A Nonadjuvanted IgG2a Monoclonal Antibody against Nucleosomes Elicits Potent T Cell-Dependent, Idiotype-Specific IgG1 Responses and Glomerular IgG1/IgG2a Deposits in Normal Mice.

Idiotypes (Ids) are unique epitopes of Ab V regions and can trigger anti-Id immune responses, but immunization with several nonadjuvanted isologous IgG mAbs has induced tolerance to their Ids. We immunized non-lupus-prone mice with 11 allotype "a" of IgG2a (IgG2a) and 4 IgG2c nonadjuvanted, isologous mAbs purified from serum-free medium. Of five IgG2a mAbs with specificity for nucleosomes, the repeating histone-DNA subunit of chromatin, four elicited an IgG1 anti-mAb response and one mAb was nonimmunogenic.

The MHC haplotype H2b converts two pure nonlupus mouse strains to producers of antinuclear antibodies.

Studies of mouse lupus models have linked the MHC H2(b) haplotype with the earlier appearance of antinuclear autoantibodies and the worsening of nephritis. However, it is unknown whether H2(b) by itself, in the context of pure nonlupus strains, is "silent" or sufficient with regard to loss of tolerance to chromatin (nucleosomes). In this study we show that, beginning approximately 6-9 mo of age, H2(b)-congenic BALB/c (denoted BALB.

[Testing of ICF core set for low back pain].

Background: International Classification of function, disability and health (ICF) is a globally accepted framework and classification system. ICF core sets for chronic health conditions have been developed to promote implementation of ICF in clinical practice. A preliminary core set for low back pain, with 78 categories, has been developed and proposed for international validation.

Antinucleosome autoantibodies bind directly to cell lines in vitro and via the FcgammaRIIB receptor to B lymphocytes in vivo: a role for immune complexes in interactions between antinucleosome IgG2a and B cells of BXSB lupus mice.

The initial novel observation of this study was that most B cells of male BXSB lupus mice bear surface IgG2a(b) of extrinsic origin. To define the surface antigen, we here examine three (NZBxBXSB)F1-derived IgG2a(b) monoclonal antibodies (mAbs) selected for binding to cell surfaces. Surprisingly, all three mAbs bound the nucleosome (nuc) particle, the fundamental unit of chromatin and an early target of autoimmunity in systemic lupus erythematosus.

Immunoglobulin heavy chain constant regions regulate immunity and tolerance to idiotypes of antibody variable regions.

Particular syngeneic adjuvant-free monoclonal antibodies are immunogenic and elicit antibody responses against the variable region idiotypes (Ids). We here study how heavy-chain constant regions (C(H)) regulate immune responses to Ids of free, uncomplexed monoclonal antibodies. To this end, we selected two hybridomas, called Id(3) and Id(A.

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.

Genes predisposing to autoimmunity augment constitutive major histocompatibility complex class II-associated presentation of the self-antigen IgG2a in vivo.

The self-antigen IgG2ab is poorly presented to a gamma2ab 435-451-reactive I-Ad-restricted T-cell hybridoma unless available in high concentrations or targeted to Fcgamma- or complement receptors. Environmental factors, probably the extent of microbial challenge, profoundly influence the constitutive gamma2ab/I-Ad presentation in IgCHb, H-2d mice. Here we report also a strong genetic impact.

Native IgG2a(b) is barely antigenic to major histocompatibility complex class II-restricted T cells owing to inefficient internalization by professional antigen-presenting cells.

Peptide epitopes derived from immunoglobulin variable regions represent tumour-specific antigens on B-cell neoplasms and can be recognized by syngeneic, major histocompatibility complex (MHC) class II-restricted T cells. Immunoglobulin peptide/MHC class II complexes may also be involved in autoimmunity and CD4+ T-cell-mediated B-cell regulation. Thus, the IgG2a(b) H-chain allopeptide gamma2a(b) 435-451 presented on I-Ad mimics the epitope implicated in herpes simplex virus-induced autoimmune stromal keratitis and is the target of T helper 1 (Th1) clones that suppress IgG2a(b) production in vivo.

The P9 peptide sidechain specificity of I-Ad.

The murine MHC class II variant I-Ad confers susceptibility to herpes simplex virus (HSV)-induced keratitis and relative protection against type 1 diabetes mellitus. The association to these autoimmune diseases appears to be largely determined by the peptide sidechain specificity of the P9 pocket, which we therefore have analyzed in detail. Assessment of T-cell responses and I-Ad binding capacity of position 446-substituted analogs of an IgG2a allotype b (IgG2a(b)) heavy chain peptide demonstrates that engagement of the P9 pocket is crucial for effective peptide presentation.

