Cold adapted variants of influenza A

The genetic and biological properties of 13 recombinant influenza A clones derived at 25°C from the A/AA/6/60-cold variant (by crosses with 4 different wild type strains) were compared with a set of 5-FU induced ts -mutants. The 5-FU mutants had previously been placed into 7 complementation-recombination groups; the A/AA/6/60-cold parent (PI-7) and the 12 cold recombinant clones which were ts were shown to share a lesion with only one of these groups. The parental strain and 5 recombinant clones were evaluated for replication in the lungs and nasal turbinates of hamsters. Each virus appeared to be attenuated; genetic stability correlated with the level of viral replication in the hamster lung, i.e., viruses which grew best showed a tendency to revert to the ts + phenotype. Characterization of the ts + revertants for the presence of the cold adaptation property revealed that these viruses exhibited a spectrum of cold adaptation properties. Two viruses, PI-7 (the parental cold variant) and the CR6 recombinant (A/Queensland/6/72) did not revert in either the lungs or nasal turbinates of hamsters.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41685/1/705_2005_Article_BF01319909.pd

The cell receptor level is reduced during persistent infection with influenza C virus

 Persistent influenza C virus infection of MDCK cells perpetuates the viral genome in a cell-associated form. Typically, virus production remains at a low level over extended periods, in the absence of lytic effects of replication. In this study, we demonstrate that persistently infected cells are very restricted in permissiveness for superinfection. By reconstitution experiments, using bovine brain gangliosides as artificial receptors, the degree of super-infection was markedly increased. Analysis of cellular receptor expression revealed reduced concentrations of sialoglycoproteins in general and a limited presentation of the major receptor gp40. Cocultures of persistently infected and uninfected cells (the latter carrying normal receptor levels) initiated a transient rise in virus titers. This kind of induction of virus synthesis appeared to be mainly receptor-linked, since a receptor-deprived subline, MDCK II, did not give rise to a similar effect. Susceptibility of MDCK II cocultures could be partly restored by ganglioside treatment. In accordance to related virus systems, these findings on influenza C virus suggest a role of cell receptor concentrations in the regulation of long-term persistence.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/42459/1/705-142-6-1155_71421155.pd

Biological characteristics of a cold-adapted influenza A virus mutation residing on a polymerase gene

The biological function of a cold-adapted (ca) mutation residing on the PB2 gene of an influenza A/Ann Arbor/6/60 (A/AA/6/60) ca variant virus in the viral replication cycle at 25° C was studied. The viral polypeptide synthesis of A/AA/6/60 ca variant at 25° C was evident approximately 6 hours earlier than the wild type (wt) virus and yielded twice as many products. The quantitative analysis of viral complementary RNA (cRNA), synthesized in the presence of cycloheximide, revealed that A/AA/6/60 ca variant and a single gene reassortant that contains only the PB2 gene of the ca variant with remaining genes of the wt virus produced equal amount of cRNA at 25° and 33° C, which was an amount approximately four fold greater than the wt virus' cRNA synthesized at 25° C. These results strongly suggest that the ca mutation residing on the PB2 gene of A/AA/6/60 ca variant affects the messenger RNA synthesis at 25° C in the primary transcription.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41692/1/705_2005_Article_BF01310893.pd

Detection of rubella antibodies in human serum by the indirect fluorescent antibody technique

Antibodies for rubella virus were detected in human serum and titrated by the indirect method of immunofluorescence using a chronically infected, continuous line of monkey kidney cells as antigen. Positive reactions were obtained with convalescent or post-exposure specimens while acute sera and those from unexposed individuals were negative.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41678/1/705_2005_Article_BF01253854.pd

Characterization of growth of rubella virus in LLC-MK2 cells

1. A characteristic CPE of rubella virus in LLC-MK 2 cells is described and its usefulness and limitations in experimental work are discussed.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41677/1/705_2005_Article_BF01253851.pd

Characterization of a nonvirulent variant of lymphocytic choriomeningitis virus

A cold-adapted, nonvirulent variant of the Armstrong strain of lymphocytic choriomeningitis virus was isolated from infected L929 cells maintained at 25° C. This variant, designated P17, was capable of replicating in the central nervous system of mice without causing disease and conferring immunity to back challenge with the parental strain.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41689/1/705_2005_Article_BF01320786.pd

Isolation and antiviral activity of the gymnemic acids

Aus den Blättern von Gymnema sylvestre wurden 4 Gymneasäuren (A, B, C und D) isoliert. Die Antivirusaktivität der Säuren A und B wurde geprüft.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/42450/1/18_2005_Article_BF02152834.pd

A Pandemic Influenza H1N1 Live Vaccine Based on Modified Vaccinia Ankara Is Highly Immunogenic and Protects Mice in Active and Passive Immunizations

The development of novel influenza vaccines inducing a broad immune response is an important objective. The aim of this study was to evaluate live vaccines which induce both strong humoral and cell-mediated immune responses against the novel human pandemic H1N1 influenza virus, and to show protection in a lethal animal challenge model.For this purpose, the hemagglutinin (HA) and neuraminidase (NA) genes of the influenza A/California/07/2009 (H1N1) strain (CA/07) were inserted into the replication-deficient modified vaccinia Ankara (MVA) virus - a safe poxviral live vector – resulting in MVA-H1-Ca and MVA-N1-Ca vectors. These live vaccines, together with an inactivated whole virus vaccine, were assessed in a lung infection model using immune competent Balb/c mice, and in a lethal challenge model using severe combined immunodeficient (SCID) mice after passive serum transfer from immunized mice. Balb/c mice vaccinated with the MVA-H1-Ca virus or the inactivated vaccine were fully protected from lung infection after challenge with the influenza H1N1 wild-type strain, while the neuraminidase virus MVA-N1-Ca induced only partial protection. The live vaccines were already protective after a single dose and induced substantial amounts of neutralizing antibodies and of interferon-γ-secreting (IFN-γ) CD4- and CD8 T-cells in lungs and spleens. In the lungs, a rapid increase of HA-specific CD4- and CD8 T cells was observed in vaccinated mice shortly after challenge with influenza swine flu virus, which probably contributes to the strong inhibition of pulmonary viral replication observed. In addition, passive transfer of antisera raised in MVA-H1-Ca vaccinated immune-competent mice protected SCID mice from lethal challenge with the CA/07 wild-type virus.The non-replicating MVA-based H1N1 live vaccines induce a broad protective immune response and are promising vaccine candidates for pandemic influenza