Essentials of Th17 cell commitment and plasticity

Muranski, Pawel; Restifo, Nicholas P.

doi:10.1182/blood-2012-09-378653

Cited by 317 publications

(341 citation statements)

References 191 publications

(99 reference statements)

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“…HLA-A2/HLA-DR1 transgenic mice immunized with D393-CD20 [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47] were CD8 T cells depleted with 500 lg of CD8 clone 2.43 and splenocytes were stimulated with D393-CD20 derived peptides and their reactivity was assessed ex vivo in an IFN-g ELISPOT (a). D393-CD20 [33][34][35][36][37][38][39][40][41] (SP) specific T cell recognition after D393-CD20 SP or D393-CD20 LP vaccination was assessed ex vivo on sorted CD8 T cells (b). D393-CD20 [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47] (LP) specific T cell recogniti...…”

Section: Discussionmentioning

confidence: 99%

“…D393-CD20 [33][34][35][36][37][38][39][40][41] (SP) specific T cell recognition after D393-CD20 SP or D393-CD20 LP vaccination was assessed ex vivo on sorted CD8 T cells (b). D393-CD20 [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47] (LP) specific T cell recognition after D393-CD20 SP or D393-CD20 LP vaccination was assessed ex vivo on CD8 T cells depleted splenocytes in an IFN-g ELISPOT (c). Five mice per group were used in each experiment.…”

Section: Discussionmentioning

confidence: 99%

“…Briefly, these mice are H-2 Class I and IA Class II knockout, and their CD8 T and CD4 T cells are restricted by the sole HLA-A*0201 and HLA-DR1*0101 molecules, respectively. 11,15 For D393-CD20 [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47] (20mer) and D393-CD20 33-41 (9mer) immunization, mice were injected twice with 100 lg of D393-CD20 20mer or 50 mg of D393-CD20 9mer were emulsified in incomplete Freund adjuvant (IFA, Sigma-Aldrich). All peptide vaccinations were done subcutaneously at the base of the tail at Days 1 and 14.…”

Section: Mouse and Vaccinationsmentioning

confidence: 99%

“…The three first amino-acids used to name these peptides are indicated in boldface. T cell lines were obtained from healthy donors' PBMCs after two rounds of in vitro stimulation in microplates with 10 lg/mL of D393-CD20 [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47] peptide. Specific responses were assessed by CD4 and IFN-g staining.…”

Section: Tumor Immunologymentioning

confidence: 99%

“…24 We then evaluated the capacity of the D393-CD20 28-47 long peptide (LP) to prime HLA-A2-restricetd CTL response. To this end, mice were immunized either with the D393-CD20 [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47] LP or with the HLA-A2-restricted D393-CD20 derived short peptide D393-CD20 [33][34][35][36][37][38][39][40][41] (SP). As shown in Figure 5b, higher IFN-g producing specific-CTL were detected ex vivo after immunization with the D393-CD20 28-47 LP than with the SP.…”

Section: Tumor Immunologymentioning

confidence: 99%

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CD20 alternative splicing isoform generates immunogenic CD4 helper T epitopes

Vauchy

Gamonet

Ferrand

et al. 2014

Intl Journal of Cancer

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Cancer-specific splice variants gain significant interest as they generate neo-antigens that could be targeted by immune cells. CD20, a membrane antigen broadly expressed in mature B cells and in B cell lymphomas, is subject to an alternative splicing named D393-CD20 leading to loss of membrane expression of the spliced isoform. D393-CD20 expression is detectable in transformed B cells and upregulated in various lymphoma B cells. In this study, we show that D393-CD20 is translated in malignant B cells and that D393-CD20 specific CD4 T cells producing IFN-c are present in B-cell lymphoma patients. Then, we have investigated whether the 20mer D393-CD20 peptide spanning the splicing site might be targeted by the immune system and we have shown that D393-CD20-specific CD4 Th1 clones could directly recognize malignant B cell lines and kill autologous lymphoma B cells indicating that D393-CD20-derived epitopes are naturally processed and presented on tumor cells. Finally, D393-CD20 peptide-based vaccination induced specific CD8 and CD4 T cell responses in HLA-humanized transgenic mice suggesting the presentation of D393-CD20 derived peptides on both HLA Class-I and -II. These findings support further investigations on the potential use of D393-CD20 directed specific immunotherapy in B cell malignancies.The effective discovery of clinically relevant tumor antigens holds a fundamental role for the development of new diagnostic tools and anticancer immunotherapies. Generally, tumor antigens are classified as unique antigens derived from point mutations or as shared antigens. The shared antigens are further divided into tumor-specific antigens, differentiation antigens and overexpressed antigens. The prioritization of cancer antigens is based on criteria such as immunogenicity, specificity, oncogenicity. 1 One mechanism that would lead to such priority targets could be alternative splicing. Alternative splicing can change the structure of mRNA by inclusion or skipping of exons, and this may alter the function, stability or binding properties of the encoded protein. 2 Aside from its role in physiological cell adaptation, alternative splicing has been shown to occur in human diseases, including cancer. 3 Particularly, the splice variants differ between cancer and normal corresponding tissues. 4,5 Cancerspecific splice variants are thus of significant interest as they may be involved in pathogenesis and may further potentially be used as biomarkers and generate novel targets for therapy. Because immune recognition of antigenic epitopes is highly specific, alternative exon splicing could provide the structural basis for expression of novel amino-acid sequences not subject to self repertoire tolerance.We previously identified an alternative transcript of the B cell lineage membrane receptor CD20. This alternative transcript lacks 168 nucleotides within Exon 3 to 7 compared to the wild-type CD20 transcript and referred thereafter as D393-CD20. 6 D393-CD20 translation gives rise to a protein lacking the extracellular domain and ...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Mouse and Vaccinationsmentioning

