ルナパスセミナー（ウイルス）
日時；2024 年 10 月 25 日(金)会場：浜松コングレスセンター4 階 41 会議室
タイムテーブル：（第 1 部）ワクチン・ウイルスベクターの最新評価方法
座長 小出 英孝 Southern Research
13:00 生ワクチン開発の神経毒力試験の実際
講師 Dr. Joseph Randall, Southern Research
13:40 ウイルスベクター治療の可能性探求
講師 Dr. Jennifer Pickens, Southern Research
14:20 最新の抗ウイルス/ワクチン評価
Human 3D culture assay
In vivo 試験のアップデート
講師 小出 英孝, Southern Research（第 2 部）進化するウイルス安全性評価
座長 安井 文彦 東京都医学総合研究所
14:40
15:10

15:30

15:50

ICH-Q5A(R2): A new era for biosafety testing
講師
DI. Jürgen Ebner, BD Manager, ViruSure
NGS: Implementation as an alternative to traditionaladventitious agent testing
講師
Dr. Andy Bailey, CEO, ViruSure
Rapid and Precise: GMP characterisation of viral vectors
using Electron Microscopy
講師
Dr. Ashley Stephen Layland, Project Director,
neotem Bioanalytics-IIT GmbH
Digital PCR testing and biosafety testing
講師
Dr. Andy Bailey, CEO, ViruSure要旨：
第 1 部）ワクチン・ウイルスベクターの最新評価方法
13:00 生ワクチン開発の神経毒力試験の実際
Neurovirulence Testing of Live Viral Vaccines
講師 Dr. Joseph Randall, Southern Research
Abstract:
Southern Research has been at the forefront of testing the safety and efficacy of novel vaccines for overtwo decades. SR has conducted many biodistribution, immunogenicity, and GLP safety studies for
SARS-CoV-2, yellow fever, influenza and many other vaccines in support of INDs and BLAs. The
development and safety of vaccines for infectious disease has received a lot of public attention in thepast few years due to the SARS-Cov2 pandemic. We have a team of experienced virologists andtoxicologists to evaluate the safety and efficacy of vaccines. We continue to invest in key capabilities toevaluate vaccine safety, including the use of rodent and an alternative to NHPs. Confirmation of
attenuation of pathogenic traits is necessary for the safe use of potentially neurotrophic vaccines.
The monkey neurovirulence test (MNVT) is the gold standard for neurovirulence safety assessment oforal polio vaccines and has been used for 75 years for lot specific release testing. The MVNT bioassay isalso required for testing new vaccine seed lots for live attenuated neurotrophic viruses when there is achange to manufacturing. The dosing procedure involves direct injection of a live virus into the central
nervous system and to evaluate neurobehavioral effects and histopathology changes in the brain.
We have validated the MVNT neurovirulence model and have demonstrated the safety of live
attenuated viral vaccines for yellow fever and dengue fever. In one such study, we evaluated theneurovirulence of master and working seed lots of 17D viruses for yellow fever in comparison to
Stamaril™ and the WHO 17D 168-73 reference vaccine (Tyagi et al, 2023). Endpoints forneurovirulence included changes in clinical scores and microscopic evaluation of the brain and spinal
cord. Viral replication in blood was detectable on days 2 and 4, but not on day 6. The yellow fever
master and working seed viruses and final vaccine were non-neurotrophic and comparable to the live
attenuated WHO 17D 168-73 vaccine.
No single species or test can predict the neurovirulence of all live viral vaccines. SR supports therefinement of animal use and to find alternatives to the MVNT. We have conducted a pilot
neurovirulence study in 4-day old suckling CD-1 mice and demonstrated neurovirulence 7-8 days after
intracranial administration of WHO 17D 168-73. The higher sensitivity of neonatal mice was
expected based on a report in the literature (Fulton and Bailey, 2021). There was 100% neurovirulencein suckling mice at a dose of 50 plaque forming units. We recommend including positive controls inrodent neurovirulence studies to demonstrate neurotrophic differences between wild type or approvedvaccines and novel live attenuated viral strains.
Rodent models of neurovirulence testing are gaining acceptance as an alternative to the MNVT assay.
For example, a transgenic knock-in mouse model of neurovirulence that expresses human polio
receptors was validated in conjunction with the World Health Organization (Dragunsky et al, 2003). In
addition, a neonatal rat model of mumps vaccine neurovirulence has been validated (Rubin et al, 2000).
The European Medicines Agency accepts neurovirulence data from mouse influenza models in lieu ofthe MVNT. Please contact the Departments of Toxicology and Infectious Disease for more information
on non-clinical vaccine safety testing.

