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T cell receptors (TCRs) expressed on the surface of T cells specifically recognize and bind to complexes of major histocompatibility complex (MHC) molecules and peptide fragments. CD4+ T cells, which include helper and regulatory T cells, recognize complexes of MHC Class II molecules and peptide.
CD4+ T cells play an important role in health and diseases. Analysis of these cells has long been hampered by a lack of suitable assays. MHC Class II tetramers have become an important tool to investigate rare antigen-specific CD4+ T cells such as CD4+CD25+ regulatory T cells. In addition to monitoring disease progression and therapeutic intervention, MHC Class II tetramers can be used to study tolerance induction, vaccination efficacy, and autoimmunity.
MHC tetramers are complexes of four MHC molecules, associated with a specific peptide and bound to a fluorochrome. Class II tetramers bind to a distinct population of CD4+ T cells.
MHC class II tetramer staining is more technically challenging than class I tetramer staining. Some reasons for this include:
Antigen-specific CD4+ T cells are more rare than antigen-specific CD8+ T cells
Affinity between the T cell receptor and MHC/peptide complex is generally lower
Acquisition of 100,000-200,000 CD4+ T cells is typically required
Exclusion gating essential
T cell expansion and/or pre-enrichment may be required
MHC Class II tetramers can be used to monitor antigen specific CD4+ T cell expansion after vaccination (Fig. 1) or after a natural immune response against a pathogen (Fig. 2).
CLIP tetramers: Negative controls in human class II tetramer assays
Negative tetramers are important in helping set the region or gate around antigen non-specific T cells to find the positive T cells. It would be especially effective if you are troubleshooting or intend to detect a small population of antigen specific T cells.
For negative control of Human class II allele, CLIP103-117 tetramers can be used. CLIP is a partial sequence of Ii (human class II associated Invariant chain). In antigen presenting cells, MHC class II molecules assemble in the ER with Ii. The CLIP segment of Ii binds the peptide biding groove of MHC class II molecules to protect the hydrophobic binding site. Following transport to the endosomal/lysosomal peptide-loading compartment, Ii is proteolytically cleaved, and the remaining CLIP peptide is exchanged with other peptides in a reaction catalyzed by HLA-DM. It is considered that there are no CD4+ T cells react with CLIP103-117 peptide.
