Baylor College of Medicine

Other

Vanderbilt University

William Marsh Rice University

Mary K Estes

David E Graham

Robert Legare Atmar

Xi Jiang

Sue Ellen Crawford

Khalil Ettayebi

Kosuke Murakami

David Y Graham

Judith M Ball

James E Crowe, Jr

Yongcheng Song

Noah F Shroyer

Robert Allen Britton

Timothy Gerald Palzkill

B Venkataram Prasad

Jennifer Dawn Rogers

Nadim Jose Ajami

Margaret E Conner

Peng Tian

Peng Tien

David Graham

Jennifer Auchtung

Tatiana Y Fofanova

Jane Grande-Allen

Joseph Petrosino

Reid Laurence Wilson

Christopher Stewart

An anaerobic chamber system to evaluate human enteric disease is described herein that can be used to test therapeutic components. In specific embodiments, the anaerobic chamber is used to determine the effect of one or more bacterial communities on ex vivo enteroid cultures. In one application, the anaerobic chamber system is used to determine the efficacy of therapeutic components in ameliorating human enteric disease using personalized medicine.

Engineering novel enteroid models for understanding human enteric disease

The present disclosure is directed to antibodies binding to and neutralizing norovirus and methods for use thereof. Thus, in accordance with the present disclosure, there is provided a method of detecting a norovirus infection in a subject comprising (a) contacting a sample from the subject with an antibody or antibody fragment having clone-paired heavy and light chain CDR sequences; and (b) detecting norovirus in the sample by binding of the antibody or antibody fragment to a norovirus antigen in the sample.

Human monoclonal antibodies that neutralize pandemic GII.4 noroviruses

Embodiments of the disclosure concern systems, methods, and/or compositions for cultivation of mammalian viruses, including at least human noroviruses and sapoviruses within the Caliciviridae family of viruses. The ex vivo culture systems include intestinal enteroids in combination with bile or a functionally active fraction or component thereof. In specific embodiments, the culture system is utilized to test inactivation compounds for therapeutic or environmental efficacy and to test contaminated comestibles and/or environmental entities for determination of the presence of infectious virus. Furthermore, antiviral compositions may be tested using systems of the disclosure, including drugs, small molecule inhibitors, and biologics such as neutralizing monoclonal antibodies.

Cultivation of human noroviruses

Embodiments of the disclosure concern systems, methods, and/or compositions for cultivation of mammalian viruses, including at least humanand sapoviruses within the Caliciviridae family of viruses. The ex vivo culture systems include intestinal enteroids in combination with bile or a functionally active fraction or component thereof. In specific embodiments, the culture system is utilized to test inactivation compounds for therapeutic or environmental efficacy and to test contaminated comestibles and/or environmental entities for determination of the presence of infectious virus. Furthermore, antiviral compositions may be tested using systems of the disclosure, including drugs, small molecule inhibitors, and biologics such as neutralizing monoclonal antibodies.

The present disclosure concerns inhibitors of Norovirus protease that are suitable for use against any genotype of Norovirus, including at least GII.4 Norovirus proteases. In particular embodiments, specific compositions are encompassed, including their use for prevention or treatment of Norovirus infection in an individual.

Structure of GII.4 norovirus protease&#x2014;design of broad-spectrum protease inhibitors

Embodiments of the disclosure concern systems, methods, and/or compositions for cultivation of mammalian viruses, including at least human noroviruses and sapoviruses within the Caliciviridae family of viruses. The ex vivo culture systems include intestinal enteroids in combination with bile or a functionally active fraction or component thereof. In specific embodiments, the culture system is utilized to test inactivation compounds for therapeutic or environmental efficacy and to test contaminated comestibles and/or environmental entities for determination of the presence of infectious virus. Furthermore, antiviral compositions may be tested using systems of the disclosure, including drugs, small molecule inhibitors, and biologies such as neutralizing monoclonal antibodies.

Embodiments of the disclosure include methods and/or compositions for the detection of viral infection, including at leastinfection. In particular embodiments, there are methods and/or compositions employing particular peptides and/or phage useful for detectingin a sample. The sample may be from an environment or from an individual. The individual may be a mammal, including a human, cow, horse, dog, cat, pig, and so forth. Certain exemplary peptides and phage that express the peptides are identified as useful for binding to. Such peptides and phage are provided to one or more samples in order to identify whether or notis present in the sample.

