Practical Considerations and Workflow in Utilizing KIR Genotyping in Transplantation Medicine

Abstract

This chapter is intended to serve as a practical guide for establishing a workflow using sequence-specific polymorphism PCR (SSP-PCR) for killer cell immunoglobulin-like receptor (KIR) genotyping in a clinical setting, especially in allogeneic hematopoietic stem cell transplantation (HSCT). As clinical evidence accumulates on the application of KIR and HLA genetics to guide donor selection in HSCT, there is an increasing need for KIR genotyping in clinical settings, and thus medical institutes may need to build this capability. Among the various KIR genotyping approaches now available, SSP-PCR methods are well-established and are the most cost-effective and will likely be the method of choice especially when expenses will be passed on to the patient. The protocol described in this chapter developed by Vilches et al. features small amplicon PCR and is suitable for KIR genotyping using FFPE-derived DNA as well as DNA extracted from blood samples. Setting up a laboratory workflow for in-house KIR genotyping is relatively straightforward; in this chapter, considerations for KIR genotyping to guide clinical decisions are discussed.In HSCT, a main objective of KIR genotyping is to apply the genetic analysis to predict donor and recipient combinations that have the most potential to produce NK cell alloresponses either through the missing-self mechanism or by action associated with activating KIR. The desired effects are reduction in acute GVHD and relapse rates and enhancement of overall survival. The information herein may also be useful to clinical laboratories considering the application of KIR genotyping in areas such as solid organ transplantation, NK cell-based treatment in other forms of cancer and autoimmune diseases, humanized antibody treatment, regenerative medicine, and reproductive medicine. Some background knowledge on KIR genetics will be necessary in managing a KIR genotyping platform. This chapter aims to address the main difficulties often encountered by physicians in understanding the KIR system, such as basic aspects of the nomenclature of KIR genes and haplotypes, genotypes, and determining presence/absence of KIR ligands in the patient and donor from the extensively diversified HLA class I allotypes. In describing the workflow, emphasis has been placed on the processes after genotype PCR and gel image acquisition: haplotype inference, generating B content scores, deduction of KIR ligands from HLA typing results, and the emerging algorithms for donor selection based on KIR and HLA genetics.