The expression of activation antigens, namely CD25, CD69, CD71, and HLA‐DR on T cells from 15 healthy individuals stimulated with different mitogens and specific antigens was evaluated by immunofluorescence assay and flow cytometric analysis and compared with cell proliferation as a function of [3H]thymidine incorporation. CD69 was the earliest expressed antigen on stimulated cells, while HLA‐DR was the latest. Regardless of the stimulus used, lymphocytes expressing CD25 and CD71 were always more numerous than cells expressing CD69 and HLA‐DR. Variations in the proportion of CD4+ and CD8+ T cells expressing each activation marker were observed with different antigenic stimuli. The expression of each activation marker showed overall agreement with the [3H]thymidine incorporation assay in discriminating between positive and negative immune response. However, no correlation was observed between the percentage of CD25‐, CD69‐, CD71‐, and HLA‐DR‐positive T cells and the amount of [3H]thymidine incorporation. Moreover, low doses of mitogens and antigens as well as short time of stimulation were sufficient to induce T cells to express activation antigens but not to proliferate. Our data show that results obtained by flow cytometry and [3H]thymidine incorporation may differ qualitatively, at least under certain conditions; this suggests that the 2 assays are complementary, and when combined, may gives a clearer understanding of events leading to efficient cell‐mediated immune response. Cytometry 27:71–76, 1997. © 1997 Wiley‐Liss, Inc.

Characterization of the human factor(s) and identification of the interacting p17 epitope(s) will improve our understanding of the mechanisms used by HIV to efficiently replicate in our organisms.

Cells capable of interferon (IFN)-gamma synthesis following mitogenic stimulation can be detected and quantified by a recently developed immunofluorescence assay and flow cytometric analysis. The production of IFN-gamma was investigated in a cohort of 20 asymptomatic human immunodeficiency virus (HIV)-seropositive patients with normal numbers of CD4+ lymphocytes, and in 10 healthy subjects. About 60% of asymptomatic stage A1 patients had increased percentages of blood lymphocytes capable of IFN-gamma synthesis, as compared to healthy subjects. The difference reflected the relatively higher numbers of CD8+ cells, in particular the CD8+ T cell subset lacking CD28 antigen expression. The strong correlation between the CD4+/CD8+ ratio and the CD8+CD28+/CD8+CD28- ratio suggests either a role for CD4+ cells in controlling the CD28+ phenotype or a role for CD8+CD28- cells in the decline of CD4+ lymphocytes. The peculiar ability of CD8+CD28- cells to produce high amounts of IFN-gamma, as compared to CD8+CD28+ cells, supports the hypothesis that the CD8+CD28- lymphocytes constitute a population that is functionally distinct from their double-positive counterparts.

Like T cells from healthy subjects, those of HIV-1-infected patients are capable of expressing activation antigens on their surface after antigenic or mitogenic stimulation, but their proliferative activity is strongly reduced or even absent, especially in patients with advanced stages of the disease. The characteristic of expressing activation antigens in response to different stimuli in the absence of cell proliferation is shared by CD4+ and CD8+ T-cell subsets from HIV-1-infected patients. The number of T cells capable of expressing CD25 and CD71 in response to HIV-1-related antigens but not of proliferating increased significantly with the progression of the disease, but the number of T cells capable of expressing the two activation antigens in response to the classic tetanus toxoid recall antigen decreased. The higher numbers of T cells capable of responding to HIV-1-related antigens in conjunction with a reduction in the number of T cells responding to recall antigens may explain the occurrence of different infections, including opportunistic microorganisms, during the more advanced stages of HIV-1 infection. Because the increase in the number of HIV-1 antigen-responding T cells (defined by CD25 and CD71 activation antigen expression) is a characteristic of symptomatic HIV-1-infected patients, expression (by flow cytometry) of these activation antigens on T cells in response to HIV-1 antigens could be used as a new marker of disease progression.

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