Autoimmunity, 1992, Vol. 13, pp. 243-247 Reprints available directly from the publisher Photocopying permitted by license only
0 1992 Harwood Academic Publishers GmbH Printed in the United Kingdom
Review MERCURIC CHLORIDE-INDUCED AUTOIMMUNITY PETER W. MATHIESON Department of Medicine, University of Cambridge, Cambridge, UK
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(Received April 22,1992; in final form June 10,1992)
INTRODUCTION The potential of mercuric chloride (HgCI2) to induce an autoimmune response was first recognised in the rat by Druet and his colleagues', and much of the subsequent work in this model has come from the same group. Most early studies concentrated on the kidney: high doses of HgCI2 lead to renal tubular damage2, but when lower doses are given over a longer period tubular toxicity is less apparent and glomerular lesions are seen. The original observations were in outbred Wistar rats, which after injections of HgCI2 for several weeks developed glomerulonephritis with deposition of immunoglobulin in a granular pattern in the glomeruli'. In a later study inbred Brown Norway (BN) rats were given similar amounts of HgCI2: linear deposition of immunoglobulin along the glomerular basement membrane (GBM) was seen, and shown to be due to anti-GBM autoantibodie?. The pattern of binding in the kidney was similar to that seen in human anti-GBM disease4, so that much of the early interest in HgCI2-induced autoimmunity was based on its use as an experimental model of anti-GBM disease. In subsequent years, chemically-induced autoimmunity has been studied most extensively in the rat, although HgC12 induces similar autoimmune phenomena in the mouses and in the rabbit6. Much information has been produced on the mechanism of action of HgCI2, and on the factors involved in the induction and regulation of the autoimmune response. Recent observations have strengthened the analogies between HgCI2induced autoimmunity and certain human autoimmune diseases. Furthermore, this experimental system may provide useful insights into the functional role of subsets of CD4' T cells, and into the genetic and other Correspondence to: Dr P.W. Mathieson, Department of Medicine, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2QQ, U.K. Tel: 0223 336741. Fax 0223 336709.
243
factors controlling the activation of these cells. The purpose of this article is to review some of the recent developments in the field, and to highlight the fact that although this is an experimental model which has been extensively studied in the past, it still has the potential to yield further important information about the immune system in health and disease. CHARACTERISTICS OF HgC12-INDUCED AUTOIMMUNITY HgCI2 is conventionally given as five subcutaneous injections, each of 1 mg/kg, over a 10-day period. In the BN rat, the autoimmune response thus induced is self-limiting, even if HgCI2 injections are continued'. Once the response has spontaneously resolved, animals are relatively resistant to rechallenge with HgCI2'. HgC12-induced autoimmunity can therefore be divided into three phases: induction, regulation and resistance.
lnduction When BN rats are given low doses of HgCI2, the spleen and lymph nodes enlarge, with a sharp increase in the number of spleen cells'. There is hypergammaglobulinaemia, with a modest rise in total IgG' and a marked elevation in total IgE". A number of IgG autoantibodies are produced, including antibodies to GBM, single- and double-stranded DNA, collagen types I and I1 and thyroglob~lin~. The specificity of the IgE response has not been determined'". The antiGBM response mainly consists of anti-laminin antibodies", although a recent study on immunoglobulin eluted from the kidneys of HgC12-treated BN rats showed that the eluted antibodies bound in Western blot analysis to monomeric and dimeric components of both rat and human GBM, in a pattern identical to
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that seen with human anti-GBM antibodiesI2. HgCI2 also induces anti-basement membrane antibodies in the rabbit‘, and in susceptible mouse strains it induces antinuclear antibodies5, and/or antinucleolar antibodies which are predominantly directed against fibrillarin, a ribonuclear protein which is a common antigenic target of autoantibodies in the human autoimmune disease s ~ l e r o d e r m a ” , ’ ~ . The ability of HgCI2 to induce polyclonal B cell activation is dependent upon T cells, both in vitro* and in vivo15. In HgCl,-treated BN rats, limiting dilution analysis has shown a high frequency of autoreactive CD4’ T which can themselves transfer the disease18. These autoreactive T cells proliferate in response to self major histocompatibility complex (MHC) class I1 molecules. Similar autoimmune phenomena can be induced in the BN rat by penicillamine and by gold salts, and HgClz shares with these agents a propensity to react with thiol groups. This has led to the suggestion that chemical modification of the class I1 molecules by HgC12 might lead to autoreactivity”. However, this cannot be the sole explanation, since the autoreactive T cells recognise normal class I1 molecules as well as those on cells exposed to mercury”. Other possible mechanisms of action of HgCI2 include chemical modification of self peptides, T cell receptors or cell-surface adhesion molecules; these putative explanations are not mutually exclusive. The mechanism by which the autoreactive T cells induce polyclonal B cell activation is not known, but the predominance of the IgE response strongly suggests that interleukin-4 (IL-4) is involved, since this cytokine is critical in the switching of B cells to IgE production*”.”