Epigenetic induction of adaptive immune response in multiple myeloma: sequential azacitidine and lenalidomide generate cancer testis antigen-specific cellular immunity

Abstract

Patients with multiple myeloma (MM) undergoing high dose therapy and autologous stem cell transplantation (SCT) remain at risk for disease progression. Induction of the expression of highly immunogenic cancer testis antigens (CTA) in malignant plasma cells in MM patients may trigger a protective immune response following SCT. We initiated a phase II clinical trial of the DNA hypomethylating agent, azacitidine (Aza) administered sequentially with lenalidomide (Rev) in patients with MM. Three cycles of Aza and Rev were administered and autologous lymphocytes were collected following the 2nd and 3rd cycles of Aza-Rev and cryopreserved. Subsequent stem cell mobilization was followed by high-dose melphalan and SCT. Autologous lymphocyte infusion (ALI) was performed in the second month following transplantation. Fourteen patients have completed the investigational therapy; autologous lymphocytes were collected from all of the patients. Thirteen patients have successfully completed SCT and 11 have undergone ALI. Six patients tested have demonstrated CTA up-regulation in either unfractionated bone marrow (n = 4) or CD138+ cells (n = 2). CTA (CTAG1B)-specific T cell response has been observed in all three patients tested and persists following SCT. Epigenetic induction of an adaptive immune response to cancer testis antigens is safe and feasible in MM patients undergoing SCT.

Conflict of interest statement

Conflict of interest disclosure

Amir A Toor: Celgene corporation, research funding. Kyle K Payne: no conflicts. Harold M Chung: no conflicts. Roy T Sabo: no conflicts. Allison F Hazlett: no conflicts. Maciej Kmieciak: no conflicts. Kimberly Sanford: no conflicts. David C Williams Jr: no conflicts. William B Clark: no conflicts. Catherine Roberts: no conflicts. John McCarty: Celgene corporation, research funding. Masoud H Manjili: Celgene corporation, research funding.

This work is dedicated to the memory of the late Taylor Roberts (1984–2003).

© 2012 Blackwell Publishing Ltd.

Figures

Fig 1

Blood T cell subset and NK cell counts (mean) following investigational therapy. Counts prior to lymphaphereses 1 and 2, following cycles 2 (C2) and 3 (C3) of Aza-Rev.

Fig 2

Disease response to investigational therapy. (A) Paraprotein levels before and after 3 cycles of Aza-Rev. (B) Percent change in paraprotein levels, before and after 3 cycles of Aza-Rev.

Fig 3

Blood T cell subset and NK Cell counts (mean) following SCT. Counts pre-ALI, 2 and 8 weeks following ALI. * Signifies statistically significant increase over earlier time point.

Fig 4

Induction of a panel of CTA expression in bone marrow of patients with MM following a 3-cycle administration of Aza-Rev. (A) Fold expression of CTA prior to (before) or following 3 cycles of Aza-Rev (after) in unfractionated bone marrow cells. Bone marrow biopsy % clonal plasma cell infiltration, pre- and post Aza-Rev in individual patients: Patient 1: 15% and 20%; Patient 2: 5–10% and 5%; Patient 3: 10% and 10%; Patient 4: 5% and 5%. (B) Fold expression of CTA in fractionated CD138− and CD138+ bone marrow cells of two patients. Data were normalized to human GAPDH.

Fig 5

Induction of CTAG1B-reactive T cell responses in patients with MM. Lymphocytes were prepared from blood samples prior to, after 1st cycle or 3rd cycles Aza-Rev administration as well as 8 weeks or 11 months after autologous lymphocyte infusion (ALI). Lymphocytes were co-cultured with autologous dendritic cells (DCs) in 2:1 ratios in the presence or absence of recombinant CTAG1B for 20 h. γ-interferon (IFNG) production was detected in the supernatant. Data are presented after subtracting background IFNG production by T cells plus DCs in the absence of the antigen. ND: not done.