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In situ chimerism after double cord blood transplantation

Bone Marrow Transplantation volume 48, page 1582 (2013)Cite this article

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Cord blood transplantation (CBT) is an alternative source of allo-SCT in patients lacking a suitable identical sibling or a well-matched unrelated donor. Although CBT may have the advantage of less-stringent HLA matching, it is limited by the number of cells leading to increased risk of nonengraftment. A strategy has been pioneered by the Minneapolis’s group in adults to combine two units from two different donors.1, 2 This strategy allows sustained engraftment but has been claimed to be associated with increased risk of GVHD.3 Furthermore, in the vast majority of such double CBT, only one unit accounts for long-term engraftment (reviewed by Eldjerou et al.4). However, initial engraftment usually relies on both units and it is currently unknown which of the 2 units, the dominant unit, or the ‘loser’ one, triggers GVHD in target organs. To address this question, we developed, in two patients, HLA allele-specific quantitative PCR and assessed in situ chimerism after laser micro dissection of mononuclear cells that infiltrate the gut at onset of acute GVHD.

We took advantage of the frequent HLA mismatches in CBT to design a multiplex quantitative PCR with sets of primers and probes specific for each UCB HLA polymorphism and for the control albumin gene. Using lymphoblastoid cell lines obtained by specific immortalization of B lymphocytes from the recipient and each unit, we verified the lack of interaction between the different primers and probes, as well as the absence of unspecific amplification with other HLA sequences; and finally, we optimized primer and probe concentrations for each to increase PCR sensitivity to be able to detect 1 in 500 cells (see Table 1). Lymphocytes were dissected from biopsy slide with the Laser Pressure Catapulting system (PALM, Bernreid, Germany) as previously described.5 This study has been approved by the review board of the Hospital Saint Louis, and biopsies were performed for clinical purposes.

Table 1 Patients and cord blood unit compatibility
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References

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Authors and Affiliations

  1. Department of Hematology/Transplantation, AP/HP, Hospital Saint Louis, Paris, France

    H Labussière & G Socié

  2. Inserm U 940, Hospital Saint Louis, Paris, France

    P Loiseau, E Clave & G Socié

  3. Immunology, Hospital Saint Louis, Paris, France

    P Loiseau

  4. Inserm U 728, Hospital Saint Louis, Paris, France

    A Janin

Authors
  1. H Labussière
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  2. P Loiseau
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  3. E Clave
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  4. A Janin
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  5. G Socié
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Correspondence to G Socié.

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The authors declare no conflict of interest.

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Labussière, H., Loiseau, P., Clave, E. et al. In situ chimerism after double cord blood transplantation. Bone Marrow Transplant 48, 1582 (2013). https://doi.org/10.1038/bmt.2013.102

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