N-terminal elongation of a peptide determinant beyond the first primary anchor improves binding to H-2 I-Ad and HLA-DR1 by backbone-dependent and aromatic side chain-dependent interactions, respectively.

The IgG2a(b) heavy chain allopeptide determinant gamma2a(b) 436-451 (Kabat numbering) presented by the major histocompatibility complex (MHC) class II molecule I-Ad is recognized by T cells which cross-react with a corneal self antigen and with the UL6 protein of the herpes simplex virus which induce autoimmune keratitis, and is the target of Th1 clones that suppress IgG2a(b) production in vivo. In the gamma2a(b) peptide/l-Ad complex, tyrosine438 is the first primary anchor (P1) and residues 440-445 encompass the T cell receptor contact residues. Amino-terminal elongation of gamma2a(b) 437-451 by a single residue (P-2) augmented the I-Ad binding capacity 10-fold and the antigenicity 55-195-fold.

A novel first primary anchor extends the MHC class II I-Ad binding motif to encompass nine amino acids.

The MHC class II molecule I-Ad has been reported to bind peptides containing a motif of six consecutive amino acids. We demonstrate that binding of the murine IgG2ab heavy chain allopeptide gamma 2ab 435-451 (Kabat numbering) to I-Ad is strongly enhanced by a novel first primary anchor (P1) three residues N-terminal to this hexamer. This is based on flow cytometric assessment of the I-Ad binding capacity of gamma 2ab peptide analogues, their antigenicity for I-Ad-restricted T cell clones and molecular modelling.

A retro-inverso analog mimics the cognate peptide epitope of a CD4+ T cell clone.

Synthetic analogs of peptide epitopes may activate specific T helper cells, antagonize their antigen receptors, or block recognition by competing for major histocompatibility complex (MHC) class II binding sites. Rationally designed peptides may therefore prove useful as vaccines and for treatment of autoimmune diseases and allergies mediated by CD4+ T cells. However, their susceptibility to proteolytic degradation limits the applicability of conventional peptides in vivo.

Engagement of the B lymphocyte antigen receptor induces presentation of intrinsic immunoglobulin peptides on major histocompatibility complex class II molecules.

By means of the clonotypic variable region, the immunoglobulin (Ig) is a tumor-specific antigen on B cell neoplasms. We report that engagement of the B cell antigen receptor (BcR) promotes presentation of peptides derived from the B cell's intrinsic Ig to major histocompatibility complex (MHC) class II-restricted T cells. Thus, anti-Ig endowed normal, ex vivo B lymphocytes from H-2d, Ig constant heavy chain allotype b (IgCHb) mice with the capacity to stimulate an I-Ad-restricted T cell clone which recognizes the gamma 2ab 435-451 allopeptide.

A syngeneic idiotype is immunogenic when borne by IgM but tolerogenic when joined to IgG.

Some syngeneic monoclonal antibodies (mAb) elicit immune responses like conventional T-dependent antigens. To find out whether the heavy chain class (isotype) plays a role for the immunogenicity of an idiotype (Id), we isolated rare subclones of an IgM mAb (termed Id3) in which the variable region of the heavy chain (VH) is associated with a new constant region (CH). The VH-Id3 gene is a member of the murine 36-60 family and probably has three replacement mutations.

A human hybridoma monoclonal antibody (TrJ11) recognizing a new HLA-DR epitope shared by DR4, DR8, DR11, and DRB1*1303.

The cytotoxic IgM lambda human hybridoma mAb TrJ11 reacts with lymphoblastoid B-cell lines expressing DR4, DR8, DR11, and DRB1*1303. However, TrJ11 was monospecific when normal B cells freshly isolated from blood served as targets in that it only killed HLA-DR4-positive cells. Thus, of 235 HLA-typed persons TrJ11 was strongly cytotoxic for normal B cells of all 90 DR4-positive individuals, but it did not react with B cells from any of the 145 DR4-negative donors.

Identification of the gene encoding a novel HLA-B39 subtype. Two amino acid substitutions on the beta-sheet out of the peptide-binding floor form a novel serological epitope.

Serological analysis suggests the existence of a novel HLA-B39 subtype (HLA-B39N) in the Japanese population. To identify this novel allele, a gene encoding HLA-B39N was cloned and the exons were sequenced. A gene encoding HLA-B39N (B*3904) and B*39011 differs by two nucleotide substitutions at codons 11 and 12 whereas B*3904 and B*39013 differ by three nucleotide substitutions at codons 11, 12, and 312.

A novel HLA-A determinant recognized by a cytotoxic human hybridoma IgG1 monoclonal antibody (TrJ14).

