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Comparative analysis of LAG3 antibodies shows differential binding patterns by flow cytometry.
Graydon, Colin G; Balasko, Allison; Kowatsch, Monika; Fowke, Keith R.
Afiliación
  • Graydon CG; Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Room 543 - 745 Bannatyne Avenue, Winnipeg, MB R3E 0J9, Canada. Electronic address: graydonc@myumanitoba.ca.
  • Balasko A; Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Room 543 - 745 Bannatyne Avenue, Winnipeg, MB R3E 0J9, Canada. Electronic address: umbalask@myumanitoba.ca.
  • Kowatsch M; Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Room 543 - 745 Bannatyne Avenue, Winnipeg, MB R3E 0J9, Canada. Electronic address: umkowats@myumanitoba.ca.
  • Fowke KR; Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Room 543 - 745 Bannatyne Avenue, Winnipeg, MB R3E 0J9, Canada; Community Health Sciences, University of Manitoba, Room S113 - 750 Bannatyne Avenue, Winnipeg, MB R3E 0W3, Canada; Medical Microbiology and Immunology, U
J Immunol Methods ; 534: 113757, 2024 Sep 12.
Article en En | MEDLINE | ID: mdl-39277073
ABSTRACT

BACKGROUND:

LAG3 is an immune checkpoint molecule with emerging therapeutic use. Expression of LAG3 is well studied on T cells, but the proportion of LAG3-expressing cells varies greatly by study and its comparative expression between non-T cells is lacking. METHODS/

OBJECTIVES:

This study uses flow cytometry to compare surface LAG3 expression between T cells, NK cells, B cells, pDCs and monocytes of healthy donors. This study also compares three monoclonal LAG3 antibodies to a commonly used polyclonal LAG3 antibody on ex vivo and PHA-blasts from healthy donors and LAG3+ and LAG3- cell lines.

RESULTS:

LAG3 was most highly expressed on classical and intermediate monocytes (25 % and 32 %, respectively), while LAG3 expression on B cells, NK cells and iNKT cells was negligible. Notably, the polyclonal antibody stained a higher proportion of all cell types than the monoclonal antibodies, which had similar staining patterns to one another. Further study using LAG3+ and LAG3- cell lines showed greater specificity and similar sensitivity of the monoclonal antibody T47-530 than the polyclonal antibody.

CONCLUSION:

Monocytes may represent an unappreciated source of LAG3 and target of LAG3 checkpoint inhibitors. Furthermore, the discrepancies between monoclonal and polyclonal LAG3 antibodies warrants consideration when designing future studies and interpreting past studies, and may explain discrepancies in the literature.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: J Immunol Methods Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: J Immunol Methods Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos