Detection of autoreactive CD4 T cells using major histocompatibility complex class II dextramers

Chandirasegaran Massilamany, Bijaya Upadhyaya, Arunakumar Gangaplara, Charles Kuszynski, N R Jayagopala Reddy

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Background: Tetramers are useful tools to enumerate the frequencies of antigen-specific T cells. However, unlike CD8 T cells, CD4 T cells - especially self-reactive cells - are challenging to detect with major histocompatibility complex (MHC) class II tetramers because of low frequencies and low affinities of their T cell receptors to MHC-peptide complexes. Here, we report the use of fluorescent multimers, designated MHC dextramers that contain a large number of peptide-MHC complexes per reagent.Results: The utility of MHC dextramers was evaluated in three autoimmune disease models: 1) proteolipid protein (PLP) 139-151-induced experimental autoimmune encephalomyelitis in SJL/J (H-2s) mice; 2) myelin oligodendrocyte glycoprotein (MOG) 35-55-induced experimental autoimmune encephalomyelitis in C57Bl/6 (H-2b) mice; and 3) cardiac myosin heavy chain (Myhc)-α 334-352-induced experimental autoimmune myocarditis in A/J (H-2a) mice. Flow cytometrically, we demonstrate that IAs/PLP 139-151, IAb/MOG 35-55 and IAk/Myhc-α 334-352 dextramers detect the antigen-sensitized cells with specificity, and with a detection sensitivity significantly higher than that achieved with conventional tetramers. Furthermore, we show that binding of dextramers, but not tetramers, is less dependent on the activation status of cells, permitting enumeration of antigen-specific cells ex vivo.Conclusions: The data suggest that MHC dextramers are useful tools to track the generation and functionalities of self-reactive CD4 cells in various experimental systems.

Original languageEnglish (US)
Article number40
JournalBMC Immunology
Volume12
DOIs
StatePublished - Jul 18 2011

Fingerprint

Major Histocompatibility Complex
T-Lymphocytes
Autoimmune Experimental Encephalomyelitis
Myosin Heavy Chains
Antigens
Cardiac Myosins
Peptides
Myocarditis
T-Cell Antigen Receptor
Autoimmune Diseases

Keywords

  • Antigen-specific CD4 cells
  • Cardiac myosin heavy chain-alpha
  • Central nervous system
  • Dextramers
  • Experimental autoimmune encephalomyelitis
  • Experimental autoimmune myocarditis
  • Heart
  • Major histocompatibility complex class II
  • Myelin oligodendrocyte glycoprotein
  • Proteolipid protein
  • Tetramers

ASJC Scopus subject areas

  • Immunology

Cite this

Detection of autoreactive CD4 T cells using major histocompatibility complex class II dextramers. / Massilamany, Chandirasegaran; Upadhyaya, Bijaya; Gangaplara, Arunakumar; Kuszynski, Charles; Reddy, N R Jayagopala.

In: BMC Immunology, Vol. 12, 40, 18.07.2011.

Research output: Contribution to journalArticle

Massilamany, Chandirasegaran ; Upadhyaya, Bijaya ; Gangaplara, Arunakumar ; Kuszynski, Charles ; Reddy, N R Jayagopala. / Detection of autoreactive CD4 T cells using major histocompatibility complex class II dextramers. In: BMC Immunology. 2011 ; Vol. 12.
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AB - Background: Tetramers are useful tools to enumerate the frequencies of antigen-specific T cells. However, unlike CD8 T cells, CD4 T cells - especially self-reactive cells - are challenging to detect with major histocompatibility complex (MHC) class II tetramers because of low frequencies and low affinities of their T cell receptors to MHC-peptide complexes. Here, we report the use of fluorescent multimers, designated MHC dextramers that contain a large number of peptide-MHC complexes per reagent.Results: The utility of MHC dextramers was evaluated in three autoimmune disease models: 1) proteolipid protein (PLP) 139-151-induced experimental autoimmune encephalomyelitis in SJL/J (H-2s) mice; 2) myelin oligodendrocyte glycoprotein (MOG) 35-55-induced experimental autoimmune encephalomyelitis in C57Bl/6 (H-2b) mice; and 3) cardiac myosin heavy chain (Myhc)-α 334-352-induced experimental autoimmune myocarditis in A/J (H-2a) mice. Flow cytometrically, we demonstrate that IAs/PLP 139-151, IAb/MOG 35-55 and IAk/Myhc-α 334-352 dextramers detect the antigen-sensitized cells with specificity, and with a detection sensitivity significantly higher than that achieved with conventional tetramers. Furthermore, we show that binding of dextramers, but not tetramers, is less dependent on the activation status of cells, permitting enumeration of antigen-specific cells ex vivo.Conclusions: The data suggest that MHC dextramers are useful tools to track the generation and functionalities of self-reactive CD4 cells in various experimental systems.

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