National Institute of Allergy and Infectious Diseases Division of

AIDS - Immunology Quality Assessment ProgramAt the Center for Laboratory Investigation - New Jersey

Medical School - Newark, NJAccredited by CAP, Licensed by NJ Dept. of Health, CLIA Approved

IQA MISSION

The IQA is a resource designed to help ACTG

Immunologists evaluate and enhance theintegrity and comparability of immunologicallaboratory determinations performed onpatients enrolled in multi-site HIV/AIDSinvestigations (therapeutic, vaccine,prevention, etc.).

IQA Program Organization

FSTRFAMHERST, NY

CENTER for LABORATORYINVESTIGATION

NEWARK, NJ

IQA PROGRAM

IQA OPERATIONS STRUCTURE

SDAC

FSTRFData Management

CLI UnitsLaboratory Support

IQA Project ManagerRaul Louzao

IQA PIThomas N. Denny

NIAIDPROJECT OFFICER

Daniella Livnat

CLI Flow Cytometry Unit• Multi Color Immunophenotyping• Sorting• Intracellular Cytokines• Flow Cytometric Evaluation Studies

(e.g.Rainbow Bead Study)

• Tetramer• Evaluation of Alternative messurements of CD4

(Facscount, Guava, Dynabeads, Cytospheres)

CLI Functional Immunology Unit

• Lymphocyte Proliferative Assays (LPA)– HIV Antigen Evaluation

• HIV p24 (Protein Science, Austral)• HIV gp120 (Protein Science, Austral)• HIV-1 MN (NCI)

• LPA Instrument Evaluation• ELISPOT• Cryopreservation

– Viability and Viable Cellular Rcovery

IQA/ACTG Projects

• CD4/CD8 QA Bi-Monthly Sendout (83 National Laboratories)– Advance Flow Evaluation

• Reviewed Histograms– Troubleshooting Problem Sites

• PICL Cryopreservation Study 3 Color Immunophenotyping (WB and PBMCs)– LPA and Flow Cytometry(Fresh PBMCs and Frozen/Thawed

PBMCs)– Freezing, Thawing and Viability Assessment

• Adult ACTG Cryopreservation Study (38 National Laboratories)

IQA/ACTG Projects

• LPA CMV QA • LPA HIV Ag QA Study

– HIV PCR RNA/ CD4 Evaluation on 15 Donors– Four Different Ag Concentrations Day 4 and Day 6– Protein Science (HIV p24, HIV gp120)– Austral (HIV p24, HIV gp 120)

• ELISPOT QA Sendouts (7 Pediatric ACTG Laboratories)• PLG (CD4 Gating Strategy)• CD4 International Validation Study (Bali 4/2002 and 4/2003)• Evaluation of Whole Blood Stabilizers• Validation of Simpler CD4 T cell Substets

Bali Results 6/11/02Received Processing Averagedate/time date/time count cell/ul count cell/ul count cell/ul cell/ul

1 020601 10/6/02, 16.00 pm 11/06/02, 8.30 am 137 152 148 164 1582 020602 10/6/02, 16.00 pm 11/06/02, 8.30 am 535 594 411 457 5263 020603 10/6/02, 16.00 pm 11/06/02, 8.30 am 101 112 99 110 1114 020605 10/6/02, 16.00 pm 11/06/02, 10.15 am 62 69 71 79 745 020606 10/6/02, 16.00 pm 11/06/02, 10.15 am 234 260 227 252 2566 020607 10/6/02, 16.00 pm 11/06/02, 10.15 am 78 87 81 90 887 020609 10/6/02, 16.00 pm 11/06/02, 11.40 am 33 37 38 42 398 020610 10/6/02, 16.00 pm 11/06/02, 11.40 am 122 136 126 140 1389 020611 10/6/02, 16.00 pm 11/06/02, 11.40 am 66 73 70 78 7610 020613 10/6/02, 16.00 pm 11/06/02, 11.40 am 88 98 99 110 10411 020614 10/6/02, 16.00 pm 11/06/02, 12.50 pm 313 348 328 364 35612 020615 10/6/02, 16.00 pm 11/06/02, 12.50 pm 65 72 81 90 8113 020617 10/6/02, 16.00 pm 11/06/02, 12.50 pm 63 70 60 67 6814 020618 10/6/02, 16.00 pm 11/06/02, 12.50 pm 173 192 164 182 18715 020619 10/6/02, 16.00 pm 11/06/02, 14.10 pm 71 79 91 101 9016 020621 10/6/02, 16.00 pm 11/06/02, 14.10 pm 81 90 77 86 8817 020622 10/6/02, 16.00 pm 11/06/02, 14.10 pm 182 202 198 220 21118 020623 10/6/02, 16.00 pm 11/06/02, 14.10 pm 64 71 69 77 7419 020625 10/6/02, 16.00 pm 11/06/02, 15.40 pm 53 59 55 61 6020 020626 10/6/02, 16.00 pm 11/06/02, 15.40 pm 285 317 321 357 33721 020627 10/6/02, 16.00 pm 11/06/02, 15.40 pm 93 103 87 97 10022 020629 10/6/02, 16.00 pm 11/06/02, 15.40 pm 51 57 43 48 5223 020630 10/6/02, 16.00 pm 11/06/02, 15.40 pm 302 336 329 366 35124 020631 10/6/02, 16.00 pm 11/06/02, 15.40 pm 76 84 72 80 82

