The human antibody repertoire: Heavy and light chain variable region gene usage in six alloantibodies specific for human HLA class I and class II alloantigens

Andris, Jennifer S.; Abraham, Sheena; Pascual, Virginia; Pistillo, Maria Pia; Mantero, Stefano; Ferrara, Giovanni Battista; Capra, J D

doi:10.1016/0161-5890(95)00071-2

Cited by 13 publications

(7 citation statements)

References 78 publications

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“…A preferential usage of certain V H genes in the generation of disease‐specific/eliciting/protective antibodies has been reported for several immunologically mediated diseases. 13–18 Recent evidence for a limited V H usage in atopy came from the observations that ε transcripts obtained from the PBMC of three atopic dermatitis patients 19 and spleen‐derived lymphocytes of an asthmatic individual 20 preferentially used the V H 5 family. In addition, molecular and structural analyses of allergens indicates that many atopic individuals are sensitized against a limited panel of cross‐reactive allergens and epitopes.…”

Section: Discussionmentioning

confidence: 99%

“…While for certain infectious, 13–14 as well as immunological, diseases a bias towards the usage of certain V H ‐gene families has been reported, 15–18 rather little information is available regarding the IgE V H ‐gene usage in atopy. Analysis of the IgE V H ‐gene usage in peripheral blood lymphocytes (PBL) from three atopic dermatitis patients 19 and from spleen‐derived lymphocytes of an asthmatic individual 20 indicated a preferential usage of the V H 5‐gene family.…”

Section: Introductionmentioning

confidence: 99%

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Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of V_H‐gene families

et al. 2000

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SUMMARYThe term`atopy' describes the genetically determined tendency to mount immunoglobulin E (IgE) antibody responses against per se harmless antigens (allergens). In this study we investigated the usage of V H families in the formation of IgE antibodies in 10 patients suffering from mucosal and/or skin manifestations of atopy. IgE antibody reactivities to exogenous allergen sources as well as to autoallergens were determined and, by immunoabsorption, it was demonstrated that allergenspeci®c IgE accounted for most of the total serum IgE levels in these patients. Using primers with speci®city for the V H 1±6 gene families and a primer speci®c for the ®rst constant region of human IgE, cDNAs coding for IgE heavy chain fragments were ampli®ed using the reverse transcription± polymerase chain reaction (RT±PCR) from peripheral blood lymphocytes of the 10 atopic individuals. Hybridization of the heavy chain-encoding cDNAs with an IgE-speci®c internal oligonucleotide probe revealed a broad usage of all V H -gene families in the atopic individuals. The spectrum of V H families used in a given atopic individual was neither associated with the type or severity of clinical symptoms nor with the number of allergens recognized. The fact that allergenspeci®c IgE antibodies in atopic individuals originate from a broad variety of B cells thus re¯ects the activation of multiple B-cell clones during allergen sensitization. This ®nding should be borne in mind if therapeutic strategies for Type I allergy are considered that aim at a clonal elimination of allergen-speci®c B cells.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of V_H‐gene families

et al. 2000

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“…The availability of HLA-specific B cell lines permits the analysis of their Ig H and L chain genes in terms of V family usage. Such analyses have been reported for specific B cells in other antigenic systems (9), but for alloimmunity these studies have been done on established hybridomas (21,22). The anti-HLA-A2 repertoire has been studied by phage display (23); however, this technology is unable to distinguish correct pairing of H and L chain genes within single B cells.…”

Section: Discussionmentioning

confidence: 99%

Identification, Isolation, and Culture of HLA-A2-Specific B Lymphocytes Using MHC Class I Tetramers

Mulder

Eijsink

Kardol

et al. 2003

The Journal of Immunology

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Characterizing the individual B cells that participate in the production of anti-HLA Abs requires isolation and culture of these cells and a suitable assay for detection of Abs produced in these B cell cultures. We previously showed that B cell precursors, programmed for anti-HLA Ab secretion, are present at measurable frequencies in peripheral blood of women immunized by pregnancy. In this study, we show that tetrameric HLA-A2, although designed for characterization of CTLs, provides a suitable affinity ligand for isolation of allospecific B cells, which subsequently can be induced to produce HLA-A2 Ab in a CD40-driven culture system. The validity of this concept was established by assaying human hybridomas, producing anti-HLA Abs, for specific tetrameric HLA-A2 binding. The availability of anti-HLA Ab-producing B cell cultures that are established without immortalization will be of value when T-B cell interaction is studied at an alloantigen-specific level.

