Anti-HLA-E mAb 3D12 mimics MEM-E/02 in binding to HLA-B and HLA-C alleles: Web-tools validate the immunogenic epitopes of HLA-E recognized by the antibodies

Ravindranath, Mepur H.; Pham, Tho D.; El‐Awar, Nadim; Kaneku, Hugo; Terasaki, Paul I.

doi:10.1016/j.molimm.2010.09.011

Cited by 36 publications

(86 citation statements)

References 21 publications

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“…The peptides inhibited both HLA-Ia and HLA-E reactivities of anti-HLA-E mAbs. 14,15 These observations were extended to the sera of nonalloimmunized males to document that the observed HLA-Ia reactivity in human sera could be due to anti-HLA-E Abs. 13 To ascertain whether immunizing humans with HLA-E would augment anti-HLA-E Abs with HLA-Ia reactivity, we examined sera of melanoma patients immunized with autologous melanoma cells grown in interferon-g (IFN-g), a cytokine known to induce overexpression of HLA-E. [16][17][18] The immunized patients not only showed the augmentation of anti-HLA-E Abs, confirming the immunogenicity of HLA-E in humans, but also showed increased reactivity with HLA-Ia (single antigen) coated onto microbeads.…”

Section: Introductionmentioning

confidence: 92%

“…19 Immunoassay with single-HLA antigen-coated microbeads To detect IgG reactivity to HLA-E and HLA-Ia alleles in IVIg/purified human IgG/murine mAbs, multiplex Luminex-based immunoassay (One L, Inc) was used, as described elsewhere. [13][14][15] IVIg and standard human IgG from commercial sources were serially diluted (from 1/2, ending in 1/256) with phosphate-buffered saline (PBS; pH 7.2) and added to the wells containing the antigen-coated microbeads. Anti-HLA-E mAbs were developed in-house (vide infra), and the supernatants collected from the Figure 1.…”

Section: Sources Of Normal Human Seramentioning

confidence: 99%

“…Using dual-laser flow cytometry (Luminex xMAP multiplex technology), the single antigen assays were performed for data acquisition and analysis of anti-HLA-Ia and anti-HLA-E Abs. [13][14][15] The LABScreen Single Antigen (One L) assay consists of a panel of color-coded microspheres (single antigen beads) coated with HLA antigens to identify antibody specificities. The single recombinant HLA-Ia antigens in LS1A04-lot 003, 004, and 005 were used for screening IVIg and human standard IgG.…”

Section: Sources Of Normal Human Seramentioning

confidence: 99%

“…The recombinant HLA-E, HLA-F, and HLA-G heavy chains are individually attached by a process of simple chemical coupling to 5.6-mm polystyrene microspheres, which are internally dyed (by One L) with infrared fluorophores. [13][14][15] The HLA-Ia microbeads have built-in control beads: positive beads that are coated with human IgG (or murine IgG when mAb was used) and negative beads that are coated with serum albumin (HSA/BSA). For HLA-E, HLA-F, and HLA-G, the control beads (both positive and negative) were added separately.…”

Section: Sources Of Normal Human Seramentioning

confidence: 99%

“…14,15 The suggestion that the HLA-Ia reactivity of anti-HLA-E mAbs could be due to the peptide sequences shared between HLA-E and HLA class-Ia alleles 14 was verified using synthetic peptides. The peptides inhibited both HLA-Ia and HLA-E reactivities of anti-HLA-E mAbs.…”

Section: Introductionmentioning

confidence: 99%

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Therapeutic preparations of IVIg contain naturally occurring anti–HLA-E antibodies that react with HLA-Ia (HLA-A/-B/-Cw) alleles

Ravindranath

Terasaki

Pham

et al. 2013

Blood

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Section: Introductionmentioning

confidence: 92%

Section: Sources Of Normal Human Seramentioning

confidence: 99%

Section: Sources Of Normal Human Seramentioning

confidence: 99%

Section: Sources Of Normal Human Seramentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Therapeutic preparations of IVIg contain naturally occurring anti–HLA-E antibodies that react with HLA-Ia (HLA-A/-B/-Cw) alleles

