Versatility of using major histocompatibility complex class II dextramers for derivation and characterization of antigen-specific, autoreactive T cell hybridomas

Bharathi Krishnan, Chandirasegaran Massilamany, Rakesh H. Basavalingappa, Rajkumar A. Rajasekaran, Charles Kuszynski, Barbara Switzer, Daniel A. Peterson, N R Jayagopala Reddy

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Antigen-specific, T cell hybridomas are useful to study the cellular, molecular and functional events, but their generation is a lengthy process. Thus, there is a need to develop robust methods to generate the hybridoma clones rapidly in a short period of time. To this end, we have demonstrated a novel approach using major histocompatibility complex (MHC) class II dextramers to generate T cell hybridomas for an autoantigen, proteolipid protein (PLP) 139-151. Using MHC class II dextramers assembled with PLP 139-151 as screening and sorting tools, we successfully obtained mono antigen-specific clones within seven to eight weeks. In conjunction with other T cell markers, dextramers permitted phenotypic characterization of hybridoma clones for their antigen specificity in a single step by flow cytometry. Importantly, we achieved successful fusions using dextramer+ cells sorted by flow cytometry as a starting population, resulting in direct identification of multiple antigen-specific clones. Characterization of selected clones led us to identify chemokine receptor, CCR4+ to be expressed consistently, but their cytokine-producing ability was variable. Our work provides a proof-of principle that the antigen-specific, CD4 T cell hybridoma clones can be generated directly using MHC class II dextramers. The availability of hybridoma clones that bind dextramers may serve as useful tools for various in vitro and in vivo applications.

Original languageEnglish (US)
Pages (from-to)86-94
Number of pages9
JournalJournal of Immunological Methods
Volume426
DOIs
StatePublished - Nov 1 2015

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Hybridomas
Major Histocompatibility Complex
Clone Cells
T-Lymphocytes
Antigens
Flow Cytometry
CD4 Antigens
Chemokine Receptors
Autoantigens
Cytokines
Population

Keywords

  • Autoreactive T cells
  • MHC class II dextramers
  • T cell hybridoma

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Versatility of using major histocompatibility complex class II dextramers for derivation and characterization of antigen-specific, autoreactive T cell hybridomas. / Krishnan, Bharathi; Massilamany, Chandirasegaran; Basavalingappa, Rakesh H.; Rajasekaran, Rajkumar A.; Kuszynski, Charles; Switzer, Barbara; Peterson, Daniel A.; Reddy, N R Jayagopala.

In: Journal of Immunological Methods, Vol. 426, 01.11.2015, p. 86-94.

Research output: Contribution to journalArticle

Krishnan, Bharathi ; Massilamany, Chandirasegaran ; Basavalingappa, Rakesh H. ; Rajasekaran, Rajkumar A. ; Kuszynski, Charles ; Switzer, Barbara ; Peterson, Daniel A. ; Reddy, N R Jayagopala. / Versatility of using major histocompatibility complex class II dextramers for derivation and characterization of antigen-specific, autoreactive T cell hybridomas. In: Journal of Immunological Methods. 2015 ; Vol. 426. pp. 86-94.
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