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HSP60 as a target of anti-ergotypic regulatory T cells.

Quintana FJ, Mimran A, Carmi P, Mor F, Cohen IR - PLoS ONE (2008)

Bottom Line: Anti-ergotypic T cells responded to T-cell HSP60 by proliferation and secreted IFNgamma and TGFbeta1.In vivo, adoptive transfer of an anti-ergotypic HSP60-specific T-cell line led to decreased secretion of IFNgamma by arthritogenic T cells and ameliorated adjuvant arthritis (AA).Thus, by functioning as an ergotope, HSP60 can control both the effector T cells and the regulatory HSP60-specific T cells that control them.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel. fquintana@rics.bwh.harvard.edu

ABSTRACT
The 60 kDa heat shock protein (HSP60) has been reported to influence T-cell responses in two ways: as a ligand of toll-like receptor 2 signalling and as an antigen. Here we describe a new mechanism of T-cell immuno-regulation focused on HSP60: HSP60 is up-regulated and presented by activated T cells (HSP60 is an ergotope) to regulatory (anti-ergotypic) T cells. Presentation of HSP60 by activated T cells was found to be MHC-restricted and dependent on accessory molecules - CD28, CD80 and CD86. Anti-ergotypic T cells responded to T-cell HSP60 by proliferation and secreted IFNgamma and TGFbeta1. In vitro, the anti-ergotypic T cells inhibited IFNgamma production by their activated T-cell targets. In vivo, adoptive transfer of an anti-ergotypic HSP60-specific T-cell line led to decreased secretion of IFNgamma by arthritogenic T cells and ameliorated adjuvant arthritis (AA). Thus, the presentation of HSP60 by activated T cells turns them into targets for anti-ergotypic regulatory T cells specific for HSP60. However, the direct interaction between the anti-ergotypic T regulators (anti-HSP60) and the activated T cells also down-regulated the regulators. Thus, by functioning as an ergotope, HSP60 can control both the effector T cells and the regulatory HSP60-specific T cells that control them.

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DNA vaccination with HSP60 induces anti-ergotypic T cells.A and B. Anti-ergotypic proliferative response of LNC from rats vaccinated with pcDNA3, pHSP65 or pHSP60 (A) or pcDNA3, pI or pII (B), taken 26 days after the induction of AA. Proliferative responses are presented as the ΔCPM±SEM of quadruplicate cultures. * p<0.05 compared to the pHSP65 (A) or the pcDNA3 (B) groups. C. Monoclonal antibodies to MHC-II/RT1.B, MHC-II/RT1.D or MHC-I were assayed for their ability to block the anti-ergotypic proliferative response. Results are presented as the percent of inhibition of proliferation±SEM of quadriplicate cultures. D. Anti-ergotypic cytokine response of LNC taken from rats vaccinated with pcDNA3, pHSP65, pHSP60, pI or pII 26 days after the induction of AA. IFNγ (IFNg), TGFβ1 (TGFb1), IL-10 and IL-4 were quantified in the culture supernatants after 72 hr of stimulation with 105 activated or resting, irradiated, A2b cells per well. The results are presented as pg/ml±SEM of triplicate cultures. Three independent experiments produced similar results.
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pone-0004026-g001: DNA vaccination with HSP60 induces anti-ergotypic T cells.A and B. Anti-ergotypic proliferative response of LNC from rats vaccinated with pcDNA3, pHSP65 or pHSP60 (A) or pcDNA3, pI or pII (B), taken 26 days after the induction of AA. Proliferative responses are presented as the ΔCPM±SEM of quadruplicate cultures. * p<0.05 compared to the pHSP65 (A) or the pcDNA3 (B) groups. C. Monoclonal antibodies to MHC-II/RT1.B, MHC-II/RT1.D or MHC-I were assayed for their ability to block the anti-ergotypic proliferative response. Results are presented as the percent of inhibition of proliferation±SEM of quadriplicate cultures. D. Anti-ergotypic cytokine response of LNC taken from rats vaccinated with pcDNA3, pHSP65, pHSP60, pI or pII 26 days after the induction of AA. IFNγ (IFNg), TGFβ1 (TGFb1), IL-10 and IL-4 were quantified in the culture supernatants after 72 hr of stimulation with 105 activated or resting, irradiated, A2b cells per well. The results are presented as pg/ml±SEM of triplicate cultures. Three independent experiments produced similar results.

Mentions: We have reported that DNA vaccination with the hsp60 gene (pHSP60) or with its N-terminal fragments – constructs pI (aa 1–130) or pII (aa 120–240)– induced HSP60-specific T cells and inhibited the development of AA [6], [7]. DNA vaccination with mycobacterial HSP65 (pHSP65) also protected rats against AA [30], but this vaccination was significantly less effective than was vaccination with self-HSP60 [6]. Does protective HSP60 vaccination activate anti-ergotypic reactivity? To approach this question, we studied the anti-ergotypic T-cell responses in rats vaccinated with pHSP60, pI, pII, pHSP65 or pcDNA3, 26 days after the induction of AA. Lymph node cells (LNC) of the vaccinated rats were incubated with irradiated activated or resting A2b T cells, and proliferative responses were measured to different numbers of A2b stimulator cells. Figure 1A shows that vaccination with pHSP60 induced a proliferative anti-ergotypic T-cell response, which was significantly (p<0.05) higher than that induced by pHSP65. Moreover, vaccination with the pI or pII constructs of HSP60 also induced a significant (p<0.05) anti-ergotypic response compared to pcDNA3 (Figure 1B). Using neutralizing antibodies, we found that the anti-ergotypic response induced by DNA vaccination included both MHC-II (RT1.B) and MHC-I restricted T cells (Figure 1C).


