Overview of Laboratory Overview of Laboratory of Bacterial of Bacterial

PolysaccharidesPolysaccharides

ByBy

Willie F. Vann, Chief LBPWillie F. Vann, Chief LBP

Description of the Laboratory of Bacterial Description of the Laboratory of Bacterial PolysaccharidesPolysaccharides

The Laboratory of Bacterial The Laboratory of Bacterial Polysaccharides investigates the Polysaccharides investigates the biochemistry, biology, chemistry, and biochemistry, biology, chemistry, and immunology of virulence factors of immunology of virulence factors of encapsulated bacteria. encapsulated bacteria.

These virulence factors include capsular These virulence factors include capsular polysaccharides, lipopolysaccharides, polysaccharides, lipopolysaccharides, and outer membrane proteins. and outer membrane proteins.

Description of the Laboratory of Bacterial Description of the Laboratory of Bacterial Polysaccharides (continued)Polysaccharides (continued)

The Laboratory of Bacterial The Laboratory of Bacterial Polysaccharides has review Polysaccharides has review responsibility for submissions related responsibility for submissions related to polysaccharide and polysaccharide to polysaccharide and polysaccharide conjugate vaccines in addition to conjugate vaccines in addition to non-capsular immunogens of non-capsular immunogens of encapsulated pathogens.encapsulated pathogens.

Chronology of Laboratory of Bacterial Chronology of Laboratory of Bacterial PolysaccharidesPolysaccharides

20022002– Last Site VisitLast Site Visit

20042004– CE Frasch steps down as Lab ChiefCE Frasch steps down as Lab Chief– MS Blake becomes Acting Lab ChiefMS Blake becomes Acting Lab Chief

20062006– WF Vann appointed Lab ChiefWF Vann appointed Lab Chief– Glycobiology Group of Lab of Bacterial Toxins Glycobiology Group of Lab of Bacterial Toxins

joined Lab of Bacterial Polysaccharidesjoined Lab of Bacterial Polysaccharides– NMR and Mass Spectrometry groups of Lab of NMR and Mass Spectrometry groups of Lab of

Biophysics joined Lab of Bacterial Biophysics joined Lab of Bacterial PolysaccharidesPolysaccharides

Current Organization of the Laboratory Current Organization of the Laboratory of Bacterial Polysaccharidesof Bacterial Polysaccharides

Labo rato ry o f Bacte r ialPo lysacch arid e s

W illie F. V ann , Ch ief

Glyco b io logyW illie F. V ann ,PI

Structu ral Bio log yDaron Freedb erg,

PI

An alyticalBio che mistry

ChaoMing Tsai, PI

Path og e ne sisMilan S. Blake, PI

Ce llu lar Immuno lo gyMu stafa Akkoyun lu ,

PI

Molecu larEpidemiology

Bioch emistry Con jugationChemistry

Vaccine Structu reJoh n Cip o llo , PI

Hug o Azurm e ndiPos tdoctoral Fe llow

Sco tt Norr isNM R S pe ctrom e trist

Structural Bio lo gyDaron Freedb erg, PI

F lora LichaaChe mist

The re sa W a ngBiologist

An alyticalBio che mistry

ChaoMing Tsai, PI

Su n ita Kansw alPos tdoctoral Fe llow

Nora Katse n e lsonBiologist

Ce llu lar Immun o logyM ustafa A k k oyunlu, PI

CURRENT RESEARCH STAFF

Glycobiology Group

Willie F. Vann, PI

Conjugate ChemistryRobert Lee, PhD, SS

Amy KuoBritany Bowen

BiochemistryEkaterina Andreishcheva

Seshu GudlavalettiJustine VionnetDwight Peterson

Molecular Epidemiology

Margaret Bash, MD, MOIvano Felippis

Craig Hammack

Vaccine StructureJohn Cipollo, PI(April, 2007)

Mass Spectrometrist(new recruit)

Postdoctoral Fellow(new recruit)