| Reactivity | Target | Conjugate | Class | Locus*Allele | Sequence | Cat. No. | Product Name |
| ms | BDC2.5 mimotope | PE | Class II | I-A*g7 | AHHPIWARMDA | MBL-TS-M727-1 | T-Select I-Ag7 BDC2.5 mimotope Tetramer-PE |
| ms | BDC2.5 mimotope | APC | Class II | I-A*g7 | AHHPIWARMDA | MBL-TS-M727-2 | T-Select I-Ag7 BDC2.5 mimotope Tetramer-APC |
| hu | Bet v 1 142-156 | PE | Class II | HLA-DRB1*15:01 | ETLLRAVESYLLAHS | MBL-TS-M818-1 | HLA-DRB1*15:01 Bet v 1 142-156 Tetramer-ETLLRAVESYLLAHS-PE |
| hu | Bet v 1 142-156 | APC | Class II | HLA-DRB1*15:01 | ETLLRAVESYLLAHS | MBL-TS-M818-2 | HLA-DRB1*15:01 Bet v 1 142-156 Tetramer-ETLLRAVESYLLAHS-APC |
| hu | CLIP 103-117 | PE | Class II | HLA-DRB1*01:01 | CLIP 103-117 | MBL-TS-M801-1 | HLA-DRB1*01:01 CLIP 103-117 Tetramer-PVSKMRMATPLLMQA-PE |
| hu | CLIP 103-117 | APC | Class II | HLA-DRB1*01:01 | CLIP 103-117 | MBL-TS-M801-2 | HLA-DRB1*01:01 CLIP 103-117 Tetramer-PVSKMRMATPLLMQA-APC |
| hu | CLIP 103-117 | BV421 | Class II | HLA-DRB1*01:01 | CLIP 103-117 | MBL-TS-M801-4 | HLA-DRB1*01:01 CLIP 103-117 Tetramer-PVSKMRMATPLLMQA-BV421 |
| hu | CLIP 103-117 | PE | Class II | HLA-DRB1*04:01 | CLIP 103-117 | MBL-TS-M809-1 | HLA-DRB1*04:01 CLIP 103-117 Tetramer-PVSKMRMATPLLMQA-PE |
| hu | CLIP 103-117 | APC | Class II | HLA-DRB1*04:01 | CLIP 103-117 | MBL-TS-M809-2 | HLA-DRB1*04:01 CLIP 103-117 Tetramer-PVSKMRMATPLLMQA-APC |
| hu | CLIP 103-117 | BV421 | Class II | HLA-DRB1*04:01 | CLIP 103-117 | MBL-TS-M809-4 | HLA-DRB1*04:01 CLIP 103-117 Tetramer-PVSKMRMATPLLMQA-BV421 |
| ms | E alpha 52-68 | PE | Class II | I-A*b | ASFEAQGALANIAVDKA | MBL-TS-M706-1 | T-Select I-Ab E alpha 52-68 Tetramer-PE |
| hu | EBV EBNA1 515-527 | PE | Class II | HLA-DRB1*01:01 | TSLYNLRRGTALA | MBL-TS-M803-1 | HLA-DRB1*01:01 EBV EBNA1 515-527 Tetramer-TSLYNLRRGTALA-PE |
| hu | EBV EBNA1 515-527 | APC | Class II | HLA-DRB1*01:01 | TSLYNLRRGTALA | MBL-TS-M803-2 | HLA-DRB1*01:01 EBV EBNA1 515-527 Tetramer-TSLYNLRRGTALA-APC |
| ms | ESAT-6 120 | PE | Class II | I-A*b | MTEQQWNFAGIEAAASAIQG | MBL-TS-M707-1 | I-Ab ESAT-6 1-20 Tetramer-PE |
| hu | Fel d 1 49-66 | PE | Class II | HLA-DRB1*01:01 | LPVVLENARILKNCVDAK | MBL-TS-M813-1 | HLA-DRB1*01:01 Fel d 1 49-66 Tetramer-LPVVLENARILKNCVDAK-PE |
| hu | Fel d 1 49-66 | APC | Class II | HLA-DRB1*01:01 | LPVVLENARILKNCVDAK | MBL-TS-M813-2 | HLA-DRB1*01:01 Fel d 1 49-66 Tetramer-LPVVLENARILKNCVDAK-APC |
| ms | FMLV 123-141 | PE | Class II | I-A*b | EPLTSLTPRCNTAWNRLKL | MBL-TS-M705-1 | I-Ab FMLV123-141 Tetramer-PE |
| hu | GAD65 555-567 | PE | Class II | HLA-DRB1*04:01 | NFFRMVISNPAAT | MBL-TS-M811-1 | HLA-DRB1*04:01 GAD65 555-567 Tetramer-NFFRMVISNPAAT-PE |
