DNA antibody idiotypes: A review of their genetic, clinical, and immunopathologic features

DNA Antibody

Idiotypes: A Review of Their Genetic, and Immunopathologic Features By Yehuda Shoenfeld

A

NTIBODIES may be identified in a variety of ways, most commonly in relation to the antigens with which they bind in in vitro assays. Examples might include anti-DNA, anti-Sm, and anti-R0 antibodies. An alternative method of distinguishing antibodies serologically involves analyzing the variable region structures that constitute the antigen binding component of the immunoglobulin molecules. These structures are referred to as idiotypes (Fig 1). Oudin and Michel’ and Kunkel et al3 were the first to show that individual immunoglobulins could be immunogenic. At least two types of idiotypic determinants may be distinguished functionally, those at the combining site (paratopes) and those adjacent to the combining site. The latter are referred to as framework-related determinants. Antibodies (anti-idiotypes) directed against combining site related determinants will inhibit the binding of antigen and antibodies bearing the relevant idiotype. In contrast, antibodies against framework determinants do not usually inhibit such interactions. In order to identify these idiotypic determinants, anti-idiotypic antibodies must be produced. This may be done by immunizing an animal (eg, a mouse) with a human monoclonal antibody (or a monoclonal antibody from any other species). The mouse’s immune system may produce an antibody directed against the antigen binding portion of the human antibody. Using an absorption technique, the irrelevant antibodies (ie, those binding to other parts of the immunoglobulin molecule) can be removed and the mouse antiserum will then function as an antiidiotypic antibody. Jerne4 proposed that any given individual possesses many thousands of idiotypes and that they reflect the multiplicity of foreign antigenic shapes that the immune system encounters. As lymphocytes are clearly able to identify a broad range of antigenic determinants, they should also be able to identify many of the idiotypes present on other lymphocytes. In this respect, it is notable that recent studies have shown that the structure of the T lymphocyte receptor is closely related to the immunoglobulin structure asso-

Clinical,

and David lsenberg

ciated with B lymphocytes (see Cooke5 for a recent review). Jerne4 postulated that immune regulation must operate at several levels. An external antigen will usually trigger a suitable immune response, with the production of an appropriate antibody, that terminates with the removal of the antigen. However, an additional response, perhaps contributing to the “damping down” of the antigenic challenge, is also generated between idiotype and anti-idiotype. The relative importance of antigen and anti-idiotype in regulating an immune response is unknown and may vary with different antigens. It was originally thought that idiotypes were unique markers of individual antibodies.2*3 However, it is now recognized that antibody molecules of the same antigen binding specificity from different individuals can possess identical or closely related idiotypes.6*7 The existence of these shared or cross-reactive idiotypes suggests that they may be derived from a restricted family of germ line genes. Shared idiotypes have been found on antibodies to foreign proteins, including tetanus toxoid’ and rye grass pollens.’ Furthermore, the sharing of idiotypes has been demonstrated on autoantibodies including anti-Sm,” anti-acetylcholine receptor antibodies,” and, as will be discussed in detail, anti-DNA antibodies. It used to be thought that an idiotype on an antibody of given antigenic specificity would be confined to those antibodies. However, it has been shown that idiotypes may be shared by antibodies with different antigen binding proper-

From the Department of Medicine D. Research Unit for Autoimmune Diseases, The Faculty of Health Sciences, Ben Gurion University, Beer-Sheva. Israel; and The Bloomsbury Rheumatology Department. The Middlesex Hospital, Arthur Stanley House, London. Yehuda Shoenfeld, MD: Professor, Department of Medicine D. Ben Gurion University, Beer-Sheva, Israel; David A. Isenberg, MD, MRCP: Senior Lecturer, The Bloomsbury Rheumatology Department, Middlesex Hospital, London. Address reprint requests to David A. Isenberg, MD, The Bloomsbury Rheumatology Department, Middlesex Hospital, Arthur Stanley House, Tottenham St, London WIP 9PG. 0 I987 by Grune & Stratton, Inc. 0049-0172/s7/1604-0002$5.0000/0

Seminars in Arthritis and Rheumatism, Vol 16. No 4 (May), 1987: pp 245-252

245

246

SHOENFELD AND ISENBERG

lymphoblastoid cell line, designated as GM 4672. The antigen binding profile of these antibodies is variable. They have been shown to bind to single and double stranded DNA (ss- and dsDNA), other combinations of polynucleotides, phospholipids, cytoskeletal proteins, and Raji anticells,‘3-‘6 and to react as lymphocytotoxic bodiesI Two of them, designated as 16/6 and 32/ 15, derived from two unrelated patients and with different antigen binding profiles, were chosen to prepare anti-idiotypic reagents.