confidence: 99%

Section: Tumor Immunologymentioning

confidence: 99%

Section: Tumor Immunologymentioning

confidence: 99%

See 3 more Smart Citations

CD20 alternative splicing isoform generates immunogenic CD4 helper T epitopes

Vauchy

Gamonet

Ferrand

et al. 2014

Intl Journal of Cancer

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Rapamycin and abundant TCR stimulation are required for the generation of stable human induced regulatory T cells

et al. 2020

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Objectives Regulatory T cells (Tregs) are a vital sub‐population of CD4+ T cells with major roles in immune tolerance and homeostasis. Given such properties, the use of regulatory T cells for immunotherapies has been extensively investigated, with a focus on adoptive transfer of ex vivo expanded natural Tregs (nTregs). For immunotherapies, induced Tregs (iTregs), generated in vitro from naïve CD4+ T cells, provide an attractive alternative, given the ease of generating cell numbers required for clinical dosage. While the combination of TGF‐β, ATRA and rapamycin has been shown to generate highly suppressive iTregs, the challenge for therapeutic iTreg generation has been their instability. Here, we investigate the impact of rapamycin concentrations and α‐CD3/CD28 bead ratios on human iTreg stability. Methods We assess iTregs generated with various concentrations of rapamycin and differing ratios of α‐CD3/CD28 beads for their differentiation, stability, expression of Treg signature molecules and T helper effector cytokines, and Treg‐specific demethylation region (TSDR) status. Results iTregs generated in the presence of TGF‐β, ATRA, rapamycin and a higher ratio of α‐CD3/CD28 beads were highly suppressive and stable upon in vitro re‐stimulation. These iTregs exhibited a similar expression profile of Treg signature molecules and T helper effector cytokines to nTregs, in the absence of TSDR demethylation. Conclusion This work establishes a method to generate human iTregs which maintain stable phenotype and function upon in vitro re‐stimulation. Further validation in pre‐clinical models will be needed to ensure its suitability for applications in adoptive transfer.

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Apoptotic cell capture by DCs induces unexpectedly robust autologous CD4⁺ T‐cell responses

et al. 2014

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Keywords: Apoptotic cells r CD4 + T-cell responses r Dendritic cellsAdditional supporting information may be found in the online version of this article at the publisher's web-site IntroductionOne of the key features of the immune system is to react to foreign pathogens and not respond to the self-antigens derived from dying cells that can be presented to the immune system in far greater amounts. Thymic selection removes the vast majority of developing thymocytes that express a T-cell receptor with high specificity for self-peptide: MHC complexes, a process referred to as central tolerance. Nevertheless, some self-reactive T cells are released into the periphery [1,2]. The mechanisms that control such potentially [25][26][27][28]. Moreover, in animal models, the use of apoptotic cells to prevent autoimmunity has also been reported [29,30]. DCs also possess the unique ability to induce proliferation of autologous T cells in the absence of exogenous antigen, a process termed "autologous mixed leukocyte reaction" [31]. T cells activated in this process display normal immune response [32] in terms of specificity and memory, and a capacity for immunosuppression [33][34][35][36]. There is evidence that, in this process, DCs present peptides derived from self-proteins [37], mostly derived from residual apoptotic cells [38,39]. However, the effect of the capture of apoptotic cells by DCs on their ability to stimulate and support the proliferation of naive autologous CD4 + T cells, and therefore to generate self-reactive responses, has never been studied in detail. Surprisingly, we found that DCs loaded with apoptotic cells induced a robust proliferation of autologous naive CD4 + T cells. These proliferating autologous CD4 + T cells exhibited suppressive phenotype and function, unless a TLR ligand mimicking concomitant infection was added to the apoptotic cell-DC culture, yielding Th17 differentiation. Results Robust autologous responses induced by apoptotic cell-loaded DCsIn order to examine the effect of apoptotic cell uptake by DCs on their ability to induce potential autologous CD4 + T-cell responses, we have set up an experimental procedure in which DCs were loaded with autologous apoptotic CD4 + T-cell blasts for 16 hours, purified, and cultured with autologous naive CD4 stimulated-CD4 + T cells were also able to induce proliferation of autologous naive CD4 + T cells, whereas DCs loaded with necrotic CD4 + T-cell blasts were not (Supporting Information Fig. 1). To understand whether this was due to a very few cells that have proliferated to high levels or whether this reflected an increase in the frequency of proliferative precursors, we used two different methods. The first was based on intracellular staining with the antibody specific for Ki67, which is strictly expressed only during cell proliferation ( Fig. 1B), but is absent from resting cells, making it a marker to determine the fraction of cells that will proliferate. This staining was performed on day 2, just before the beginning of the proliferation, and ...

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Essentials of Th17 cell commitment and plasticity

Cited by 317 publications

References 191 publications

CD20 alternative splicing isoform generates immunogenic CD4 helper T epitopes

CD20 alternative splicing isoform generates immunogenic CD4 helper T epitopes

Rapamycin and abundant TCR stimulation are required for the generation of stable human induced regulatory T cells

Apoptotic cell capture by DCs induces unexpectedly robust autologous CD4⁺ T‐cell responses

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Essentials of Th17 cell commitment and plasticity

Cited by 317 publications

References 191 publications

CD20 alternative splicing isoform generates immunogenic CD4 helper T epitopes

CD20 alternative splicing isoform generates immunogenic CD4 helper T epitopes

Rapamycin and abundant TCR stimulation are required for the generation of stable human induced regulatory T cells

Apoptotic cell capture by DCs induces unexpectedly robust autologous CD4+ T‐cell responses

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Apoptotic cell capture by DCs induces unexpectedly robust autologous CD4⁺ T‐cell responses