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“…12 The sequence of nucleotides specifying the C was as expected until codon 187, where there was a 2-bp deletion (GA) that resulted in an extended, aberrant product due to loss of the normal TAG stop codon 215. Notably, this frameshift mutation was not present in the patient's gDNA (Figure 2) where the deduced protein product consisted of the anticipated 106 amino acids, including cysteines at positions 194 and 214.…”

Section: Resultsmentioning

confidence: 99%

“…First-strand DNA was amplified by forward and reverse primers specifying the first and last 7 amino acid residues of a prototypic 1 light chain. 12 The polymerase chain reaction (PCR) products were cloned using the perfectly blunt cloning kit with the pSTBlue-1 vector (Novagen, Madison, WI) and the colonies screened by PCR. Plasmids were isolated from candidate clones using the Quantum Miniprep kit (Bio-Rad, Richmond, CA), further screened by EcoRI digestion to confirm insert size, and sequenced in both directions.…”

Section: Rna Preparation and Rt-pcr Amplificationmentioning

confidence: 99%

A molecular basis for nonsecretory myeloma

Coriu

Weaver

Schell

et al. 2004

Blood

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IntroductionNonsecretory myeloma (NSM) is defined by the absence of serum or urinary (or both) monoclonal immunoglobulins in patients who otherwise manifest features typically found in multiple myeloma (MM). 1,2 This condition, which occurs in about 1% to 5% of patients with MM, results either from the inability of malignant plasma cells to synthesize immunoglobulin (ie, nonproducer) or, more commonly, the failure of such components to be exported from the cells (ie, nonsecretor). In the latter case, it has been assumed that the protein undergoes intracellular proteolysis due to a structural defect in the immunoglobulin and, as a consequence, is not excreted. However, there is only limited information regarding the molecular factors that might account for this phenomenon. [3][4][5][6][7] We now report our finding that the plasma cells of an individual with NSM indeed synthesized aberrant monoclonal light chains that resulted from a somatic mutation in the gene encoding the constant (C L ) region of the molecule. Our studies provide the first conclusive evidence that an abnormality in this portion of the light chain may be implicated in the pathogenesis of this disorder. Study design Protein analysesSerum IgG, IgA, and IgM proteins were quantitated with the Synchron LX20 Clinical System (Beckman Coulter, Fullerton, CA) and the concentration of free and light chains determined by enzyme-linked immunosorbent assay (ELISA), using our highly specific monoclonal antibodies (mAbs), 8 as well as by nephelometry (Free Lite, The Binding Site, Birmingham, United Kingdom) with polyclonal reagents. 9 For detection of monotypic immunoglobulins, serum was diluted 1:10 and subjected to immunofixation electrophoresis using the Paragon system (Beckman, Norcross, GA), according to the procedure specified by the manufacturer. Undiluted samples also were examined in similar fashion, as were unconcentrated urine specimens and 24-hour collections that had been dialyzed extensively against distilled water, lyophilized, and reconstituted to a protein concentration of 100 mg/mL. 10 ImmunocytochemistryBone marrow plasma cells were isolated and immunostained with our murine mAbs specific for human light-chain variable region (V L ) subgroups 11 and for total (heavy chain-bound) and free (unbound) and polypeptides, 8 as well as with mouse antihuman plasma cell (Dako, Carpinteria, CA) and rabbit antihuman ␥, ␣, , and ␦ heavy-chain antisera (Biosource, Camarillo, CA). RNA preparation and RT-PCR amplificationTotal RNA was extracted from plasma cells with the PURESCRIPT RNA isolation kit (Gentra, Minneapolis, MN) and transcribed with both oligo (dT) 15 and random primers, using reverse transcriptase (RT; Promega, Madison, WI) in a single reaction. First-strand DNA was amplified by forward and reverse primers specifying the first and last 7 amino acid residues of a prototypic 1 light chain. 12 The polymerase chain reaction (PCR) products were cloned using the perfectly blunt cloning kit with the pSTBlue-1 vector (Novagen, Madison, WI) and the co...

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The human antibody repertoire: Heavy and light chain variable region gene usage in six alloantibodies specific for human HLA class I and class II alloantigens

Cited by 13 publications

References 78 publications

Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of V_H‐gene families

Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of V_H‐gene families

Identification, Isolation, and Culture of HLA-A2-Specific B Lymphocytes Using MHC Class I Tetramers

A molecular basis for nonsecretory myeloma

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The human antibody repertoire: Heavy and light chain variable region gene usage in six alloantibodies specific for human HLA class I and class II alloantigens

Cited by 13 publications

References 78 publications

Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of VH‐gene families

Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of VH‐gene families

Identification, Isolation, and Culture of HLA-A2-Specific B Lymphocytes Using MHC Class I Tetramers

A molecular basis for nonsecretory myeloma

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Product

Resources

About

Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of V_H‐gene families

Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of V_H‐gene families