Idiotypic Cascades Associated with the CD4-HIV gpl20 Interaction: Principles for Idiotype-Based Vaccines

Idiotypes (Id) are antigenic determinants expressed on the variable (V) region of the immunoglobulin molecule. Id-bearing antibodies, or Ab-1, are produced upon stimulation with a given antigen. Ab-1 may elicit the production of anti-idiotypic antibodies (anti-Id) or Ab-2. The anti-Id also expresses Id determinants and may in turn elicit the production of anii-anti-Id or Ab-3. The production of Ab-1, Ab-2, and Ab-3 responses resulting from stimulation with the antigen is representative of components within an Id cascade. The existence of this Id cascade is the basis for the development of Id based strategies for controlling the immune response to infectious agents and tumors. In this review we will focus on several aspects regarding the Id cascades that may be operational during the immune response to the human immunodeficiency virus (HIV). In light of several studies which suggest the existence of Id-anti-Id interactions operating during the course of HIV infection, we will discuss the potential applications of Id based strategies in manipulating the immune response to HIV.     

Administration of noninternal image monoclonal anti-idiotypic antibodies induces idiotype-restricted responses specific for human immunodeficiency virus envelope glycoprotein epitopes.

A mouse monoclonal anti-idiotypic antibody (anti-Id), designated MC1, was generated against chimpanzee antibodies specific for a synthetic peptide corresponding to a native epitope associated with gp41 of human immunodeficiency virus (HIV). This anti-Id recognized a shared idiotope/idiotype (Id) on a second chimpanzee anti-gp41 peptide preparation but failed to detect this Id on rabbit and mouse anti-gp41 peptide antibodies induced by immunization with the gp41 synthetic peptide. The chimpanzee Id-MC1 reaction was not inhibited by either synthetic peptide or recombinant gp160 suggesting that MC1 exhibits noninternal image, Ab-2 alpha-like characteristics. Immunization of syngeneic Balb/c mice with MC1 induced an antigen-positive (Ag+) response capable of binding the synthetic peptide, recombinant gp160, and gp41, whereas MC1-immunized rabbits did not produce any detectable anti-peptide and/or anti-HIV envelope glycoprotein antibody response. The MC1-induced anti-Id response (Ab-3) in both mice and rabbits expressed a similar Id with the Ab-1, which is not normally expressed in the anti-gp41 peptide antibody response induced by the nominal antigen in Balb/c mice and in rabbits. Together, these studies indicate that a mouse monoclonal anti-Id of the Ab-2 alpha class can induce an anti-HIV response specific for a gp41 epitope defined by a synthetic peptide, which does not cross species barriers.     

Characterization of private and cross-reactive idiotypes associated with human antibodies to hepatitis B surface antigen.

Four mouse monoclonal anti-idiotypic antibodies (anti-Id) were generated against human monoclonal and polyclonal antibodies to hepatitis B surface antigen (HBsAg). These monoclonal anti-Id, along with a polyclonal anti-Id raised in rabbits, were used to characterize the idiotype (Id) specificity of the human antibody response to HBsAg (anti-HBs). The anti-Id reagents identified distinct private and cross-reactive Id expressed on monoclonal and polyclonal human anti-HBs preparations respectively. The anti-Id recognized both HBsAg combining site and non-combining site related private Id, and HBsAg combining site related cross-reactive Id. The Id specificities recognized by two of the monoclonal anti-Id were associated with the H chain alone, whereas two of the monoclonal anti-Id, along with the polyclonal anti-Id appeared to recognize Id determinants associated with both isolated H and L chains. These data suggest that Id heterogeneity exists within the human antibody response to HBsAg. The knowledge that Id heterogeneity exists is of importance in understanding the observed variability in the immune response during hepatitis B virus infection.     

Genetic control of the immune response to staphylococcal nuclease. XI. Effects of in vivo administration of anti-idiotypic antibodies