. The role of IL-4 has been difficult to study in the rat until recently because of lack of suitable reagents, but now that rat IL-4 has been cloned22 it will be possible to address this issue. In the mouse, mRNA for IL-4 is increased in CD4’ T cells after HgCI2 treatrnentl9, and in vivo administration of an anti-IL4 monoclonal antibody leads to complete abrogation of the IgE response to HgCI2”. There has been considerable recent progress in the characterisation of functional subsets of CD4’ T cells. In the mouse, most CD4’ T cells can be divided on the basis of the cytokines they secrete into two discrete subsets, denoted T h l and Th223. T h l cells predominantly produce IL-2 and gamma interferon and mediate delayed-type hypersensitivity, and Th2 cells produce mainly IL-4, IL-5 and IL-6 and provide B cell help. There is preliminary evidence that similar subsets exist in the rat22and in man24.There are reciprocal interactions betwen T h l and Th2 cells, so that in any immune response there is a balance between these two cellular compartments. In view of the polyclonal B cell activation which characterises HgClz-induced autoimmunity, and the apparent importance of IL-4
therein, an attractive hypothesis is that HgCI2 preferentially activates the Th2 subset of CD4’ T cells”. In the rat, CD4’ T cells which provide B cell help can be distinguished by their lack of reactivity with the monoclonal antibody OX-22, which is directed against a high molecular weight isoform of CD452s. There is preliminary evidence that depletion of 0X-22h’ghT cells exacerbates the tissue injury in HgC12-treated BN rats (author’s unpublished observations), suggesting that this subset has an important regulatory role in this model. If indeed HgCI2 preferentially activates “Th2” (i.e. OX22’””) cells then removal of “Th I ” (i.e. OX22high)cells would be expected to exacerbate the resulting autoimmune response”. Susceptibiity to HgC1,-induced autoimmunity is genetically controlled, with evidence that both MHC and background genes are i n v o l ~ e d ’ ~The ~ ~ ~MHC . haplotype which confers susceptibility in the rat is RT-lnZ7.In resistant strains of rat HgCI2 induces immunosuppression, such that induction of organ specific autoimmune diseases, e.g. experimental allergic encephalomyelitis (EAE)”’ or Heymann nephritis3’, in rat strains which are normally fully susceptible, is prevented by prior treatment with HgCI2. Susceptibility to HgClz-induced immunosuppression is also under the influence of the MHC, with RT-I’ being the key haplotype”. A hypothesis which would fit with the importance of the balance between T h l and Th2 cells is that in susceptible strains Th2 cells are preferentially activated by HgCI2, whereas in resistant strains it is a T h l response which predominates’’. In the mouse, the MHC genotype may determine whether T h l or Th2 cells are activated by a particular antigen, as has been shown for CD4’ T cell responses to human type IV collagen, where H-2’ mice produce a T h l response whereas H-2b mice produce a Th2 response”. The response to HgClz is mainly a Thl-type response in H-2d mice and a Th2type response in H-2s mice”. Responsiveness to HgCI2 is linked to the MHC class I1 A’ molecule, and coexpression of the other class 11 molecule (the E molecule) in an A‘ strain decreases responsiveness”. Thus it is clear that MHC genes play a major role in the response to HgCI2, and that this may, at least in part, be mediated by differential activation of T h l or Th2 cells. Regulation
In the rat, most of the manifestations of HgC12induced autoimmunity spontaneously resolve, even if HgClz is The IgE response and the autoantibodies to GBM and thyroglobulin peak at the end of the second week and return to basal levels after about five weeksy~l”~3s, whereas the IgG response and the anti-DNA antibodies persist beyond this time’. In the mouse, antinucleolar antibodies also persist”. A
MERCURIC CHLORIDE-INDUCED AUTOIMMUNITY
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role for anti-idiotypic antibodies in this down-regulation has been proposed", and such antibodies have been directly demonstratedq7.However, their immunoregulatory importance remains uncertain3x.A role for CD8' "suppressor" cells has also been suggested'.''), but recent work from two groups has shown that the down-regulation is unaffected by depletion of CD8' T cells in V ~ I ~ O ' ~ .In ~ " .EAE, which has a similar pattern of immunoregulation, the spontaneous resolution coincides with a surge of adrenal cortico~teroids~' and a similar mechanism could occur in HgCI2-treated rats. Alternatively, the down-regulation might reflect the changing balance between T h l and Th2 cells alluded to earlier, with varying cytokine profiles responsible for the termination of some aspects of the autoimmune response.
