Coulter Epics XL™ and XL-MCL™ Flow Cytometer

Rare event cytometry faces the challenge of abundancy of non-target events and heterogeneity that may overlap with hallmark features of the target cells. However, this challenge can be resolved by using highly discriminative antibody panels with high-recovery gating strategies, robust plausibility checks and effective gate purification steps. Such a methodology is demonstrated in the application note below.

Peripheral blood dendritic cells (PBDC) can be subdivided into two main cell types, namely myeloid DC (MDC) or DC1, and plasmacytoid DC (PDC) or DC2. PBDC are defined as lineage negative cells, i.e lacking markers for T lymphocytes, B lymphocytes, NK cells and monocytes/macrophages, while expressing high levels of HLADR. However, the lack of consensus on what markers should be used in this complicated “lineage” cocktail is an issue for comparison among studies, and raises the need for a more consensual approach to PBDC identification. This poster demonstrates a simple approach for MDC and PDC identification in whole blood samples using the restricted expression pattern of the ILT3 molecule.

Cytotoxic effector lymphocytes (CTL) are the premier line of defense in viral infection or tumor infiltration. Granzyme B is the main weapon in the CTL arsenal. Granzyme B is a serine protease found primarily in natural killer (NK) cells and CTL and it acts by means of inducing apoptosis in target cells. Contact between CTL and target cells results in directed exocytosis of the CTL granules and penetrance into target by means perforin-polymerized target-cell pores. This poster assesses whether measurement of granzyme B may be a more accurate assessment of cytotoxic T cell and NK activation, and thus more indicative of an acute cytotoxic response.

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