The effects of prior treatment with heterologous anti-idiotypic antibodies on the response to staphylococcal nuclease (Nase) have been examined. Previous studies have shown that 100% of A/J mice treated with Nase in completes Freund's adjuvant produce anti-Nase antibodies possessing a characteristic idiotype (Id). Mice treated with anti-Id antibodies followed by Nase produced levels of Id equal to or greater than those of control animals treated with Nase alone. The appearance of Id in treated mice preceded the appearance of anti-Nase activity, and animals treated with anti-Id alone produced high levels of Id without detectable anti-Nase activity. Id expression in such animals could be detected using anti-Id reagents produced in several different species suggesting that it represented true idiotope expression rather than unrelated molecules reactive only with the anti-Id reagent used for initial treatment. Isolation of the nonantigen-binding Id-bearing molecules (Id') showed them to be immunoglobulins bearing the same idiotopes as do anti-Nase antibodies. However, quantitative comparisons of Id levels vs. amount of Id or Id'-bearing immunoglobulin suggested that the nonantigen-binding immunoglobulins bore fewer idiotopes per molecule than did anti-Nase antibodies. Evidence was also obtained for the production of some nonantigen-binding Id-bearing molecules during the normal immune response Nase. These findings are therefore consistent with the existence of a network of Id-anti-Id interactions in the immune response to Nase.     

Moloney leukemia virus-induced cell surface antigen mimicry by monoclonal antibodies

We have investigated antigen-independent modulation of immune responses by monoclonal antibodies directed against both viral and nonviral antigens. BALB/c mice were immunized with monoclonal IgM (i.e. Ab1) specific for either Moloney murine leukemia virus-induced cell surface antigen (MCSA) or the hapten 2,4-dinitrophenyl (DNP). Injection with either Ab1 activated a functional idiotypic (Id) network as evidenced by production of both anti-Id (Ab2) antibodies and anti-anti-Id (Ab3) antibodies. A subset of induced Ab3 (designated Ab1), exhibited specificity for antigen (virus or DNP). In mice immunized with anti-Id antibodies (Ab2), production of Ab3 and Ab1′ was also observed. In the MCSA system, antibody-induced Ab1′ responses were effective in protecting mice from tumor development upon subsequent challenge with live virus. Furthermore, antigen-independent modulation of immunity to both viral and nonviral antigens was found to be thymus-dependent. Similar findings in other viral systems suggest that antibody-induced activation of Id networks may prove a viable alternative vaccine strategy that can elicit antigen-specific responses, and in some cases protection, in the apparent absence of exposure to antigen.     

Study of idiotypes expressed by monoclonal antibodies to the 35 kD and 12 kD antigens of Mycobacterium leprae.

Rabbit antisera were raised against four monoclonal antibodies (MoAb) binding with the 35 kD protein and four MoAb binding with the 12 kD protein antigen of Mycobacterium leprae. Antisera showed idiotype (Id) specificity following cross-absorption with normal mouse globulin. One Id on a single MoAb and another Id shared between three MoAb were identified for each group. Functional studies were carried out with the Rb04 anti [anti-35 kD] specificity. The expression of this Id and paratope in antigen immunized mice was associated with Igh alleles. Inoculation of mice with anti-Id Rb04 induced an 'Ab3' serum response of corresponding Id specificity only when the anti-Id was given in emulsion with incomplete Freund's adjuvant (IFA). Conversely, prior injection of soluble anti-Id inhibited the subsequent Ab3 response to Rb04/IFA. Moreover, the suppressive effect of soluble anti-Id was abrogated by prior injection of 50 mg/kg cyclophosphamide. These results indicate that regulatory mechanisms similar to those involved in antigenic stimulation may explain the stimulatory or suppressive potency of anti-Id antibodies. Finally, the Ab3 responses to the two tested anti-Ids did not contain any antigen binding activity.     

Insight into the potential for DNA idiotypic fusion vaccines designed for patients by analysing xenogeneic anti-idiotypic antibody responses

DNA vaccines induce immune responses against encoded proteins, and have clear potential for cancer vaccines. For B-cell tumours, idiotypic (Id) immunoglobulin encoded by the variable region genes provides a target antigen. When assembled as single chain Fv (scFv), and fused to an immunoenhancing sequence from tetanus toxin (TT), DNA fusion vaccines induce anti-Id antibodies. In lymphoma models, these antibodies have a critical role in mediating protection. For application to patients with lymphoma, two questions arise: first, whether pre-existing antibody against TT affects induction of anti-scFv antibodies; second, whether individual human scFv fusion sequences are able to fold consistently to generate antibodies able to recognize private conformational Id determinants expressed by tumour cells. Using xenogeneic vaccination with scFv sequences from four patients, we have shown that pre-existing anti-TT immunity slows, but does not prevent, anti-Id antibody responses. To determine folding, we have monitored the ability of nine DNAscFv-FrC patients' vaccines to induce xenogeneic anti-Id antibodies. Antibodies were induced in all cases, and were strikingly specific for each patient's immunoglobulin with little cross-reactivity between patients, even when similar VH or VL genes were involved. Blocking experiments with human serum confirmed reactivity against private determinants in 26-97% of total antibody. Both immunoglobulin G1 (IgG1) and IgG2a subclasses were present at 1.3 : 1-15 : 1 consistent with a T helper 2-dominated response. Xenogeneic vaccination provides a simple route for testing individual patients' DNAscFv-FrC fusion vaccines, and offers a strategy for production of anti-Id antibodies. The findings underpin the approach of DNA idiotypic fusion vaccination for patients with B-cell tumours.     