Resistance
24.5
dence of tissue injury has been reported to be lacking in this model". One study reported the infiltration of kidney, salivary glands and tongue by activated T cells, and increased expression of MHC class I1 antigens by epithelial cells in these tissues4'. In the similar syndrome induced in BN rats by treatment with D-penicillamine, one group observed "dermatitis" and also reported granulomatous lesions -in liver and spleen4'. We have recently reported47that HgC12treated BN rats develop widespread tissue injury, with lesions in skin, liver and gut in a pattern resembling acute graft versus host disease (GVHD), and also a necrotizing leukocytoclastic vasculitis particularly affecting the gut. In addition these animals develop autoantibodies to myeloperoxidase (MPO)4X.In man, anti-MPO antibodies are closely associated with systemic vasculitis (SV)"'.'", in which the characteristic tissue lesion is a necrotizing leukocytoclastic vasculitis". Anti-MPO antibodies may coexist with antiGBM antibodies in man", as they do in HgCI2-treated BN rats. Another observation of interest in the rats was the prevention of vasculitis by pre-treatment with broad-spectrum antibiotic^^^. In SV, tissue injury is exacerbated by infection'?, and antibiotics alone may have useful therapeutic effectss4. Our observations in the BN rat have been highly reproducible, using rats from two commercial sources as well as animals bred in our own colony. Another group have confirmed the finding of anti-MPO antibodies in HgC12-treated BN rats (E. Brouwer, personal communication). Thus HgCI2-induced autoimmunity may provide a useful animal model of SV in which the role of anti-MPO antibodies in the pathogenesis of vasculitis can be studied, although the fact that there is polyclonal B cell activation is a major limitation.
After the resolution of the HgC1,-induced response, BN rats are relatively resistant to the induction of a further autoimmune response by rechallenge with HgCI?'. This relative resistance to rechallenge can be transferred to naive animals by spleen cells from HgCI?-treated animals, but if CD8' cells are removed prior to cell transfer the ability to transfer resistance is lost7. The hypothesis that CDX' T cells mediate the resistance to rechallenge is supported by recent studies in which animals depleted in viiw of their CD8' T cells were no longer resistant to rechallenge'x. There is an analogy here with human anti-GBM disease, in which the autoantibody response also resolves with time, even without immunosuppressive therapy"?, and relapse is very rare4'. Whether this apparent re-establishment of immunoregulatory control in man is also mediated by CD8' cells remains unknown. In the rat, a similar state of resistance to rechallenge can be induced by pre-treatment with low doses of HgCI2, insufficient in themselves to induce autoimmunity7. I n CONCLUSIONS vi\w depletion studies suggest that this phenomenon is also mediated by CD8' T cells3x.It is noteworthy that The immunological response to HgCl? is genetically there is thus a dissociation between the factors respon- determined, and the influence of the MHC may be at sible for the spontaneous resolution of the auto- least partly via preferential activation of either T h 1 or immune response and the factors which mediate trans- Th2 cells. In rodents which develop autoimmunity after HgCI?, IL-4 seems to be a key cytokine, and this ferable resistancejx. may reflect a predominant Th2-type response. CD8' cells are not required for the spontaneous resolution of Tissue injury in HgC12-induced uuroimmunity HgCI,-induced autoimmunity in the rat, but are Although anti-GBM antibodies are deposited in the important in the mediation of resistance to rechalkidney in HgCI2-treated BN rats and these animals lenge. There are close analogies between HgCI2develop proteinuria, there is no overt glomeru- induced autoimmunity and several human autolonephritis'. Anti-basement membrane antibodies are immune diseases. Further study of this experimental deposited in other tissues, but are not associated with system is likely to yield valuable information about evidence of tissue injury". Circulating immune com- the induction and regulation of the autoimmune plexes are detectable during the autoimmune response, and as well as being of fundamental response", and may be deposited in the walls of blood immunological importance this may have clinical vessels in various organsJ4. However, histological evi- relevance.
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Acknowledgements I would like to thank David Oliveira for helpful comments on the manuscript. The author is a Medical Research Council Clinician Scientist Fellow.