Preparation of Anti-Idiotypic Anti-DNA Antibodies Fig 1. Representation of a human Ig molecule showing the variable W) and constant (Cl regions, heavy (H) and light (L) chains, the location of the carbohydrate (CHO) molecules, and carboxy terminal regional (COOHI. Enzymatic cleavage sites are indicated. The darkly shaded areas represent the hypervariable regions in which the idiotypic determinants are located. (Reprinted with permission.‘)

ties. For example, Oudin and Cazenave12 reported that rabbits immunized with ovalbumin produced immunoglobulins with shared idiotypes, only some of which bound to ovalbumin. We will be describing other examples of this phenomenon in relation to DNA antibody idiotypes. The apparent broad diversity among autoantibodies detectable in the sera of patients with systemic lupus erythematosus (SLE) has been studied by analyses of both their antigen binding and idiotypic repertoires. In this report, we will concentrate on DNA antibody idiotypes, notably one designated as 1616 (1616 Id). Studies of these idiotypes have indicated that lupus antibodies are more restricted than had been thought, have contributed to an increased understanding of the immunopathology of lupus, have suggested some new therapeutic strategies, and have revealed important implications concerning the relationship between infection and autoimmunity.

Derivation of DNA Antibody Monoclonal Antibodies Using a human-human hybridoma technique, 60 IgM monoclonal anti-DNA antibodies were produced from the peripheral blood lymphocytes of seven unrelated patients.13*14 The peripheral blood lymphocytes were fused with a human

The 1616 and 32115 monoclonal antibodies were injected into rabbits and mice. Following extensive absorptions on IgG/IgM sepharose columns and many control experiments, three specific anti-idiotypic reagents were prepared; two polyclonal antibodies in the rabbits (R-anti 16/6 and R-anti 32/ 15)) and one monoclonal in the mouse (M-anti 32/15).18 More than 50% of the binding of R-anti 32/ 15 to plates coated with 32/15 itself was inhibited by ssDNA and two synthetic polynucleotides, poly thymidylic acid (poly [dT]) and poly inosinic acid (poly [I]). These polynucleotides consist of repeating units of thymidine 5’-phosphate and inosine 5’-phosphate, respectively. Similarly, the binding of M-anti 32/l 5 to 32/ 15 was more than 50% inhibited by poly (dT) and poly (I), but only 30% with ssDNA. In contrast, R-anti 16/6 binding to 16/6 was more than 50% inhibited by poly (1) and approximately 40% by ssDNA and poly (dT). Thus, it is evident that both the 16/6 and 32/l 5 idiotypes are at, or very close to, the antigen binding sites on DNA antibodies.

Idiotypic Analysis of Human Monoclonal Anti-DNA Antibodies Using these three anti-idiotypic antibodies, the 60 original human hybridoma derived monoclonal anti-DNA antibodies were screened for cross-reacting idiotypes. Twenty of these 60 antibodies failed to react with any of the three anti-idiotypic reagents and were thus defined as having “private” idiotypes.‘* In contrast, 30 monoclonal antibodies (originating from four unrelated patients) reacted with R-anti 16/6, 22 (from three patients) reacted with R-anti 32/ 15,

DNA ANTIBODY IDIOTYPES: A REVIEW

and 26 (from six patients) reacted with M-anti 32/15. Among the 40 monoclonal antibodies showing evidence of binding to these antiidiotypes, 15 reacted with one anti-idiotype only, ten reacted with two anti-idiotypes, and 15 reacted with all three. Thus, the 16/6 and 32/ 15 idiotypes may be thought of as common, dominant, or public. These results are in agreement with other studies showing idiotypic cross-reactivity among monoclonal anti-DNA antibodies’9V20 as well as other autoantibodies, including anti-acetylcholine receptor antibodies” and anti-Sm antibodies.”