Polyclonal anti-idiotypic antibodies exhibit antigenic mimicry of limited type 1 fimbrial proteins of Escherichia coli.

Polyclonal anti-idiotypic antibodies (anti-Ids)(fim) developed against idiotypes on antibodies (Ab-1s) that specifically bind structural, organelle fimbrial proteins of Escherichia coli were able to modulate immune function in anti-Id(fim)-immunized mice. Proliferation or suppression of splenic lymphoid cell responses by polyclonal anti-Ids in tissue culture appeared to be dose dependent. Anti-Ids were able to induce a dose-dependent T-cell-mediated immunity specific for type 1 fimbrial antigen(s) in immunized animals when assessed in vitro, but they failed to elicit in vivo positive ear-swelling skin reactions. Anti-Ids were unable to induce protective immunity against an in vivo infectious challenge with E. coli in anti-Id-immunized adult animals, but they stimulated a specific, secondary antibody response in anti-Id-challenged mice. Anti-Ids stimulated the development of anti-anti-Ids (Ab-3s) specifically binding a fimbrial antigen(s) and revealed the presence of antibody idiotypes binding E. coli adhesin proteins in the 27- to 29-kilodalton range. Results suggest discrete, but subtle, immunomodulatory effects of the anti-Ids and potential vaccinoid properties capable of stimulating a specific humoral and cellular response in vivo.     

Specific antigen/antibody complexes induce the in vivo production of a parallel set of nonantigen-binding idiotype-positive antibodies

Immune complexes prepared with the polysaccharide antigen (PnC) extracted from Streptococcus pneumoniae R36a and two different PnC-specific antibodies were found to differ in their regulatory properties depending on the isotype of the antibody. Thus, complexes formed in antibody excess with TEPC15 (IgA) were suppressive whereas complexes formed with 96-G (IgG3) antibodies enhanced the IgM response to PnC. During the course of these studies, we found that little or no PnC-specific IgG antibody was induced during the response to PnC coupled to sheep red blood cells (PnC-SRBC). Interestingly, however, immunization with 96-G/PnC complexes either alone or with PnC-SRBC resulted in the induction of IgG3 antibodies that express the T15 idiotype (Id) but which do not bind PnC. This unique IgG3 response occurred after injection of 96-G/PnC complexes formed in antibody excess but not when complexes were formed in antigen excess. The Id+ nonspecific IgG3 response peaked on day 5 and could be activated with 96-G/PnC complexes but not with free PnC antigen. The Id+ nonspecific response was not due to polyclonal activation of IgG3 production since there was no difference in IgG3 levels in mice injected with 96-G/PnC complexes with those injected with PnC-SRBC. Finally, mice that had been suppressed for expression of the T15 Id by neonatal injection of anti-Id antibody were able to produce Id+-unspecific IgG3 antibody after immunization with 96-G/PnC complexes, further suggesting that Id+ IgG3 was produced by different clones than those that usually comprise the antibody response to PnC. The results suggest that the formation of IgG immune complexes during an immune response may result in stimulation of idiotypically related clones thus resulting in degeneracy of the immune response.     

Serologic and molecular characterization of the T15 idiotype--I. Topologic mapping of idiotopes on TEPC15

The present study was designed to determine the minimum number of idiotopic determinants (Id) comprising the T15 idiotype and to construct a topological map of Id on the TEPC15 immunoglobulin molecule. Seven monoclonal anti-Id antibodies were used to map determinants by solid phase competition radioimmunoassay. The binding of some anti-Id to T15 was inhibited by phosphorylcholine-protein conjugates, while other anti-Id were not, suggesting that these anti-Id bind to different portions of T15 relative to the paratope. In addition, Id were mapped relative to one another in reciprocal competition assays between the anti-Id for binding to T15 coated plates. The topologic relationships between six Id were discerned by this method. A seventh binding site was identical or so close that it could not be discriminated. These studies suggest that the T15 idiotype is composed of at least six discrete overlapping Id whose locations range from the antigen binding region to the CH1 domain on TEPC15.     