TrJ14 is a cytotoxic human IgG1 lambda hybridoma mAb that recognized a novel HLA-A epitope expressed by lymphoblastoid B cells that are homo- or heterozygous for A2, A3, A11, A30, A31, A33, A68 and A69. Based on these results, the HLA type of cell line TEM (10w9057) was retyped as A66. When peripheral blood T cells isolated freshly from 265 HLA-typed normal individuals served as targets, TrJ14 killed cells expressing two TrJ14-positive HLA-A alleles, as well as the majority of cells having one TrJ14-positive and one TrJ14-negative HLA-A antigen.

A cytotoxic human hybridoma monoclonal antibody (TrJ6) defining an epitope expressed by HLA-DQ4 and -DQ5.

We have generated a cytotoxic human hybridoma monoclonal IgM lambda antibody, designated TrJ6, that is specific for a new epitope shared by HLA-DQ4 and -DQ5. TrJ6 strongly killed all ten DQ4-bearing cells and weakly killed all four DQ5-bearing cell lines. In contrast, none of the 36 cell lines lacking DQ4 and DQ5 antigens was recognized by TrJ6.

Th1 clones that suppress IgG2ab specifically recognize an allopeptide determinant comprising residues 435-451 of gamma 2ab.

We have previously reported that gamma 2ab/I-A(d)-specific Th1 clones from BALB/c mice (gamma 2aa, H-2d) mediated a long-lasting, selective suppression of serum IgG2ab levels when transferred to newborn (BALB/c x B10.D2)F1 (gamma 2a/b, H-2d) mice (Bartnes, K. and Hannestad, K.

A cytotoxic human hybridoma monoclonal antibody (TrJ5) specific for HLA-B38(16) and -B39(16).

We have generated a cytotoxic IgM lambda human hybridoma mAb (TrJ5), that is specific for HLA-B38(16) and -B39(16). Among a panel of 42 HLA-defined cell lines, TrJ5 killed all five B38-positive and both B16-positive cell lines, as well as the single B39-positive cell line, but not any cell lines lacking these antigens. It could be ruled out that the TrJ5 epitope is located on the alpha 1 domain because TrJ5 did not react with cells bearing HLA-B14, the alpha 1 domain of which is identical to alpha 1 of B39.

A human monoclonal hybridoma antibody (TrJ1) specific for HLA-DQ2.

TrJ1 is a cytotoxic human hybridoma mAb (IgM lambda). Its reaction pattern with a panel of 42 HLA-defined lymphoblastoid B-cell lines correlated precisely with expression of DQ2. By flow cytometry it was shown that the binding of TrJ1 to DQ2 was efficiently blocked by the murine anti-DQ2 mAb 358.

Mapping of an epitope defined by a human hybridoma antibody (TrD3): a new HLA-B supertype associated with a subset of HLA-Bw6.

The new human-human hybridoma TrD3 secretes a cytotoxic IgM mAb, which reacted with 28 of a panel of 56 HLA-typed lymphoblastoid cells. All 28 TrD3+ cells expressed the HLA-B supertype Bw6, whereas 10 Bw6+ cells were not recognized by the mAb. None of the 17 Bw4 homozygous cells were positive with TrD3.

A cytotoxic monoclonal human hybridoma antibody (TrJ3) against HLA-B44(12) and -B45(12).

We have generated a human monoclonal cytotoxic IgM lambda antibody (TrJ3) that reacted specifically with all lymphoblastoid B-cell lines expressing HLA-B44(12) and B45(12). TrJ3 hybridoma supernatant was suitable for HLA-B12 typing of freshly isolated blood mononuclear cells. Analysis of available amino acid sequences of HLA-B molecules indicated that the alpha 1 domain does not contain the TrJ3 serological epitope.

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; lambda 2) 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; lambda isotype. All mAb purified from supernatants containing fetal bovine serum had formed spontaneous complexes with bovine serum albumin possibly by way of disulfide interchange.

Igh-1b-specific CD4+CD8- T cell clones of the Th1 subset selectively suppress the Igh-1b allotype in vivo.

The demonstration of major histocompatibility complex (MHC)-restricted T helper (Th) cells specific for peptides from the variable (V) regions of syngeneic immunoglobulin (Ig) (idiopeptides) opens the possibility that Th cells regulate B cell functions via idiopeptide-based cognate T-B interactions. As a model for such interactions we investigated the influence of Ig allotype-specific T cells on the differentiation of H-2-syngeneic B cells expressing that particular Ig allotype. We established a BALB/c (H-2d, Iga) CD4+CD8- T cell line and clones of the Th1 subset (interleukin 2+, interleukin 4-, interferon-gamma+, tumor necrosis factor-alpha+) that recognized Igh-1 (IgG2a) of the b allotype (Igh-1b) together with I-Ad.