Document 3071
 DOCN  M94A3071
 TI    A semi-automated 4-color method for T-cell subset enumeration.
 DT    9412
 AU    Mandy F; Bergeron M; Shenkin M; Horton A; Mylvaganan R; Rud E; LCDC,
       HPB, Health Canada, Ottawa, ON.
 SO    Int Conf AIDS. 1994 Aug 7-12;10(1):160 (abstract no. PB0068). Unique
       Identifier : AIDSLINE ICA10/94369504
 AB    Precise, rapid and cost effective determination of T-cell subsets for
       initiation and maintenance of HIV+ patient therapy is a continual
       challenge for clinical immunologists. Guidelines released by US
       agencies, such as AIDS Clinical Trial Group (ACTG) of NIH, and the
       Center for Disease Control and Prevention, both recommend gating
       protocols that ensure high recovery and purity of lymphocytes within the
       selected gate. To meet these requirements, a panel of 12 monoclonal
       antibodies (MAbs), in 6 tubes, are recommended. A new immunophenotyping
       protocol that takes advantage of the simultaneous 4-color capabilities
       of the Coulter XL clinical flow cytometer (CFC), was compared with a
       conventional 2-color method. 25 healthy and 25 HIV+ volunteers were
       evaluated with both protocols using the same instrument. MAbs used for
       the 4-color protocol were: CD45FITC, CD3CyC, CD4RD1, CD8ECD and for the
       2-color protocol they were: CD45FITC/CD14RD1, CD3FITC/CD4RD1,
       CD3FITC/CD8RD1. The two protocols were evaluated side-by-side. The
       4-color in 1-tube protocol incorporated 2 heterogeneous gating
       strategies, taking advantage of cell lineage specific properties of
       CD45+ and CD3+ cell clusters. Linear regression (LR) equations were
       generated for both, HIV+ and HIV- T-subsets. LR analysis indicated that
       the simultaneous 4-color compared well with conventional 2-color
       protocol. For the CD4+ T-cell subsets, the R value range was between
       0.956-0.992 with slopes of 0.949 to 1.001. These preliminary results
       indicate that a single laser CFC equipped with 4-color capabilities can
       analyze 4 different antibodies simultaneously from a single whole blood
       sample tube. This approach may provide more precise reproducible
       clinical data at lower cost.
 DE    Antibodies, Monoclonal/DIAGNOSTIC USE  Comparative Study  Flow
       Cytometry/*INSTRUMENTATION  Human  HIV Infections/*IMMUNOLOGY  HIV
       Seropositivity/*IMMUNOLOGY  Immunophenotyping/*INSTRUMENTATION
       Leukocyte Count/*INSTRUMENTATION  T-Lymphocyte Subsets/*IMMUNOLOGY
       MEETING ABSTRACT

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