Areas of ResearchAreas of Research

Structure and conformation of capsular Structure and conformation of capsular polysaccharidespolysaccharides

Biosynthesis of capsular polysaccharidesBiosynthesis of capsular polysaccharides Role of non-capsular antigens in Role of non-capsular antigens in

protectionprotection Interaction of the capsular Interaction of the capsular

polysaccharides with the immune systempolysaccharides with the immune system Development of methodology for Development of methodology for

analysis of conjugate vaccinesanalysis of conjugate vaccines

Relevance of Research Program to Relevance of Research Program to CBER MissionCBER Mission

The Laboratory of Bacterial The Laboratory of Bacterial Polysaccharides has regulatory Polysaccharides has regulatory responsibility for vaccines against responsibility for vaccines against encapsulated bacteria and products encapsulated bacteria and products containing bacterial polysaccharides containing bacterial polysaccharides

The overall goal of the research program The overall goal of the research program of the LBP is to understand the virulence of the LBP is to understand the virulence factors that are components of vaccines factors that are components of vaccines against bacterial pathogens. against bacterial pathogens.

Relevance of Research Program to Relevance of Research Program to CBER Mission (continued)CBER Mission (continued)

The research program of the Laboratory of The research program of the Laboratory of Bacterial Polysaccharides is directed Bacterial Polysaccharides is directed toward understanding the physical, toward understanding the physical, chemical, and immunological properties of chemical, and immunological properties of bacterial polysaccharides, and bacterial polysaccharides, and polysaccharide conjugate vaccines polysaccharide conjugate vaccines

The knowledge and expertise gained in The knowledge and expertise gained in this research endeavor provide a scientific this research endeavor provide a scientific basis for our decisions related to the basis for our decisions related to the review of manufacturing, purity, potency, review of manufacturing, purity, potency, and safety of carbohydrate containing and safety of carbohydrate containing vaccines. vaccines.

Some Significant AchievementsSome Significant Achievements Development of Efficient Method for Development of Efficient Method for

Meningococcal Group A Conjugate Meningococcal Group A Conjugate Vaccine SynthesisVaccine Synthesis– Vaccine in MVP/WHO Sponsored Phase II Vaccine in MVP/WHO Sponsored Phase II

Clinical TrialClinical Trial

Meningococcal Group A Conjugate Vaccine ProjectMeningococcal Group A Conjugate Vaccine Projectmanaged by CE Frasch - CBER & M. LaForce – MVP managed by CE Frasch - CBER & M. LaForce – MVP

SerologyCDC

Margaret Bash

Conjugate TechnologyRobert Lee/DBPAP

Manufacturing Serum Institute of India

FundingPATH /MVP

Men A VaccineProject

Daron Freedberg and Scott Norris contributed to analysis during development

Gates Foundation

Some Significant AchievementsSome Significant AchievementsAnalytical Biochemistry – ChaoMing Tsai, PIAnalytical Biochemistry – ChaoMing Tsai, PI

Developed HPAEC method for quantitation Developed HPAEC method for quantitation of phosphate and acetylation in of phosphate and acetylation in polysaccharide vaccinespolysaccharide vaccines

Characterized the Characterized the lgtlgtH gene of H gene of NeisseriaNeisseria LOS gene clusterLOS gene cluster

Demonstrated that the LOS of commensal Demonstrated that the LOS of commensal N. polysacchareaN. polysaccharea is similar to LOS of is similar to LOS of meningococcal pathogenmeningococcal pathogen

Some Significant AchievementsSome Significant AchievementsMolecular Epidemiology – Margaret BashMolecular Epidemiology – Margaret Bash

Developed and applied molecular Developed and applied molecular methods to study PorB diversitymethods to study PorB diversity

Horizontal genetic exchange (mosaicism) Horizontal genetic exchange (mosaicism) predominatespredominates

Persistence of PorB variable region sequence Persistence of PorB variable region sequence types indicates diversification is constrainedtypes indicates diversification is constrained

Identified survival advantages associated Identified survival advantages associated with PorB typeswith PorB types

Relevant to development and evaluation of Relevant to development and evaluation of broadly protective OMP vaccines broadly protective OMP vaccines

Some Significant AchievementsSome Significant AchievementsCellular Immunology-Mustafa Akkoyunlu, PICellular Immunology-Mustafa Akkoyunlu, PI

Interactions of bacterial capsular polysaccharides with innate immune system.