| hu | GAD65 555-567 | APC | Class II | HLA-DRB1*04:01 | NFFRMVISNPAAT | MBL-TS-M811-2 | HLA-DRB1*04:01 GAD65 555-567 Tetramer-NFFRMVISNPAAT-APC |
| hu | HIV gag 295-307 | PE | Class II | HLA-DRB1*01:01 | DYVDRFYKTLRAE | MBL-TS-M802-1 | HLA-DRB1*01:01 HIV gag 295-307 Tetramer-DYVDRFYKTLRAE-PE |
| hu | HIV gag 295-307 | APC | Class II | HLA-DRB1*01:01 | DYVDRFYKTLRAE | MBL-TS-M802-2 | HLA-DRB1*01:01 HIV gag 295-307 Tetramer-DYVDRFYKTLRAE-APC |
| hu | HIV gag 295-307 | BV421 | Class II | HLA-DRB1*01:01 | DYVDRFYKTLRAE | MBL-TS-M802-4 | HLA-DRB1*01:01 HIV gag 295-307 Tetramer-DYVDRFYKTLRAE-BV421 |
| hu | human CLIP 103-117 | PE | Class II | HLA-DRB1*11:01 | PVSKMRMATPLLMQA | MBL-TS-M807-1 | HLA-DRB1*11:01 human CLIP103-117 Tetramer-PVSKMRMATPLLMQA-PE |
| hu | human CLIP 103-117 | PE | Class II | HLA-DRB1*15:02 | PVSKMRMATPLLMQA | MBL-TS-M817-1 | HLA-DRB1*15:02 human CLIP103-117 Tetramer-PVSKMRMATPLLMQA-PE |
| hu | human CLIP 103-117 | APC | Class II | HLA-DRB1*15:02 | PVSKMRMATPLLMQA | MBL-TS-M817-2 | HLA-DRB1*15:02 human CLIP103-117 Tetramer-PVSKMRMATPLLMQA-APC |
| hu | human CLIP 103-117 | PE | Class II | HLA-DRB1*04:05 | PVSKMRMATPLLMQA | MBL-TS-M805-1 | HLA-DRB1*04:05 human CLIP 103-117 Tetramer-PVSKMRMATPLLMQA-PE |
| hu | human CLIP 103-117 | APC | Class II | HLA-DRB1*04:05 | PVSKMRMATPLLMQA | MBL-TS-M805-2 | HLA-DRB1*04:05 human CLIP 103-117 Tetramer-PVSKMRMATPLLMQA-APC |
| hu | human CLIP 103-117 | APC | Class II | HLA-DRB1*11:01 | PVSKMRMATPLLMQA | MBL-TS-M807-2 | HLA-DRB1*11:01 human CLIP103-117 Tetramer-PVSKMRMATPLLMQA-APC |
| hu | human CLIP 103-117 | PE | Class II | HLA-DRB1*15:01 | PVSKMRMATPLLMQA | MBL-TS-M816-1 | HLA-DRB1*15:01 human CLIP103-117 Tetramer-PVSKMRMATPLLMQA-PE |
| ms | human CLIP 103-117 | PE | Class II | I-A*d | PVSKMRMATPLLMQA | MBL-TS-M720-1 | T-Select I-Ad human CLIP103-117 Tetramer-PE |
| ms | human CLIP 103-117 | APC | Class II | I-A*d | PVSKMRMATPLLMQA | MBL-TS-M720-2 | T-Select I-Ad human CLIP103-117 Tetramer-APC |
| ms | human CLIP 103-117 | PE | Class II | I-A*g7 | PVSKMRMATPLLMQA | MBL-TS-M717-1 | T-Select I-Ag7 human CLIP103-117 Tetramer-PE |
| ms | human CLIP 103-117 | APC | Class II | I-A*g7 | PVSKMRMATPLLMQA | MBL-TS-M717-2 | T-Select I-Ag7 human CLIP103-117 Tetramer-APC |
| ms | human CLIP103-117 | PE | Class II | I-A*b | PVSKMRMATPLLMQA | MBL-TS-M715-1 | T-Select I-Ab human CLIP103-117 Tetramer-PE |
| ms | human CLIP103-117 | APC | Class II | I-A*b | PVSKMRMATPLLMQA | MBL-TS-M715-2 | T-Select I-Ab human CLIP103-117 Tetramer-APC |