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References I . Bariety J, Druet P, Laliberte F, Sapin C. Glomerulonephritis with gamma and beta I C globulin deposits induced in rats by mercuric chloride. Am J Purhol 1971; 65: 293-302 2. Gritzka TL, Trump BF. Renal tubular lesions caused by mercuric chloride. Electron microscopic observations: degeneration of the pars recta. AmJPuthol 1968; 52: 1225-1277 3. Sapin C, Druet E, Druet P. Induction of anti-glomerular basement membrane antibodies in the brown Norway rat by mercuric chloride. Clin Exp Immunol 1977; 28: 173-179 4. Scheer RL, Grossman MA. Immune aspects of the glomerulonephritis associated with pulmonary haemorrhage. Ann Intern Med 1964; 60: 1009-1021 5 . Hultman P, Enestrom S. Mercury induced antinuclear antibodies in mice: characterization and correlation with renal immune complex deposits. Clin Exp Inimunol 1988; 71: 269-274 6. Roman-Franco AA, Turiello M, Albini B, Ossi E. Milgrom F, Andres GA. Anti-basement membrane antibodies and antigenantibody complexes in rabbits injected with mercuric chloride. Clin lmmunol Imniunuputhol 1978; 9 : 4 6 4 4 8 1 7. Bowman C, Mason DW, Pusey CD, Lockwood CM. Autoregulation of autoantibody synthesis in mercuric chloride nephritis in the Brown Norway rat. 1. A role for T suppressor cells. Eur J . lmmunol 1984; 14: 4 6 4 4 7 0 8. Hirsch F, Couderc J , Sapin C, Fournie G, Druet P. Polyclonal effect of HgCI? in the rat, its possible role in an experimental autoimmune disease. Eur J Immunol 1982; 12: 620-625 9. Pusey CD, Bowman C, Morgan A, Weetman AP, Hartley B, Lockwood CM. Kinetics and pathogenicity of autoantibodies induced by mercuric chloride in the brown Norway rat. Clin E s p lmmunol 1990; 81: 76-82 10. Prouvost-Danon A, Abadie A, Sapin C . Barin H, Druet P. Induction of IgE synthesis and potentiation of anti-ovalbumin IgE antibody response by HgCI? in the rat. J Immunol 1981; 126: 699-702 II GuCry J-C, Druet E, Clotz D, e f ul. Specificity and crossreactive idiotypes of anti-glomerular basement membrane autoantibodies in HgCI,-induced autoimmune glomerulonephritis. E u r J l n i m u n o l 1990; 20: 93-100 12 Makker SP, Kanalas J J . Renal antigens in mercuric chloride induced, anti-GBM autoantibody glomerular disease. Kidney lnr 1990; 37: 64-7 1 13. Reuter R. Tessars G, Vohr H-W, Gleichmann E, Liihrmann R. Mercuric chloride induces autoantibodies against U3 small ribonucleoprotein in susceptible mice. Proc Nurl A u d Sci USA 1989; 86: 237-241 14. Hultman P, Enestrom S, Pollard KM, Tan EM. Antifibrillarin autoantibodies in mercury-treated mice. C l i n E.\p lmniunol 1989; 78: 4 7 0 4 7 7 15. Pelletier L, Pasquier R. Vial M-C. er ul. Mercury-induced autoimmune glomerulonephritis: requirement for T cells. Nephrol DiulTrunsplunr 1987; 1: 21 1-218 16. Pelletier L, Pasquier R. Hirsch F, Sapin C , Druet P. Autoreactive T cells in mercury-induced autoimmune disease: i n vitro demonstration. J Imniunol 1986; 137: 2548-2554 17. Rossert J, Pelletier L, Pasquier R, Druet P. Autoreactive T cells in mercury-induced autoimmunity. Demonstration by limiting dilution analysis. Eur JImmunol 1988; 18: 1761-1766 18. Pelletier L , Pasquier R. Rossert J . Vial M-C, Mandet C , Druet
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P. Autoreactive T cells in mercury-induced autoimmunity: ability to induce the autoimmune disease. J Immunol 1988; 140: 750-754 Goldman M, Druet P, Gleichmann E. Th2 cells in systemic autoimmunity: insights from allogeneic diseases and chemically-induced autoimmunity. Immunol Today 1991; 12: 223-227 Doutrelepont JM, Moser M, Leo 0, et a/. Hyper-lgE in stimulatory graft-versus-host disease: role of interleukin 4. Clin El-p Irnmunol 1991: 83: 133-136 Ochel M, Vohr HW, Pfeiffer C, Gleichmann E. IL4 is required for the IgE and IgGl increase and IgGl autoantibody formation in mice treated with mercuric chloride. J Immunol 1991; 146: 3006-301 I McKnight AJ, Barclay AN, Mason DW. Molecular cloning of rat interleukin 4 cDNA and analysis of the cytokine repertoire of subsets of CD4’ T cclls. Eur J Imniunol 1991; 21: 1187-1 194 Mosmann TR, Coffman RL. Heterogeneity of cytokine secretion patterns and functions of helper T cells. Adv Inimunol 1989; 46: I 1 1-147 Salgame P, Abrams JS, Clayberger C, e t a / . Differing lymphokine profiles of functional subsets of human CD4 and CD8 T cell clones. Science 1991; 254: 279-282 Spickett GP, Brandon MR, Mason DW, Williams AF, Woolett GR. MRC OX-22, a monoclonal antibody that labels a new subset of T lymphocytes and reacts with the high molecular weight form of the leukocyte common antigen. J E.up Med 1983; 158: 795-810 Powrie F, Mason D. OX-22 CD4’ T cell induce wasting disease with multiple organ pathology: prevention by the OX-22 low subset../ Exp Med 1990; 172: 1701-1708 Druet E, Sapin C. Gunther E, Feingold N, Druet P. Mercuric chloride-induced anti-glomerular basement membrane antibodies in the rat. Genetic control. Eur ./ lmniunol 1977: 7: 348-35 1 Druet E, Sapin C , Fournie G, Mandet C, Gunther E, Druet P. Genetic control of susceptibility to mercury-induced immune nephritis in various strains of rat. Clin Imniunol Inimunoputho/ 1982; 25: 203-212 Mirtcheva J. Pfeiffer C, Bruijn JA, Jaquesmart F, Gleichmann E. Immunological alterations inducible by mercury compounds 111. H-2A acts as an immune response and H-2E as an immune “suppression” locus for HgCI2-induced antinucleolar autoantibodies. Eur. J lmmunol 1989; 19: 2257-2261 Pelletier L, Rossert J, Pasquier R, er a/. Effect of HgC12 on experimental allergic encephalomyelitis in Lewis rats. HgC12induced down-modulation of the disease. Eur J Immunol 1988: 18: 243-247 Pelletier L, Galceran M, Pasquier R, er ul. Down modulation of Heymann’s nephritis by mercuric chloride. Kidney I n ! 