Ravindranath

Terasaki

Pham

et al. 2013

Blood

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Monoclonal antibodies to HLA‐E bind epitopes carried by unfolded β₂m‐free heavy chains

Tremante¹,

Monaco²,

Ingegnere³

et al. 2015

Eur J Immunol

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Since HLA-E heavy chains accumulate free of their light β 2 -microglobulin (β 2 m) subunit, raising mAbs to folded HLA-E heterodimers has been difficult, and mAb characterization has been controversial. Herein, mAb W6/32 and 5 HLA-E-restricted mAbs (MEM-E/02, MEM-E/07, MEM-E/08, DT9, and 3D12) were tested on denatured, acid-treated, and natively folded (both β 2 m-associated and β 2 m-free) HLA-E molecules. Four distinct conformations were detected, including unusual, partially folded (and yet β 2 m-free) heavy chains reactive with mAb DT9. In contrast with previous studies, epitope mapping and substitution scan on thousands of overlapping peptides printed on microchips revealed that mAbs MEM-E/02, MEM-E/07, and MEM-E/08 bind three distinct α1 and α2 domain epitopes. All three epitopes are linear since they span just 4-6 residues and are "hidden" in folded HLA-E heterodimers. They contain at least one HLA-E-specific residue that cannot be replaced by single substitutions with polymorphic HLA-A, HLA-B, HLA-C, HLA-F, and HLA-G residues. Finally, also the MEM-E/02 and 3D12 epitopes are spatially distinct. In summary, HLA-E-specific residues are dominantly immunogenic, but only when heavy chains are locally unfolded. Consequently, the available mAbs fail to selectively bind conformed HLA-E heterodimers, and HLA-E expression may have been inaccurately assessed in some previous oncology, reproductive immunology, virology, and transplantation studies.Keywords: Conformation r HLA-E r HLA-A r -B r -C r mAbs r β 2 -microglobulin (β 2 m) Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionBeing the ligand of both the conserved CD94/NKG2A immune receptor and the variable TCR, the oligomorphic HLA-E molecule bridges classical and nonclassical HLA class I antigens [1]. Often classified among the latter, HLA-E may rather be considered an inbetweener. This definition also applies to HLA-C, since this is the least polymorphic among classical (HLA-A, -B, -C) Correspondence: Dr. Patrizio Giacomini e-mail: giacomini@ifo.it class I antigens, and shares with HLA-E several unorthodox properties, such as selective peptide binding, poor association with β 2 m [2-4], and massive accumulation in a β 2 m-free form [5,6].Although often unappreciated, unfolded and β 2 m-free heavy chains may share epitopes with fully folded HLA class I heterodimers. These "intermediate" folds may confound the assignment of antibody specificity. For instance, a mAb called Q1/28 binds HLA-B but not HLA-C when heavy chains are β 2 massociated, and HLA-C but not HLA-B when heavy chains are β 2 m-free [7]. Likewise, mAbs MEM-E/07 and MEM-E/08 bind β 2 m-free HLA-E, but cross-react with HLA-A, -B, -C heavy chainswww.eji-journal.eu Eur. J. Immunol. 2015. 45: 2356-2364 Molecular immunology 2357 associated with β 2 m [6]. Therefore, mAbs to inbetweeners require thorough characterization, including their systematic testing on both the β 2 m-associated and the β 2 m-free heavy chains specified by ...

