenovirus pneumonia that permits investigation of the viral gene products required to produce the disease and the molecular mechanisms effecting the damage. This study was carried out to test the hypothesis that early region 3 (E3) of the adenovirus genome plays a critical role in pathogenesis of the virus's disease process even though none of its gene products are essential for its replication. Mutants whose E3 region is largely deleted (i.e., H2dl801 and H5dl327) replicated like wild-type virus in the cotton rats' lungs, but the lymphocyte and macrophage/monocyte inflammatory response was markedly increased. Viruses containing mutations that ablated production of the 19-kDa glycoprotein had the same effect as H2dl801 and H5dl327. However, mutants with deletions in the other E3 open reading frames, some of which encode known proteins, did not differ from wild-type virus in their pathogenic properties. The 19-kDa glycoprotein markedly reduces expression of the class I major histocompatibility complex antigens on the surface of infected cells. A complete correlation was found between those mutants that had increased pathogenic effects and those that lost the ability to reduce transport of the class I major histocompatibility complex antigens to surface of infected cells (i.e., all mutants unable to express the 19-kDa glycoprotein). H5sub304, which has a deletion between 83.2 and 85.1 map units in the E3B region and expresses the 19-kDa glycoprotein, did not increase the extent of pneumonia but qualitatively changed the inflammatory response in that increased numbers of polymorphonuclear leukocytes accumulated, often in small foci.

Adenoviruses, Human↗
PubMed1989

Immunosuppression of the antibody response to respiratory syncytial virus (RSV) by pre-existing serum antibodies: partial prevention by topical infection of the respiratory tract with vaccinia virus-RSV recombinants.

Immunization strategies to prevent respiratory syncytial virus (RSV) disease will involve immunization of infants less than 2 months of age who possess maternally derived RSV antibodies. Vaccinia-RSV recombinant viruses are useful tools for defining parameters important in immunization against RSV and also are being considered as live virus vaccines for use in humans. Previous studies demonstrated that passively acquired RSV antibodies can suppress the immune response and the protective efficacy of vaccinia-RSV recombinants administered by the intradermal route. The present study demonstrates that the suppressive effects of passively acquired antibody on immunity induced by intradermally administered vaccinia-RSV recombinants in cotton rats can be partially overcome by administration of the recombinants by the intranasal route.

Animals↗
PubMed1989

Mice immunized with recombinant vaccinia virus expressing dengue 4 virus structural proteins with or without nonstructural protein NS1 are protected against fatal dengue virus encephalitis.

We have constructed vaccinia virus recombinants expressing dengue virus proteins from cloned DNA for use in experimental immunoprophylaxis. A recombinant virus containing a 4.0-kilobase DNA sequence that codes for three structural proteins, capsid (C), premembrane (pre-M), and envelope (E), and for nonstructural proteins NS1 and NS2a produced authentic pre-M, E, and NS1 in infected CV-1 cells. Mice immunized with this recombinant were protected against an intracerebral injection of 100 50% lethal doses of dengue 4 virus. A recombinant containing only genes C, pre-M, and E also induced solid resistance to challenge. Deletion of the putative C-terminal hydrophobic anchor of the E glycoprotein did not result in secretion of E from recombinant-virus-infected cells. Recombinants expressing only the E protein preceded by its own predicted N-terminal hydrophobic signal or by the signal of influenza A virus hemagglutinin or by the N-terminal 71 amino acids of the G glycoprotein of respiratory syncytial virus produced glycosylated E protein products of expected molecular sizes. These vaccinia virus recombinants also protected mice.

Animals↗
PubMed1988

Current approaches to the development of vaccines effective against parainfluenza and respiratory syncytial viruses.

Vaccines against parainfluenza (PIV) and respiratory syncytial viruses (RSV) that are currently being developed include both live and subunit vaccines. Candidate live PIV vaccines that have been found to be attenuated and efficacious in rodents or primate models are (1) cold-adapted, temperature-sensitive mutants of PIV-type 3 that have been serially passaged at low temperature (20 degrees C) in simian kidney tissue culture; (2) protease-activation mutants (PIV-1-Sendai), which have mutations that decrease the cleavability of their F glycoprotein by host cell protease; (3) an animal virus, bovine PIV-3 virus, which is antigenically related to the human PIV-3 virus, and (4) vaccinia recombinant viruses bearing RSV or PIV-3 glycoproteins. Subunit RSV and PIV-3 viruses are being produced and evaluated as immunogens. A major concern with these vaccines is the possibility of disease potentiation following virus infection as occurred previously with formalin-inactivated measles and RSV vaccines. Studies indicate that PIV-3 and RSV glycoprotein vaccines are immunogenic and efficacious in animals but insufficient data exist to estimate their capacity to potentiate disease. However, since a cotton rat model is available to detect potentiated disease resulting from infection of cotton rats previously immunized with formalin-inactivated RSV vaccine, it is now possible to systematically evaluate new vaccines in experimental animals for disease potentiation before studies are initiated in humans. It is likely within the next several years that one or more of these PIV or RSV vaccines will be tested in humans for safety and immunogenicity.

