A CD28 superagonistic antibody elicits 2 functionally distinct waves of T cell activation in rats
J. Clin. Invest. Nora Müller, et al. 118:1405 doi:10.1172/JCI32698 [Go to this article.]

Figure 1
Administration of JJ316 induces T lymphopenia and leads to the redistribution of CD4⁺ T cells. (A) Blood samples from rats i.v. injected with 1.0 mg, 0.2 mg, or 0.04 mg JJ316 were taken at the indicated time points and analyzed for the percentages of circulating T cells (left panel) and granulocytes (right panel) by flow cytometry. n = 6; mean ± standard error of the mean. Statistical analysis refers to the individual comparison of each time course to untreated control rats by 1-way ANOVA. (B) Leukocytes were isolated from lymph nodes, spleen, lung, and liver 10 and 72 hours after injection of 1.0 mg JJ316 or PBS as a control. Subsequently, the total number CD4⁺ T cells was determined by microscopic counting followed by flow cytometric analysis of CD4 and TCRβ expression. n = 3; mean ± standard error of the mean. (C) The relative frequencies of CD4⁺ naive (CD45RC⁺RT6.1⁺), activated (CD45RC^–RT6.1^–), and memory T cells (CD45RC^–RT6.1⁺/CD45RC⁺RT6.1^–) was determined 10 and 72 hours after application of JJ316 or PBS. Thy-1⁺ RTEs (recent thymic emigrants) were excluded from the analysis. n = 3; mean ± standard error of the mean. (D) CFSE-labeled lymphocytes (4 × 10⁷) were transferred into recipient rats i.v., and 3 days later 0.2 mg JJ316 or PBS were administered. After 10 hours the frequencies of CFSE⁺CD4⁺ T cells in blood, liver, lung, and spleen were determined by microscopic counting and flow cytometry. n = 3; mean ± standard error of the mean. *P < 0.05; **P < 0.01; ***P < 0.001.