The Canadian Immunology Quality Assessment Program (CIQAP)

CD4 T-cell enumeration is the single most frequently utilized clinical surrogate marker available for measurement of disease progression and immune reconstitution in HIV/AIDS patients. A reproducible and accurate measurement of T-cell subsets, such as CD4, CD8 and CD3, is critical in monitoring immune reconstitution and the efficacy of drug interventions such as HAART (highly active antiretroviral therapy).

Since 1989, the National Laboratory for HIV Immunology (NLHI) has been providing a federal Quality Assessment Program for T-cell subset enumeration. Developed in close collaboration with the Canadian HIV Trials Network (CTN) and other national and international agencies, its objective is to support, nation-wide, the clinical laboratories belonging to the CTN by ensuring reproducible and accurate T-cell subset measurements by flow cytometry.

The primary activities of NLHI that support the CIQAP for T-cell enumeration include the following

• Proficiency testing
• Method validation and development
• Technology transfer and training
• National and international reference services
• Collaboration with provincial proficiency programs
• Organization of the annual Biotechnology Transfer Meeting

The National Laboratory for HIV Immunology oversees the entire CIQAP and is responsible for providing all remedial action and assistance to the laboratories when required.

 

Six times a year, NLHI sends out three fresh EDTA whole blood specimens for the enumeration of T-cell subsets by flow cytometry. Both HIV+ and HIV- specimens are included in these sendouts. The time between sample shipment and receipt of the comprehensive lab report by a participating lab is 4 weeks or less - the fastest turnaround time for any program of its kind.

 

Participation in the program is aimed at increasing accuracy, reproducibility, and between-laboratory comparability in order to ensure the reliability of laboratory results for CTN participating labs. More than 50 Canadian flow cytometry laboratories, which monitor CD4 T-cells for HIV therapies, currently participate in the program. The success of the program is evidenced by the following chart which displays data showing how ongoing participation in the CIQAP has resulted in an overall improvement in the variability of CD4/CD8 results reported by the member labs.

NLHI's current aim is to improve laboratory performance by converting laboratories to single platform technology for absolute counts. The goal is to ensure all participating laboratories generate absolute and %CD4 T-cell counts utilizing only their flow cytometers (single platform).

Method validation and development

As part of our mandate, NLHI is continually developing and validating new technologies, protocols, and instrumentation that show potential in enabling clinicians to monitor disease progression in HIV/AIDS patients in a more efficient and/or cost-effective manner. These projects are detailed in "Current Projects".

Technology transfer and training

NLHI has a commitment to provide participating Canadian laboratories with leading-edge technology. NLHI works closely with Canadian and foreign focus groups and technical committees involved in the evaluation and development of biotechnology-based assays for monitoring immune status. Technology transfer is achieved through workshops conducted as part of the annual Biotechnology Transfer Meeting, and through specialized workshops on new technologies that are to be introduced into the CIQAP on a trial basis. Topics of recent workshops include the introduction of bead based reference material for uniform instrument calibration, and the introduction of single platform absolute count technology for enumeration of CD4 T-cells. In special circumstances, individualized training is also available for members of the CIQAP.

National and international reference services

• NLHI must provide and maintain national reference services and act as a national resource in the field of HIV/AIDS related clinical flow cytometry. These reference services are provided in the form of expert advice, training, organization of annual meetings, and providing expertise and exchanging information as members of national and international advisory committees. NLHI facilities in Ottawa are also often used as test sites for prototype methods and instruments, software and reagents. This provides NLHI with a unique appreciation for future directions and trends in new technologies.

• To maintain expertise and ensure a high performance standard NLHI is also enrolled in a number of national and international accreditation programs. Performance evaluations are conducted through several assessment programs: NIH's NIAID DAIDS Immunophenotyping QAP, UK NEQAS, Australia's RCPA QA Program for Immunology (Immunophenotyping), and Ontario's Laboratory Proficiency Testing Program (LPTP) for Immunophenotyping and CD34 Stem Cell enumeration. The Immunology Advisory Committee (IAC) is responsible for the second component of the peer review system. IAC, and its sub-committees, which include the Immune Reconstitution Focus Group, the Technical Advisory Group, and the CD4 and Viral Load Guidelines Group, oversee the operations of NLHI on a monthly basis via conference calls, and meet each year at the annual Biotechnology Transfer Meeting.

• Aside from certification, cooperation and collaboration with international organizations such as ISAC, NIH, CDC, NEQAS, PAHO, UNAIDS, and WHO is essential. The measurable outcomes of collaboration with these organizations are joint publications, development of guidelines, and jointly organized workshops at international meetings.

• To maintain an active role in biotechnological development, NLHI must also have visible representation at national and international meetings to promote the exchange of information and share expertise with other members of the national and international HIV/AIDS analytical cytology community; negotiate significant corporate contributions for Bureau projects; and build and maintain important cost share partnerships with the provinces and private sector industries.

Collaboration with provincial proficiency programs

NLHI actively supports several provincial core flow cytometry facilities. This role is maintained through exchange of information and performance and method validation. The Canadian National Reference Services for Flow Cytometry (CNRFC) has funding available only for HIV related projects, hence NLHI only supports activities related to HIV work. The measurable outcomes of these activities are acceptance of NLHI immunophenotyping guidelines, instrument system evaluations and jointly organized workshops/seminars at the national level.

Organization of the biannual Biotechnology Transfer Meeting

The biannual Biotechnology Transfer Meeting brings national and international leaders in the field of HIV/AIDS analytical cytology together with members of the participating laboratories of the QAP in a bilingual meeting dedicated to biotechnology transfer and information exchange. In 1998, for the first time, NLHI invited corporate sponsorship, which allowed companies in the flow cytometric and related field to display their wares, which included instrumentation and reagents, new technologies, and software. In addition to sharing financial support of the meeting with corporate sponsors, NLHI also is solely responsible for organizing all aspects of the annual meeting, which will continue to take place in a different Canadian city every second year.