IMMUNOLOGY CORE AND

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IMMUNOLOGY CORE AND SPECIALTY LABORATORIES Stev
en D. Douglas, MD The Childrens Hospital of
Philadelphia for the International Maternal
Pediatric Adolescent AIDS Clinical Trials Group
(IMPAACT) July 26, 2006
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Immunology Core Lab

• End Point Single Platform CD4
• Multiparameter Flow Cytometry
• LPA
• PBMC Cryopreservation
• Assays in studies
• HIV-specific Immunity
• Cytokines/Chemokines
• Innate Immunity

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SUMMARY OF WORKLOAD EFFECTIVE 6/19/06 FOR
EXISTING PACTG PROTOCOLS

• Immunophenotyping
• LPA
• ELISA
• Cell processing and cryopreservation for TREC
• Processing and cryopreservation
• Cell processing and
• cryopreservation for ELISPOT
• TREC
• ELISPOT
• On saliva samples stored at UCSF

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• Immunophenotyping E. Processing and
  cryopreservation
• LPA F. Cell processing and cryopreservation for
  ELISPOT
• C. ELISA G. TREC
• Cell processing and H. ELISPOT
• cryopreservation for TREC

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• Immunophenotyping
• LPA
• ELISA
• Cell processing and cryopreservation for TREC
• Processing and cryopreservation
• Cell processing and
• cryopreservation for ELISPOT
• TREC
• ELISPOT

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Cross-Network Lab Issues Advanced Flow Cytometry
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Three Color, Flow Cytometric Evaluation of
Natural Cytotoxicity Receptor (NKp30) Expression
on Human Whole Blood-Derived Mononuclear Cells
Showing Restriction to CD3-/CD56 Natural Killer
Cells
0.83 0.79 26
72
94 0.28 3.63
2.24
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Four Color, Flow Cytometric Evaluation of CCR5
Expression on Human Whole Blood-Derived Myeloid
and Plasmacytoid Dendritic Cells
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Three Color, Flow Cytometric Evaluation of
PHAPMA-Induced CD3CD8- Single Cell IL-4
Production
0.24 8.54
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Flow Cytometric Bead Assay for the Measurement of
Antigen and Mitogen-Induced Soluble Cytokines
Background Tetanus PHA
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Three Color Flow Cytometric Evaluation of PHA
Induced T Cell Subset Proliferation Based on
Carboxyfluorescein Diacetate, Succinimidyl Ester
(CFSE) Staining
1.62
1.89
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Three Color Flow Cytometric Evaluation of PHA
Induced T Cell Subset Proliferation Based on
Carboxyfluorescein Diacetate, Succinimidyl Ester
(CFSE) Staining
94
82
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Four Color Flow Cytometric Evaluation of
Spontaneous Proliferation of T Helper Memory
Cells Based on Ki-67 Expression
0.08 0.12
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Differentiation of Apoptotic and Necrotic Cells
based on Propidium Iodide and Annexin V Staining
3.73 3.7
3.5 6.74