https://southernresearch.org/services/toxicology/
ANIMAL MODELSоGolden Syrian Hamstersо

Rodents and Ferrets

о

New Zealand White Rabbits

о

Non-Human Primates

IN VIVO SERVICESо

Biodistribution and Immunogenicity

оGeneral Toxicology After Repeated Dosingо

Neurovirulence Testing in Mice or NHPs

о

Evaluation of Effects on Embryofetal
Development

Stereotaxic injections into brain for neurovirulence and oncolytic viruses

Bilateral Stereotaxic Injections into the Thalamus

13:40 ウイルスベクター治療の可能性探求
Exploring the Therapeutic Potential of Viral Vectors
講師 Dr. Jennifer Pickens, Southern Research
Abstract:
There has been a surge in vaccine development research in the wake of recent health threats that include
the COVID-19 pandemic, the 2022 Monkey Pox outbreak, and H5N1 highly pathogenic influenza
zoonotic transmission this year (2024). The research is aimed at identifying vaccine candidates thatexhibit optimal safety profiles across a wide age range; elicit robust and durable cellular and humoral
immunity; and are easy, low cost, and scalable to manufacture/distribute. Vaccines come in manyvarieties from killed, live-attenuated, mRNA, DNA, and viral vectored to name a few. The primary
vaccinology approach centers around creating a vaccine that immunologically protects against disease,
reduces transmission, and establishes herd immunity. First genetically engineered in 1972, viral vectors
remain a promising vaccine platform because they are capable of heterologous expression of antigens
without replication, thus inducing a robust and durable humoral and cellular immune response. Various
viruses have been used as viral vectors, including but not limited to influenza, lentivirus, retrovirus,
measles virus, vesicular stomatitis virus (VSV), and adenovirus. Southern Research (SR) has been
instrumental as a preclinical CRO, partnering with government and commercial clients to assess thetherapeutic potential of their viral vectored vaccines in small and large animal models. In this seminar,
SR will review the preclinical approach to better understand the therapeutic advantage of viral vectoredvaccines and review current case studies and data to support advancing these from concept to the clinic.

14:20 最新の抗ウイルス/ワクチン評価
Human 3D culture assay
In vivo 試験のアップデート
講師 小出英孝, Southern Research
Abstract:
SR, chartered in 1941, is one of the oldest independent, not-for-profit research institutes in the UnitedStates. For more than 80 years, SR’s scientists have addressed many of the most urgent and challenging
technological problems of our age. By responding to the needs of private industry and addressing global
issues in health, national defense, and the environment, SR has contributed greatly to providing technicalsolutions in a broad range of areas.
To support its mission of providing sponsors and clients with the highest quality research, SR has
disinvested from its Engineering and Energy & Environment divisions, as well as its Fredrick, Maryland,location, raising more than $150 million to invest in the construction of a 70,000-square-foot flagshipbuilding and the renovation of 40,000 square feet of the organization’s existing facility, including new
biosafety level 3 (BSL-3) space. SR focuses mainly on life science research through drug discovery anddrug development services.
SR operates in full compliance with federal Good Laboratory Practice (GLP) and animal welfare
regulations, and its animal facilities are accredited by the Association for Assessment and Accreditationof Laboratory Animal Care International (AAALAC).
SR has supported US government Operation Warp Speed program under Biomedical Advanced Researchand Development Authority (BARDA) funding, entitled “Development of Animal Models for Evaluationof Medical Countermeasures (MCMs) for SARS COV-2” and supported the federal countermeasuresacceleration effort. SR has conducted more than $30 million in coronavirus research over the past three
years, including support of Phase 1-3 COVID-19 vaccine clinical trials, as well as conducting thousandsof HTS screening and refining the underlying chemistry of critical treatments.
Here, we present the update on our latest infectious disease services including Omicron XBB.1.5, Highly
Pathogenic Avian Influenza (HPAI) and human metapneumovirus (HPMV) models.

第 2 部）進化するウイルス安全性評価
14:40 ICH-Q5A(R2): A new era for biosafety testing
講師 DI. Jürgen Ebner, BD Manager, ViruSure
Abstract:
The revised ICH Q5A(R2) guideline, a cornerstone for virus safety evaluation of biotechnology products,
signals a transformative phase in biosafety testing. This presentation will explore the key updates,
including the incorporation of new products, new technologies, innovative approaches in biosafety
testing for cell line characterization and bulk harvest testing, and advancements in virus clearance
studies.

