Quantitative determination of CD4/CD8 molecules by a cell marker ELISA

Abstract

Determination of percentages of CD4+ and CD8+ T cells from patients with human immunodeficiency virus infection is usually done by flow cytometric analysis. We compared a cell marker ELISA with flow cytometry for quantitation of CD4 and CD8 molecules on T lymphocytes, and correlated the values both with the number of CD4+ and CD8+ T lymphocytes and with clinical data. Results by cell marker ELISA (y) correlated well with those by flow cytometric analysis (x); r = 0.69, P < 0.001 (y = 0.01x + 3.9) for CD4; r = 0.81, P < 0.001 (y = 0.03x + 5.4) for CD8; n = 343. The ELISA detected changes in numbers of CD8 molecules on the cells earlier than flow cytometry recognized changes in CD8+ T-cell counts. The advantages of the ELISA are the small sample volume required (0.5 mL of blood), its internal standardization by a CD4+/CD8+ cell line, and its simple and fast performance. The cell marker ELISA appears to be an efficient alternative to flow cytometry.

Evaluation of a quantitative determination of CD4 and CD8 molecules as an alternative to CD4 + and CD8 + T lymphocyte counts in Africans

“…We found a significant correlation between the Capcellia assay and the reference method. The coefficient of correlation was similar to that previously reported in a Caucasian population ( Franke et al . 1994 ).…”

“…We found a significant correlation between the Capcellia assay and the reference method. The coefficient of correlation was similar to that previously reported in a Caucasian population (Franke et al 1994). Carrière et al (1994) suggested an equivalence between pmol and number of cells: for CD4, 1 pmol ϭ 50 ϫ 10 6 cells/l and for CD8, 1 pmol/l ϭ 25 cells ϫ 10 6 /l.…”

“…1997 ). In other immunodeficiency models, follow‐up studies showed good agreement betwen the two methods ( Franke et al . 1994 ).…”

“…Previous studies suggested that modifications in Capcellia values should be earlier or more sensitive than modifications in CD4 counts during the follow-up of HIV seropositive patients (Choi et al 1993;Carrière et al 1996;Pinilla Moraza et al 1997). In other immunodeficiency models, follow-up studies showed good agreement betwen the two methods (Franke et al 1994). Nevertheless, as the spectrum of HIV-associated diseases in our African setting differs from the occidental World (Ledru et al 1998), a prospective study should be done to confirm the reliability of Capcellia assays for the monitoring of HIV-infected people, with special regard to the prevention of opportunistic diseases.…”

Effects of Vitamin B6 Deficiency on the Composition and Functional Potential of T Cell Populations

“…To investigate the trend of T lymphocyte differentiation influenced by the different doses of vitamin B6 in diet after immune stimulation, the proliferation of the CD3+ cells was estimated to represent that of total T lymphocytes, and percentages of two main T- lymphocyte cell subsets associated with immunomodulation, CD4+ (helper T lymphocytes) and CD8+ (cytotoxic T lymphocytes), were estimated by detecting their cell surface specific glycoproteins CD4 and CD8 molecule, respectively, using an ELISA assay kit (Shanghai XinRan Biological, Shanghai, China) according to Franke et al's description [ 25 ] with some modifications. All kit reagents were allowed to reach room temperature prior to usage.…”