Control 464 516 409 454 485

Sample IDNo. Count 1 Count 2 Count 3

IQA Results Over 3 day period

CD4 % Abs # Lymphs Abs# CD4 CD4 % Abs # Lymphs Abs# CD4 CD4 % Abs # Lymphs Abs# CD4 020601 19 624 120 18 609 109 34 542 182

020602 51 1250 632 49 1210 597 54 1990 1081

020603 21 1030 218 21 1070 222 21 968 208

020605 19 607 114 17 881 148 33 837 277

020606 49 1390 680 52 1380 712 52 1500 787

020607 20 1110 221 20 1050 212 22 843 182

020609 19 472 92 19 929 177 37 556 204

020610 51 1580 811 52 1280 662 53 1300 691

020611 21 986 207 22 836 182 22 933 207

020613 20 704 144 15 640 97 35 551 192

020614 49 1270 621 50 1070 531 53 1380 728

020615 20 722 147 22 915 198 22 853 184

020617 20 676 136 15 980 146 33 392 131

020618 51 1270 651 50 1160 585 52 1460 765

020619 20 943 189 20 902 184 21 759 163

020621 18 364 65 16 632 98 31 441 139

020622 50 1580 793 51 1100 561 52 1350 707

020623 21 1060 220 21 944 197 22 880 191

020625 18 938 168 14 883 126 33 410 133

020626 52 1370 714 50 1110 551 54 1333 724

020627 22 981 216 19 1020 199 24 960 231

020629 19 1030 191 14 582 82 31 510 159

020630 49 1450 704 50 1330 671 51 1310 669

020631 22 827 183 21 879 181 22 944 205

IQA Counts

IQA Day 1 6/4/02 IQA Day 3 6/6/02 IQA Day 7 6/10/02Sample ID

BALI/IQA Comparison

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ACTG Statistical Results4/11/2003

Sample Number CD4% CD8% WBC %Lymph Total Lymph CD4 # CD8#

30401 46.95 18.11 3040 0.483 1468 689 44630405 44.7 17.27 2990 0.481 1438 643 43030413 45.89 17.8 3180 0.484 1539 706 48930409 46.24 17.4 3190 0.492 1569 726 501

SD 0.94 0.38 100 0.00 61 35 34Mean 45.95 17.65 3100 0.49 1504 691 467Median 46.07 17.60 3110 0.48 1504 698 468

4/14/03Sample Number CD4% CD8% WBC %Lymph Total Lymph CD4 # CD8#

30401 47.08 17.04 2750 0.434 1194 562 32830409 45.84 16.63 1670 0.693 1157 531 19330413 45.55 18.01 1780 0.765 1362 620 24230405 47.49 17.62 1770 0.778 1377 654 244

SD 0.94 0.61 507 0.16 113 56 56Mean 46.49 17.33 1993 0.67 1272 592 252Median 46.46 17.33 1775 0.73 1278 591 243

4/16/03Sample Number CD4% CD8% WBC %Lymph Total Lymph CD4 # CD8#

30401 45.06 17.84 2640 0.391 1032 465 27330405 44.89 16.94 1550 0.777 1204 541 18730409 43.2 16.58 1770 0.778 1377 595 24430413 43.77 17.49 2670 0.439 1172 513 313

SD 0.89 0.56 582 0.21 142 54 53Mean 44.23 17.21 2158 0.60 1196 528 254Median 44.33 17.22 2205 0.61 1188 527 258

Bali/IQA 4/2003 Comparison

Bland-Altman NJ/Prodia April 2003

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Prodia and NJ CD4 results, April 2003

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IQA –International Projects

• CD4 International Validation Study• Evaluation of Whole Blood Stabilizers• Validation of Simpler CD4 T cell Subsets• Designing laboratories for resource poor countries

– Training personnel – Suggesting equipment purchases– Validating laboratory

InternationalGeorgetown Hospital

Complex• Evaluate locations for laboratory • Recommend equipment• Establish SOP and Job descriptions• Design Laboratory layout• Establish timeline for training • Develop training schedule

The top floor is the lab space. These pictures are taken from different angles.

Top 2 pictures represents the mail lab area of the building

Housing for the electrical generator (outside shot and inside shot on the right)

Laboratory

Court Yard