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Annals of the Rheumatic Diseases logoLink to Annals of the Rheumatic Diseases
. 2003 Oct;62(10):983–990. doi: 10.1136/ard.62.10.983

IFNγ deficient C57BL/6 (H-2b) mice develop collagen induced arthritis with predominant usage of T cell receptor Vß6 and Vß8 in arthritic joints

C Chu, Z Song, L Mayton, B Wu, P Wooley
PMCID: PMC1754310  PMID: 12972478

Abstract

Background: Transgenic deficiency in interferon γ (IFNγ) or IFNγ receptor makes resistant strains of mice bearing H-2b or H-2d susceptible to collagen induced arthritis (CIA).

Objective: To determine whether the escape from regulation of disease susceptibility at the major histocompatibility complex level involves a new use of autoimmune T cells expressing T cell receptor (TCR) Vß that vary from the cell populations previously identified within arthritic joints.

Methods: Arthritis was induced by a standard protocol with type II bovine collagen (CII) in complete Freund's adjuvant. Clinical features, histopathology, immunological responses, and TCR profile in arthritic joints in IFNγ knockout C57BL/6 (B6.IFNγ KO) mice (H-2b) were compared directly with those in DBA/1 mice (H-2q).

Results: 60–80% of B6.IFNγ KO mice developed a progressive arthritis with a similar clinical course to classical CIA in DBA/1 mice. The affected joints in B6.IFNγ KO mice had an erosive form of arthritis with similar features to joint disease in DBA/1 mice. B6.IFNγ KO mice produced significantly higher levels of IgG2b and IgG1 autoantibodies to murine CII and showed increased proliferative response to CII compared with B6 mice. Comparable levels of interleukin 1ß and tumour necrosis factor α expression were detected in arthritic joints from ß6.IFNγ KO and DBA/1 mice. B6.IFNγKO mice used predominantly TCR Vß6 and Vß8 in arthritic joints. This TCR Vß profile is similar to that found in DBA/1 mice with CIA.

Conclusions: C57BL/6 mice deficient in IFNγ production can develop arthritis that resembles classical CIA. These data suggest that IFNγ is a key factor mediating susceptibility to CIA.

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Figure 1.

Figure 1

Arthritis in B6.IFNγ KO mice immunised with bovine CII in CFA. (A) An arthritic hind paw of a B6.IFNγ KO mouse at day 3 of arthritis with a clinical score 1. There is substantial erythematous oedema from the ankle through the digits. Note the marked swollen tarsal and metatarsal joints. This is the typical sign of an arthritic paw seen in CIA in DBA/1 mice. (B) Normal appearance of a non-arthritic hind paw of a B6 mouse immunised with bovine CII in CFA.

Figure 2.

Figure 2

Natural clinical course of arthritis in B6.IFNγ KO mice. The average clinical score of arthritic mice reflects the clinical severity of an individual arthritic limb and the number of affected limbs. DBA/1 mice exhibited a typical monophasic progressive disease. Similarly, B6.IFNγ KO mice showed a progressive type of arthritis. Most of the affected joints progressed from initial swelling to distortion and some affected joints progressed to ankylosis. The lower clinical score at the late stage of disease in B6.IFNγ KO mice reflects the smaller number of limbs affected and less severity of the affected limbs than in DBA/1 mice (*p<0.05). B6 mice had a mild and transient arthritis affecting only one limb in each arthritic mouse.

Figure 3.

Figure 3

Histopathology of arthritis in B6.IFNγ KO mice. (A) Section of a hind paw with arthritis at the day of onset showing inflamed synovium with massive infiltrate of mononuclear cells. There is decreased number of chondrocytes in the articular cartilage. (B) Section of a hind paw with arthritis at day 14 showing severe synovitis, pannus formation, and erosion and some visible destruction of articular cartilage. (C) Section of a hind paw with arthritis at day 35 showing massive synovitis, erosion of cartilage and subchondral bone, and destruction of joint structure. (D) Normal appearance of a section of a hind paw from a non-arthritic CII immunised B6 mouse. (E) Section of a remitted arthritic hind paw from a B6 mouse showing normal joint appearance. (F) Histological assessment of arthritis in affected limbs in B6.IFNγ KO mice (open bars) at the end of the experiments (day 35 of arthritis) in comparison with that in DBA/1 mice (filled bars). S, synovitis; P, pannus formation; E, erosion; A, architectural changes; O, overall histological score. *p<0.01; **p<0.05.

Figure 4.

Figure 4

Levels of anti-heterologous and autologous CII antibodies in B6.IFNγ KO mice immunised with bovine CII in CFA (n=10 mice in each group). *p<0.01 (B6.IFNγ KO v B6); **p<0.05 (B6.IFNγ KO v B6).

Figure 5.

Figure 5

Enhanced proliferation of lymphocytes from CII immunised B6.IFNγ KO mice. Results are representative of three experiments; n=3 in each group.

Figure 6.

Figure 6

Expression of cytokines in arthritic joints in B6.IFNγ KO mice. IL1ß and TNFα gene expression was quantified using a quantitative real-time RT-PCR. Levels of IL1ß and TNFα gene expression in arthritic joints in B6.IFNγ KO are comparable with those in DBA/1 mice (p>0.05). Affected and remitted joints in B6 mice had negligible levels of expression of IL1ß and TNFα (p<0.01) compared with those in DBA/1 or B6.IFNγ KO mice.

Figure 7.

Figure 7

Predominant TCR Vß6 and Vß8 usage in arthritic joints in B6.IFNγ KO mice. Expression of TCR Vß gene expression was determined by quantitative real-time RT-PCR. This composite graph shows individual arthritic mouse samples from both DBA/1 (top, n=9) and B6.IFNγ KO (bottom, n=12) mice. The Vß gene expression was expressed as a relative quantity in reference to the Cß expression in the same sample. In DBA/1 mice, Vß6 had a predominant expression over the other Vß genes in most of the samples. In B6.IFNγ KO mice, Vß6 had a two- to 50-fold increased expression over other Vß genes in 10 of 12 samples. In addition, the Vß8 gene family exhibited the second most increased expression in most Vß6 predominant samples. In the two samples in which Vß6 was not predominant, Vß8.1 and Vß8.2 outnumbered Vß6 as the predominantly expressed Vß genes.

Selected References

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