Animals↗
PubMed1988

Evaluation in non-human primates of the safety, immunogenicity and efficacy of recombinant vaccinia viruses expressing the F or G glycoprotein of respiratory syncytial virus.

It has been shown previously that immunization with recombinant vaccinia viruses expressing the F or G envelope glycoprotein of human respiratory syncytial virus (RSV) strain A2 induced a protective immune response in the lower respiratory tract of cotton rats against live RSV challenge. As a continuation of these studies, the safety, immunogenicity and efficacy of these recombinant vaccinia viruses was evaluated in non-human primates. Rhesus and patas monkeys were each inoculated intradermally at separate sites with the vaccinia-A2-F or vaccinia-A2-G recombinant or the parental vaccinia virus WR strain and the dermal lesion sizes were compared. Vaccinia-A2-F and vaccinia-A2-G recombinants produced lesions that were 5- to 15-fold smaller in area than vaccinia-WR. These studies indicated that insertion of either RSV gene into the thymidine kinase (TK) gene of vaccinia-WR significantly attenuated the virus for rhesus and patas monkeys. The immunogenicity of vaccinia-A2-F and vaccinia-A2-G was evaluated in squirrel, rhesus, African green, owl and patas monkeys. In four of the five species tested, the vaccinia-RSV recombinants stimulated levels of RSV serum-neutralizing antibodies considered to be protective for the lower respiratory tract of human infants and cotton rats. Interestingly, the level of RSV serum-neutralizing antibodies correlated with the size of the lesion. A boost in RSV serum-neutralizing antibody titres was not observed following a second inoculation. Owl monkeys inoculated with a single intradermal dose of vaccinia-A2-F and vaccinia-A2-G were completely resistant to infection of the lower respiratory tract with live RSV.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Live viral vaccines for respiratory and enteric tract diseases.

In its programme for accelerated development of vaccines for viral respiratory and enteric tract diseases the WHO has assigned a very high priority to respiratory syncytial virus (RSV), parainfluenza viruses and rotaviruses. There is also some interest in alternative approaches to immunization against influenza viruses because of the failure of inactivated vaccines to provide complete and reasonably durable immunity. Current attempts to develop satisfactorily attenuated viruses for use in prevention of disease caused by the above viral pathogens are described.

Gastrointestinal Diseases↗
PubMed1988

Site-directed ELISA identifies a highly antigenic region of the simian immunodeficiency virus transmembrane glycoprotein.

The transmembrane glycoprotein (gp32) of the simian immunodeficiency virus (SIV) contains a highly antigenic region that includes amino acid residues 606-628. A synthetic peptide representing this region was highly immunoreactive with sera from SIV-infected primates in a site-directed enzyme-linked immunosorbent assay (ELISA). This reactivity extended across four primate species from three genera and identified infection with at least two distinct isolates of SIV. This site-directed ELISA represents a simple, accessible method with broad specificity for screening large numbers of primates for antibodies against SIV.

Amino Acid Sequence↗
PubMed1988

Human-rhesus reassortant rotavirus vaccines: safety and immunogenicity in adults, infants, and children.

Human-rhesus reassortant rotavirus vaccines derived from rhesus rotavirus and human rotavirus serotypes 1, 2, or 4 were administered to adults, children, and infants after they had been given sodium bicarbonate buffer. Over 70% of infants and children developed antibody responses to or shed the candidate vaccine viruses. Individuals with prevaccination serum neutralization antibody titers less than or equal to 1:160 had significantly (P less than .0001) higher response rates than did individuals with prevaccination titers greater than or equal to 1:320. Similarly, shedding of vaccine viruses occurred significantly (P = .03) more often in children than adults and was inversely correlated with prevaccination antibody titers. No illnesses were observed in adults, and the vaccines were well tolerated in children. An increased rate of low-grade, transient fever (38.0-38.8 C) was noted only in children given serotype 2 vaccine, compared with controls (P = .006). No significant differences in the average number of unformed stools passed during the seven days after vaccination were noted in vaccinees versus controls. These vaccine strains are currently undergoing further evaluation in expanded clinical trials.

Adult↗