Naslov
Naïve and memory CD4+ T cell frequency in mice lacking ganglioside synthesis – a preliminary study

Autori
Tokić, Stana ; Viljetić, Barbara ; Mihalj, Martina ; Mišković Špoljarić, Katarina ; Štefanić, Mario ; Glavaš-Obrovac, Ljubica ; Heffer, Marija

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Congress of the Croatian Society of Biochemistry and Molecular Biology, Book of Abstracts / Katalinić, Maja ; Dulić, Morana ; Stuparević, Igor - Zagreb, 2019, 129-129

ISBN
978-953-95551-7-5

Skup
Congress of the Croatian Society of Biochemistry and Molecular Biology "Crossroads in Life Sciences" (HDBMB2019)

Mjesto i datum
Lovran, Hrvatska, 25.09.2019. - 28.09.2019

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
B4GALNT1 ; knockout mice ; T-cells ; naive ; memory

Sažetak
Sialyated glycosphingolipids known as gangliosides are essential elements of lipid enriched membrane microdomains or lipid rafts, which facilitate proper cross-talk, adhesion and molecular signalling of immune cells. Several asialo- (GM1, GM1b), monosialo- (GM1a, GM3) and disialocylated (GD1b, GD3) glycosphingolipids, members of the o-, a- and b-series of gangliosides, have been previously implicated in the structural organisation and function of T cell immunological synapse. Moreover, sequential expression of GM1, GM1a, GM1b gangliosides, regulated via modulation of β-1, 4-N- acetylgalactosaminyltransferase 1 (B4GALNT1) levels, have been associated with specific T cell subtype differentiation. The relevance of such ganglioside alterations in phenotype or function of naïve and memory T cell compartments is however still unclear. We thus aimed to evaluate and compare relative frequencies of naïve (Tnaive, CD62L+CD44-), effector/effector memory (TEff/EM, CD62L-CD44+) and central memory (TCM, CD62L+CD44+) CD4+ T cells in peripheral (PLN) and mesenteric lymph nodes (MLN), spleen and peripheral blood (PB) of B4GALNT1 null and C57BL/6 wild type mice. Blood was collected in tubes containing EDTA, and lymphocytes isolated by mechanical tearing and filtering of dissected organs through 70-μm pore-size strainer. Single- cell aliquots (106 cells/100 μl PBS) were immunophenotyped (FACS Canto II) using anti-CD3- APC/Cy7, anti-CD4-PerCp/Cy5.5, anti-CD62L-APC and anti-CD44-PE mixture of monoclonal antibodies. Dead cells were excluded according to their light scattering properties, and flow cytometry data analysed with Flowlogic software. Compared to wild type, B4GALNT1 null mice had higher frequency of naïve T cells, accompanied by reciprocal decrease in TEff/EM abundance in PLN and MLN tissue. Opposite frequency patterns, with reduced naïve and increased TEff/EM cells, were however observed in PB of B4GALNT1 null mice, whereas B4GALNT1 spleen was enriched in TCM. No difference in CD3+CD4+ T cell frequency was observed between wild type and B4GALNT1 mice. Our preliminary results support important role of B4GALNT1-regulated ganglioside synthesis in development of naïve and antigen challenged T cells, and warrant further analysis of T cell phenotype, number and function.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Temeljne medicinske znanosti

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