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The immunological impact of 5-Azacytidine was evaluated on a diverse cohort of MDS, AML and CMML patients.

Peripheral blood was collected and analyzed before and throughout therapy.

Together, these data support a strategy for combining 5-Azacytidine treatment with immune therapy for potential clinical benefit.5-Azacytidine is a cytosine analog and a potent DNA methyltransferase inhibitor, previously shown to induce DNA demethylation.

Treatment with 5-Azacytidine (Vidaza, Celgene Corporation, Boudry, Switzerland) is used for patients with higher-risk myelodysplastic syndrome (MDS), They are of special interest because of their very restricted expression pattern in healthy tissues, involving primarily immune-privileged sites, such as testis, placenta and during fetal development.

In particular, the Natural Killer (NK)-cell subset has previously been of interest in relation to the development and prognosis of AML and MDS.

The absolute counts and activity of NK cells are reduced in leukemic patients, and low NK cell counts are associated with poor prognosis.

A number of other chemotherapeutic regiments has been shown to modulate the immune system in a favorable manner to increase antitumor immunity.

To potentially combine 5-Azacytidine with immune therapy, it is essential to understand any functional impact of 5-Azacytidine directly on immune stimulatory and inhibitory cell subsets.

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Induced immune recognition of tumor cells and increased CTA-specific T-cell responses during therapy would speak for the combination of 5-Azacytidine and CTA-specific immune therapeutic strategies.All analyses were conducted on cryopreserved material.Manual cell counting was done using trypan blue (Sigma-Aldrich) staining.Together these results signify the feasibility of combining 5-Azacytidine with immune therapeutic strategies. 5-azacytidine daily for 5 days every fourth week for at least three cycles at University Hospital Herlev, Denmark.Distance between cycles could be increased on the treating physicians’ request due to treatment toxicity or slow bone marrow recovery.

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