| hu | Influenza HA 306-318 | PE | Class II | HLA-DRB1*01:01 | HA 306-318 | MBL-TS-M804-1 | HLA-DRB1*01:01 Influenza HA 306-318 Tetramer-PKYVKQNTLKLAT-PE |
| hu | Influenza HA 306-318 | APC | Class II | HLA-DRB1*01:01 | HA 306-318 | MBL-TS-M804-2 | HLA-DRB1*01:01 Influenza HA 306-318 Tetramer-PKYVKQNTLKLAT-APC |
| hu | Influenza HA 306-318 | BV421 | Class II | HLA-DRB1*01:01 | HA 306-318 | MBL-TS-M804-4 | HLA-DRB1*01:01 Influenza HA 306-318 Tetramer-PKYVKQNTLKLAT-BV421 |
| hu | Influenza HA 306-318 | PE | Class II | HLA-DRB1*04:01 | HA 306-318 | MBL-TS-M810-1 | HLA-DRB1*04:01 Influenza HA 306-318 Tetramer-PKYVKQNTLKLAT-PE |
| hu | Influenza HA 306-318 | APC | Class II | HLA-DRB1*04:01 | HA 306-318 | MBL-TS-M810-2 | HLA-DRB1*04:01 Influenza HA 306-318 Tetramer-PKYVKQNTLKLAT-APC |
| hu | Influenza HA 306-318 | BV421 | Class II | HLA-DRB1*04:01 | HA 306-318 | MBL-TS-M810-4 | HLA-DRB1*04:01 Influenza HA 306-318 Tetramer-PKYVKQNTLKLAT-BV421 |
| hu | Influenza HA 306-318 | PE | Class II | HLA-DRB1*04:05 | PKYVKQNTLKLAT | MBL-TS-M806-1 | HLA-DRB1*04:05 Influenza HA 306-318 Tetramer-PKYVKQNTLKLAT-PE |
| hu | Influenza HA 306-318 | APC | Class II | HLA-DRB1*11:01 | PKYVKQNTLKLAT | MBL-TS-M806-2 | HLA-DRB1*04:05 Influenza HA 306-318 Tetramer-PKYVKQNTLKLAT-APC |
| hu | Influenza HA306-318 | PE | Class II | HLA-DRB1*11:01 | PKYVKQNTLKLAT | MBL-TS-M808-1 | HLA-DRB1*11:01 Influenza HA306-318 Tetramer-PKYVKQNTLKLAT-PE |
| hu | Influenza HA306-318 | APC | Class II | HLA-DRB1*11:01 | PKYVKQNTLKLAT | MBL-TS-M808-2 | HLA-DRB1*11:01 Influenza HA306-318 Tetramer-PKYVKQNTLKLAT-APC |
| ms | Influenza NP311-325 | PE | Class II | I-A*b | QVYSLIRPNENPAHK | MBL-TS-M716-1 | T-Select I-Ab Influenza NP311-325 Tetramer-PE |
| ms | Influenza NP311-325 | APC | Class II | I-A*b | QVYSLIRPNENPAHK | MBL-TS-M716-2 | T-Select I-Ab Influenza NP311-325 Tetramer-APC |
| hu | Lol p 1 105-117 | PE | Class II | HLA-DRB1*04:01 | APYHFDLSGHAFG | MBL-TS-M814-1 | HLA-DRB1*04:01 Lol p 1 105-117 Tetramer-APYHFDLSGHAFG-PE |
| hu | Lol p 1 105-117 | APC | Class II | HLA-DRB1*04:01 | APYHFDLSGHAFG | MBL-TS-M814-2 | HLA-DRB1*04:01 Lol p 1 105-117 Tetramer-APYHFDLSGHAFG-APC |
| ms | MOG 35-55 | PE | Class II | I-A*b | MEVGWYRSPFSRVVHLYRNGK | MBL-TS-M704-1 | T-Select I-Ab MOG35-55 Tetramer-PE |
| ms | MOG 35-55 | APC | Class II | I-A*b | MEVGWYRSPFSRVVHLYRNGK | MBL-TS-M704-2 | T-Select I-Ab MOG 35-55 Tetramer-APC |
| ms | mouse 2W1S | PE | Class II | I-A*b | EAWGALANWAVDSA | MBL-TS-M722-1 | T-Select I-Ab mouse 2W1S Tetramer-PE |
| ms | mouse 2W1S | APC | Class II | I-A*b | EAWGALANWAVDSA | MBL-TS-M722-2 | T-Select I-Ab mouse 2W1S Tetramer-APC |