A Murine Monoclonal Anti-Idiotypic Antibody Detects a Common Idiotope on Human, Mouse and Rabbit Antibodies to Allergen Lol p IV

A syngeneic mouse monoclonal anti-idiotypic antibody (anti-Id), designated as B1/1, was generated against a monoclonal antibody (MoAb 91) specific for Ryegrass pollen allergen Lol p IV. This anti-Id recognized an idiotope (Id) that was also present on other monoclonal antibodies with the same specificity as MoAb 91. Observations that (i) the anti-Id inhibited the binding of MoAb 91 to Lol p IV and (ii) the Id-anti-Id interaction could be inhibited by Lol p IV indicated that the Id was located within or near the antigen combining site. These properties served to characterize B1/1 as an internal image anti-Id. Evidence that an immune response in different species to Lol p IV elicits the formation of antibodies which express a common Id was provided by the observations that (i) the Id-anti-Id interactions could be inhibited by mouse, human and rabbit antisera to Lol p IV and (ii) the binding of these antisera to Lol p IV could be inhibited by the anti-Id. Interestingly, the internal image anti-Id B1/1 also recognized an Id on a monoclonal antibody which was directed to an epitope of Lol p IV, different from that recognized by MoAb 91.     

Methods for generating reagents to examine idiotype networks within antiviral immune responses

The use of anti-idiotypic antibodies to examine and/or modulate the immune response to various viral antigens has the potential to be of use in many diverse systems. This paper details the method and immunologic parameters used in our laboratory to generate and characterize anti-idiotypic antibodies (anti-Id or Ab-2) with specificity for antibodies directed against viral antigens. These anti-Id reagents have been used in our laboratory for studies involving the immune responses to hepatitis B virus and simian virus 40, which we describe here, as well as herpes simplex virus, and the human immunodeficiency virus. We have utilized these anti-Id reagents to examine the fine specificity of the idiotypes on antiviral antibodies in these systems and have attempted to modulate or induce specific antiviral immune responses. It is anticipated that the methods described herein will be helpful in analyzing the immune response in other viral systems including studies involving viral-receptor interactions.     

Anti-idiotypic immune responses against adjuvant-free isologous IgM monoclonal antibodies and their augmentation by complex formation between IgM and albumin in bovine serum.

In previous studies we demonstrated that the hypermutated isologous myeloma protein MOPC 315 (isotype IgA; λ2) is recognized by T helper cells like an ordinary foreign protein antigen. To what extent can an immune system recognize and respond to V domains from the primary (pre-immune) repertoire? To study this question we made 21 BALB/c hybridoma anti-2, 4, 6 trinitrophenyl monoclonal antibodies (mAb) of IgM; λ isotype. All mAb purified from supernatants containing fetal bovine serum had formed spontaneous complexes with bovine serum albumin possibly by way of disulfide interchange. Twenty of twenty-one mAb from this source elicited IgG₁ anti-idiotypic (Id) Ab when given as a single adjuvant-free dose of 200 μg. For 12 of them even 10 μg was sufficient. This indicated that BSA augmented the anti-Id responses by a carrier effect. Three of the mAb were therefore purified from ascites fluid and from serum-free medium. Only one of them then induced humoral anti-Id responses in BALB/c mice when given as a single adjuvant-free dose of 100 μg. The other two became immunogenic when emulsified in Freund's complete adjuvant. The results indicate that some IgM mAb exist whose Id determinants alone can elicit substantial anti-Id responses because they are recognized like ordinary foreign protein antigens. Since albumin in fetal bovine serum forms complexes with IgM and greatly augments its immunogenicity, serum-free medium should be used for production of human or humanized therapeutic IgM mAb. A possible role for Id of IgM Ab as cardinal autoantigens is discussed.     

Human immune response to group A streptococcal carbohydrate (A-CHO). III. Comparison of the efficiencies of anti-idiotopic antibody and of nominal antigen in the induction of IgM anti-A-CHO-producing B cells