15:10 NGS: Implementation as an alternative to traditional adventitious
agent testing
講師 Dr. Andy Bailey, CEO, ViruSure
Abstract:
Revision 2 of the ICH Q5A guideline on Viral Safety Evaluation of Biotechnology Products Derived from
Cell Lines of Human or Animal Origin discusses in depth the expectations for meeting virus safety
requirements in the modern era. This presentation will explore some of the practical implications andstrategies that can be applied when endeavouring to meet these virus safety requirements. Using casestudies, it will explore various aspects, including how to best control risk in raw materials and biologicalmaterials as well as introduce how next generation sequencing (NGS) can be effectively integrated into
these control strategies.

15:30 Rapid and Precise: GMP characterisation of viral vectors using
Electron Microscopy
講師 Dr. Ashley Stephen Layland, Project Director, neotem
Bioanalytics-IIT GmbH
Abstract:
The advent of transmission electron microscopy (TEM) marked a pivotal moment in the history of
microbiology, as it became the first technique capable of visualising viruses in their natural state. Thisgroundbreaking discovery has had a profound impact on our understanding of these enigmaticparticles. In the present era, TEM continues to be a cornerstone of the biopharmaceutical industry,
playing a pivotal role in ensuring the safety of biologics.
This presentation will highlight the critical role of TEM in GMP-quality assessment, where its
unparalleled resolution allows for a rapid and detailed investigation of vaccine products and viral
vectors, including Adeno-Associated Viruses (AAV) ensuring product safety and efficacy. By facilitating
the ultrastructural investigation of viral vectors and enabling the determination of key parameters suchas particle integrity, morphology, distribution and filling grade (full/empty ratio), TEM continues to playa crucial role in the comprehensive characterisation and quality assurance of viral vectors.

15:50 Digital PCR testing and biosafety testing
講師 Dr. Andy Bailey, CEO, ViruSure
Abstract:
Digital PCR (dPCR) is an advanced molecular biology technique used for the precise and absolute
quantification of nucleic acids (DNA or RNA) by end-point analysis and Poisson statistics. Unlike qPCR
assays, which provide relative quantification in relation to a standard curve, dPCR divides the sample
into thousands of individual partitions. Each partition is analyzed for presence (positive) or absence(negative) of a fluorescent signal, allowing for the detection and counting of single DNA or RNA
molecules. With its high sensitivity and precision, coupled with reduced susceptibility to inhibitors, it canbe an essential tool in biosafety testing: viral load quantification, residual host cell DNA quantification,
copy number variation, genetic stability and titre determination in cell and gene therapy products. Digital
PCR provides precise and absolute quantification of nucleic acids. Therefore, its usage in biosafety
testing ensures consistently higher qualities in final products and this presentation will highlight how andwhere this testing can used effectively to support product safety.

演者紹介：
Joseph Chase Randall, PhD.
Southern Research
Joseph has more than 25 years of drug development experience inthe design, conduct and reporting of regulated toxicology studies insupport of IND and NDA applications for pharmaceutical andbiotechnology companies. Areas of therapeutic expertise include
the safety assessment of antivirals, vaccines, and drugs to treat
cancer and neurodegenerative diseases. He has developed liposomal
nanoparticle formulations and vascular targeting agents inoncology. His current focus is on central nervous safety of live viral
vaccines and oncolytic viruses and the reproductive safety testing ofnovel vaccines and genetic therapies for rare diseases.

Jennifer Pickens, PhD
Director of Infectious Disease Research, Southern Research
Dr. Pickens has over 15 years scientific leadership experience, where
she is an infectious disease and preclinical drug development subject
matter expert (SME) with a focus on the advancement of cuttingedge

andfirst-in-classvaccineandtherapeutic

medicalcountermeasures in the nonclinical and clinical setting. Before
joining Southern Research in 2019 and again in 2022, she earned
her Ph.D. in Infectious Diseases from the College of Veterinary
Medicine at the University of Georgia in 2011 under the mentorshipof S. Mark Tompkins and later completed postdoctoral fellowships(2012-2017) within the laboratories of James E. Crowe, Jr at the
Vanderbilt Vaccine Center and Ralph A. Tripp at the University
Georgia. Her career has focused on managing preclinical research
programs aimed at identifying and assessing the efficacy of novel
antiviral therapeutics against potentially lethal respiratory viruses(i.e.

respiratorysyncytial

virus(RSV),

metapneumoviruses(hMPV), SARS-CoV-2 and influenza). During the COVID-19
pandemic, her work focused predominately on developing thenonclinical commercial and government A/BSL-3 SAR-CoV-2
program at Southern Research before joining the Biomedical
Advanced Research & Development Authority (BARDA) in 2021 asa nonclinical and infectious disease SME/Biologist. She returned toSouthern Research in 2022 and is the current Director of Infectious
Disease Research. She works alongside commercial and government
clients providing scientific and operational guidance through thelifecycle of their pre-IND pipelines.