Serologic Studies of Anti-DNA Antibody Idiotypes

Many analyses of DNA antibody idiotypes in the lupus prone mouse models have been reported and are reviewed elsewhere.21 In general, they have all reported substantial idiotype sharing among both monoclonal and serum antiDNA antibodies. A study of 98 SLE patients revealed increased serum levels of the 16/6 Id in 46 cases, compared with four slightly elevated levels in 96 healthy controls.22 When the patients were divided into those with and without active disease, 40 of 74 patients (54%) with active disease were found to have increased levels compared with six of 24 patients (25%) whose disease was in remission. Studies of the same patients with M-anti 32/ 15 showed elevated serum levels of the 32/15 Id in 21 of 74 clinically active patients (28%) compared with one of 24 patients (4%) with inactive disease. In studies of serially collected specimens, notable correlation of the 16/6 Id level with disease activity was found in eight of 12 patients studied. In both this and subsequent studies,23 it was noted that the 16/6 Id level sometimes reflected disease activity better than the DNA binding (Fig 2). We also have evidence that the 16/6 idiotype is not confined to anti-DNA antibodies. This is, in part, based on experiments showing that after immuno-absorption of serum antiDNA antibodies, the 16/6 Id could still be detected.24 These results support the idea that anti-DNA antibodies are of restricted diversity. The results are broadly in agreement with those of Diamond and Solomon, who showed that approximately

247

”

SEP

DCT

ND”

DEC JAN

FEB MAR APR

MAY

,“N

JUL

AUC

Fig 2. In patient EM, the 16/6 Id and ssDNA levels are high during both periods of clinical relapse in contrast to the dsDNA levels. (Reprinted with permission.“)

80% of their SLE patients with anti-DNA activity expressed an idiotype designated as 31.25This idiotype was not associated with the DNA binding site. However, the TOF anti-DNA antibody idiotype identified by Zouali and Eyquem was binding site related and was detected in 31 of 34 patients tested.26 These observations, showing the existence of common anti-DNA antibody idiotypes in humans, imply that a restricted number of germ line genes must exist among the lupus population. Genetic Preponderance of the 16/6 Idiotype

Two recent reports have demonstrated the presence of cross-reactive idiotypes in the first degree relatives of lupus patients. Halpern et alz7 found that 15 of 19 family members had high titered reactivity with an anti-idiotypic antibody measuring the 31 idiotype. In our own study, we found idiotypes 16/6 and 32115 in 24% and 7%, respectively, of 147 first degree relatives of 48 lupus patients.*’ Since these two idiotypes were first identified on antibodies that bind primarily to ssDNA, we also examined the sera of lupus relatives for an idiotype, designated as 134, which was found on antibodies binding to dsDNA antibodies.29 In this study, 45% of the patients and 30% of their first degree relatives were found to have the 134 idiotype. These observations suggest that the common anti-DNA antibody idiotypes cannot be automatic pathogenic factors on their own. However, in a family with an inherited complement C4 deficiency, we observed one asymptomatic relative of a lupus patient with a striking elevation of the serum

248

16/6 Id level who subsequently SLE.30

SHOENFELD AND ISENBERG

developed

overt

Relationship Between Common Anti-DNA Antibody Idiotypes in “Mice And Men” Two other recent studies have shown that common anti-DNA antibody idiotypes may be shared by both lupus prone mice and human lupus patients.3’.32 The 16/6 Id, for example, was found on four of 15 mouse monoclonal anti-DNA antibodies tested, using a mouse monoclonal 16/6 antibody. These observations imply notable evolutionary conservation of V genes that code for antibodies binding to DNA, and that perhaps become diversified by somatic mutation.

Involvement of 1616 Idiotype in Tissue Damage Studies have also been performed to demonstrate that cross-reactive anti-DNA antibody idiotypes can be found on tissue bound immunoglobulins from kidney and skin biopsies of SLE patients.33*34 In the renal study, 11 of 26 biopsies (42%) had the 16/6 Id demonstrable in the glomerular basement membrane (Fig 3) focal tuft proliferation, or in the mesangium (Fig 4). In five of these biopsies (19%), the 32/ 15 Id was also detected. In contrast, neither of the idiotypes could be detected on immunoglobulins deposited in the kidneys of 24 disease control biopsies. Using four polyclonal and two monoclonal anti-idiotypic reagents to screen skin biopsies from 24 SLE patients, up to 45% of the immunoglobulins at the dermal epidermal junction were found to share idiotypes (Fig 5). In comparison, up to 30% of 23 discoid lupus biopsies also shared these idiotypes, unlike any of the 15 immunoglobulin positive disease controls.

Fig 3. 16/6 Id identified in the glomerular basement membrane of a lupus patient using a polyclonal rabbit anti-idiotypic antibody.

1616 Id identified in the mesangium of a lupus Fig 4. patient using a polyclonal rabbit anti-idiotypic antibody.

The demonstration of these common idiotypes at the site of the lupus renal lesions strongly suggests that they are involved in the immunopathology of the disease. However, the same cannot be said of the skin, in which the idiotypes (and immunoglobulins in general) were found in both lesional and non-lesional skin. The immunopathology of the skin lesions in SLE remains obscure. For example, it has never been demonstrated unequivocally that the immunoglobulins at the dermal-epidermal junction contain antiDNA antibodies. In Table 1, the frequency with which the 16/6 Id and 32115 Id have been found among human monoclonal anti-DNA antibodies, serum lupus antibodies, and tissue bound antibodies from lupus patients is shown.

Fig 6. Binding of R anti-16/6 to the dermal-epidermal junction (arrowed) in a skin biopsy from a patient with SLE. There is also non-specific binding in the epidermis (staining with normal rabbit serum gives the latter, but not former, staining pattern).

249

DNA ANTIBODY IDIOTYPES: A REVIEW

Table 1. H”lm”

Monoclo”al And-DNA

IdiotVpe

Antibodies

(%I

Human

LUPUS

LUPUS

LUPUS

scna(%)

Renal 9iisv (%I

Biopsy (%) 46

Skin

16/8

52

47

42

R-anti 32115

39

4

NT

17

M-anti

32115

43

22

19

NT

Abbreviation:

NT, not tested.

R-anti

Common Anti-DNA Antibody Idiotypes, Infection, and Autoimmunity

Atkinson et a13’ and Naparstek et al36 have demonstrated marked similarities between human-hybridoma derived antibodies bearing the 16/6 Id and a monoclonal IgM (WEA) that was derived from a patient with Waldenstrom’s macroglobulinaemia. The lupus antibodies and WEA bound DNA and the Klebsiella polysaccharide K30, as well as possessing the 16/6 Id. A striking similarity in the amino acid sequences was demonstrated in the light chains of these antibodies (though to a lesser extent in the heavy chains). We have followed up these interesting data in two recent studies. In the first, an experiment was carried out in which peripheral blood mononuclear cells (PMN) from six normal subjects were stimulated with five polyclonal activators (Epstein-Barr virus, pokeweed mitogen, group-A Streptococcus, Staphlyococcus, and Klebsiel!a) .37 After seven days of incubation, significantly increased levels of the 16/6 Id (measured by enzyme linked immunosorbent assay [ELISA]) secreted by the cells, compared with unstimulated cells, were only recorded by the Klebsiella stimulated cells. These results are complimented by the second observation that an increased incidence of high titers of 16/6 Id (and anti-polynucleotide activity) was found in the sera of patients with Klebsiella infections, compared to patients with other gram negative infections and normal controls.38 Mycobacteria may also be involved in autoimmunity. Using ELISA assays, approximately 30% of patients with untreated tuberculosis were found to have anti-DNA antibodies.39 Monoclonal anti-DNA antibodies derived from mice and patients with lupus have been found to bind the three glycolipid fractions of mycobacteria cell walls, and mouse monoclonal anti-mycobacteria tuberculosis antibodies reacted with ssDNA and other polynucleotidesa Further-

more, one of the anti-mycobacterial tuberculosis antibodies was found to possess the common anti-DNA 16/6 idiotype. In another study, 34 of 57 sera (60%) with active untreated pulmonary tuberculosis had an increased level of this idiotype.4’ In an appropriate autoimmune setting, clearly a complex of immunogenetic, hormonal, and dietary factors,42,43 it now seems reasonable to postulate that either Klebsiella infection itself or the presence of an internal image of the appropriate Klebsiella antigen could be one of the trigger factors associated with a flare of the disease. 16/6 IDIOTYPE IN NORMAL SUBJECTS