In this study the efficiencies of a monoclonal anti-idiotopic (Id) antibody (anti-Id498) and of various preparations of nominal antigen in the induction of an antigen-specific human B cell response in vitro were compared. Anti-Id498 recognizes a recurrent, binding site-related Id present on IgM antibodies with specificity for N-acetyl-D-glucosamine, the immunodominant group of streptococcal group A carbohydrate (A-CHO). We have previously shown that anti-Id498 induces IgM anti-A-CHO secretion from B cells of donors that possess Id-498+ antibodies in their serum. A-CHO was presented to B cells either in soluble or insoluble form, i.e. coupled to beads or as intact bacteria. Purified blood B cell populations from three Id+ healthy donors with high numbers of circulating anti-A-CHO B cells were used and antibody-producing B cells are enumerated in a single-cell assay (spot ELISA). The data show that anti-Id498 was superior in the induction of IgM anti-A-CHO-secreting B cells in two donors (factor 4.6 and 13.5 as compared to the most efficient antigenic stimulation). In the third donor antigen stimulation was slightly more efficient than anti-Id but only with Sepharose-bound A-CHO and not with soluble A-CHO or intact bacteria. The increase of specific B cells induced after stimulation with anti-Id498 could be abolished after addition of autologous T cells in two donors. On the contrary, an enhancement of the specific response was observed after addition of autologous T cells in antigen-stimulated cultures. Neither suppression nor enhancement were induced by addition of irradiated T cells.     

ABO-blood-group-related idiotypic network: mimicry of oligosaccharide epitope by rabbit antiidiotypic antibodies to murine monoclonal anti-A antibody.

The idiotypy of antibodies (Ab) specific for oligosaccharide determinants of blood groups of the human ABO system was studied through a cascade. Xenogenic antiidiotypic Ab (Ab2) raised in rabbits to the murine monoclonal anti-A61 (Ab1) were screened for reactivity with various anti-ABH Ab. Three anti-A and three anti-A,B monoclonal antibodies (mAb) which were developed in the same mouse strain as that producing Ab1, as well as a human polyclonal anti-A, were found to share cross-reactive idiotopes (CRI) with Ab1. CRI on murine mAb could be due to a Biozzi recurrent Id on anti-A Ab reacting with anti-Id "à la Oudin", while CRI on human anti-A Ab suggested the presence of paratope-induced anti-Id. Inhibition by Ab2 of haemagglutination of A, B or O human red blood cells by many murine anti-ABH mAb, and by polyclonal or monoclonal human anti-A, strongly supported the occurrence of anti-Id mimicking ABH epitopes belonging to type 2 determinants carried by human erythrocytes. Furthermore, a rabbit immunized with Ab2 produced a potent Ab3 response characterized by anti-H-type-2 specificity. Altogether, these results are consistent with the first successful production of anti-Id Ab that mimics the tridimensional shape of a well defined and strictly carbohydrate epitope, eliciting a haemagglutinating Ab3.     

Heterogeneity of anti-idiotypic antibodies to anti-DNA antibodies in humans.

Serum antibodies in some patients with systemic lupus erythematosus (SLE) were found to have specificity to idiotypes (Id) of 0-81 (human monoclonal anti-single-stranded DNA (ssDNA) antibody) but not to Id of NE-1 (human monoclonal anti-double-stranded DNA (dsDNA) antibody) or pooled human IgM. The interaction of the antibodies and 0-81 was blocked by the co-existence of free ssDNA. Some of SLE sera also showed preferential binding to Id determinants of NE-1, which included the antigen-binding sites of the dsDNA antibody. Some other SLE sera reacted with both Id of 0-81 and NE-13. Thus, there was heterogeneous population among human anti-Id autoantibodies to anti-DNA antibodies. The anti-Id activity was commonly detected in inactive SLE sera, and less frequently in normal controls, suggesting some regulatory role for anti-Id antibodies in the production of autoantibodies.     

ABO-blood-group-related idiotypic network: Mimicry of oligosaccharide epitope by rabbit antiidiotypic antibodies to murine monoclonal anti-A antibody

The idiotypy of antibodies (Ab) specific for oligosaccharide determinants of blood groups of the human ABO system was studied through a cascade. Xenogenic antiidiotypic Ab (Ab2) raised in rabbits to the murine monoclonal anti-A61 (Ab1) were screened for reactivity with various anti-ABH Ab. Three anti-A and three anti-A,B monoclonal antibodies (mAb) which were developed in the same mouse strain as that producing Ab1, as well as a human polyclonal anti-A, were found to share cross-reactive idiotopes (CRI) with Ab1. CRI on murine mAb could be due to a Biozzi recurrent Id on anti-A Ab reacting with anti-Id “à la Oudin”, while CRI on human anti-A Ab suggested the presence of paratope-induced anti-Id. Inhibition by Ab2 of haemagglutination of A, B or O human red blood cells by many murine anti-ABH mAb, and by polyclonal or monoclonal human anti-A, strongly supported the occurence of anti-Id mimicking ABH epitopes belonging to type 2 determinants carried by human erythrocytes. Furthermore, a rabbit immunized with Ab2 produced a potent Ab3 response characterized by anti-H-type-2 specificity. Altogether, these results are consistent with the first successful production of anti-Id Ab that mimics the tridimensional shape of a well defined and strictly carbohydrate epitope, eliciting a haemagglutinating Ab3.     