HSP60 as a target of anti-ergotypic regulatory T cells.

Quintana FJ, Mimran A, Carmi P, Mor F, Cohen IR - PLoS ONE (2008)

DNA vaccination with HSP60 induces anti-ergotypic T cells.A and B. Anti-ergotypic proliferative response of LNC from rats vaccinated with pcDNA3, pHSP65 or pHSP60 (A) or pcDNA3, pI or pII (B), taken 26 days after the induction of AA. Proliferative responses are presented as the ΔCPM±SEM of quadruplicate cultures. * p<0.05 compared to the pHSP65 (A) or the pcDNA3 (B) groups. C. Monoclonal antibodies to MHC-II/RT1.B, MHC-II/RT1.D or MHC-I were assayed for their ability to block the anti-ergotypic proliferative response. Results are presented as the percent of inhibition of proliferation±SEM of quadriplicate cultures. D. Anti-ergotypic cytokine response of LNC taken from rats vaccinated with pcDNA3, pHSP65, pHSP60, pI or pII 26 days after the induction of AA. IFNγ (IFNg), TGFβ1 (TGFb1), IL-10 and IL-4 were quantified in the culture supernatants after 72 hr of stimulation with 105 activated or resting, irradiated, A2b cells per well. The results are presented as pg/ml±SEM of triplicate cultures. Three independent experiments produced similar results.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2602852&req=5

pone-0004026-g001: DNA vaccination with HSP60 induces anti-ergotypic T cells.A and B. Anti-ergotypic proliferative response of LNC from rats vaccinated with pcDNA3, pHSP65 or pHSP60 (A) or pcDNA3, pI or pII (B), taken 26 days after the induction of AA. Proliferative responses are presented as the ΔCPM±SEM of quadruplicate cultures. * p<0.05 compared to the pHSP65 (A) or the pcDNA3 (B) groups. C. Monoclonal antibodies to MHC-II/RT1.B, MHC-II/RT1.D or MHC-I were assayed for their ability to block the anti-ergotypic proliferative response. Results are presented as the percent of inhibition of proliferation±SEM of quadriplicate cultures. D. Anti-ergotypic cytokine response of LNC taken from rats vaccinated with pcDNA3, pHSP65, pHSP60, pI or pII 26 days after the induction of AA. IFNγ (IFNg), TGFβ1 (TGFb1), IL-10 and IL-4 were quantified in the culture supernatants after 72 hr of stimulation with 105 activated or resting, irradiated, A2b cells per well. The results are presented as pg/ml±SEM of triplicate cultures. Three independent experiments produced similar results.
Mentions: We have reported that DNA vaccination with the hsp60 gene (pHSP60) or with its N-terminal fragments – constructs pI (aa 1–130) or pII (aa 120–240)– induced HSP60-specific T cells and inhibited the development of AA [6], [7]. DNA vaccination with mycobacterial HSP65 (pHSP65) also protected rats against AA [30], but this vaccination was significantly less effective than was vaccination with self-HSP60 [6]. Does protective HSP60 vaccination activate anti-ergotypic reactivity? To approach this question, we studied the anti-ergotypic T-cell responses in rats vaccinated with pHSP60, pI, pII, pHSP65 or pcDNA3, 26 days after the induction of AA. Lymph node cells (LNC) of the vaccinated rats were incubated with irradiated activated or resting A2b T cells, and proliferative responses were measured to different numbers of A2b stimulator cells. Figure 1A shows that vaccination with pHSP60 induced a proliferative anti-ergotypic T-cell response, which was significantly (p<0.05) higher than that induced by pHSP65. Moreover, vaccination with the pI or pII constructs of HSP60 also induced a significant (p<0.05) anti-ergotypic response compared to pcDNA3 (Figure 1B). Using neutralizing antibodies, we found that the anti-ergotypic response induced by DNA vaccination included both MHC-II (RT1.B) and MHC-I restricted T cells (Figure 1C).

Bottom Line: Anti-ergotypic T cells responded to T-cell HSP60 by proliferation and secreted IFNgamma and TGFbeta1.In vivo, adoptive transfer of an anti-ergotypic HSP60-specific T-cell line led to decreased secretion of IFNgamma by arthritogenic T cells and ameliorated adjuvant arthritis (AA).Thus, by functioning as an ergotope, HSP60 can control both the effector T cells and the regulatory HSP60-specific T cells that control them.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel. fquintana@rics.bwh.harvard.edu

ABSTRACT
The 60 kDa heat shock protein (HSP60) has been reported to influence T-cell responses in two ways: as a ligand of toll-like receptor 2 signalling and as an antigen. Here we describe a new mechanism of T-cell immuno-regulation focused on HSP60: HSP60 is up-regulated and presented by activated T cells (HSP60 is an ergotope) to regulatory (anti-ergotypic) T cells. Presentation of HSP60 by activated T cells was found to be MHC-restricted and dependent on accessory molecules - CD28, CD80 and CD86. Anti-ergotypic T cells responded to T-cell HSP60 by proliferation and secreted IFNgamma and TGFbeta1. In vitro, the anti-ergotypic T cells inhibited IFNgamma production by their activated T-cell targets. In vivo, adoptive transfer of an anti-ergotypic HSP60-specific T-cell line led to decreased secretion of IFNgamma by arthritogenic T cells and ameliorated adjuvant arthritis (AA). Thus, the presentation of HSP60 by activated T cells turns them into targets for anti-ergotypic regulatory T cells specific for HSP60. However, the direct interaction between the anti-ergotypic T regulators (anti-HSP60) and the activated T cells also down-regulated the regulators. Thus, by functioning as an ergotope, HSP60 can control both the effector T cells and the regulatory HSP60-specific T cells that control them.

Show MeSH
Related in: MedlinePlus