1987; 32: 227-232 Aten J , Veninga A, de Heer E, er ul. Susceptibility to the induction of either autoimmunity or immunosuppression by mercuric chloride is related to the major histocompatibility complex class I1 haplotype. Eur JIniniunol 1991; 21: 61 1-616 Murray JS. Madri J , Tite J, Carding SR. Bottomly K. MHC control of CD4+ T cell subset activation. J E,vp Med 1989; 170: 2 135-2 140 Bellon B, Capron M. Druet E, et ul. Mercuric chloride induced autoimmune disease in Brown-Norway rats: Sequential search for anti-basement membrane antibodies and circulating immune complexes. Eur J Cliri I n i ~ s t1982; 12: 127-133 Bowman C, Lockwood CM, Amos N. Peters DK. Circulating anti-GBM antibody and immune complexes in mercuric chloride induced nephritis in the brown Norway rat. Kidney I n / 1981; 20: 686 (Abstract) Chalopin JM, Lockwood CM. Autoregulation of autoantibody synthesis in mercuric chloride nephritis in the Brown Norway rat. 11. Presence of antigen-augmentable plaque-forming cells
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in the spleen is associated with humoral factors behaving as auto-anti-idiotypic antibodies. Eur J Irnmunol 1984; 14: 470475 Gutry J-C, Druet P. A spontaneous hybridoma producing autoanti-idiotypic antibodies that recognize a Vk-associated idiotope in mercury-induced autoimmunity. Eur J lmniunol 1990; 20: 1027-1031 Mathieson PW, Stapleton KJ, Oliveira DBG, Lockwood CM. Immunoregulation of mercuric chloride-induced autoimmunity in Brown Norway rats: a role for CD8' T cells revealed by in vivo depletion studies. EurJlmmunol 1991; 21: 2105-2109 Bowman C, Green C, Borysiewicz L, Lockwood CM. Circulating T-cell populations during mercuric chloride-induced nephritis in the Brown Norway rat. Immunology 1987; 61: 515-520 Pelletier L, Rossert J, Pasquier R, Vial M-C, Druet P. Role of CD8' cells in mercury-induced autoimmunity or immunosuppression in the rat. Scand J Immuno2 1990 31: 65-74 MacPhee IAM, Antoni FA, Mason DW. Spontaneous recovery of rats from experimental allergic encephalomyelitis is dependent on regulation of the immune system by endogenous adrenal corticosteroids. J Exp Med 1989; 169: 43 1 4 4 5 Flores JC, Taube D, Savage COS, et a/. Clinical and immunological evolution of oligoanuric anti-GBM nephritis treated by haemodialysis. Lancef 1986 i: 5-8 Bowman C, Ambrus K , Lockwood CM. Restriction of human IgG subclass expression in the population of auto-antibodies to glomerular basement membrane. Clin Exp Immunol 1987; 69: 34 1-349 Bernaudin JF, Druet E, Belair MF, Pinchon MC, Sapin C, Druet P. Extrarenal immune complex type deposits induced by mercuric chloride in the Brown Norway rat. Clin Exp lmmunol 1979; 38: 265-273 Aten J, Bosman CB, Rozing J, Stijnen T, Hoedemaeker PJ, Weening JJ. Mercuric chloride-induced autoimmunity in the
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brown Norway rat: cellular kinetics and major histocompatibility complex antigen expression. Am J Pathol 1988; 133: 127-138 Donker AJ, Venuto RC, Vladutiu AO, Brentjens JR, Andres GA. Effects of prolonged administration of D-penicillamine or captopril in various strains of rats. Brown Norway rats treated with D-penicillamine develop autoantibodies, circulating immune complexes, and disseminated intravascular coagulation. Clin Immunol Immunopathol 1991; 30: 142-155 Mathieson PW, Thiru S , Oliveira DBG. Mercuric chloridetreated Brown Norway rats develop widespread tissue injury including necrotizing vasculitis. Lab Invest 1992 (in press) Esnault VLM, Mathieson PW, Thiiu S, Oliveira DBG, Lockwood CM. Autoantibodies to myeloperoxidase in brown Norway rats treated with mercuric chloride. Lab Invest 1992 (in press) Van der Woude FJ, Daha MR, Van Es LA. The current status of neutrophil cytoplasmic antibodies. Clin Exp Immunol 1989; 78: 143-148 Jennette JC, Falk RJ. Antineutrophil cytoplasmic autoantibodies and associated diseases: a review. Am J Kidney Dis 1990; 15: 517-529 Lie JT. Members and consultants of the American College of Rheumatology. Illustrated histopathologic classification criteria for selected vasculitis syndromes. Arthritis Rheum 1990; 33: 1074-1087 Jayne DRW, Marshall PD, Jones SJ, Lockwood CM. Autoantibodies to GBM and neutrophil cytoplasm in rapidly progressive glomerulonephritis. Kidney Int 1990; 37: 965-970 Pinching AJ, Rees AJ, Pussell BA, Lockwood CM, Mitchison RS, Peters DK. Relapses in Wegener's granulomatosis: the role of infection. Br M e d J 1980: 281: 836-838 Deremee RA. The treatment of Wegener's granulomatosis with trimethoprim/sulfamethoxazole: illusion or vision'? Arthritis Rheum 1989; 31: 1068-1073.