The primary IgM antibody repertoire: a source of potent idiotype immunogens.

A widely held view is that, to elicit adaptive immune responses, most protein antigens must be given with adjuvants that activate the innate immune system. It has also been proposed that the immune system is tolerant to idiotypes (Id) of the syngeneic primary antibody (Ab) repertoire. We now show that among 73 purified noncomplexed secretory IgM monoclonal antibodies (mAb), 4 (5.5%) elicited high levels of IgG Ab against the Id even though no adjuvant was added. The responses were controlled by H2-linked immune response genes. IgG1, but no IgG2a or IgG2b, anti-Id Ab were detected, indicating involvement of T helper type 2 (Th2) cells. All 4 IgM mAb are likely germ-line gene-encoded, and 1 was shown to represent a recurrent Id. After endotoxin depletion the most potent immunogen of the 4 still provoked robust humoral anti-Id responses. The results suggest that a natural protein of the primary IgM Ab repertoire can be immunogenic without an adjuvant.     

Anti-idiotypic humoral and cellular responses to antibody to hepatitis B surface antigen in hepatitis B viral infections.

In order to investigate regulatory significance of humoral and cellular responses to the idiotypic (Id) determinants on the antibody to hepatitis B surface antigen (anti-HBs), they were studied in acute hepatitis B and in chronic HBV infection. The results were compared with humoral and cellular responses of the same patients to hepatitis B surface antigen (HBsAg). In acute hepatitis B, the responses to HBsAg, were delayed until 3-4 weeks after the onset of clinical symptoms. However, the leucocyte migration inhibition (LMI) and the lymphocyte transformation (LTT) responses to affinity purified anti-HBs were found to be evolved very early in the course of acute hepatitis B, though anti-Id antibodies were absent. The majority of chronic HBV carriers showed a poor humoral and cellular response to HBsAg. Ten out of 38 chronic carriers showed anti-Id antibodies which recognized a major cross-reactive idiotype (CRI) on the anti-HBs molecule. Twenty-five out of 38 chronic carriers also showed LMI response to the Id determinants on the anti-HBs. LMI response induced by anti-HBs could be blocked by a specific Balb/c anti-Id antibody which also recognized the CRI. Thus, in both acute and chronic HBV infections, the anti-Id humoral and cellular responses correlated with poor humoral and cellular responses to HBsAg, indicating regulatory significance.     

Anti-Idiotypes to HLA and their Role in Transplantation

The network theory proposed by Jerne is based on the finding that variable regions of T and B cell antigen receptors are structurally diverse and express unique variable region determinants. We have postulated that idiotypes present on the V region of the anti-HLA antibody molecule (Ab1) can elicit the production of anti-anti-HLA antibodies or Ab2 and that such Ab2 may play a role in the suppression of anti-HLA antibody responses. We first tested the validity of this concept in pregnancy, natures most perfect model of allogeneic tolerance. Our studies revealed that anti-Id antibodies were present during pregnancy at times when HLA antibodies had disappeared from the circulation (1,2). This inverse correlation between Ab1 and Ab2 suggested that Ab2 suppresses the production of Ab1. Once the validity of this concept was substantiated in the model of pregnancy, we tried to determine whether anti-Id antibodies to HLA also play a role in the down regulation of the alloimmune response to transfusions and transplantation. For this, we monitored patients for the development of anti-anti-HLA antibodies following donor specific blood transfusion (3). We found that most patients developed Ab2 to the mismatched HLA class II antigens of the blood donor two weeks following transfusion and were associated with successful transplantation. Furthermore, we found a positive correlation between the presence of anti-anti-HLA antibodies and tolerance to the graft in patients with a history of presensitization to HLA antigens of the donor (4). The role of anti-Id antibodies to HLA was also evaluated in renal and heart transplantation (5,6,7). In these studies, patients were monitored following transplantation for the production of Ab2. We found that 5 year actuarial survival of heart and kidney allografts was significantly higher in patients developing Ab2, compared to patients without Ab2. Taken together, our data suggest that anti-Id antibodies to HLA play an important role in suppressing the immune response to HLA and that such anti-Id may be of therapeutic interest in transplantation.