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Browsing Public Health by Subject "Desert hedgehog"
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Publication Open Access Desert hedgehog is a negative regulator of CD44-CD25+ double negative T lymphocytes developmental stage in thymic differentiation(International Journal of Research in Medical Sciences, 2018-03) Kariuki, Stephen M.The process of thymic proliferation and differentiation in mammals is indispensable. The role of hedgehog family of proteins has been extensively studied in the recent past years. Specifically, scientists have carried out substantial amount of work on Sonic hedgehog (Shh) and Indian hedgehog (Ihh) and published on their roles on either T-cell development or peripheral T-cell activation. However, the role of Desert hedgehog (Dhh) the third member of the hedgehog family of proteins, in T-cell development has not been clearly understood. In this work, we aimed to investigate the role of Desert hedgehog in thymic differentiation, particularly in the double negative T cell developmental stages. Methods: Thymuses of three Dhh-/- mice, three Dhh+/- and three Dhh+/+ were obtained by killing the mice using A prepared suspension of cells was analyzed by a three-color flow Cytometry and the acquired data analyzed using flow jo, a tree star software for flow cytometry data analysis. To establish the statistical significance of the findings, data was subjected to student t-test and significance reported at critical p-value of 0.05. Results: The total number of thymic cells was found to be higher in Dhh KO mice relative to the WT control. Analysis of thymic subsets using flow cytometry showed that double negative CD4-CD8- thymocytes were found to be relatively higher in Dhh-/- mice compared to Dhh+/+ control. In particular the percentage representation of CD44-CD25+ DN3 thymocytes were significantly higher (p=0.03) in Dhh KO mice relative to the WT controls. Conclusions: The findings of this study suggest that Dhh signal could be a negative regulator during early thymic differentiation. The data shown here is representative of three separate experiments. To get much clearer and replicative findings, these experiments can be repeated with a much larger sample size.Publication Open Access Desert hedgehog knockout mice show a reduced ratio of CD4+:CD8+ in thymic developing T cells(International Journal of Scientific Reports, 2018-04) Kariuki, Stephen MwauraThymic differentiation is important and determines the strength of adaptive immunity in mammals in their later days in life. There are many factors that have been found to influence the development of T lymphocytes in the thymus and these include the effect of hedgehog signalling family of proteins. Immunologists and other basic science researchers have established the role of Indian hedgehog and sonic hedgehog in thymocytes development in the recent past, but the role of hedgehog is not well known. The aim of this study was to determine the influence of Desert hedgehog in CD4:CD8 ratio in developing thymocytes of mice. Methods: Smashed thymocytes from mice deficient of Desert hedgehog and those with an intact gene coding for this protein were prepared in a cell suspension using standard procedures. The cell suspensions were stained using fluorochrome-labelled monoclonal anti: CD4-PE, CD8-TRI, CD3-FITC, CD5-FITC, CD44-PE and CD25-FITC (e-Bioscience). Samples were analyzed using a three-color flow cytometry. The flow cytometry-generated data was analyzed using flowjo (Treestar, USA). Statistical significance of the findings was determined using paired t-test and reported at p<0.05. Results: There was a general upward trend on CD4+CD8+ double positive thymocytes in Desert hedgehog mice relative to WT controls. An analysis of CD4:8 ratio revealed a reduced ratio in Dhh KO mice compared to that of WT controls attributable to the finding that there might have been a preferential differentiation of DP CD4+CD8+ to SP CD4+ in Dhh knockout mice as demonstrated by percentage of thymic subsets. The results of this study were not statistically significant and this was blamed on the fewer number of animals used in the study. Conclusions: Dhh might have a role arresting the DP cell subjects from differentiating preferentially to CD4+ T cells. To get statistically significant findings, these experiments could be repeated with a larger animal sample.