The Human Leukocyte Antigen (HLA) Complex
HLA Complex
HLA Class I HLA Class II Peptide Binding Cleft
Expressed on all α2 α1 β1 α1 Expressed on nucleated cells antigen presenting including: cells (APCs): • T cells • Dendritic cells • B cells • Macrophages -microgobulin • Monocytes α3 β2 β2 α2 • B cells • Granulocytes
Cell Membrane
HLA Genes on Chromosome 6
Long Arm Short Arm
HLA Region HLA Class I 6p21.3-6p22.2 MICA, MICB - Class I Chain-Related Proteins HLA Class II HLA Class III C4,Bf,C2 - Complement Components B C E A G F TNF - Tumor Necrosis Factors Bf C2 C4 TNF DOB DRA TAP1 DOA DPB1 DMB TAP2 DRB1 DPA1 DMA MICB DRB3 DRB5 DRB4
DQB1 LMP2, LMP7 - Proteasome-Like Subunits LMP2 LMP7 MICA DQA1 TAP1, TAP2 - Peptide-Transporter Subunits DP DO DM DO DQ DR Complement Components
Class II Class III Class I 0.9 Mb 0.8 Mb 2 Mb
Nomenclature
Hyphen used to SUFFIX EXPRESSION STATUS separate gene name Field from HLA prefix Separator Separators L Low cell surface expression when compared to normal levels.
HLA-DRB4*01:03:01:02N S Expressed as a soluble, secreted molecule but is not present on the cell surface.
HLA Prefix Gene Field 1 Field 3 Suffix C Alleles that produce proteins that are present in the cytoplasm and not on the cell surface. Allele Group used to show used to a synonymous denote DNA substitution changes in A Aberrant expression where there is some doubt as to whether a protein is actually expressed. within the expression coding region Q Expression of an allele is questionable, given that the mutation seen in the allele has Field 2 Field 4 been shown to affect normal expression levels in other alleles. Specific used to show HLA Protein differences in a non-coding region N Alleles that have been shown not to be expressed (null).
HLA Typing and Crossmatching
PURPOSE ASSAYS TECHNIQUE PURIFIED LYMPHOCYTES STEMCELL TECHNOLOGIES IMPROVE ASSAY? CELL ISOLATION KITS (CATALOG #)
Determine DNA-Based HLA Typing • Sequence-Specific Oligonucleotide Probes (SSOP) NO Not Applicable HLA Type • Sequence-Specific Primer (SSP) • Next Generation Sequencing (NGS)
Serological HLA Typing • Typing Trays YES T Cells: 15061, 15061HLA1 • Microcytotoxicity Assays B Cells: 15064, 15064HLA1
Detect Donor Patient-Donor Crossmatch • Flow Cytometry Crossmatch (FCXM) Assay YES Total Lymphocytes: 19655, 19961HLA Specific Antibodies • Complement-Dependent Cytotoxicity (CDC) T Cells: 19671, 896712 (DSA) Crossmatch Assay B Cells: 19684, 896842
Anti-HLA Antibody Screening • Solid Phase Assays (Luminex®) NO Not Applicable • Enzyme-Linked Immunosorbent Assay (ELISA)
1. This kit has been CE-marked as a Class I In Vitro Diagnostic (IVD) device in the European Union according to the 98/79/EC In Vitro Diagnostic Device Directive and complies with its requirements. 2. This kit carries the CE mark and is available as a Class I In Vitro Diagnostic (IVD) device in the European Union and Australia. To learn more about the regulatory status of this product in your specific region, contact [email protected].
References: 1. Howell WM et al. (2010) J Clin Pathol 63(5): 387–90. | 2. Lima-Junior J da C & Pratt-Riccio LR. (2016) Front Immunol 7: 13. | 3. Marsh SGE et al. (2010) Tissue Antigens 75(4): 291–455. | 4. Robinson J et al. (2015) Nucleic Acids Res 43 (Database issue): D423-31. | 5. Sasazuki T et al. (2016) Adv Immunol 129: 175–249. | 6. Tinckam KJ. (2012) In: Chandraker A, MH Sayegh, AK Singh (Eds.) Core Concepts in Renal Transplantation. Boston, MA: Springer US.: pp21–42. | 7. Zhang GL et al. (2014) Front Immunol 5: 597.
Fast and Easy Cell Isolation for HLA Laboratories
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