The developing immune system is an adaptive system, primed by antigens, responsive to infectious pathogens, and can be affected by other aspects of the early rearing environment, including deviations from the normal provision of parental care. We investigated whether early rearing in an institutional setting, even when followed by years living in supportive and well-resourced families, would be associated with a persistent shift in T cell profiles. Immunophenotyping was used to enumerate CD4+ CD57+ and CD8+ CD57+ subsets, with gating strategies employed to differentiate naïve, central-memory, effector-memory, and terminally differentiated EM cells expressing CD45RA (TEMRA). Blood samples were collected from 96 adolescents, and PBMC isolated via Ficol gradient, followed by an optimized immunophenotypic characterization. CMV antibody titers were determined via ELISA. Adopted adolescents had lower CD4/CD8 ratios than did the control adolescents. Early rearing had a significant effect on the T cells, especially the CD8+ CD57+ CM, EM, and TEMRA cells and the CD4+ CD57+ EM cells. Adolescents who had spent their infancy in institutions before adoption were more likely to be seropositive for CMV, with higher antibody titers. CMV antibody titers were significantly correlated with the percentages of all CD8+ CD57+ cell subsets. In the statistical modeling, CMV antibody titer also completely mediated the relationship between institutional exposure and the ratio of CD4-to-CD8 cells, as well as the percentages of CD4+ CD57+ and CD8+ CD57+ subsets. These findings demonstrate that persistent immune differences are still evident even years after adoption by supportive American families. The shift in the T cells was associated with being a latent carrier of CMV and may reflect the role of specific T cell subsets in Herpes virus containment. In older adults, sustained CMV antigen persistence and immunoregulatory containment ultimately contributes to an accumulation of differentiated T cells with a decreased proliferative capacity and to immune senescence.

BackgroundHigh mammographic density has been correlated with a 4-fold to 6-fold increased risk of developing breast cancer, and is associated with increased stromal deposition of extracellular matrix proteins, including collagen I. The molecular and cellular mechanisms responsible for high breast tissue density are not completely understood.MethodsWe previously described accelerated tumor formation and metastases in a transgenic mouse model of collagen-dense mammary tumors (type I collagen-α1 (Col1α1)tm1Jae and mouse mammary tumor virus - polyoma virus middle T antigen (MMTV-PyVT)) compared to wild-type mice. Using ELISA cytokine arrays and multi-color flow cytometry analysis, we studied cytokine signals and the non-malignant, immune cells in the collagen-dense tumor microenvironment that may promote accelerated tumor progression and metastasis.ResultsCollagen-dense tumors did not show any alteration in immune cell populations at late stages. The cytokine signals in the mammary tumor microenvironment were clearly different between wild-type and collagen-dense tumors. Cytokines associated with neutrophil signaling, such as granulocyte monocyte-colony stimulated factor (GM-CSF), were increased in collagen-dense tumors. Depleting neutrophils with anti-Ly6G (1A8) significantly reduced the number of tumors, and blocked metastasis in over 80 % of mice with collagen-dense tumors, but did not impact tumor growth or metastasis in wild-type mice.ConclusionOur study suggests that tumor progression in a collagen-dense microenvironment is mechanistically different, with pro-tumor neutrophils, compared to a non-dense microenvironment.Electronic supplementary materialThe online version of this article (doi:10.1186/s13058-016-0703-7) contains supplementary material, which is available to authorized users.

Background: Flow cytometric analysis of human P2X 7 pore activity segregates variant from common P2RX7 genotypes and may serve as a biomarker for cancer, pain, inflammation, and immune responses to infection. Standardization is needed to accommodate variable sample age and instrumentation differences in a multicenter clinical trial.Methods: CD14-PE-stained whole blood samples were treated with YO-PRO-1 combined with a P2X 7 agonist (BzATP) or control, followed by the addition of PI after closure of the P2X 7 pore. Recalled instrument settings from previous publications were used to adapt a standardized fluorescent particle-adjusted set-up method. Experiments were performed to compare the two methods while evaluating components of systematic variability and facilitating reliable processing of samples with varied ages.Results: The median YO-PRO-1 fluorescence of BzATP-treated samples had less variability when collected by the bead-adjusted method and was less influenced by the compensation strategy used. The average day-to-day coefficient of variance for assessments of P2X 7 pore activity by this method was 0.11 6 0.04, and the exclusion of nonviable cells was found to accommodate samples aged up to 4 days after phlebotomy. The bead-adjusted set-up method produced measurements differing by only 2.0% 6 1.5% on two analog cytometers, and within similar decades when comparing analog to digital instruments.Conclusions: These results provide a standardized method for quantitative flow cytometric analysis of P2X 7 receptor phenotypes in blood monocytes with minimal intralaboratory variation and potential for interlaboratory comparisons that can greatly facilitate multicenter functional genomic clinical studies. q 2008