0 1992 Harwood Academic Publishers GmbH Printed in the United Kingdom
Review MERCURIC CHLORIDE-INDUCED AUTOIMMUNITY PETER W. MATHIESON Department of Medicine, University of Cambridge, Cambridge, UK
Autoimmunity Downloaded from informahealthcare.com by UB Kiel on 12/28/14 For personal use only.
(Received April 22,1992; in final form June 10,1992)
INTRODUCTION The potential of mercuric chloride (HgCI2) to induce an autoimmune response was first recognised in the rat by Druet and his colleagues', and much of the subsequent work in this model has come from the same group. Most early studies concentrated on the kidney: high doses of HgCI2 lead to renal tubular damage2, but when lower doses are given over a longer period tubular toxicity is less apparent and glomerular lesions are seen. The original observations were in outbred Wistar rats, which after injections of HgCI2 for several weeks developed glomerulonephritis with deposition of immunoglobulin in a granular pattern in the glomeruli'. In a later study inbred Brown Norway (BN) rats were given similar amounts of HgCI2: linear deposition of immunoglobulin along the glomerular basement membrane (GBM) was seen, and shown to be due to anti-GBM autoantibodie?. The pattern of binding in the kidney was similar to that seen in human anti-GBM disease4, so that much of the early interest in HgCI2-induced autoimmunity was based on its use as an experimental model of anti-GBM disease. In subsequent years, chemically-induced autoimmunity has been studied most extensively in the rat, although HgC12 induces similar autoimmune phenomena in the mouses and in the rabbit6. Much information has been produced on the mechanism of action of HgCI2, and on the factors involved in the induction and regulation of the autoimmune response. Recent observations have strengthened the analogies between HgCI2induced autoimmunity and certain human autoimmune diseases. Furthermore, this experimental system may provide useful insights into the functional role of subsets of CD4' T cells, and into the genetic and other Correspondence to: Dr P.W. Mathieson, Department of Medicine, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2QQ, U.K. Tel: 0223 336741. Fax 0223 336709.
243
factors controlling the activation of these cells. The purpose of this article is to review some of the recent developments in the field, and to highlight the fact that although this is an experimental model which has been extensively studied in the past, it still has the potential to yield further important information about the immune system in health and disease. CHARACTERISTICS OF HgC12-INDUCED AUTOIMMUNITY HgCI2 is conventionally given as five subcutaneous injections, each of 1 mg/kg, over a 10-day period. In the BN rat, the autoimmune response thus induced is self-limiting, even if HgCI2 injections are continued'. Once the response has spontaneously resolved, animals are relatively resistant to rechallenge with HgCI2'. HgC12-induced autoimmunity can therefore be divided into three phases: induction, regulation and resistance.