Fusataka Koide
Senior Director, Global Client Engagement, Southern Research
Fusataka Koide, M.S., is the Director of Global Client Engagementat SR and managing the groups of Proposal writes, Business
Development and Marketing team. His responsibilities include
tracking and oversight of proposal activity and progress, market
assessment,

development

of

the

strategic

businessplan,

incorporating marketing and sales strategy for each capability area,and building robust processes that allow the division to meet its
financial goals.
Mr. Koide also serving as the vaccine and animal model subject
matter expert for SR.

Mr. Koide has over 20 years of experiencein vaccine animal model development including 14 years of direct
experience with nonhuman primate (NHP) infection andvaccine/antiviral testing. His experience in NHPs and small animalmodels for evaluation of vaccine covers a wide range of infectious
diseases including SARS-CoV-2, Monkeypox (MPXV), Rabbitpox(RPXV), HIV/SIV, Chikungunya, MRSA, plague, HPAI, Dengue,
Zika, and Yellow Fever. Specific examples of the vaccine studies
include efficacy evaluation of live attenuated tetravalent dengue
vaccine in Cynomolgus and Rhesus Macaques (GLP and nonGLP), Dengue vaccine neurovirulence and vaccine viscerotropism
studies in Cynomolgus Macaques (WHO TRS protocols) andefficacy

evaluationof protein-based

Smallpox

Vaccinein

Cynomolgus monkeys challenged with Monkeypox Virus (FDA
Animal Rule). His work has included analysis of humoral and cellmediated immune responses in vaccinated and/or infected animals
with an emphasis on attempting to identify correlates of immune
protection against disease.

Many of these efforts have been

supported by the NIH’s NIAID/DMID. Mr. Koide was a PI forsupporting NIAID COVID-19 Task Order B08 (Contract No.
HHSN272201000022I) entitled “Development and Use of a NonHuman Primate Model of SARS-CoV-2 Infection”.

He has also

served as PI for another COVID-19 Task Order for BARDA BIOAMT

1021(Contract

No. HHSO100201700018I)

entitled“Development of Animal Models for Evaluation of Medical
Countermeasures for SARS-CoV-2”.
As the leader of Southern Research’s vaccine development
program, Mr. Koide provides support to many clients in the formof preclinical efficacy testing and reports that support IND
applications submitted to the U.S. FDA.

Jürgen Ebner
BD Manager, ViruSure
Jürgen Ebner studied food- and biotechnology at the University of
Natural Resources and Life Sciences. He joined ViruSure in 2009and gained a lot of experience in biosafety testing of
biopharmaceuticals. His area of expertise comprises the designand execution of Viral- and Prion Clearance Studies.

Andy Bailey, PhD
CEO, ViruSure
Originally a chemist/biochemist, Andy later specialised in Virology
serving for 9 years at the MRC Virology Unit in Glasgow, Scotland,
studying Herpesviruses and Adenoviruses. In 1995, he moved to theindustry sector, initially as Director of Virus Validation services
with Q-One Biotech Ltd, and later at the Pathogen Safety group of
Baxter Healthcare in Vienna, Austria.
Over the last 25 years Andy has been actively involved in the virus
and prion safety field, presenting at numerous regulatory agencies
and conferences either in support of product registration or as aninvited speaker at expert workshops and conferences. This has
included presentations at the UK MHRA, German PEI, French
AFFSAPS, US FDA, EMEA and PMDA (Japan) supporting
regulatory submissions for various products. He has extensive
experience in regulatory affairs and virus safety issues. He currently
serves as an external expert for the EU SCENIHR committee on
emerging human health risks, and continues to be a frequent
presenter at international biopharmaceutical conferences.
In 2005 he founded ViruSure with the goal of providing a highquality testing service to the biopharmaceutical industry, aphilosophy which remains a key focal point of the company.

Ashley Stephen Layland, PhD
Project Director, neotem Bioanalytics-IIT GmbH
Graduating as a research assistant in 2011, Ashley Stephen Layland
began his career in cell culture technology at Xell AG (which is now
part of Sartorius) where he acquired eight years of experience in
upstream processing and cultivation. From 2012, Ashley thengainedfurther

in-depthexperienceinqualityassurance

management, virology and electron microscopy from Charles River
Laboratories and the Centre for Ultrastructural Diagnostics locatedat the University of Bielefeld. Fast forward to 2022, Ashley assumed
the position of project director at neotem Bioanalytics and now
leads a GMP-certified company, applying electron microscopy forthe viral safety assessment of biologics and the characterisation ofvirus products.