Aging in the immune system results in tendency to proinflammatory responses. 1-way analysis of variance), we used GraphPad Prism version 6 for Windows (San Diego, CA, www.graphpad.com). = 0.0002 and <0.0001), the direct comparison of adult and aged mice showed no significant differences for the antibody levels found after DNA or peptide immunizations (Fig. 1). Peptide-immunized mice had antibody levels (standard deviation) of 907.4 (130.1) g/mL plasma in the aged mice and 844.4 (51.02) g/mL in the adult mice; DNA-immunized mice had antibody levels of 53.9 (85.74) g/mL in the aged mice and 29.94 (22.43) g/mL in the adult mice. For prime-boost immunizations, we tested 4 groups of aged mice and combined the data in Fig. 1. High A42 antibody levels were found in 18-month-old mice which had received the prime-boost immunizations. Both boost immunizations, DNA and peptide, were effective in increasing the antibody levels. After 3 DNA prime immunizations, aged mice had antibody levels of 19.31 (16.41) g/mL which increased to 623.2 (313.9) g anti-A42 IgG antibodies per mL plasma in the DNA prime and peptide-boost groups. Three times peptide-immunized aged mice had 116.4 (13.31) g/mL anti-A antibodies which increased to 487.2 (266.0) g/mL in the peptide prime and DNA-boost groups (= 0.0016, unpaired t test). Although there was no significant difference in the percentages of T-effector cells (CD4+CD25+Foxp3?) and Tregs in the DNA-immunized mice, the A42 peptide-immunized mice has significantly increased numbers of Teffs in the comparison with Treg numbers in the same mice (< 0.0001, unpaired t test), and in the comparison with Teff numbers in the parallel analyzed DNA immunized mice GR 38032F (= 0.0016, unpaired t test, Fig. 2A). Fig. 2 Analyses of the regulatory immune response in adult and aged mice. This figure shows the analyses of CD4+CD25+Foxp3+ cells by flow cytometry and IL-10 secreting cells measured by ELISPOT from 2 different mouse strains. In A, percentages of CD4+CD25+Foxp3+ ... We confirmed these findings in another mouse strain, Balb/c-Foxp3-EGFP, in which all cells expressing the fork head transcription factor Foxp3 are genetically labeled and show enhanced green fluorescence so that we do not have to use the intracellular staining protocol and which allows a direct comparison of cell numbers. In regard to the expected higher numbers of memory T cells (CD4+CD44+) in the aged mice, we analyzed a group of adult (8-month-old) and aged mice (2-month-old) which had received 4 DNA A42 immunizations GR 38032F for expression of CD44 and MHC class II on CD4+CD25+Foxp3+ T cells. The mean percentage of CD4+CD25+Foxp3+ cells was significantly increased in the aged DNA-immunized mice GR 38032F in the comparison with adult DNA-immunized mice (0.0286, Mann-Whitney, Fig. 2B). Although we found no differences in the mean fluorescence intensities between adult and aged mice for CD44 as a memory T-cell marker on CD4+, CD4+Foxp3+, and CD8+ T cells, we found significant differences in the percentage of CD4+CD44+Foxp3+ cells with higher numbers in the aged mice (p = 0.0286, Mann-Whitney, Fig. 2C). In regard to IL-10 secretion on A42 peptide restim-ulation in culture, we Rabbit Polyclonal to hnRNP L. found no significant difference between the 2 age groups in an ELISPOT assay: 179 37.5 cytokine secreting cell spots per 106 cells in the adult mice and 143 47.15 spots in the aged mice (= 0.2000, Mann-Whitney, Fig. 2D). Very low numbers of cytokine secreting cells were found by ELISPOT for the parallel examined cytokines IL-4, IL-17, and IFN in these 2 mouse groupings (data not proven). Whenever we likened the percentages of Compact disc4+Foxp3+ MHC course II+ cells between your adult and.

Aging in the immune system results in tendency to proinflammatory responses.

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