As with other autoantibodies,” it is evident that far from being forbidden, those antibodies bearing the 16/6 and other common anti-DNA antibody idiotypes are an integral part of the normal physiology of the immune system. It is not suprising that hybridomas derived from normal individuals4’ and healthy animals46 have been shown to possess ligand (antigen) binding and idiotypic profiles similar to monoclonal antibody 16/6 and/or other monoclonal anti-DNA antibodies derived from SLE patients or lupus mouse models. As alluded to earlier, it has become apparent that although the 16/6 Id was first identified on IgM anti-DNA antibodies, it is not confined to them. In addition to its detection in some normal sera in which no DNA binding was evident, the 16/6 Id has also been identified in the sera of 23 of 265 patients (8.7%) with monoclonal gammopathies.47 An important observation in this study was that the 16/6 Id was found on IgG immunoglobulins as well as IgM. Lymphocytotoxic antibodies from SLE patients have also been found to bear the 16/6 idiotype.17*48Finally, Datta et a149 also reported the in vitro production of 16/6 Id by lymphocytes of normal subjects and lupus patients. Using biosynthetic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis, their findings indicate that the 1616 Id exists on two distinct populations of antibodies. The first population, conserved in normal individuals, is of uncertain antigenic specificity (though it is tempting to suggest that they are directed against common environmental pathogens), and it dominates the 16/6 Id set that appears after PWM stimulation of normal indi-

SHOENFELD AND ISENBERG

250

viduals. In contrast, the second population is found on antibodies binding to nucleic acids and becomes prominent during a clinical relapse of SLE. If the presence of self-recognizing lymphocytes and autoantibodies is a normal phenomenon, what is the difference between healthy individuals and those suffering from autoimmune disease? It is conceivable that the difference is quantitative. However, several studies have shown the existence of large amounts of autoantibodies in various situations without any overt disease, those individuals with benign monoclonal gammopathies already referred to, for example.47

Immunomodulation

of the 1616 Idiotype

Schattner et also have shown that the production of the 16/6 Id from PWM-stimulated normal lymphocytes requires T cells and is preferentially enhanced by a interferon (though not by y interferon). They have also found that in some SLE patients and their first degree relatives the expression of the 16/6 Id was strongly correlated with abnormal T suppressor cell function (unpublished observations, June 1986). Whether a naturally occurring anti- 16/6 idiotypic antibody is important in regulating the 16/6 Id awaits the development of a reliable assay for measuring the anti-idiotype. These observations allow the concept shown in Fig 6 to be postulated. Thus, we envisage that under normal circumstances, the 16/6 Id is concerned with immunoregulation and protection against bacterial infection, but that during active SLE, a shift occurs and the idiotype is then found on DNA binding antibodies, which may well be concerned with the immunopathology of the disease.

Implications for Therapeutic Strategies It is tempting to speculate that down regulation of antibodies bearing common idiotypes might have a beneficial effect for patients with SLE. Hahn and Ebling” reasoned that immunizing lupus prone mice with a syngeneic (from the same species) monoclonal anti-DNA antibody bearing a high frequency idiotype would induce the production of an immunoregulatory antiidiotype antibody. Thus, they administered repeated injections of an IgG 2a monoclonal anti-DNA antibody to MZB/NZW fl female mice from the age of 6 weeks. The treatment

Fig 6.

Suggested

role of 1616 in health and disease.

resulted in the temporary suppression of circulating anti-DNA antibodies, a reduction in proteinuria, a reduction in the deposition of anti-DNA antibodies in the glomeruli, and fewer deaths from nephritis. More recently, Zouali et al’* showed that by administering a syngeneic antiDNA IgG together with a synthetic immunoadjuvant, a major reduction of both idiotype expression and total DNA antibody levels could be achieved. In a form of reciprocal experiment, Hahn and Ebling’3 were also able to show temporary suppression of antibodies to DNA and significantly prolonged survival due to a delay in the onset of nephritis by innoculating NZB/NZW fl mice with a monoclonal anti-idiotypic antibody. However, as Bluestone et als4 have commented, the in vivo administration of xenogeneic (from a different species) anti-idiotypes has generally been found to induce the expression of idiotypic molecules rather than to suppress them. For example, anti-idiotypic therapy in MRLlpr/lpr mice, which was directed against the high frequency idiotype H 130, resulted in the augmentation of H 130 idiotype and anti-DNA antibody levels.” In an effort to overcome this problem while using the valuable targeting abilities of anti-idiotypes, Saski et als6 were able to selectively eliminate anti-DNA antibody producing cells in in vivo experiments by conjugating an anti-idiotypic antibody with the cytotoxic agent, neocarzinostatin. Having determined that the serum 16/6 Id titer correlates with disease activity in a patient with SLE, it is interesting to postulate whether passing their serum over a column containing an anti- 16/6 antibody might be of some benefit. SUMMARY