lnduction When BN rats are given low doses of HgCI2, the spleen and lymph nodes enlarge, with a sharp increase in the number of spleen cells'. There is hypergammaglobulinaemia, with a modest rise in total IgG' and a marked elevation in total IgE". A number of IgG autoantibodies are produced, including antibodies to GBM, single- and double-stranded DNA, collagen types I and I1 and thyroglob~lin~. The specificity of the IgE response has not been determined'". The antiGBM response mainly consists of anti-laminin antibodies", although a recent study on immunoglobulin eluted from the kidneys of HgC12-treated BN rats showed that the eluted antibodies bound in Western blot analysis to monomeric and dimeric components of both rat and human GBM, in a pattern identical to
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P. W. MATHIESON
that seen with human anti-GBM antibodiesI2. HgCI2 also induces anti-basement membrane antibodies in the rabbit‘, and in susceptible mouse strains it induces antinuclear antibodies5, and/or antinucleolar antibodies which are predominantly directed against fibrillarin, a ribonuclear protein which is a common antigenic target of autoantibodies in the human autoimmune disease s ~ l e r o d e r m a ” , ’ ~ . The ability of HgCI2 to induce polyclonal B cell activation is dependent upon T cells, both in vitro* and in vivo15. In HgCl,-treated BN rats, limiting dilution analysis has shown a high frequency of autoreactive CD4’ T which can themselves transfer the disease18. These autoreactive T cells proliferate in response to self major histocompatibility complex (MHC) class I1 molecules. Similar autoimmune phenomena can be induced in the BN rat by penicillamine and by gold salts, and HgClz shares with these agents a propensity to react with thiol groups. This has led to the suggestion that chemical modification of the class I1 molecules by HgC12 might lead to autoreactivity”. However, this cannot be the sole explanation, since the autoreactive T cells recognise normal class I1 molecules as well as those on cells exposed to mercury”. Other possible mechanisms of action of HgCI2 include chemical modification of self peptides, T cell receptors or cell-surface adhesion molecules; these putative explanations are not mutually exclusive. The mechanism by which the autoreactive T cells induce polyclonal B cell activation is not known, but the predominance of the IgE response strongly suggests that interleukin-4 (IL-4) is involved, since this cytokine is critical in the switching of B cells to IgE production*”.”. The role of IL-4 has been difficult to study in the rat until recently because of lack of suitable reagents, but now that rat IL-4 has been cloned22 it will be possible to address this issue. In the mouse, mRNA for IL-4 is increased in CD4’ T cells after HgCI2 treatrnentl9, and in vivo administration of an anti-IL4 monoclonal antibody leads to complete abrogation of the IgE response to HgCI2”. There has been considerable recent progress in the characterisation of functional subsets of CD4’ T cells. In the mouse, most CD4’ T cells can be divided on the basis of the cytokines they secrete into two discrete subsets, denoted T h l and Th223. T h l cells predominantly produce IL-2 and gamma interferon and mediate delayed-type hypersensitivity, and Th2 cells produce mainly IL-4, IL-5 and IL-6 and provide B cell help. There is preliminary evidence that similar subsets exist in the rat22and in man24.There are reciprocal interactions betwen T h l and Th2 cells, so that in any immune response there is a balance between these two cellular compartments. In view of the polyclonal B cell activation which characterises HgClz-induced autoimmunity, and the apparent importance of IL-4
therein, an attractive hypothesis is that HgCI2 preferentially activates the Th2 subset of CD4’ T cells”. In the rat, CD4’ T cells which provide B cell help can be distinguished by their lack of reactivity with the monoclonal antibody OX-22, which is directed against a high molecular weight isoform of CD452s. There is preliminary evidence that depletion of 0X-22h’ghT cells exacerbates the tissue injury in HgC12-treated BN rats (author’s unpublished observations), suggesting that this subset has an important regulatory role in this model. If indeed HgCI2 preferentially activates “Th2” (i.e. OX22’””) cells then removal of “Th I ” (i.e. OX22high)cells would be expected to exacerbate the resulting autoimmune response”. Susceptibiity to HgC1,-induced autoimmunity is genetically controlled, with evidence that both MHC and background genes are i n v o l ~ e d ’ ~The ~ ~ ~MHC . haplotype which confers susceptibility in the rat is RT-lnZ7.In resistant strains of rat HgCI2 induces immunosuppression, such that induction of organ specific autoimmune diseases, e.g. experimental allergic encephalomyelitis (EAE)”’ or Heymann nephritis3’, in rat strains which are normally fully susceptible, is prevented by prior treatment with HgCI2. Susceptibility to HgClz-induced immunosuppression is also under the influence of the MHC, with RT-I’ being the key haplotype”. A hypothesis which would fit with the importance of the balance between T h l and Th2 cells is that in susceptible strains Th2 cells are preferentially activated by HgCI2, whereas in resistant strains it is a T h l response which predominates’’. In the mouse, the MHC genotype may determine whether T h l or Th2 cells are activated by a particular antigen, as has been shown for CD4’ T cell responses to human type IV collagen, where H-2’ mice produce a T h l response whereas H-2b mice produce a Th2 response”. The response to HgClz is mainly a Thl-type response in H-2d mice and a Th2type response in H-2s mice”. Responsiveness to HgCI2 is linked to the MHC class I1 A’ molecule, and coexpression of the other class 11 molecule (the E molecule) in an A‘ strain decreases responsiveness”. Thus it is clear that MHC genes play a major role in the response to HgCI2, and that this may, at least in part, be mediated by differential activation of T h l or Th2 cells. Regulation
In the rat, most of the manifestations of HgC12induced autoimmunity spontaneously resolve, even if HgClz is The IgE response and the autoantibodies to GBM and thyroglobulin peak at the end of the second week and return to basal levels after about five weeksy~l”~3s, whereas the IgG response and the anti-DNA antibodies persist beyond this time’. In the mouse, antinucleolar antibodies also persist”. A
MERCURIC CHLORIDE-INDUCED AUTOIMMUNITY
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role for anti-idiotypic antibodies in this down-regulation has been proposed", and such antibodies have been directly demonstratedq7.However, their immunoregulatory importance remains uncertain3x.A role for CD8' "suppressor" cells has also been suggested'.''), but recent work from two groups has shown that the down-regulation is unaffected by depletion of CD8' T cells in V ~ I ~ O ' ~ .In ~ " .EAE, which has a similar pattern of immunoregulation, the spontaneous resolution coincides with a surge of adrenal cortico~teroids~' and a similar mechanism could occur in HgCI2-treated rats. Alternatively, the down-regulation might reflect the changing balance between T h l and Th2 cells alluded to earlier, with varying cytokine profiles responsible for the termination of some aspects of the autoimmune response.
