T cell responses to a non-glycosylated epitope predominate in type II collagen-immunised HLA-DRB1*0101 transgenic mice
- Alexei von Delwig1,
- Daniel M Altmann2,
- Fraser G Charlton3,
- Norman McKie1,
- John D Isaacs1,
- Rikard Holmdahl4,
- John H Robinson1
- 1Institute of Cellular Medicine, Newcastle University, Framlington Place, Newcastle upon Tyne, UK
- 2Human Immunogenetics, Hammersmith Hospital, London, UK
- 3Royal Victoria Infirmary, Newcastle upon Tyne, UK
- 4Department of Cell and Molecular Biology, Lund University, Sweden
- Correspondence to:
Dr Alexei von Delwig
Institute of Cellular Medicine, Newcastle University, Framlington Place, Newcastle upon Tyne, NE2 4HH, UK;
- Accepted 2 November 2006
- Published Online First 17 November 2006
Aim: To study collagen-induced arthritis in human leucocyte antigen (HLA)-DR1 transgenic mice lacking endogenous major histocompatibility complex class II molecules (MHC-II) and to determine T cell specificity against the arthritogenic CII259–273 epitope of type II collagen either unmodified or post-translationally glycosylated at Lys264.
Methods: Arthritis was induced by immunisation with human type II collagen in complete Freund’s adjuvant and measured by footpad swelling, clinical score and histology. T cell responses were assessed by proliferation of spleen and lymph node cells and in antigen presentation assays, using T cell hybridomas specific for the glycosylated and non-glycosylated CII259–273 epitope.
Results: The incidence of arthritis was 50% in DR1-transgenic mice lacking endogenous MHC-II molecules. Recall T cell responses in draining lymph nodes and spleen were consistently greater against the non-glycosylated epitope than to the glycosylated CII259–273. Most of the T cell hybridomas generated from CII-immunised mice recognised the non-glycosylated CII epitope and this form of the epitope was also presented with 100-fold higher efficiency and 1 h faster kinetics by both macrophages and dendritic cells.
Conclusion: This study shows that T cell responses to the non-glycosylated epitope of heterologous (human) CII are dominant in HLA-DR1 transgenic mice lacking MHC-II, which could contribute to the pathogenicity of autoimmune arthritis.
- APC, antigen presenting cells
- CFA, complete Freund’s adjuvant
- CIA, collagen-induced arthritis
- DC, dendritic cells
- FITC, fluorescein isothiocyanate
- HLA, human leucocyte antigen
- IFA, incomplete Freund’s adjuvant
- MHC-II, major histocompatibility complex class-II
- PBS, phosphate-buffered saline
- PE, phycoerythrin
- RA, rheumatoid arthritis
- TCR, T cell receptor
- TG, transgenic
Published Online First 17 November 2006
Competing interests: None declared.