The initial studies of anti-DNA antibody idiotypes we performed, along with those of our

DNA ANTIBODY

IDIOTYPES:

A REVIEW

251

colleagues and other groups, focused on the narrow question of their relevance to lupus autoantibodies. The subsequent studies in this report have forced us to examine a much broader range of issues. It is evident that despite the great advances in understanding the structure and function of antibodies, lymphocytes, and receptors, our knowledge of many fundamental elements in autoimmune disease is woefully incomplete. We are still unsure whether the germ line gene sequences controlling antibody production have evolved solely in response to exposure to new foreign antigens. Alternatively, these anti-

bodies (and the idiotypes they bear) may have developed largely in response to changes in the internal environment. Superficially, it can be argued that “self reactivity” associated with the clinical expression of a disease results from a combination of immunologic, genetic, hormonal, and environmental elements. For example, the expression of the 16/6 Id in an appropriate setting may have pathogenetic consequences for some individuals. However, our knowledge of the precise sequence of events that result in devastating disease for some but minimal disease for others is just one of the remaining mysteries.

REFERENCES 1. Klein J: Immunology: The Science Discriminators. New York, Wiley, 1982

of Self-Non-Self

2. Oudin J, Michel M: Une nouvelle forme d’allotypie des globulines du serum de lapin, apparement liee a la fonction et a la specificite anticorps. CR Seances Acad Sci 257:805-808, 1963 3. Kunkel HG, Mannick M, Williams RC: Individual antigenic specificity of isolated antibodies. Science 140: 12181219, 1963 4. Jerne NK: Towards a network therapy system. Ann Immunol 125373-389, 1974

of the immune

5. Cooke A: Idiotypes, anti-idiotypes and autoimmune disease. Plasma Ther Trans Tech 7: 16l- 170, 1986 6. Male lmmunol65:

DK: Idiotypes I-9, 1986

and

autoimmunity.

7. Zanetti M: The idiotype network cess. Immunol Today 6:299-302, 1985

Clin

in autoimmune

Exp pro-

8. Altevogt P, Wigzell H: A VH-associated idiotype in human anti-tetanus antibodies. Stand J Immunol 17:183192, 1983 9. Bose R, Marsh DC, Duchateau J, et al: Demonstration of auto anti-idiotypic antibody cross-reacting with public idiotypic determinants in the serum of rye-sensitive allergic patients. J Immunol 135:2474-2478,1984 10. Pisetsky

DS, Semper

analysis of a monoclonal 133:2085-2089, 1984

KF, Eisenberg anti-Sm

antibody.

RA: Idiotypic J Immunol

11. Lefvert AK, James RW, Alliod C, et al: A monoclonal anti-idiotypic antibody against anti receptor antibodies from myasthenia sera. Eur J Immunol 12:790-792, 1982

