Diversity in the HLA-I Recognition of HLA-F Monoclonal Antibodies: HLA-F or HLA-Ib Monospecific, HLA-E or HLA-G Bispecific Antibodies with or without HLA-Ia Reactivity
Previous investigators have used various anti-HLA-F monoclonal antibodies (mAbs) to demonstrate that the tissue distribution of HLA-F is highly restricted. Notably, these mAbs differed in their immunodiagnostic capabilities. Specifically, mAbs Fpep1.1 and FG1 detected HLA-F intracellularly in B cell...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-01-01
|
| Series: | Antibodies |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4468/13/1/8 |
| _version_ | 1797242261005664256 |
|---|---|
| author | Mepur H. Ravindranath Narendranath M. Ravindranath Carly J. Amato-Menker Fatiha El Hilali Edward J. Filippone |
| author_facet | Mepur H. Ravindranath Narendranath M. Ravindranath Carly J. Amato-Menker Fatiha El Hilali Edward J. Filippone |
| author_sort | Mepur H. Ravindranath |
| collection | DOAJ |
| description | Previous investigators have used various anti-HLA-F monoclonal antibodies (mAbs) to demonstrate that the tissue distribution of HLA-F is highly restricted. Notably, these mAbs differed in their immunodiagnostic capabilities. Specifically, mAbs Fpep1.1 and FG1 detected HLA-F intracellularly in B cells but not on the cell surface, whereas mAb 3D11 detected HLA-F on the cell surface. The presence of HLA-F on T cells was recognized by mAb FG1 but not by mAb Fpep1.1. mAb 3D11 detected HLA-F on the cell surface of activated B cells and on peripheral blood lymphocytes, but not on the normal cells. Importantly, mAb 3D11 revealed that HLA-F exists as a heavy chain (HC) monomer, rather than as an HC associated with B2m. Although these mAbs are believed to be specific to HLA-F, their monospecificity has not been formally established, which is critical for immunodiagnostic and therapeutic purposes. Previously, we investigated the diversity of HLA class I reactivities of anti-HLA-E mAbs using HLA-I coated multiplex bead assays on a Luminex platform. We reported that more than 80% of the HLA-E mAbs were cross-reactive with other HLA-I molecules, with exceptionally few truly HLA-E-monospecific mAbs. In the present investigation, we generated IgG mAbs against HCs of HLA-F in Balb/C mice and examined the cross-reactivity of anti-HLA-F mAbs with other HLA-I alleles using a multiplex bead assay on the Luminex platform. Beads coated with an array of HLA homo- and heterodimers of different HLA-Ia (HLA-A, HLA-B, and HLA-C) and Ib (HLA-E, HLA-F, and HLA-G) alleles were used to examine the binding of the anti-HLA-F mAbs. Only two mAbs were HLA-F monospecific, and five were HLA-Ib restricted. Several anti-HLA-F mAbs cross-reacted with HLA-E (<i>n</i> = 4), HLA-G (<i>n</i> = 3), HLA-Ia alleles (<i>n</i> = 9), HLA-G and HLA-Ia (<i>n</i> = 2), and HLA-Ib and HLA-Ia (<i>n</i> = 6). This monospecificity and polyreactivity were corroborated by the presence of HLA-F monospecific and HLA-I-shared sequences. This study emphasizes the need to monitor the mono-specificity of HLA-F for reliable immunodiagnostics and passive immunotherapy. |
| first_indexed | 2024-04-24T18:36:24Z |
| format | Article |
| id | doaj.art-2d95fcd1c97d4429945aadd15821fbeb |
| institution | Directory Open Access Journal |
| issn | 2073-4468 |
| language | English |
| last_indexed | 2024-04-24T18:36:24Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibodies |
| spelling | doaj.art-2d95fcd1c97d4429945aadd15821fbeb2024-03-27T13:18:18ZengMDPI AGAntibodies2073-44682024-01-01131810.3390/antib13010008Diversity in the HLA-I Recognition of HLA-F Monoclonal Antibodies: HLA-F or HLA-Ib Monospecific, HLA-E or HLA-G Bispecific Antibodies with or without HLA-Ia ReactivityMepur H. Ravindranath0Narendranath M. Ravindranath1Carly J. Amato-Menker2Fatiha El Hilali3Edward J. Filippone4Department of Hematology and Oncology, Children’s Hospital, Los Angeles, CA 90027, USANorris Dental Science Center, Herman Ostrow School of Dentistry, University of Southern California, Los Angeles, CA 90089, USADepartment of Microbiology, Immunology and Cell Biology, School of Medicine, West Virginia University, Morgantown, WV 26506, USAMedico-Surgical, Biomedicine and Infectiology Research Laboratory, The Faculty of Medicine and Pharmacy of Laayoune & Agadir, Ibnou Zohr University, Agadir 80000, MoroccoDivision of Nephrology, Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA 19145, USAPrevious investigators have used various anti-HLA-F monoclonal antibodies (mAbs) to demonstrate that the tissue distribution of HLA-F is highly restricted. Notably, these mAbs differed in their immunodiagnostic capabilities. Specifically, mAbs Fpep1.1 and FG1 detected HLA-F intracellularly in B cells but not on the cell surface, whereas mAb 3D11 detected HLA-F on the cell surface. The presence of HLA-F on T cells was recognized by mAb FG1 but not by mAb Fpep1.1. mAb 3D11 detected HLA-F on the cell surface of activated B cells and on peripheral blood lymphocytes, but not on the normal cells. Importantly, mAb 3D11 revealed that HLA-F exists as a heavy chain (HC) monomer, rather than as an HC associated with B2m. Although these mAbs are believed to be specific to HLA-F, their monospecificity has not been formally established, which is critical for immunodiagnostic and therapeutic purposes. Previously, we investigated the diversity of HLA class I reactivities of anti-HLA-E mAbs using HLA-I coated multiplex bead assays on a Luminex platform. We reported that more than 80% of the HLA-E mAbs were cross-reactive with other HLA-I molecules, with exceptionally few truly HLA-E-monospecific mAbs. In the present investigation, we generated IgG mAbs against HCs of HLA-F in Balb/C mice and examined the cross-reactivity of anti-HLA-F mAbs with other HLA-I alleles using a multiplex bead assay on the Luminex platform. Beads coated with an array of HLA homo- and heterodimers of different HLA-Ia (HLA-A, HLA-B, and HLA-C) and Ib (HLA-E, HLA-F, and HLA-G) alleles were used to examine the binding of the anti-HLA-F mAbs. Only two mAbs were HLA-F monospecific, and five were HLA-Ib restricted. Several anti-HLA-F mAbs cross-reacted with HLA-E (<i>n</i> = 4), HLA-G (<i>n</i> = 3), HLA-Ia alleles (<i>n</i> = 9), HLA-G and HLA-Ia (<i>n</i> = 2), and HLA-Ib and HLA-Ia (<i>n</i> = 6). This monospecificity and polyreactivity were corroborated by the presence of HLA-F monospecific and HLA-I-shared sequences. This study emphasizes the need to monitor the mono-specificity of HLA-F for reliable immunodiagnostics and passive immunotherapy.https://www.mdpi.com/2073-4468/13/1/8HLA-IaHLA-IbHLA-EHLA-FHLA-Gheavy chain |
| spellingShingle | Mepur H. Ravindranath Narendranath M. Ravindranath Carly J. Amato-Menker Fatiha El Hilali Edward J. Filippone Diversity in the HLA-I Recognition of HLA-F Monoclonal Antibodies: HLA-F or HLA-Ib Monospecific, HLA-E or HLA-G Bispecific Antibodies with or without HLA-Ia Reactivity Antibodies HLA-Ia HLA-Ib HLA-E HLA-F HLA-G heavy chain |
| title | Diversity in the HLA-I Recognition of HLA-F Monoclonal Antibodies: HLA-F or HLA-Ib Monospecific, HLA-E or HLA-G Bispecific Antibodies with or without HLA-Ia Reactivity |
| title_full | Diversity in the HLA-I Recognition of HLA-F Monoclonal Antibodies: HLA-F or HLA-Ib Monospecific, HLA-E or HLA-G Bispecific Antibodies with or without HLA-Ia Reactivity |
| title_fullStr | Diversity in the HLA-I Recognition of HLA-F Monoclonal Antibodies: HLA-F or HLA-Ib Monospecific, HLA-E or HLA-G Bispecific Antibodies with or without HLA-Ia Reactivity |
| title_full_unstemmed | Diversity in the HLA-I Recognition of HLA-F Monoclonal Antibodies: HLA-F or HLA-Ib Monospecific, HLA-E or HLA-G Bispecific Antibodies with or without HLA-Ia Reactivity |
| title_short | Diversity in the HLA-I Recognition of HLA-F Monoclonal Antibodies: HLA-F or HLA-Ib Monospecific, HLA-E or HLA-G Bispecific Antibodies with or without HLA-Ia Reactivity |
| title_sort | diversity in the hla i recognition of hla f monoclonal antibodies hla f or hla ib monospecific hla e or hla g bispecific antibodies with or without hla ia reactivity |
| topic | HLA-Ia HLA-Ib HLA-E HLA-F HLA-G heavy chain |
| url | https://www.mdpi.com/2073-4468/13/1/8 |
| work_keys_str_mv | AT mepurhravindranath diversityinthehlairecognitionofhlafmonoclonalantibodieshlaforhlaibmonospecifichlaeorhlagbispecificantibodieswithorwithouthlaiareactivity AT narendranathmravindranath diversityinthehlairecognitionofhlafmonoclonalantibodieshlaforhlaibmonospecifichlaeorhlagbispecificantibodieswithorwithouthlaiareactivity AT carlyjamatomenker diversityinthehlairecognitionofhlafmonoclonalantibodieshlaforhlaibmonospecifichlaeorhlagbispecificantibodieswithorwithouthlaiareactivity AT fatihaelhilali diversityinthehlairecognitionofhlafmonoclonalantibodieshlaforhlaibmonospecifichlaeorhlagbispecificantibodieswithorwithouthlaiareactivity AT edwardjfilippone diversityinthehlairecognitionofhlafmonoclonalantibodieshlaforhlaibmonospecifichlaeorhlagbispecificantibodieswithorwithouthlaiareactivity |