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Gastric cancer progression may involve a shift in HLA‐E profile from an intact heterodimer to β2‐microglobulin‐free monomer

Sasaki

Ravindranath

Terasaki

et al. 2013

Intl Journal of Cancer

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Phenotypic expression of human leukocyte antigen (HLA)-E on the surface of tumor lesions includes intact heterodimer [HLA-E heavy chain and b2-microglobulin (b2m)] and b2m-free monomer. Anti-HLA-E monoclonal antibodies (mAbs), MEM-E=02 or 3D12 bind to the peptide sequences in b2m-free HLA-E, which is common and shared with HLA-Ia monomers. A newly developed monospecific anti-HLA-E mAb (TFL-033) recognizes HLA-E-restricted peptide sequences on a1 and a2 helices away from b2-m-site. Tumor progression may involve shedding of b2-m from HLA-E or overexpression of b2m-free monomers. There is a need to identify and distinguish the different phenotypic expression of HLA-E, particularly the intact heterodimer from the b2m-free monomer on the surface of tumor lesions. Because of the unique peptide-binding affinities of the mAbs, it is hypothesized that TFL-033 and MEM-E=02 may distinguish the phenotypic expressions of cell surface HLA-E during stages of tumor progression. Only TFL-033 stained diffusely the cytoplasm of normal mucosa. The incidence and intensity of TFL-033 staining of the cell surface in early stages, poorly or undifferentiated and non-nodal lesions and in diffuse carcinoma is greater than that of MEM-E=02. Whereas MEM-E=02 stained terminal stages, adenocarcinoma and lymph node metastatic lesions intensely, either owing to increased expression of b2m-free HLA-E with tumor progression or owing to expression of HLA-Ia molecules. Our study evaluates the relative diagnostic potential of HLA-E-monospecific TFL-033 and the HLA-Ia-reactive MEM-E=02 for determining the specific distribution and immunodiagnosis of different phenotypic expression HLA-E in tumor lesions, and the structural and functional alterations undergone by HLA-E during tumor progression.Structural and functional alterations of human leukocyte antigens (HLA) are implicated in tumor immune escape. 1 The escape mechanism involves loss or downregulation of classical HLA class I antigens (HLA-A, -B and -C) and=or aberrant overexpression of nonclassical HLA class I antigens (HLA-E and -G). Consequently, these changes in HLA classes lead to decreased recognition or destruction of tumor cells by immune cytotoxic effectors, mainly cytotoxic lymphocytes (CTL) and natural killer T (NKT) cells. 1

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Anti-HLA-E mAb 3D12 mimics MEM-E/02 in binding to HLA-B and HLA-C alleles: Web-tools validate the immunogenic epitopes of HLA-E recognized by the antibodies

Cited by 36 publications

References 21 publications

Therapeutic preparations of IVIg contain naturally occurring anti–HLA-E antibodies that react with HLA-Ia (HLA-A/-B/-Cw) alleles

Therapeutic preparations of IVIg contain naturally occurring anti–HLA-E antibodies that react with HLA-Ia (HLA-A/-B/-Cw) alleles

Monoclonal antibodies to HLA‐E bind epitopes carried by unfolded β₂m‐free heavy chains

Gastric cancer progression may involve a shift in HLA‐E profile from an intact heterodimer to β2‐microglobulin‐free monomer

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Anti-HLA-E mAb 3D12 mimics MEM-E/02 in binding to HLA-B and HLA-C alleles: Web-tools validate the immunogenic epitopes of HLA-E recognized by the antibodies

Cited by 36 publications

References 21 publications

Therapeutic preparations of IVIg contain naturally occurring anti–HLA-E antibodies that react with HLA-Ia (HLA-A/-B/-Cw) alleles

Therapeutic preparations of IVIg contain naturally occurring anti–HLA-E antibodies that react with HLA-Ia (HLA-A/-B/-Cw) alleles

Monoclonal antibodies to HLA‐E bind epitopes carried by unfolded β2m‐free heavy chains

Gastric cancer progression may involve a shift in HLA‐E profile from an intact heterodimer to β2‐microglobulin‐free monomer

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Monoclonal antibodies to HLA‐E bind epitopes carried by unfolded β₂m‐free heavy chains