Site Visit Presentation February 25, 2005

1
Site Visit Presentation February 25, 2005

• Basil Golding M.D.
• PI - Immunology Section
• Lab. of Plasma Derivatives, DH
• OBRR/CBER

2
Projects

• Toll-like receptors
• Avoiding undesirable immune responses to
  replacement factors, e.g. Factor VIII
• Polyclonal antibodies against anthrax
• HIV-Brucella abortus - immunotherapy

3
Immune Globulin Intravenous(LPD)

• Public Health Impact
• Product Contamination
• Infusion of IGIV causes adverse events 25
• Sterile filtration of final product does not
  remove microbial components, e.g. LPS, DNA.
• Microbial contaminants ? proinflammatory
  cytokines ? side effects

4
Immune Globulin Intravenous Product Contamination

• Regulatory and Scientific Challenges
• Need for rapid high throughput assays to detect
  microbial components
• Adapt cell lines expressing Toll-like receptors
  as a detector system

5
Toll-Like Receptors (TLRs) and Their Ligands
Pathogen Recognition
NF-??B-luciferase
NF?-B
Nucleus
Nucleus
Takeda and Akira Inf. Imm. 2005
6
HEK-293 Cells Transfected with TLRs and a
Reporter Gene Respond to Microbial Ligands
Huang et al, J. Immunol. 171(3)1441-6. 2003
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IGIV Outcomes

• Cell lines expressing multiple TLRs and a
  reporter gene can detect microbial components.
• These cell lines will be used to develop a rapid
  high throughput system for testing IGIVs and
  other CBER products for microbial contaminants.

8
Toxins of B. anthracis
And Or
Lethal Factor
Edema Factor
Protective Antigen (PA)
Protective Antigen (PA) heptamers
Cell lysis Cytokines
LT
Edema
ET
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Collaboration with industry-Hematech

• Cooperative Research And Development Agreement
  (CRADA)
• Generation and characterization of bovine and
  human anti-anthrax antibodies in cows
• Novel transchromsomal system to generate human
  antibodies in a cow (Tc cattle)

10
Various cow-derived anthrax IG protect A/J mice
from a lethal Sterne spore (1x106) IP
challenge (avg. results of 2 experiments)
HAC cow antibodies
20
18
16
14
12
10
Number of surviving mice
8
6
4
2
0
1
2
3
4
5
6
7
8
9
10
11
12
13
Time post challenge (days)
Antibodies (25 mg/kg) given day of challenge,
once and Cipro (10mg/kg) given day after
challenge, daily
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Anthrax IG Future Plans

• Characterize HUMAN antibodies against anthrax
  generated in the Tc cattle by
• Potency Assays
• In vitro binding
• In vitro neutralization
• Sterne strain mouse protection assays
• Virulent Ames strain challenges in collaboration
  with USAMRIID
• measuring binding affinity/avidity of the human
  antibodies using surface plasmon resonance
  technology Biacore