lupus antibodies. Clin Immunol Immunopathol 31:261-271, 1984 16. Shoenfeld Y, Smorodinsky I, Lavie Cl, et al: Human lupus autoantibodies bind to Raji cells. Immunol Lett 11:121-126, 1985 17. Shoenfeld Y, Zamir R, Joshua H: Human monoclonal anti-DNA antibodies react as lymphocytotoxic antibodies. Eur J Immunol 15:1024-1028, 1985 18. Shoenfeld Y, Isenberg DA, Rauch J, et al: Idiotypic cross-reactions of monoclonal human lupus autoantibodies. J Exp Med 158:7 18-730, 1983 19. Marion TN, Lawton AR III, Kearney JR, et al: Anti-DNA autoantibodies in (NZB x NZW) Fl mice are clonally heterogeneous, but the majority share a common idiotype. J Immunol 128:668-674, 1982 20. Rauch J, Murphy E, Roths JB, et al: A high frequency idiotypic marker of anti-DNA autoantibodies in MRL-lpr/ Ipr mice. J Immunol 129:236-241, 1982 21. Isenberg DA, Shoenfeld Y, Schwartz RS: The use of hybridomas to analyse autoimmunity. Br Med Bull 40:262266,1984 22. Isenberg DA, Shoenfeld Y, Madio MP, et al: AntiDNA antibody idiotypes in systemic lupus erythematosus. Lancet 2:4 17-422, 1984 23. Isenberg DA, Colaco CB, Dudeney C, et al: A study of the relationship between anti-DNA antibody idiotypes and anticardiolipin antibodies with disease activity in systemic lupus erythematosus. Medicine 65:46-55, 1986 24. Madaio MP, Schattner A, Schattner M, et al: Lupus serum are normal human serum containing anti-DNA antibodies with the same idiotypic marker. J Immunol 137:25352560, 1986

12. Oudin J, Cazenave PA: Similar idiotypic specificities in immunoglobulin fractions with different antibody functions or even without detectable antibody function. Proc Natl Acad Sci USA 68:26 16-2620, 197 1

25. Diamond B, Solomon G: A monoclonal antibody that recognizes anti-DNA antibodies in patients with systemic lupus erythematosus. NY Acad Sci 418:379-385, 1983

13. Shoenfeld Y, Hsu-Lin SC, Gabriels LE, et al: Production of autoantibodies by human-human hybridomas. J Clin Invest 70:205-208, 1982

26. Zouali M, Eyquem A: Idiotypic restriction in human autoantibodies to DNA in systemic lupus erythematosus. Immunol Lett 7:187-193, 1984

14. Shoenfeld Y, Rauch J, Massicote H, et al: Polyspecificity of monoclonal lupus autoantibodies produced by human-human hybridomas. N Engl J Med 308:414-420, 1983

27. Halpern R, Davidson A, Lazo A, et al: Familial systemic lupus erythematosus. Presence of a cross-reactive idiotype in healthy family members. J Clin Invest 76:731736, 1985

15. Andre-Schwartz Binding of cytoskeletal

28. Isenberg DA, Shoenfeld Y, Walport tion of cross-reactive anti-DNA antibody

J, Datta SK, Shoenfeld Y, et al: proteins by monoclonal anti-DNA

M, et al: Detecidiotypes in the

SHOENFELD AND ISENBERG

252

serum of patients and their relatives.

who have systemic lupus erythematosus Arthritis Rheum 24:256-262, 1985

29. Dudeney C, Shoenfeld Y, Rauch J, et al: A study of anti-poly(ADP-ribose) antibodies and an anti-DNA antibody idiotype and other immunological abnormalities in lupus family members. Ann Rheum Dis 45:502-507, 1986 30. Shoenfeld Y, Brill S, Weinberger A, et al: High levels of a common anti-DNA idiotype (16/6), a genetic marker for SLE. Acta Haematol 76, 107-109, 1986 3 1. Morgan A, lsenberg DA, Naparstek Y, et al: Shared idiotypes are expressed on mouse and human anti-DNA antibodies. Immunology 56:393-400, 1983 32. Eilat D, Fischel R, Zlotnick A: A central anti-DNA idiotype in human and murine systemic lupus erythematosus. Eur J Immunol 15:368-375, 1985 33. lsenberg DA, Collins C: Detection of cross-reactive anti-DNA antibody idiotypes on renal tissue bound immunoglobulins from lupus patients. J Clin Invest 76:287-294, 1985 34. lsenberg DA, Dudeney C, Wojnaruska F, et al: Detection of cross-reactive anti-DNA antibody idiotypes on tissue bound immunoglobulins from skin biopsies of lupus patients. J lmmunol 135:261-264.1985 35. Atkinson PM, Lampman GW, Furie BC, et al: Homology of the NHZ-terminal amino acid sequences of the heavy and light chains of human monoclonal lupus autoantibodies containing the dominant l6/6 idiotype. J Clin Invest 75:1138-1143, 1985 36. Naparstek Y, Duggan D, Schattner A, et al: Immunochemical similarities between monoclonal antibacterial Waldenstrom’s macroglobulins and monoclonal anti-DNA lupus autoantibodies. J Exp Med 161:1525-1538, 1985 37. El-Roiey A, Gross WL, Ludemann J, et al: Preferential secretion of a common anti-DNA idiotype (16/6 Id) and anti-polynucleotide antibodies by normal mononuclear cells following stimulation with klebsiella pneumoniae. lmmunol Lett 12:313-319, 1986 38. El-Roiey A, Sela 0, Isenberg DA, et al: The sera of patients with klebsiella infections contain a common antiDNA idiotype (16/6 Id) and polynucleotide activity. Clin Exp lmmunol (in press) 39. lsenberg DA, Maddison P, Swana G, et al: Profile of autoantibodies in the serum of patients with tuberculosis, klebsiella and other gram negative infections. Clin Exp lmmunol (in press) 40. Shoenfeld Y, Vilner Y, Coates ARM, et al: Monoclonal anti-TB antibodies react with DNA and monoclonal anti-DNA autoantibodies react with mycobacterium tuberculosis. Clin Exp lmmunol66:255-261, 1986 41. Sela I, El-Roiey

