National Laboratory for HIV Immunology (NLHI)

Who We Are

The National Laboratory for HIV Immunology (NLHI) is one of three national laboratories within the National HIV/AIDS Laboratories. The mission of NLHI is to serve as a national resource facility for HIV Immunology. By dissemination of innovative, cutting-edge technical information, the laboratory aims to meet current and future needs of Canadian clinicians facing the fight against HIV and AIDS. With a broad knowledge base and unparalleled facilities, our lab continually evaluates, assesses, adapts and develops new leading-edge technologies that may be suitable for monitoring HIV/AIDS disease progression. NLHI is responsible for developing standard techniques and guidelines for immunological monitoring of HIV/AIDS patients and provides a National Proficiency Program to monitor markers of disease progression prior to and during therapy.

Scientific and Financial Accountability

In any large organization, there is a tendency to make unilateral decisions based on departmentally defined mandates. To prevent these priorities from being in conflict with what is in the public's best interest, NLHI's service delivery operation and performance are overseen by three external review processes, each addressing a different aspect of NLHI's work.

• External international expert panels are summoned from time to time to assess the public health role of the NLHI in a national and international context.
  
• The Canadian Immunology Advisory Committee (IAC) oversees the scientific/clinical merits and priorities of all NLHI projects. IAC is an external peer review group that provides feedback to NLHI on a monthly basis. It's ten members represent clinical immunologists, flow cytometry technologists and members of the HIV-infected community.
  
• A review of the fiscal accountability of NLHI is conducted by the Strategic Planning and Review Directorate (SPRD), of the Policy and Consultative Branch of Health Canada.

To provide leadership in a complex and often changing biotechnology-based discipline, it is important to maintain, and provide proof of, technical competence. As such, NLHI is regularly evaluated by several external Quality Assessment Programs (QAP).

• NLHI participates in the Immunology proficiency testing program, specifically for lymphocyte phenotyping by flow cytometry, which is operated by the Ontario Laboratory Proficiency Testing Program (LPTP).
• Recently , NLHI began participating in the proficiency testing program for CD34 Stem Cell enumeration, also conducted by the Ontario LPTP. This new service helps to maintain and regulate the quality of transfusion medicine in Ontario.
• NLHI has participated in the National Institutes of Health's (NIH) NIAID DAIDS Quality Assessment Program for Flow Cytometric Immunophenotyping since 1993. Participation in this program is a requirement in the U.S. for laboratories that wish to participate in clinical trials conducted by the AIDS Clinical Trials Group (ACTG).
• Participation in the Leucocyte Immunophenotyping scheme (immune monitoring ), which is conducted by the United Kingdom National External Quality Assessment Schemes (UK NEQAS), ensures that NLHI is evaluated by the most extensively used quality assessment program for immunophenotyping in Europe.
• The most significant international quality assessment system for the Pacific rim nations is the Royal College of Pathologists of Australasia (RCPA) Quality Assurance Programs. NLHI has participated in this Immunology QAP (Immunophenotyping) for almost a decade.

Facilities

As a national reference laboratory, NLHI is equipped with a wide range of flow cytometric instrumentation and support equipment. 

Flow Cytometers

Beckman Coulter EPICS XL/MCL
Equipped with a 488 nm Argon laser, it has four-colour immunofluorescence capability and a Multi Carousel Loader (MCL), allowing automated measurement of multiple samples. 

BD FACSCalibur
The FACSCalibur is a modular instrument equipped with a 488 nm Argon laser and a 635 nm red diode laser. A fourth PMT allows four-colour applications and the FACS Loader provides automated presentation of multiple prepared samples. The additional Sorting option allows sorting of specific cell subsets. 

BD FACScan
This instrument is equipped with a single 488 nm Argon laser and is used for routine 3-colour applications. 

BD FACSCount
A small, fully-automated benchtop instrument, the FACSCount uses a specifically designed BD reagent kit to provide absolute counts of CD4, CD8, and CD3 T lymphocytes in unlysed whole blood. 

Beckman Coulter EPICS Elite
This versatile flow cytometer, equipped with sense-in-quartz flow cell and enhanced optics, is capable of up to 8-parameter analysis and sorting. The Autoclone option provides for single cell sorting into microwell plates. Currently, the instrument is equipped with an air-cooled 488 nm Argon ion laser and two He-Ne lasers emitting at 543 nm and 633 nm, respectively.

Luminex 100 (Luminex Corporation)
The Luminex 100 system is a fully digital processing flow cytometer designed for microbead based multiplex bio-assays. It consists of a compact flow analyzer, incorporating a 635 nm diode laser and a 532 nm frequency doubled Nd-YAG laser, as well as avalanche photodiodes and a high sensitivity photomultiplier tube for signal detection. It is controlled by a PC with proprietary acquisition and analysis software. The computer also controls an XY plate handler, which is used for processing samples in 96-well microtiter plates.

Flowmetrix (Luminex Corporation)
The instrument is well suited for microbead based multiplex bio-assays in a single tube format. It uses a flow cytometer (FACScan or FACSCalibur, Becton Dickinson) and a PC with proprietary digital signal processor board and acquisition software.

Super Mot (Howard M. Shapiro, M.D., P.C.)
This benchtop, research grade flow cytometer is the newest of our instruments. It incorporates a 20 mW frequency doubled Nd-YAG laser emitting at 532 nm and a 635 nm diode laser . This instrument is used for high sensitivity antibody quantitation. 

Support Equipment

Beckman Coulter MaxM Hematology Analyzer
A quantitative, automated, differential cell counter that generates WBC count and differential, RBC count, Hgb concentration and platelet counts from EDTA whole blood.

Beckman Coulter Multi-Q-Prep, TQ-Prep and Prep Plus
The lab has several of these instruments for rapid, automated RBC lysis and preparation of whole blood leucocyte suspensions for quantitative immunofluorescence. Designed for use with the Coulter ImmunoPrep reagent kit, they have 32-tube capacity removable carousels for multiple sample preparation. 

Computing Environment: Local Area Network (LAN)
Our laboratory uses a diverse set of computers (PCs and Macs) for control of flow cytometers and data acquisition. There is also a number of workstations which are used for data analysis and reporting. All instrument control computers and user workstations are connected by Windows NT network. Experimental data is stored on a file server, whose disks are partitioned to accommodate DOS and Macintosh files. At regular intervals data is backed up to high capacity tapes.