1.Detection of Avian Antigen-Specific T Cells Induced by Viral Vaccines.
Dalgaard TS1, Norup LR2, Juul-Madsen HR3. Methods Mol Biol. 2016;1404:77-88. doi: 10.1007/978-1-4939-3389-1_5.
Live attenuated viral vaccines are widely used in commercial poultry production, but the development of new effective inactivated/subunit vaccines is needed. Studies of avian antigen-specific T cells are primarily based on analyses ex vivo after activating the cells with recall antigen. There is a particular interest in developing robust high-throughput assays as chicken vaccine trials usually comprise many individuals. In many respects, the avian immune system differs from the mammalian, and T cell assessment protocols must be adjusted accordingly to account for, e.g., differences in leukocyte subsets.The carboxyfluorescein succinimidyl ester (CFSE) method described in this chapter has been adapted to chicken cells. In this test, cells of interest are stained with CFSE. The succinimidyl ester group covalently binds to cellular amines forming fluorescent conjugates that are retained in the cells even throughout division. This leads to daughter cells containing half the fluorescence of their parents.
2.Functional Role of FcγRIIB in the Regulation of Mesenchymal Stem Cell Function.
Zhu T1, Chen R2, Li Z3, Tian J4, Deng C5, Zhang X5, Zhang K6, Tong L7, Yu Y8, Bai C5. Int J Med Sci. 2016 Feb 5;13(2):154-60. doi: 10.7150/ijms.13649. eCollection 2016.
Mesenchymal stem cells (MSCs) derived from bone marrow are plural-potent stem cells with immune regulatory functions. We aimed to evaluate role of FcγRIIB in the regulation of bone marrow-derived MSC function. MSCs were prepared from mouse bone marrow derived from wild-type (WT) or FcγRIIB-deficient (FcγRIIB-/-) mice. MSCs were co-cultured with bone marrow-derived dendritic cells (BMDCs), and BMDC maturation and function were evaluated by flow cytometric analysis and carboxyfluorescein succinimidyl ester-labeled OT-II T-cell addition. An acute asthma model was established by aeresol ovalbumin challenge in mice. Mice received WT or FcγRIIB-/- MSC therapy. Lung function was evaluated by histological examination and cytokine production measurement. mRNA and protein expression levels of target genes were examined by real-time quantitative polymerase chain reactionor western blotting. We found that MSCs derived from bone marrow exhibit a high level of FcγRIIB expression.
3.Growth suppression effect of human mesenchymal stem cells from bone marrow, adipose tissue, and Wharton's jelly of umbilical cord on PBMCs.
Ayatollahi M1, Talaei-Khozani T2, Razmkhah M3. Iran J Basic Med Sci. 2016 Feb;19(2):145-53.
OBJECTIVES: Immunosuppressive property of mesenchymal stem cells (MSCs) has great attraction in regenerative medicine especially when dealing with tissue damage involving immune reactions. The most attractive tissue sources of MSCs used in clinical applications are bone marrow (BM), adipose tissue (AT), and Wharton's jelly (WJ) of human umbilical cord. The current study has compared immunomodulatory properties of human BM, AT, and WJ-MSCs.
4.Simultaneous flow cytometric measurement of antigen attachment to phagocytes and phagocytosis.
Laopajon W1, Takheaw N1, Kasinrerk W1,2, Pata S1,2. J Immunoassay Immunochem. 2016 Mar 28. [Epub ahead of print]
The current available assays cannot differentiate the stages of phagocytosis. We, therefore, established methods for concurrent detection antigen attachment and engulfment by phagocyte using latex beads coated with lipopolysaccharide, rabbit IgG, and carboxyfluorescein diacetate succinimidyl ester. The generated beads were incubated with whole blood at 37°C for 1 hr and stained with PE-Cy5.5 anti-rabbit IgG antibody. By flow cytometry, attachment and phagocytic processes could be detected, simultaneously. The established method is a valuable tool for diagnosis of phagocytic disorder and study of molecules involved in phagocytosis.
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