A, Isenberg

DA, et al: A common

anti-DNA idiotype in the sera of patients with active pulmonary tuberculosis. Arthritis Rheum (in press) 42. Shoenfeld Y, Schwartz RS: Immunologic and genetic factors in autoimmune diseases. N Engl J Med 311:10191029, 1984 43. Morrow WJW, Ohashi Y, Hall J, et al: Dietary fat and immune function in (NZB/NZW) Fl mice. J Immunol 135:3857-3863, 1985 44. Graber P: Autoantibodies and the physiologic role of immunoglobulins. Immunol Today 4:337-341, 1983 45. Cairns EJ, Block J, Bell DA: Anti-DNA autoantibody producing hybridomas of normal human lymphoid cell origin. J Clin Invest 74:880-887, 1984 46. Monier JC, Brochier J, Moreira A, et al: Generation of hybridoma antibodies to double-stranded DNA from nonautoimmune Balb/c strain: Studies on anti-idiotype. Immunol Lett 8:6 I-68, 1984 47. Shoenfeld Y, Ben-Yehuda 0, Naparstek Y, et al: The detection of a common idiotype of anti-DNA antibodies in the sera of patients with monoclonal gammopathies. J Clin Immunol6:194-203, 1986 48. Lydyard PM, lsenberg DA, De Sousa B, et al: A cross reactive idiotypc on anti-DNA and lymphocytotoxic antibodies. Clin Exp lmmunol (in press) 49. Datta SK, Naparstek Y, Schwartz RS: In vitro production of an anti-DNA idiotype lymphocytes of normal subjects and patients with systemic lupus erythematosus. Clin Immunol lmmunopathol 38:302-308, 1986 50. Schattner A, Duggan D, Naparstek Y, et al: Effects of interferon on the expression of a lupus idiotype in normal humans, Clin lmmunol Immunopathol38:327-336, 1986 51. Hahn BH, Ebling FM: Suppression of NZB/NZW murine nephritis by administration of a syngeneic monoclonal antibody to DNA. J Clin Invest 71:1728-1736, 1983 52. Zouali M, Jolivet M, Leclerc C, et al: Suppression of murine lupus autoantibodies to DNA by administration of muramyl dipeptide and syngeneic anti-DNA IgG. J Immunol 135:1091-1096, 1985 53. Hahn BH, Ebling FM: Suppression of murine lupus nephritis by administration of an anti-idiotypic antibody to anti-DNA. J lmmunol 132:187-190, 1984 54. Bluestone JA, Leo 0, Epstein SL, et al: Idiotypic manipulation of immune response to transplantation antigens. Immunol Rev 90:5-27, 1986 55. Teitelbaum D, Rauch J, Stollar BD, et al: In vivo effects of antibodies against a high frequency idiotype of anti-DNA antibodies in MRL mice. J Immunol 132:12821285,1984 56. Saski T, Muryoi T, Tamate E, et al: Selective elimination of anti-DNA antibody producing cells by antidiotypic antibody conjugated with neocarzinostatin. J Clin Invest 77:1382-1386, 1986