Identification and characterization of a peptide affinity reagent for the detection of noroviruses in clinical samples

Double-stranded cDNA was synthesized from nucleic acid extracted from Norwalk virus purified from stool specimens of volunteers. One clone was isolated from a cDNA library constructed in a pUC-13 vector after amplification of the cDNA. The specificity of this cDNA (pUCNV-953) was shown by hybridization assays. The cDNA reacted with post (but not pre-) infection stool samples from Norwalk volunteers and with highly purified Norwalk virus, but not with other common enteric viruses such as hepatitis A virus and rotavirus. Finally, the probe detected virus in the same fractions of CsCl gradients in which viral antigen was detected using a specific Norwalk virus radioimmunoassay, and particles were detected by immune electron microscopy. Single-stranded RNA probes derived from the DNA clone after subcloning into an in vitro transcription vector were also used to show that the Norwalk virus contains a ssRNA genome of about 8 kb in size. The original clone was also used to detect additional cDNAs which represent at least 7 kb of nucleic acid of the Norwalk genome. The availability of a Norwalk-specific cDNA and the first partial genome sequence information allow rapid cloning of the entire genome and of establishment of sensitive diagnostic assays. Such assays can be based on detection of Norwalk virus nucleic acid or Norwalk viral antigen using polyclonal or monoclonal antibodies to proteins expressed from the cDNA or to synthetic peptides made based on the knowledge of the genome sequence. Assays using proteins deduced from the Norwlk virus genome and produced in expression systmes can measure antibody responses. Vaccines made by recombinant DNA technology are now feasible.

Methods and reagents to detect and characterize norwalk and related viruses

The invention relates to an improved rotavirus vaccine for man and animals and methods of using them. The invention comprises a method of immunizing humans, particularly children, and animals against rotavirus infections by parenteral immunization. The immunization may be carried out in a series of injections using live or inactivated vaccines, alone or in combination with each other or in combination with a rotavirus subunit vaccine or oral vaccine.

Parenteral immunization against rotavirus

The rotavirus nonstructural glycoprotein, NSP4, performs multiple functions in the virus replication cycle, especially during viral morphogenesis. Specifically, NSP4 is an intracellular receptor that mediates the acquisition of a transient membrane envelope during the budding of newly formed subviral particles into the endoplasmic reticulum (ER). The present invention relates to NSP4 and methods of use. More particulary, it relates to the use of NSP4 and fragments thereof (NSP4 114-135, NSP4 120-147, NSP4 112-174, or NSP4 112-150) as a prevention and/or treatment of rotaviral disease.

Rotavirus enterotoxin NSP4 and methods of using same

Recombinant proteins and peptides of Norwalk virus are claimed. Also claimed are vaccines against Norwalk virus and methods of immunizing against Norwalk virus using recombinant Norwalk virus antigens and derivatives thereof. Also claimed are vaccines for non-Norwalk virus agents including a portion of the Norwalk virus capsid as a carrier.

Methods and reagents to detect and characterize Norwalk and related viruses

This invention relates to a method of potentiating an immune response by administering a viral enterotoxin or derivative as an adjuvant. More particularly it relates to administering a viral enterotoxin or derivative as an adjuvant and an antigen to a mucosal surface of a mammal.

Rotavirus enterotoxin adjuvant

Methods of immunization against rotavirus infection or rotavirus disease by administering to a subject a peptide NSP4 114-135, a peptide NSP4 120-147, or a toxoid thereof are disclosed.

Double-stranded cDNA was synthesized from nucleic acid extracted from Norwalk virus purified from stool specimens of volunteers. One clone was isolated from a cDNA library constructed in a pUC-13 vector after amplification of the cDNA. The specificity of this cDNA (pUCNV-953) was shown by hybridization assays. The cDNA reacted with post- (but not pre-) infection stool samples from Norwalk volunteers and with highly purified Norwalk virus, but not with other common enteric viruses such as hepatitis A virus and rotavirus. Finally, the probe detected virus in the same fractions of CsCl gradients in which viral antigen was detected using a specific Norwalk virus radioimmunoassay, and particles were detected by immune electron microscopy. Single-stranded RNA probes derived from the DNA clone after subcloning into an in vitro transcription vector were also used to show that the Norwalk virus contains a ssRNA genome of about 8 kb in size. The original clone was also used to detect additional cDNAs which represent at least 7 kb of nucleic acid of the Norwalk genome. The availability of a Norwalk-specific cDNA and the first partial genome sequence information allow rapid cloning of the entire genome and of establishment of sensitive diagnostic assays. Such assays can be based on detection of Norwalk virus nucleic acid or Norwalk viral antigen using polyclonal or monoclonal antibodies to proteins expressed from the cDNA or to synthetic peptides made based on the knowledge of the genome sequence. Vaccines made by recombinant DNA technology are now feasible.

Polyclonal and monoclonal antibodies to Norwalk virus and methods for

A method to express rotavirus genes in a baculovirus system. Different clones are used to express rotavirus genes for all of the viral proteins. These proteins are isolated in their native conformation. Some of these proteins show antigenic properties and are used to vaccinate human, agricultural animals and pet animals against diarrheal disease. The antigenic proteins are also used to detect the presence of the viral infectious agent either by themselves or in conjunction with antibodies produced against the antigenic proteins.

Growing community of inventors

Houston, TX, United States of America

Mary K Estes