– Neisseria meningitidis type C polysaccharide binding to CD14 and LBP inhibits meningococcal LPS mediated cell activation

Modulation of BAFF/APRIL system molecules by microbial products.

– Decreased expression of TACI on newborn mouse B cells may to be responsible for the impaired response of newborns to polysaccharide antigens.

– Toll-like receptor agonists, CpG DNA and LPS, strongly upregulate TACI expression on B cells.

Some Significant AchievementsSome Significant Achievements Structural Biology-Daron Freedberg, PIStructural Biology-Daron Freedberg, PI

1. 1. On cell NMR: On cell NMR: In-vivoIn-vivo antigen characterization by NMR antigen characterization by NMRMening B PSMening B PS structure structure on cellson cells = = Mening B PSMening B PS structure in vaccine structure in vaccine

2. Carbohydrate 3D Structure: Sucrose

m=monomer

p=polymer

HOO

H

H

HO

H

OHHH

OH

O

H

HO

H

H

OH

O

H

H

HO

OH

HOO

H

H

HO

H

OHHH

OH

O

HHO

H

H

OHO

H

H

OH

HO

HOO

H

H

HO

H

OHHH

OH

O

H

HO

H H

OH

O

H

HHO

HO

Licensed Product ResponsibilityLicensed Product ResponsibilityVaccine ClassVaccine Class ProductProduct ManufacturerManufacturer

PolysaccharidePolysaccharide Pneumococcal 23-valentPneumococcal 23-valent WyethWyeth

MerckMerck

Meningococcal tetravalentMeningococcal tetravalent Sanofi PasteurSanofi Pasteur

Typhoid ViTyphoid Vi Sanofi PasteurSanofi Pasteur

ConjugateConjugate Haemophilus influenzaeHaemophilus influenzae type b type b WyethWyeth

Sanofi PasteurSanofi Pasteur

Merck (2)Merck (2)

Pneumococcal heptavalentPneumococcal heptavalent WyethWyeth

Meningococcal tetravalentMeningococcal tetravalent Sanofi PasteurSanofi Pasteur

Regulatory ResponsibilitiesRegulatory Responsibilities

Lot ReleaseC-M Tsai

(Formerly CJ Lee)

Theresa Wang Performs Lot Release Tests on Fraction of Submitted Samples

Approximate 400 Samples and Protocols Submitted

Per Year

Regulatory AccomplishmentsRegulatory Accomplishments

Licensing Tetravalent Meningococcal Licensing Tetravalent Meningococcal Conjugate Diphtheria Toxoid Vaccine Conjugate Diphtheria Toxoid Vaccine – against groups A,C, Y, and W-135– against groups A,C, Y, and W-135– Reviewers included Frasch, Tsai, Lee, Reviewers included Frasch, Tsai, Lee,

Bash, Lynn, BlakeBash, Lynn, Blake Significant Changes in Analytical Significant Changes in Analytical

Methodology for Lot ReleaseMethodology for Lot Release– Reviewers included Tsai, FreedbergReviewers included Tsai, Freedberg

Other Regulatory AccomplishmentsOther Regulatory Accomplishments

IND Supplement Reviewed – 350IND Supplement Reviewed – 350 BLA & BLA Supplement Review -85BLA & BLA Supplement Review -85 Participated in International and Participated in International and

CBER Policy Working GroupsCBER Policy Working Groups Distributed Reference Material for Distributed Reference Material for

Haemophilus and Pneumococcal Haemophilus and Pneumococcal Antibody AssaysAntibody Assays