Our Research
Tetramer Core Laboratory
The Tetramer Core Laboratory provides MHC class II tetramer reagents for collaborators both within and outside BRI. These tetramers are synthetic protein conjugates that allow the direct detection of antigen specific T cells by flow cytometry. Tetramers produced in our facility have been used to detect CD4+ (helper) T cells and have facilitated the study of viral immunity, autoimmune diseases, allergic responses, and cancer. The class II MHC proteins are produced using stable insect cell cultures and purified by affinity chromatography. The purified proteins are loaded with exogenous peptide and conjugated using PE labeled streptavidin. In some cases we are able to provide T cell clones for use as experimental positive controls.
Manager:
Eddie James, PhD
Availability:
Our intention is to distribute tetramer reagents as widely as possible. To receive materials from the Tetramer Core, users must complete a material transfer agreement. Unless alternative arrangements are made, all users must provide peptide solution (20-50 mg/mL in DMSO) for each tetramer to be delivered. A modest service fee ($3.50 per test) will be assessed to defray production costs.
Proteins available:
DRB1*01:01, DRB1*01:02, DRB1*03:01, DRB1*04:01, DRB1*04:02, DRB1*04:03, DRB1*04:04, DRB1*04:05, DRB1*07:01, DRB1*09:01, DRB1*10:01, DRB1*11:01, DRB1*11:04, DRB1*12:01, DRB1*13:01, DRB1*13:02, DRB1*14:01, DRB1*15:01, DRB3*01:01, DRB3*02:02, DRB4*01:01, DRB5*01:01, DPB1*04:01, DPB1*05:01, and DQB1*06:02
Pricing:
The cost for 100 staining tests is $500. Custom peptide biding assays are $500 per plate.
Contact:
For more information or to obtain a request form, contact Eddie James (ejames@benaroyaresearch.org or 206-625-7373 ext.68176). To discuss a potential collaborative project contact Bill Kwok (bkwok@benaroyaresearch.org or 206-583-6517).
To see video:
Visualizing Antigen Specific CD4+ T Cells using MHC Class II Tetramers
For more information:
Recent Publications:
Differentiation stage determines pathologic and protective allergen-specific CD4(+) T-cell outcomes during specific immunotherapy. Wambre E, Delong JH, James EA, Lafond RE, Robinson D, Kwok WW. J Allergy Clin Immunol. 2011 Oct 3. [Epub ahead of print]
Human CD8+ and CD4+ T Cell Memory to Lymphocytic Choriomeningitis Virus Infection. Kotturi MF, Swann JA, Peters B, Arlehamn CL, Sidney J, Kolla RV, James EA, Akondy RS, Ahmed R, Kwok WW, Buchmeier MJ, Sette A. J Virol. 2011 Nov;85(22):11770-80. Epub 2011 Sep 7.
Uveitis-associated epitopes of retinal antigens are pathogenic in the humanized mouse model of uveitis and identify autoaggressive T cells. Mattapallil MJ, Silver PB, Mattapallil JJ, Horai R, Karabekian Z, McDowell JH, Chan CC, James EA, Kwok WW, Sen HN, Nussenblatt RB, David CS, Caspi RR. J Immunol. 2011 Aug 15;187(4):1977-85. Epub 2011 Jul 15.
Papillomavirus-specific CD4+ T cells exhibit reduced STAT-5 signaling and altered cytokine profiles in patients with recurrent respiratory papillomatosis. James EA, DeVoti JA, Rosenthal DW, Hatam LJ, Steinberg BM, Abramson AL, Kwok WW, Bonagura VR. J Immunol. 2011 Jun 1;186(11):6633-40. Epub 2011 Apr 29.
Ara h 1-reactive T cells in individuals with peanut allergy. DeLong JH, Simpson KH, Wambre E, James EA, Robinson D, Kwok WW. J Allergy Clin Immunol. 2011 May;127(5):1211-8.e3. Epub 2011 Apr 2.
Differences in self-peptide binding between T1D-related susceptible and protective DR4 subtypes. Ge X, James EA, Reijonen H, Kwok WW. J Autoimmun. 2011 Mar;36(2):155-60. Epub 2011 Jan 22.
T-cell responses in two unrelated hemophilia A inhibitor subjects include an epitope at the factor VIII R593C missense site. James EA, van Haren SD, Ettinger RA, Fijnvandraat K, Liberman JA, Kwok WW, Voorberg J, Pratt KP. J Thromb Haemost. 2011 Apr;9(4):689-99. doi: 10.1111/j.1538-7836.2011.04202.x.