| ms | Mtb Ag85B 240-254 | PE | Class II | I-A*b | FQDAYNAAGGHNAVF | MBL-TS-M719-1 | I-Ab Mtb Ag85B240-254 Tetramer-FQDAYNAAGGHNAVF-PE |
| ms | Mtb Ag85B 240-254 | APC | Class II | I-A*b | FQDAYNAAGGHNAVF | MBL-TS-M719-2 | I-Ab Mtb Ag85B240-254 Tetramer-FQDAYNAAGGHNAVF-APC |
| ms | OVA 323-339 | PE | Class II | I-A*d | ISQAVHAAHAEINEAGR | MBL-TS-M703-1 | T-Select I-Ad OVA323-339 Tetramer-PE |
| ms | OVA 323-339 | APC | Class II | I-A*d | ISQAVHAAHAEINEAGR | MBL-TS-M703-2 | T-Select I-Ad OVA323-339 Tetramer-APC |
| ms | OVA 323-339 | PE | Class II | I-A*b | ISQAVHAAHAEINEAGR | MBL-TS-M710-1 | T-Select I-Ab OVA 323-339 Tetramer-PE |
| ms | OVA 323-339 | APC | Class II | I-A*b | ISQAVHAAHAEINEAGR | MBL-TS-M710-2 | T-Select I-Ab OVA 323-339 Tetramer-APC |
| hu | QuickSwitch™ | PE | Class II | HLA-DRB1*01:01 | Custom | MBL-TS-7500-1R | QuickSwitch™ Quant HLA-DRB1*01:01 Tetramer Kit-PE |
| hu | QuickSwitch™ | APC | Class II | HLA-DRB1*01:01 | Custom | MBL-TS-7500-2R | QuickSwitch™ Quant HLA-DRB1*01:01 Tetramer Kit-APC |
| hu | QuickSwitch™ | BV421 | Class II | HLA-DRB1*01:01 | Custom | MBL-TS-7500-4R | QuickSwitch™ Quant HLA-DRB1*01:01 Tetramer Kit-BV421 |
| hu | QuickSwitch™ | PE | Class II | HLA-DRB1*04:01 | Custom | MBL-TS-7502-1R | QuickSwitch™ Quant HLA-DRB1*04:01 Tetramer Kit-PE |
| hu | QuickSwitch™ | APC | Class II | HLA-DRB1*04:01 | Custom | MBL-TS-7502-2R | QuickSwitch™ Quant HLA-DRB1*04:01 Tetramer Kit-APC |
| hu | QuickSwitch™ | BV421 | Class II | HLA-DRB1*04:01 | Custom | MBL-TS-7502-4R | QuickSwitch™ Quant HLA-DRB1*04:01 Tetramer Kit-BV421 |
| hu | QuickSwitch™ | PE | Class II | HLA-DRB1*15:01 | Custom | MBL-TS-7506-1R | QuickSwitch™ Quant HLA-DRB1*15:01 Tetramer Kit-PE |
| hu | QuickSwitch™ | APC | Class II | HLA-DRB1*15:01 | Custom | MBL-TS-7506-2R | QuickSwitch™ Quant HLA-DRB1*15:01 Tetramer Kit-APC |
| hu | QuickSwitch™ | BV421 | Class II | HLA-DRB1*15:01 | Custom | MBL-TS-7506-4R | QuickSwitch™ Quant HLA-DRB1*15:01 Tetramer Kit-BV421 |
| ms | T. gondii (CD4Ag28m) 605-619 | PE | Class II | I-A*b | AVEIHRPVPGTAPPS | MBL-TS-M723-1 | T-Select I-Ab T. gondii CD4Ag28m605-619 Tetramer-PE |
| ms | T. gondii (CD4Ag28m) 605-619 | APC | Class II | I-A*b | AVEIHRPVPGTAPPS | MBL-TS-M723-2 | T-Select I-Ab T. gondii CD4Ag28m605-619 Tetramer-APC |
| hu | TT p2829-844 | PE | Class II | HLA-DRB1*11:01 | MQYIKANSKFIGITEL | MBL-TS-M812-1 | HLA-DRB1*11:01 TT p2829-844 Tetramer-MQYIKANSKFIGITEL-PE |
| hu | TT p2829-844 | APC | Class II | HLA-DRB1*11:01 | MQYIKANSKFIGITEL | MBL-TS-M812-2 | HLA-DRB1*11:01 TT p2829-844 Tetramer-MQYIKANSKFIGITEL-APC |