Resistance
24.5
dence of tissue injury has been reported to be lacking in this model". One study reported the infiltration of kidney, salivary glands and tongue by activated T cells, and increased expression of MHC class I1 antigens by epithelial cells in these tissues4'. In the similar syndrome induced in BN rats by treatment with D-penicillamine, one group observed "dermatitis" and also reported granulomatous lesions -in liver and spleen4'. We have recently reported47that HgC12treated BN rats develop widespread tissue injury, with lesions in skin, liver and gut in a pattern resembling acute graft versus host disease (GVHD), and also a necrotizing leukocytoclastic vasculitis particularly affecting the gut. In addition these animals develop autoantibodies to myeloperoxidase (MPO)4X.In man, anti-MPO antibodies are closely associated with systemic vasculitis (SV)"'.'", in which the characteristic tissue lesion is a necrotizing leukocytoclastic vasculitis". Anti-MPO antibodies may coexist with antiGBM antibodies in man", as they do in HgCI2-treated BN rats. Another observation of interest in the rats was the prevention of vasculitis by pre-treatment with broad-spectrum antibiotic^^^. In SV, tissue injury is exacerbated by infection'?, and antibiotics alone may have useful therapeutic effectss4. Our observations in the BN rat have been highly reproducible, using rats from two commercial sources as well as animals bred in our own colony. Another group have confirmed the finding of anti-MPO antibodies in HgC12-treated BN rats (E. Brouwer, personal communication). Thus HgCI2-induced autoimmunity may provide a useful animal model of SV in which the role of anti-MPO antibodies in the pathogenesis of vasculitis can be studied, although the fact that there is polyclonal B cell activation is a major limitation.
After the resolution of the HgC1,-induced response, BN rats are relatively resistant to the induction of a further autoimmune response by rechallenge with HgCI?'. This relative resistance to rechallenge can be transferred to naive animals by spleen cells from HgCI?-treated animals, but if CD8' cells are removed prior to cell transfer the ability to transfer resistance is lost7. The hypothesis that CDX' T cells mediate the resistance to rechallenge is supported by recent studies in which animals depleted in viiw of their CD8' T cells were no longer resistant to rechallenge'x. There is an analogy here with human anti-GBM disease, in which the autoantibody response also resolves with time, even without immunosuppressive therapy"?, and relapse is very rare4'. Whether this apparent re-establishment of immunoregulatory control in man is also mediated by CD8' cells remains unknown. In the rat, a similar state of resistance to rechallenge can be induced by pre-treatment with low doses of HgCI2, insufficient in themselves to induce autoimmunity7. I n CONCLUSIONS vi\w depletion studies suggest that this phenomenon is also mediated by CD8' T cells3x.It is noteworthy that The immunological response to HgCl? is genetically there is thus a dissociation between the factors respon- determined, and the influence of the MHC may be at sible for the spontaneous resolution of the auto- least partly via preferential activation of either T h 1 or immune response and the factors which mediate trans- Th2 cells. In rodents which develop autoimmunity after HgCI?, IL-4 seems to be a key cytokine, and this ferable resistancejx. may reflect a predominant Th2-type response. CD8' cells are not required for the spontaneous resolution of Tissue injury in HgC12-induced uuroimmunity HgCI,-induced autoimmunity in the rat, but are Although anti-GBM antibodies are deposited in the important in the mediation of resistance to rechalkidney in HgCI2-treated BN rats and these animals lenge. There are close analogies between HgCI2develop proteinuria, there is no overt glomeru- induced autoimmunity and several human autolonephritis'. Anti-basement membrane antibodies are immune diseases. Further study of this experimental deposited in other tissues, but are not associated with system is likely to yield valuable information about evidence of tissue injury". Circulating immune com- the induction and regulation of the autoimmune plexes are detectable during the autoimmune response, and as well as being of fundamental response", and may be deposited in the walls of blood immunological importance this may have clinical vessels in various organsJ4. However, histological evi- relevance.
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Acknowledgements I would like to thank David Oliveira for helpful comments on the manuscript. The author is a Medical Research Council Clinician Scientist Fellow.
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