Characterization of two human monoclonal antibodies recognizing HLA‐A30 and HLA‐A3+A31, respectively

Mazzoleni, Osvaldo; Pistillo, Maria Pia; Falco, Michela; Tazzari, Pier Luigi; Ferrara, Giovanni Battista

doi:10.1111/j.1399-0039.1991.tb01901.x

Cited by 10 publications

(3 citation statements)

References 12 publications

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“…No A*30‐specific antibody was available for FACS analysis and the HLA class I‐specific monoclonal W6/32 ( 12), was used for detection of HLA class I expression. Subsequently, more specific confirmation of expression was made through serological typing of the transfectants, using A*30‐specific monoclonal antibodies ( 13) in the complement‐mediated lymphocytotoxicity assay ( 14, 15).…”

Section: Methodsmentioning

confidence: 99%

Definition of peptide binding motifs amongst the HLA‐A*30 allelic group

Krausa¹,

Münz²,

Keilholz³

et al. 2000

Tissue Antigens

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HLA class I molecules present endogenously processed peptide ligands for surveillance by the T-cell receptor. This potentially immunogenic surface of HLA and peptide is a consequence of the polymorphism found within the HLA molecule and its preference for ligand binding together with peptide conformation within the binding groove. To investigate the relation between the polymorphic differences between some closely related HLA alleles and their effect on peptide preference, transfectants were established, each containing one of four allelic variants of HLA-A*30. Peptides from all four transfectants were eluted, and both individual ligands and peptide pools were sequenced. The data shows two distinct peptide motifs which distinguish A*3001 from the other three known A*30 variants. Differences in preferences at minor positions within the peptide sequence were noted between A*3002, A*3003 and A*3004, providing additional evidence of the implications of sequence polymorphism to HLA function.

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Section: Methodsmentioning

confidence: 99%

Definition of peptide binding motifs amongst the HLA‐A*30 allelic group

Krausa¹,

Münz²,

Keilholz³

et al. 2000

Tissue Antigens

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“…As a source for this purpose, alloantisera from multiparous woman, mouse monoclonal antibodies (mAb) and human mAb are used. In the last few years, a series of specific human anti-HLA class I mAb was published (9)(10)(11)(12)(13)(14)(15)(16). The uniformly used protocol for producing these mAb was Epstein-Barr-Virus (EBV) transformation of peripheral B lymphocytes and fusing of anti-HLA class I antibody-producing lymphoblastoid cell lines (LCLs) with myeloma cells (17).…”

Section: Introductionmentioning

confidence: 99%

Generation and characterization of a human monoclonal IgG antibody specific for HLA‐B12

Siemoneit

Wölpl

Wegener³

et al. 1994

Tissue Antigens

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We describe here the generation and characterization of a human monoclonal IgG antibody (UL/F14) specific for HLA-B12. The antibody is suitable for complement-dependent lysis on lymphoblastoid cell lines and stimulated peripheral blood mononuclear cells. UL/F14 competes for antigen binding with an HLA-B12 human monoclonal IgM antibody and with a specific alloantiserum. Protein chemistry shows that the monoclonal antibody UL/F14 can precipitate solubilized HLA-B12 molecules.

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“…It was also noted that anti-IgM FITC-labelled cells given an additional wash after incubation with the labelling antibody showed a marked reduction in fluorescence not observed with the same use of an anti-IgG FITC label. A human IgM anti-class I monoclonal antibody (MP9) with specificity for HLA-A3/31 [8] showed a cytotoxic endpoint of 1 : 64 against CLL cells, whereas binding could only be detected flow cytometrically at 1 : 8. A serum sample with a specificity for HLA B8, removable by treatment with DTT, gave a cytotoxic endpoint of 1 : 16.…”

Section: Relative Sensitivity Of Flow Cytometric Assay For Igg Hla Anmentioning

confidence: 99%

The flow cytometric detection of alloantibodies in screening for renal transplantation

et al. 1995

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Flow cytometric screening of sera using pooled chronic lymphocytic leukaemia (CLL) cells has previously been reported as a quick method for detecting HLA antibodies of the IgG class. In this study we investigated the sensitivity of this method in the detection of IgG and IgM alloantibodies, and its performance in serum screening when compared to conventional microlymphocytotoxic screening. Results indicate that flow cytometric screening is more sensitive in the measurement of IgG alloantibodies by up to five doubling dilutions, whereas the converse is true for IgM. IgM autoantibodies were found not to be detectable by flow cytometry. By testing a large number of sera by both methods in parallel, we have found that a significant proportion of sera exhibiting no activity of IgM activity alone on cytotoxic screening contain IgG antibodies detectable with a pool of CLL cells on the flow cytometer.

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Characterization of two human monoclonal antibodies recognizing HLA‐A30 and HLA‐A3+A31, respectively

Cited by 10 publications

References 12 publications

Definition of peptide binding motifs amongst the HLA‐A*30 allelic group

Definition of peptide binding motifs amongst the HLA‐A*30 allelic group

Generation and characterization of a human monoclonal IgG antibody specific for HLA‐B12

The flow cytometric detection of alloantibodies in screening for renal transplantation

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